Specifications and Standards for Food, Food Additives, etc. Ministry of Health and Welfare Notification No. 370, 1959

Provisional Translation from the Japanese Original 1/100 Specifications and Standards for Food, Food Additives, Etc. Ministry of Health and Welfare Noti f icat ion No. 370, 1959 The Minis ter of Heal th and Welfare has es tabl ished the specif ica t ions and s tandards for food, food addi t ives g iven below, based on Art ic le 10 and Art ic le 11 Paragraph 1 of the Food Sani ta t ion Law (Law No. 233, 1947) . Part I Food Sect ion A General Composit ional Standards for Food 1. Foods shal l not conta in any ant ib iot ics or chemical ly synthes ized ant ibacter ia l substances (substances obtained by inst igat ing chemical react ions to e lements and/or compounds through chemical methods , except for decomposi t ion; th is appl ies hereinaf ter in th is paragraph) , except for the fol lowing cases : (1) When the substance concerned is ident ical to the food addi t ive determined by the Minis ter of Heal th , Labour and Welfare as having no potent ial to cause damage to human heal th under Art ic le 10 of the Food Sani ta t ion Law (Law No. 233, 1947, here inaf ter “ the Law.”) (2) When composi t ional s tandards are se t for th in 5 , 6 , 7 , 8 or 9 below for the substance concerned. (3) When the food product concerned has been manufactured or processed us ing a food ingredient that meets the composi t ional s tandards given in 5 , 6 , 7 , 8 or 9 below (except for foods conta ining ant ib iot ics or chemical ly synthes ized ant ibacter ia l substances for which composi t ional s tandards are not se t for th in 5 , 6 , 7 , 8 or 9 below.) 2 .~4. (Omit ted) 5 . Foods shal l not conta in substances ( including substances produced by chemical t ransformat ion: th is appl ies here inaf ter in this paragraph) used as ingredients of agr icul tura l chemicals and other chemical substances l i s ted in the table in (1) below. The agr icul tura l chemicals and other chemical substances s ta ted above, here and a lso la ter in th is paragraph, refer to substances used for purposes des ignated by the Agricul ture , Fores t ry and Fisher ies Minis ter ia l Ordinance according to the provis ion Provisional Translation from the Japanese Original 2/100 of Paragraph 3 of Art ic le 2 of the Law Concerning Safety Assurance and Qual i ty Improvement of Agricul tura l Chemicals and Feeds (Law No. 35, 1953) , which is s t ipula ted under Paragraph 1 of Art ic le 1-2 of the Agricul tural Chemicals Regulat ion Law (Law No. 82, 1948) , wi th such a ims as adding to , mixing wi th , or inf i l t ra t ing in to feeds ( the feeds s t ipula ted under Paragraph 2 of Art ic le 2 of Law No. 35) , or medical products to be used for animals , which are s t ipula ted in Paragraph 1 of Art ic le 2 of the Pharmaceut ical Affa i rs Law (Law No. 145, 1960) . In associa t ion wi th th is regulat ion, a sample of the foods l i s ted in the “foods” column in the table in (2) below shal l be tes ted us ing the part l i s ted in the “samples” column in the table by the tes t ing methods descr ibed in (3) to (16) below. No ingredients of agricul tura l chemicals or o ther chemical substances shal l be detected in these tes ts . (1) Substances used as ingredients of agr icul tura l chemicals and other chemical substances that are s t ipula ted to be “Not detected” in foods 1 . 2 , 4 , 5-T 2 . Azocyclot in and cyhexat in 3 . Amit ro l 4 . Captafol 5 . Carbadox 6 . Coumaphos 7 . Chloramphenicol 8 . Chlorpromazine 9 . Diethylst i lbest rol 10. Dimetr idazole 11. Daminozide 12. Ni t rofurans 13. Propham 14. Malachi te green 15. Metronidazole 16. Ronidazole Provisional Translation from the Japanese Original 3/100 (2) Samples Food Sample mater ia l Bar ley and buckwheat Threshed seeds Wheat and rye Husked seeds Rice (brown r ice) Husked seeds Corn (maize) Seeds wi th the husks , the s i lk and the cores removed Other cereal gra ins Threshed seeds Peas , beans (dry)( including but ter beans , cowbeans ( red beans) , lent i l , l ima beans , pegia , su l tani , sul tapya and white beans)* , broad beans and soybeans (dry)) Seeds wi thout the pods Peanuts With the shel ls removed Other legumes/pulses Seeds wi thout the pods Apricot , mume plum, cherry, Japanese plum ( including prune) and nectar ine With the peduncle and the seeds removed Peach With the skins and the seeds removed Orange ( including navel orange) , grapefrui t , c i t rus natsudaidai (whole) , l ime and lemon Whole f rui t Ci t rus natsudaidai (pulp) and unshu orange (pulp) With the peels removed Ci t rus natsudaidai , peels With the calyxes removed Other c i t rus f rui ts Whole f rui t Pear, Japanese pear, quince and apple With the blossom scars , the cores and the peduncles removed Loquat With the peduncles , the skins and the seeds removed Avocado and mango With the seeds removed Kiwifrui t With the skins removed Guava With the calyxes removed Date With the calyxes and the seeds removed Provisional Translation from the Japanese Original 4/100 P ineapple With the tops removed Pass ion f rui t and papaya Whole f ru i t Banana With the pedicels removed Strawberry, cranberry, huckleberry, b lackberry and blueberry With the calyxes removed Raspberry Whole f rui t Other berr ies With the calyxes removed Japanese pers immon With the calyxes and the seeds removed Watermelon, makuwauri melon and melons With the r inds removed Grape With the peduncles removed Other f rui ts Edible por t ions Turnip ( roots) and Japanese radish ( roots , including radish) With the dir t l ight ly r insed off wi th water Turnip ( leaves) , watercress , kale , Japanese radish ( leaves , including radish) , and brussels sprouts With the decayed leaves removed Caul i f lower and broccol i With the leaves removed Cabbage and Chinese cabbage A sample consis t ing of one por t ion f rom each of four heads , wi th each head equal ly cut into four por t ions , wi thout the decayed outer leaves and the cores Kyona and komatsuna ( Japanese mustard spinach) With the roots and the decayed leaves removed Horseradish Roots wi th the di r t l ight ly r insed off wi th water Qing-geng-cai and other cruci ferous vegetables Edible por t ions Sweet potato , konjac , taro , pota to, yam and other pota toes With the dir t l ight ly r insed off wi th water Pumpkin ( including squash) , cucumber ( including gherkin) and or iental pickl ing melon (vegetable) With the vines removed Other cucurbi taceous vegetables Edible por t ions Art ichoke, endive and chicory With the decayed leaves removed Provisional Translation from the Japanese Original 5/100 Burdock and sa ls i fy A sample th inly s l iced then ground wi th a meat gr inder, the leaves having been removed and the d i r t having been l ight ly r insed off with water Shungiku With the roots and the decayed leaves removed Let tuce ( including cos le t tuce and leaf le t tuce) With the decayed outer leaves and the cores removed Other composi te vegetables Edible por t ions Shi i take mushroom, but ton mushroom and other mushrooms Edible por t ions Celery, pars ley and mitsuba With the roots and the decayed leaves removed Carrot and parsnip With the dir t l ight ly r insed off wi th water Other umbel l i ferous vegetables Edible por t ions Tomato, egg plant and pimiento (sweet pepper) With the calyxes removed Other solanceous vegetables Edible por t ions Asparagus Stems Onion, gar l ic , welsh ( including leek) and mul t ip lying onion With the outer skins and the root hai r removed Nira and other l i l iaceous vegetables Edible por t ions Green soybeans , k idney beans ( immature , wi th pods) and peas ( immature , wi th pods) With the pedicels removed Okra With the calyxes removed Sugarcane With the husks removed Ginger With the leaves removed, and wi th the di r t l ight ly r insed off wi th water Sugar beet With the dir t l ight ly r insed off wi th water Spinach With red roots lef t on, and wi th the root hai r and the decayed leaves removed Bamboo shoots and other vegetables Edible por t ions Provisional Translation from the Japanese Original 6/100 Sesame seeds , rapeseeds , sunf lower seeds , saff lower seeds , cot ton seeds and other oi l seeds Seeds only Almond, g inkgo nut , ches tnut , walnut , pecan and other nuts With the husks removed Cacao beans and coffee beans Beans wi thout the pods Tea Tea leaves Hop Dried f lowers Other spices and other herbs Edible por t ions (3) 2 , 4 , 5-T analyt ical method 1 . Apparatus A gas chromatograph wi th an e lec t ron capture detector (GC-ECD) and a gas chromatograph/mass spect rometer (GC/MS or GC/MS/MS). 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Reagents des ignated as “specia l grade” in th is sect ion must meet the requirements for “specia l grade” specif ied in the Japan Indust r ia l Standards for the reagents . Acetoni t r i le : Use a ro tary vacuum evaporator on 300 mL of acetoni t r i le to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Acetone: Use a ro tary vacuum evaporator on 300 mL of acetone to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Ether : Use a ro tary vacuum evaporator on 300 mL of e ther to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion in to a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Sodium chlor ide: Sodium chlor ide (specia l grade) . I f the reagent i s found to include any substance that may in terfere wi th the analysis of Provisional Translation from the Japanese Original 7/100 substances to be analyzed f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Synthet ic magnesium s i l ica te (Flor is i l ) for column chromatography: Heat f lor is i l (150-250 μm in par t ic le s ize) a t 130°C for 12 hours or longer. Cool down to room temperature in a des iccator. Diatomaceous ear th : Diatomaceous earth for chemical analys is . . Ethyl acetate : Use a ro tary vacuum evaporator on 300 mL of e thyl aceta te to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion in to a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11g . Reagent for butyl es ter i f ica t ion: Dissolve 10 g of boron t r i f luor ide e ther complex in 25 ml of n-butanol . n-Hexane: Use a ro tary vacuum evaporator on 300 mL of n -hexane to evaporate unt i l 5 mL is lef t . For analys is , in ject 5 μL of the solut ion in to a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11g . Water : Dis t i l led water. I f the dis t i l led water i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Sodium sul fa te (anhydrous) : Sodium sulfa te (anhydrous)(specia l grade) . I f the reagent i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tural chemicals , wash wi th a solvent such as n -hexane before use . Methanol : Use a ro tary vacuum evaporator on 300 mL of methanol to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . 3 . Reference s tandard 2 ,4 ,5-T: This product conta ins not less than 98% of 2 ,4 ,5-T, and i t s mel t ing point i s 156°C. 4 . Procedure a . Extract ion Provisional Translation from the Japanese Original 8/100 i . Cereal gra ins , legumes/pulses and seeds Weigh 10.0 g of the tes t sample , previously ground so as to pass through a s tandard mesh s ieve (420 µm). Add 20 ml of water to the obtained sample and leave i t to s tand for two hours . Then, add 100 ml of acetone and 5 ml of 4 mol / l hydrochlor ic acid , and homogenize the mixture for three minutes . Fi l ter the homogenized sample by suct ion in to a ro tary vacuum evaporator through a f i l ter paper covered wi th a 1-cm thick layer of d ia tomaceous ear th . Col lect the res idue on the surface of the f i l ter paper. Add 50 ml of acetone and homogenize the mixture for three minutes . Repeat the above procedure and combine the f i l t ra te into the rotary vacuum evaporator, and concentra te the mixture to approximate ly 30 ml a t 40°C or lower. Transfer the concentra ted solut ion to a 300-ml separa t ing funnel a l ready conta ining 100 ml of 10% sodium chlor ide solut ion. Wash the eggplant-shaped f lask of the above rotary vacuum evaporator wi th 100 ml of e thyl aceta te and add the washings to the separat ing funnel above. Shake the mixture v igorously for f ive minutes using a shaker and leave the funnel to s tand, and then t ransfer the e thyl aceta te layer to a 300-ml conical f lask . Add 50 ml of e thyl acetate to the aqueous layer and repeat the above procedure , and then add the e thyl acetate layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occas ional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of e thyl aceta te and wash twice the res idue on the surface of the f i l ter paper wi th the washings . Add the washings in to the rotary vacuum evaporator and concentrate the mixture to approximately 1 ml a t 40°C or lower, and then evaporate the solvent to near dryness in a n i t rogen s t ream at room temperature . Add 30 ml of n-hexane to the dr ied res idue and t ransfer the mixture to a 100-ml separa t ing funnel , and then add 30 ml of n-hexane sa tura ted acetoni t r i le . Shake the mixture vigorously for f ive minutes us ing a shaker and leave the funnel to s tand, and then t ransfer the acetoni t r i le layer to a 200-ml separa t ing funnel . Add 30 ml of n-hexane-satura ted acetoni t r i le to the n-hexane layer and repeat the above procedure twice, and then combine the acetoni t r i le Provisional Translation from the Japanese Original 9/100 layer to the separa t ing funnel above. Add 50 ml of n-hexane sa tura ted acetoni t r i le in to the separat ing funnel and shake l ight ly, and then leave the funnel to s tand. Transfer the acetoni t r i le layer in to a ro tary vacuum evaporator and concentra te the mixture to approximate ly 1 ml a t 40°C or lower, and then evaporate the solvent to near dryness in a n i t rogen st ream at room temperature . i i . Frui t , vegetables , matcha and hops In case of f ru i t and vegetables , weigh accurate ly about 1 kg of the tes t sample and add an appropr ia te amount of water to the sample , i f required. Homogenize the mixture and mesure out a sample equivalent to 20.0 g . In case of matcha, weigh 5 .00 g of the tes t sample and add 20 ml of water, and then leave i t to s tand for two hours . In case of hops , weigh 5 .00 g of the tes t sample , previously ground, and add 20 ml of water, and then leave i t to s tand for two hours . Then, add 100 ml of acetone and 5 ml of 4 mol / l hydrochlor ic acid , and homogenize the mixture for three minutes . Fi l ter the homogenized sample by suct ion in to a ro tary vacuum evaporator through a f i l ter paper covered wi th a 1-cm thick layer of d ia tomaceous ear th . Col lect the res idue on the surface of the f i l ter paper and add 50 ml of acetone, and then homogenize the mixture for three minutes . Repeat the above procedure and combine the f i l t ra te in to the rotary vacuum evaporator, and then concentra te the mixture to approximate ly 30 ml a t 40°C or lower. Transfer the concentra ted solut ion to a 300-ml separat ing funnel a l ready conta ining 100 ml of 10% sodium chlor ide solut ion. Wash the eggplant-shaped f lask of the above rotary vacuum evaporator wi th 100 ml of e thyl aceta te and add the washings to the separa t ing funnel above. Shake the mixture v igorously for f ive minutes us ing a shaker and leave the funnel to s tand, and then t ransfer the e thyl aceta te layer to a 300-ml conical f lask . Add 50 ml of e thyl aceta te to the aqueous layer and repeat the above procedure twice , and then combine the e thyl aceta te layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occasional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of e thyl aceta te and wash twice the residue on the Provisional Translation from the Japanese Original 10/100 surface of the f i l ter paper wi th the washings . Add the washings in to the rotary vacuum evaporator and concentra te the mixture to approximate ly 1 ml a t 40°C or lower, and then evaporate the solvent to near dryness in a n i t rogen st ream at room temperature . i i i . Teas other than matcha Weigh 9 .00 g of the tes t sample and soak i t in 540 ml of water a t 100°C, and then leave i t to s tand a t room temperature for f ive minutes . Transfer 360 ml of the cooled f i l t ra te in to a 500-ml conical f lask and add 18 g of sodium chlor ide and 4 mol / l hydrochlor ic acid to adjus t the pH to 1 or lower. Transfer the solut ion to a 1 ,000-ml separa t ing funnel a l ready conta ining 100 ml of e thyl aceta te . Shake the mixture v igorously for f ive minutes us ing a shaker and leave the funnel to s tand, and then t ransfer the e thyl aceta te layer to a 300-ml conical f lask . Add 100 ml of e thyl aceta te to the aqueous layer and repeat the above procedure twice , and combine the e thyl acetate layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occas ional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of e thyl aceta te and wash twice the res idue on the surface of the f i l ter paper wi th the washings . Add the washings in to the rotary vacuum evaporator and concentra te the mixture to approximate ly 1 ml a t 40°C or lower, and then evaporate the solvent to near dryness in a n i t rogen s t ream at room temperature . iv. Foods other than those l i s ted in i to i i i above Obtain the extracts according to the methods descr ibed in i or i i . b . Hydrolysis Dissolve the res idue obta ined by the ext ract ion descr ibed in 4-a in 20 ml of methanol and t ransfer the mixture to a 100-ml eggplant-shaped f lask , and then add 10 ml of 1 .5 mol / l sodium hydroxide solut ion. At tach a ref lux condenser to the f lask and heat the mixture for 30 minutes in a water bath a t 80°C, and then a l low i t to cool . Transfer the solut ion in to a rotary vacuum evaporator and remove most of the methanol a t 40°C or lower. Fi l ter the res idue by suct ion in to a 300-ml separat ing funnel ( I ) through a glass f i l ter (pore s ize G3) . Wash the res idue on the g lass f i l te r wi th a smal l amount of acetone and water and add the washings to the separa t ing funnel above, and then add 50 Provisional Translation from the Japanese Original 11/100 ml of e ther and 100 ml of 10% sodium chlor ide solut ion. Shake the mixture v igorously for f ive minutes us ing a shaker and leave i t to s tand. Transfer the aqueous layer to a 300-ml separa t ing funnel (I I ) and add 4 mol / l hydrochlor ic ac id to adjus t the pH to 1 or lower. Add 50 ml of e thyl acetate to the adjus ted solut ion and shake the mixture v igorously for f ive minutes us ing a shaker, and then leave i t to s tand. Transfer the e thyl aceta te layer to a 300-ml conical f lask . Add 50 ml of e thyl aceta te to the aqueous layer and repeat the above procedure , and then combine the e thyl aceta te layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occasional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of e thyl aceta te and wash twice the res idue on the surface of the f i l ter paper wi th the washings . Add the washings in to the rotary vacuum evaporator and concentra te the mixture to approximate ly 1 ml a t 40°C or lower. c . Butyl es ter i f ica t ion Transfer the solut ion obtained by the hydrolysis descr ibed in 4-b to a 20-ml eggplant-shaped f lask and then evaporate the solvent to near dryness in a n i t rogen s t ream at room temperature . After the des iccat ion, add 1 ml of a butyl ester i f ica ted agent to the res idue. At tach a ref lux condenser to the eggplant-shaped f lask descr ibed above and heat the mixture for 30 minutes in a water bath at 90°C, and a l low i t to cool . Transfer the mixture to a 200-ml separa t ing funnel a l ready conta ining 50 ml of 10% sodium chlor ide solut ion and 50 ml of n-hexane and shake the mixture v igorously for f ive minutes us ing a shaker, and then leave i t to s tand. Transfer the n-hexane layer to a 200-ml conical f lask and add 50 ml of n-hexane to the aqueous layer and repeat the above procedure , and then combine the n-hexane layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occas ional shaking. Fi l ter the content of the f lask in to a rotary vacuum evaporator. Wash the f lask wi th 10 ml of n-hexane and wash the res idue on the surface of the f i l ter paper wi th the washings . Add the washings to the rotary vacuum evaporator and concentra te the mixture to approximate ly 2 ml a t 40°C or lower. d . Clean-up Provisional Translation from the Japanese Original 12/100 Add 5 g of f lor is i l for column chromatography suspended in n-hexane in to a chromatograph tube (15mm in inner d iameter, 300 mm in length) and add over approximate ly 5 g of sodium sulfa te (anhydrous) in to the column. Spi l l out the n-hexane unt i l only a smal l amount remains on the packing of the column and pour the solut ion obtained by the butyl es ter i f icat ion descr ibed in 4-c in to the column. Pour 50 ml of e ther /n-hexane (1 :19) in to the column and discard the eff luent . Pour 150 ml of e ther /n-hexane (3 :17) in to the column and col lect the e luate in to a ro tary vacuum evaporator. Concentra te the mixture to approximate ly 1 ml a t 40°C or lower and evaporate the solvent to near dryness in a n i t rogen s t ream at room temperature . Dissolve the res idue in n-hexane to make exact ly 10 ml of solut ion, which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard under the procedure descr ibed in 4-c . Test ing condi t ions Column: A s i l ica te g lass capi l lary column (0 .25 mm in inner d iameter, 30 m in length) coated wi th 5% phenyl methyl s i l icone for gas chromatography to a th ickness of 0 .25 µm. Column temperature : Hold the column temperature a t 50°C for one minute , fo l lowed by an increase of 25°C every minute unt i l reaching 125°C, af ter which increase the temperature by 10°C every minute unt i l reaching 300°C, and hold for f ive minutes . In le t temperature : 260°C Detector : Operate at 300°C. Gas f low ra te : Use ni t rogen or hel ium as the carr ier gas . Adjust the f low ra te so that n-butyl (2 ,4 ,5- t r ichlorophenoxy) acetate f lows out in approximate ly 15 minutes . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. c . Conf i rmat ion tes ts Perform gas chromatography/mass spect rometry under the same Provisional Translation from the Japanese Original 13/100 condi t ions as those descr ibed in 5-a . Test resul ts obta ined for the reference s tandard must be the same as those obta ined under the procedure descr ibed in 4-c . Determine the quant i ty using e i ther the peak height or peak area method, i f required. (4) Analyt ica l method for azocyclot in and cyhexat in 1 . Apparatus A gas chromatograph wi th a f lame photometr ic detector (GC-FPD, in terference f i l ter for t in determinat ion, wavelength: 610nm) and a gas chromatograph/mass spectrometer (GC/MS or GC/MS/MS). 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Reagents des ignated as “specia l grade” in th is sect ion must meet the requirements for “specia l grade” specif ied in the Japan Indust r ia l Standards for the reagents . Acetoni t r i le : Use a ro tary vacuum evaporator on 300 mL of acetoni t r i le to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Acetone: Use a ro tary vacuum evaporator on 300 mL of acetone to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion in to a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . 3 mol / l e thylmagnesium bromide-ethereal solut ion: 3 mol / l e thyl magnesium bromide-ethereal solut ion. Ether : Use a ro tary vacuum evaporator on 300 mL of e ther to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Sodium chlor ide: Sodium chlor ide (specia l grade) . I f the reagent i s found to include any substance that may in ter fere wi th the analysis of substances to be analyzed f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Synthet ic magnesium s i l icate (Flor is i l ) for column chromatography: Provisional Translation from the Japanese Original 14/100 Heat f lor is i l (150-250 μm in par t ic le s ize) a t 130°C for 12 hours or longer. Cool down to room temperature in a des iccator. Diatomaceous ear th : Diatomaceous ear th for chemical analys is . Cyhexat in s tandard solut ion: Add acet ic acid/e thyl aceta te (1 :99) to 10.0 mg of cyhexat in to make a 100-ml solut ion. Take out 10 ml of the solut ion and add n-hexane to make 100 ml of mix. Sodium dodecyl sul fa te : Sodium dodecyl sulfa te wi th a pur i ty of 85% or h igher. n-Hexane: Use a ro tary vacuum evaporator on 300 mL of n -hexane to evaporate unt i l 5 mL is lef t . For analys is , in ject 5 μL of the solut ion in to a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11g . Water : Dis t i l led water. I f the dis t i l led water i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Sodium sul fa te (anhydrous) : Sodium sulfa te (anhydrous)(specia l grade) . I f the reagent i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tural chemicals , wash wi th a solvent such as n -hexane before use . 3 . Reference s tandard Cyhexat in : This product conta ins not less than 99% of cyhexat in , and i t s mel t ing point i s 195-198°C. 4 . Procedure a . Extract ion i . Legumes/pulses and seeds Weigh 10.0 g of the tes t sample , previously ground so as to pass through a s tandard mesh s ieve (420 µm). Add 20 ml of water to the obtained sample and leave i t to s tand for two hours . Then, add 100 ml of acetone/acet ic ac id (99:1) and homogenize the mixture for three minutes . Fi l ter the homogenized sample by suct ion in to a ro tary vacuum evaporator through a f i l ter paper covered wi th a 1-cm thick layer of dia tomaceous ear th . Col lec t the res idue on the surface of the f i l ter paper and add 50 ml of acetone/acet ic acid (99:1) , and then homogenize the mixture for three minutes . Repeat the above procedure and combine the f i l t ra te in to the rotary vacuum Provisional Translation from the Japanese Original 15/100 evaporator, and then concentra te the mixture to approximate ly 30 ml a t 40°C or lower. Trasfer the concentra ted solut ion to a 500-ml separa t ing funnel a l ready conta ining 200 ml of 10% sodium chlor ide solut ion. Wash the eggplant-shaped f lask of the above rotary vacuum evaporator wi th 100 ml of n-hexane and add the washings to the separa t ing funnel above. Shake the mixture v igorously for f ive minutes using a shaker and leave i t to s tand, and then t ransfer the n-hexane layer to a 300-ml conical f lask . Add 50 ml of n-hexane to the aqueous layer and repeat the above procedure , and then add the n-hexane layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occas ional shaking. Fi l ter the content of the f lask in to a rotary vacuum evaporator. Wash the f lask wi th 20 ml of n-hexane and wash twice the res idue on the surface of the f i l ter paper wi th the washings . Add the washings into the ro tary vacuum evaporator and remove the n-hexane at 40°C or lower. Add 20 ml of n-hexane to the residue and t ransfer the mixture to a 100-ml separa t ing funnel , and then add 40 ml of n-hexane-sa tura ted acetoni t r i le . Shake the mixture v igorously for f ive minutes us ing a shaker and leave i t to s tand, and then t ransfer the acetoni t r i le layer in to the ro tary vacuum evaporator. Add 40 ml of n-hexane-satura ted acetoni t r i le to the n-hexane layer and repeat the above procedure twice , and then combine the acetoni t r i le layer in to the rotary vacuum evaporator above. Remove the acetoni t r i le a t 40°C or lower. Dissolve the res idue in n-hexane to make exact ly 5 ml of solut ion. i i . Cereal gra ins , f ru i t and vegetables In case of cereal gra ins , weigh 10.0 g of the tes t sample , previously ground so as to pass through a s tandard mesh s ieve (420 µm). Add 20 ml of water to the obtained sample and leave i t to s tand for two hours . In case of f ru i t and vegetables , weigh accurate ly 1 kg of the test sample and add an appropr ia te amount of water to the sample , i f required. Homogenize the mix ture and measure out a sample equivalent to 20.0 g . Then, add 100 ml of acetone/acet ic acid (99:1) and homogenize the mixture for three minutes . Fi l ter the homogenized sample by suct ion Provisional Translation from the Japanese Original 16/100 in to a ro tary vacuum evaporator through a f i l ter paper covered wi th a 1-cm thick layer of dia tomaceous ear th . Col lec t the residue on the surface of the f i l ter paper and add 50 ml of acetone/acet ic acid (99:1) , and then homogenize the mixture for three minutes . Repeat the above procedure and combine the f i l t ra te in to the rotary vacuum evaporator, and then concentra te the mixture to approximately 30 ml a t 40°C or lower. Transfer the concentra ted solut ion to a 500-ml separa t ing funnel a l ready conta ining 200 ml of 10% sodium chlor ide solut ion. Wash the eggplant-shaped f lask of the above rotary vacuum evaporator wi th 100 ml of n-hexane and add the washings to the separa t ing funnel above. Shake the mixture v igorously for f ive minutes using a shaker and leave i t to s tand, and then t ransfer the n-hexane layer to a 300-ml conical f lask . Add 50 ml of n-hexane to the aqueous layer and repeat the above procedure twice , and then combine the n-hexane layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occasional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of n-hexane,and wash twice the res idue on the surface of the f i l te r paper wi th the washings . Add the washings in to the rotary vacuum evaporator and remove the n-hexane a t 40°C or lower, and then dissolve the res idue in n-hexane to make exact ly 10 ml of solut ion. i i i . Matcha and hops In case of matcha, weigh 5 .00 g of the tes t sample and add 20 ml of water, and then leave i t to s tand for two hours . In case of hops , weigh 5 .00 g of the tes t sample , previously ground, and add 20 ml of water, and then leave i t to s tand for two hours . Then, add 100 ml of acetone/acet ic acid (99:1) and homogenize the mixture for three minutes . Fi l ter the homogenized sample by suct ion in to a ro tary vacuum evaporator through a f i l ter paper covered wi th a 1-cm thick layer of dia tomaceous ear th . Col lec t the residue on the surface of the f i l ter paper and add 50 ml of acetone/acet ic acid (99:1) , and then homogenize the mixture for three minutes . Repeat the above procedure and combine the f i l t ra te in to the rotary vacuum evaporator, and then concentra te the mixture to approximately 30 ml a t 40°C or lower. Provisional Translation from the Japanese Original 17/100 Transfer the concentra ted solut ion to a 500-ml separa t ing funnel a l ready conta ining 200 ml of 10% sodium chlor ide solut ion. Wash the eggplant-shaped f lask of the above rotary vacuum evaporator wi th 100 ml of n-hexane and add the washings to the separa t ing funnel above. Shake the mixture v igorously for f ive minutes using a shaker and leave i t to s tand, and then t ransfer the n-hexane layer to a 300-ml conical f lask . Add 50 ml of n-hexane to the aqueous layer and repeat the above procedure twice , and then combine the n-hexane layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occasional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of n-hexane and wash twice the res idue on the surface of the f i l ter paper wi th the washings . Add the washings in to the rotary vacuum evaporator and concentra te the mixture to approximate ly 5 ml a t 40°C or lower, and then add n-hexane to make exact ly 10 ml of solut ion. iv. Teas other than matcha Weigh 9 .00 g of the tes t sample and soak i t in 540 ml of water a t 100°C, and then leave i t to s tand a t room temperature for f ive minutes . Transfrer 360 ml of the cooled f i l t ra te in to a 500-ml separa t ing funnel and add 30 g of sodium chlor ide , 2 ml of 2% sodium dodecyl sul fa te solu t ion and 100 ml of n-hexane. Shake the mixture v igorously for f ive minutes us ing a shaker and leave i t to s tand, and then t ransfer the n-hexane layer to a 300-ml conical f lask . Add 100 ml of n-hexane to the aqueous layer and repeat the above procedure twice , and then combine the n-hexane layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes with occasional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of n-hexane and wash twice the res idue on the surface of the f i l ter paper wi th the washings . Add the washings in to the rotary vacuum evaporator and remove n-hexane a t 40°C or lower. Dissolve the res idue in n-hexane to make exact ly 6 ml of solut ion. v. Foods other than those l i s ted in i to iv above Obtain the extracts according to the methods descr ibed in i , i i , or Provisional Translation from the Japanese Original 18/100 i i i . b . Ethylat ion Transfer 1 ml (2 ml for cereal gra ins , teas and hops) of the solut ion obtained by the ext rac t ion descr ibed in 4-a to a 50-ml tes t tube wi th a g lass s topper and add 1 ml (2 ml for cereal gra ins , teas and hops) of a 3 mol / l e thylmagnesium bromide-ethereal solut ion, and then leave i t to s tand for 20 minutes a t room temperature . Then, add 10 ml of 0 .5 ml / l su l fur ic acid gradual ly and add 10 ml of water to be mixed. Add 10 ml of n-hexane to the mixture and shake the mixture v igorously for one minute , and then leave i t to s tand. Transfer the n-hexane layer to a 50-ml conical f lask. Add 5 ml of n-hexane to the aqueous layer and repeat the above procedure , and then combine the n-hexane layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occas ional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 5 ml of n-hexane and wash the res idue on the surface of the f i l ter paper wi th the washings . Add the washings to the ro tary vacuum evaporator and concentra te the mixture to 2 ml a t 40°C or lower. c . Clean-up Add 5 g of f lor is i l for column chromatography suspended in n-hexane in to a chromatograph tube (15 mm in inner d iameter, 300 mm in length) and add over approximate ly 5 g of sodium sulfa te (anhydrous) in to the column. Spi l l out the n-hexane unt i l only a smal l amount remains on the packing of the column and pour the solut ion obtained by the e thyla t ion descr ibed in 4-b in to the column. Then, wash the eggplant-shaped f lask of the ro tary vacuum evaporator wi th 15 ml of n-hexane and t ransfer rhe washings to the column, and then col lec t the e luate in to the rotary vacuum evaporator. Then, pour 50 ml of e ther /n-hexane (1:99) in to the column and col lect the e luate in the ro tary vacuum evaporator, and then remove the e ther and n-hexane a t 40°C or lower. Dissolve the res idue in n-hexane to make exact ly 2 ml of solut ion, which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the cyhexat in Provisional Translation from the Japanese Original 19/100 reference s tandard under the procedure descr ibed in 4-b . Azocyclot in i s modif ied by e thyla t ion in to the same substance as cyhexat in . Test ing condi t ions Column: A s i l ica te g lass capi l lary column (0 .32 mm-0.53 mm in inner d iameter, 30 m in length) coated wi th 5% phenyl methyl s i l icone for gas chromatography to a th ickness of 1 .5 µm. Column temperature : Hold the column temperature a t 120°C for two minutes , fo l lowed by an increase of 10°C every minute unt i l reaching 200°C, af ter which increase the temperature by 20°C every minute unt i l reaching 300°C, and hold for f ive minutes . In le t temperature : 280°C Detector : Operate at 300°C. Gas f low ra te : Use hel ium as the carr ier gas . Adjust the f low ra te so that cyhexat in f lows out in approximate ly 13-15 minutes . Adjus t the f low ra tes of a i r and hydrogen to opt imal condi t ions . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the same condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. c . Conf i rmat ion tes ts Perform gas chromatography/mass spect rometry under the same condi t ions descr ibed in 5-a . Test resul ts obta ined for the reference s tandard must be same as those obta ined under the procedure descr ibed in 4-b. Determine the quant i ty us ing e i ther the peak height or peak area method, i f required. (5) Amitrole analyt ica l method 1 . Apparatus A high-performance l iquid chromatograph wi th a f luorescence detector (HPLC-FL) and a l iquid chromatograph/mass spectrometer (LC/MS or LC/MS/MS). 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Acetone: Use a ro tary vacuum evaporator on 300 mL of acetone to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Provisional Translation from the Japanese Original 20/100 Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Ethanol : Use a ro tary vacuum evaporator on 300 mL of e thanol to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion in to a gas chromatograph equipped wi th an e lec t ron capture detector. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Diatomaceous ear th : Diatomaceous ear th for chemical analys is . Acet ic acid buffer solu t ion: Add 0 .05 mol / l sodium aceta te solut ion to 800 ml of 0 .05 mol / l acet ic acid solut ion to make a 1 ,000-ml solut ion. Weakly acidic cat ion exchange res in: Wash the weakly acidic cat ion exchange res in produced for column choromatography wi th 1 mol / l hydrochlor ic ac id , secondly wi th a 2 .8% aqueous ammonia , and th i rdly wi th 1 mol / l hydrochlor ic ac id again . Thereaf ter, wash the washings wi th water unt i l they become neutra l . F luorescamin: A reagent wi th a pur i ty of 98% or h igher. Water : Dis t i l led water. I f the dis t i l led water i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Phosphate buffer solut ion: Add 10% phosphor ic acid solut ion to a 0 .05 ml/ l monosodium phosphate solut ion to adjus t the pH to 3 .0. 3 . Reference s tandard Amitrole : This product conta ins not less than 98% of amit role , and i t s mel t ing point i s 157-159°C. 4 . Procedure a . Extract ion i . Cereal gra ins , legumes/pulses , seeds , f ru i t , vegetables , matcha and hops In case of cereal grains , legumes/pulses and seeds , weigh 30.0 g of the tes t sample , previously ground so as to pass through a s tandard mesh s ieve (420 µm). In case of f ru i t and vegetables , weigh accurate ly about 1 kg.of the tes t sample and add an appropr ia te amount of water to the sample , i f required. Homogenize the mix ture and measure out a sample equivalent to 30.0 g . Provisional Translation from the Japanese Original 21/100 In case of matcha, weigh 30.0 g of the tes t sample . In case of hops , weigh 30.0 g of the tes t sample , previously ground. Then, add 80 ml of e thanol and homogenize the mixture for three minutes . Fi l ter the homogenized sample by suct ion in to a ro tary vacuum evaporator through a f i l ter paper covered wi th a 1-cm thick layer of d ia tomaceous ear th , and then t ransfer the f i l t ra te to a 200-ml graduated cyl inder. Col lec t the res idue on the surface of the f i l ter paper and add 40 ml of 60% ethanol , and then homogenize the mixture for three minutes . Repeat the above procedure and combine the f i l t ra te to the graduated cyl inder above, and then measure the amount of f i l t ra te . Transfer 10 ml of the above f i l t ra te to a 200-ml round bot tom f lask and add 1 ml of hydrogen peroxide solut ion. At tach a ref lux condenser to the f lask and heat the mixture for 30 minutes in a water bath a t 75°C, and then a l low i t to cool . i i . Teas except matcha Weigh 10.0 g of tes t sample and soak i t in 600 ml of water a t 100°C, and then leave i t to s tand a t room temperature for f ive minutes . Transfer 12 ml of the cooled f i l t ra te in to a 200-ml round bot tom f lask and add 1 ml of hydrogen peroxide solut ion. At tach a ref lux condenser to the f lask and heat the mixture for 30 minutes in a water bath a t 75°C, and then a l low i t to cool . i i i . Foods except those l i s ted in i and i i above Obtain the extracts according to the methods descr ibed in i . b . Clean-up Add 1 ml of s t rongly ac idic cat ion exchange res in (0 .063-0.156 µm in par t ic le s ize) suspended in water in to a chromatograph tube (10 mm in inner d iameter, 300 mm in length) and discard the water unt i l only a smal l amount remains on the packing of the column. Pour 5 ml of water in to the column and discard the eff luent . Pour the solut ion in to the column obtained by the ext rac t ion described in 4-a . Then, wash the above round bot tom f lask wi th 10 ml of water and t ransfer the washings in to the column, and then discard the eff luent . Pour 12 ml of 2 .8% aqueous ammonia into the column and col lec t the e luate in a ro tary vacuum evaporator. Add 30 ml of n-propanol and remove the water and n-propanol a t 45°C or lower. Dissolve the res idue in 5 ml of water. Provisional Translation from the Japanese Original 22/100 Add 5 ml of weakly ac idic ca t ion exchange res in (0 .33-0 .50 µm in par t ic le s ize) suspended in water in to a chromatograph tube (10 mm in inner d iameter, 300 mm in length) and discard the water unt i l only a smal l amount remains on the packing of the column. Pour 10 ml of water in to the column and discard the eff luent . Pour the above solut ion in to the column and discard the eff luent . Pour 50 ml of water in to the column and discard the eff luent . Pour 35 ml of 2 .8% aqueous ammonia in to the column and col lect the e luate in a ro tary vacuum evaporator. Add 100 ml of n-propanol and remove the aqueous ammonia and n-propanol a t 45°C or lower. c . Der ivat iza t ion Add 2 ml of acet ic ac id buffer solut ion to the res idue to d issolve i t wel l . Add 100 µl of 0 .25% f luorescamin-acetone solut ion to 1 ml of the above solut ion and shake the mixture wel l , and then leave i t to s tand for one hour. Add 0 .5 ml of 0 .05 mol / l sodium borate solut ion to use as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard under the procedure descr ibed in 4-c . Test ing condi t ions Column packing: Octadecyls i lane-bonded s i l ica gel (5 µm in par t ic le s ize) . Column: A s ta in less tube (4 .6 mm in inner d iameter, 150 mm in length) . Column temperature : 40°C Detector : Operate wi th an exci ta t ion wavelength of 380 nm and a f luorescent wavelength of 484 nm. Mobile phase: Use acetoni t r i le /phosphate buffer so lut ion (3:7) . Adjust the f low ra te so that amit role f lows out in approximately 15 minutes . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the same condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. c . Conf i rmat ion tes ts Provisional Translation from the Japanese Original 23/100 Perform l iquid chromatography/mass spect rometry under the same condi t ions descr ibed in 5-a . Test resul ts obta ined for the reference s tandard must be the same as those obta ined under the procedure descr ibed in 4-c . Determine the quant i ty us ing e i ther the peak height or peak area method, i f required. (6) Captafol analyt ica l method 1 . Apparatus A gas chromatograph wi th an e lec t ron capture detector (GC-ECD) and a gas chromatograph/mass spect rometer (GC/MS or GC/MS/MS). 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Reagents des ignated as “specia l grade” in th is sect ion must meet the requirements for “specia l grade” specif ied in the Japan Indust r ia l Standards for the reagents . Acetoni t r i le : Use a ro tary vacuum evaporator on 300 mL of acetoni t r i le to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Acetone: Use a ro tary vacuum evaporator on 300 mL of acetone to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in jec t 5 μL of the solut ion in to a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Sodium chlor ide: Sodium chlor ide (specia l grade) . I f the reagent i s found to include any substance that may in ter fere wi th the analysis of substances to be analyzed f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Synthet ic magnesium s i l icate (Flor is i l ) for column chromatography: Heat f lor is i l (150-250 μm in par t ic le s ize) a t 130°C for 12 hours or longer. Cool down to room temperature in a des iccator. Diatomaceous ear th : Diatomaceous ear th for chemical analys is . Ethyl acetate : Use a ro tary vacuum evaporator on 300 mL of e thyl aceta te to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion in to a GC-ECD. Peaks other than that of n -hexane in the resul t ing Provisional Translation from the Japanese Original 24/100 chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11g . n-Hexane: Use a ro tary vacuum evaporator on 300 mL of n -hexane to evaporate unt i l 5 mL is lef t . For analys is , in ject 5 μL of the solut ion in to a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11g . Water : Dis t i l led water. I f the dis t i l led water i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Sodium sul fa te (anhydrous) : Sodium sulfa te (anhydrous)(specia l grade) . I f the reagent i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tural chemicals , wash wi th a solvent such as n -hexane before use . 3 . Reference s tandard Captafol : This product conta ins not less than 98% of captafol , and i t s mel t ing point i s 159-161°C. 4 . Procedure a . Extract ion i . Cereal gra ins , legumes/pulses and seeds Weigh 10.0 g of the tes t sample , previously ground so as to pass through a s tandard mesh s ieve (420 µm). Add 20 ml of 3% phosphor ic ac id solut ion to the obta ined sample and leave i t to s tand for two hours . Then, add 100 ml acetone and homogenize the mixture for three minutes Fi l ter the homogenized sample by suct ion in to a rotary vacuum evaporator through a f i l ter paper covered wi th a 1-cm thick layer of d ia tomaceous ear th . Col lec t the residue on the surface of the f i l ter paper and add 50 ml of acetone, and then homogenize the mixture for three minutes . Repeat the above procedure and combine the f i l t ra te in to the rotary vacuum evaporator, and then concentra te the mixture to approximate ly 30 ml a t 40°C or lower. Transfer the concentra ted solut ion to a 300-ml separa t ing funnel a l ready conta ining 100 ml of 10% sodium chlor ide solut ion. Wash the eggplant-shaped f lask of the above rotary vacuum evaporator wi th 100 ml of n-hexane,and add the washings to the separa t ing Provisional Translation from the Japanese Original 25/100 funnel above. Shake the mixture v igorously for f ive minutes us ing a shaker and leave i t to s tand, and then t ransfer the n-hexane layer to a 300-ml conical f lask . Add 50 ml of n-hexane to the aqueous layer and repeat the above procedure , and add the n-hexane layer to the conical f lask above. Add an adequate amount of sodium sul fa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occas ional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of n-hexane and wash twice the res idue on the surface of the f i l ter paper wi th the washings . Add the washings in to the ro tary vacuum evaporator and remove the n-hexane at 40°C or lower. Add 30 ml of n-hexane to the residue and t ransfer the mixture to a 100-ml separa t ing funnel , and then add 30 ml of n-hexane-satura ted acetoni t r i l . Shake the mixture v igorously for f ive minutes us ing a shaker and leave i t to s tand, and then t ransfer the acetoni t r i le layer in to the ro tary vacuum evaporator. Add 30 ml of n-hexane-satura ted acetoni t r i le to the n-hexane layer and repeat the above procedure twice , and then combine the acetoni t r i le layer in to the rotary vacuum evaporator above. Remove the acetoni t r i le a t 40°C or lower and dissolve the res idue in 5 ml of n-hexane. i i . Frui t , vegetables , matcha and hops In case of f ru i t and vegetables , weigh accurate ly 1 kg of the test sample and add 500 ml of 10% phosphor ic ac id solut ion. Homogenize the mixture and mesure out a sample equiva lent to 20 .0 g . In case of matcha, weigh 5 .00 g of the te t sample and add 20 ml of 3% phosphor ic ac id solut ion, and then leave i t to s tand for two hours . In case of hops , weigh 5 .00 g of the tes t sample , previously ground, and add 20 ml of 3% phosphor ic ac id solut ion, and then leave i t to s tand for two hours . Then, add 100 ml of acetone and homogenize the mixture for three minutes . Fi l ter the homogenized sample by suct ion in to a ro tary vacuum evaporator through a f i l ter paper covered wi th a 1-cm thick layer of d ia tomaceous ear th . Col lec t the residue on the surface of the f i l ter paper and add 50 ml of acetone, and then homogenize the mixture for three minutes . Rpeat the above procedure and combine the f i l t ra te in to the rotary vacuum evaporator, and then concentra te Provisional Translation from the Japanese Original 26/100 the mixture to approximate ly 30 ml a t 40°C or lower. Transfer the concentra ted solut ion to a 300-ml separa t ing funnel a l ready conta ining 100 ml of 10% sodium chlor ide solut ion. Wash the eggplant-shaped f lask of the above rotary vacuum evaporator wi th 100 ml of n-hexane and add the washings to the separa t ing funnel above. Shake the mixture v igorously for f ive minutes using a shaker and leave i t to s tand, and then t ransfer the n-hexane layer to a 300-ml conical f lask . Add 50 ml of n-hexane to the aqueous layer and repeat the above procedure twice , and then combine the n-hexane layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occasional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of n-hexane and wash twice the res idue on the surface of the f i l ter paper. Add the washings into the ro tary vacuum evaporator and remove the n-hexane at 40°C or lower. Dissolve the res idue in 5 ml of n-hexane. i i i . Teas except matcha Weigh 9 .00 g of the tes t sample and soak i t in 540 ml of water a t 100°C, and then leave i t to s tand at room temperature for f ive minutes . Transfer 360 ml of the cooled f i l t ra te in to a 500-ml conical f lask . Add 30 ml of phosphor ic ac id , 100 ml of acetone and 2 ml of sa tura ted lead aceta te solut ion to the above conical f lask , and then leave the mixture to s tand for one hour at room temperature . Fi l ter the mixture by suct ion in to a 1 ,000-ml separa t ing funnel through a f i l ter paper covered wi th a 1-cm thick layer of d ia tomaceous ear th . Wash the above conical f lask with 50 ml of acetone and wash the res idue on the surface of the f i l ter paper wi th the washings . Add the washings in to the above separa t ing funnel and a lso add 30 g of sodium chlor ide and 100 ml of n-hexane. Shake the mixture v igorously for f ive minutes us ing a shaker and leave i t to s tand, and then t ransfer the n-hexane layer to a 300-ml conical f lask . Add 100 ml of n-hexane to the aqueous layer and repeat the above procedure, and then add the n-hexane layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occas ional shaking. Fi l ter the Provisional Translation from the Japanese Original 27/100 content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of n-hexane and wash twice the res idue on the surface of the f i l ter paper wi th the washings . Add the washings in to the rotary vacuum evaporator and remove the n-hexane a t 40°C or lower. Dissolve the res idue in 5 ml of n-hexane. iv. Foods except those l i s ted in i to i i i above Obtain the extracts according to the methods descr ibed in i or i i . b . Clean-up Add 5 g of f lor is i l for column chromatography suspended in n-hexane in to a chromatograph tube (15 mm in inner d iameter, 300 mm in length) and add over approximate ly 5 g of sodium sulfa te (anhydrous) in to the column. Spi l l out the n-hexane unt i l only a smal l amount remains on the packing of the column and pour the solut ion obtained by the ext rac t ion descr ibed in 4-a in to the column. Then, pour 100 ml of n-hexane in to the column and discard the eff luent . Pour 150 ml of e thyl acetate /n-hexane (1:9) in to the column and col lec t the eluate in to a ro tary vacuum evaporator, and then romove the e thyl acetate and n-hexane a t 40°C or lower. Dissolve the res idue in n-hexane to make exact ly 5 ml of solut ion, which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard. Tes t ing condi t ions 1 Column: A s i l icate g lass capi l lary column (0 .25 mm inner d iameter, 10-30 m in length) coated wi th methyl s i l icone for gas chromatography to a th ickness of 1 .5 µm. Column temperature : Hold the column temperature a t 50°C for one minute , fo l lowed by an increase of 25°C every minute unt i l reaching 175°C, af ter which increase the temperature by 10°C every minute unt i l reaching 300°C, and hold for f ive minutes . In le t temperature : 230°C Detector : Operate at 300°C Gas f low ra te : Use hel ium as the carr ier gas . Adjust the f low ra te to the opt imal condi t ion. Tes t ing condi t ions 2 Provisional Translation from the Japanese Original 28/100 Column: A s i l ica te g lass capi l lary column (0 .25 mm in inner d iameter, 10-30 m in length) coated wi th 5% phenyl methyl s i l icone for gas chromatography to a th ickness of 0 .25 µm. Column temperature : Hold the column temperature a t 50°C for one minute , fo l lowed by an increase of 25°C every minute unt i l reaching 125°C, af ter which increase the temperature by 10°C every minute unt i l reaching 300°C, and hold for three minutes . In le t temperature : 230°C Detector : Operate at 300°C Gas f low ra te : Use hel ium as the carr ier gas . Adjust the f low ra te to the opt imal condi t ion. b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the same condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. c . Conf i rmat ion tes ts Perform gas chromatography/mass spect rometry under the same condi t ions descr ibed in 5-a . Tes t resul ts obtained must be the same as those obta ined for the reference s tandard. Determine the quant i ty us ing e i ther the peak height or peak area method, i f required. (7) Carbadox analyt ica l method Analyze quinoxal ine-2-carboxyl ic ac id . 1 . Apparatus A high-performance l iquid chromatograph wi th an ul t raviole t spect rophotometr ic detector (HPLC-UV) and a l iquid chromatograph/ mass spectrometer (LC/MS or LC/MS/MS) 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Reagents des ignated as “specia l grade” in th is sect ion must meet the requirements for “specia l grade” specif ied in the Japan Indust r ia l Standards for the reagents . Octadecylsi lane-bonded s i l ica gel car t r idge column (500 mg): A polyethylene column of 8-9 mm in inner d iameter packed wi th 500 mg of octadecyls i lane-bonded s i l ica gel , or a column equivalent to the speci f ied one in separa t ion capabi l i ty. St rongly bas ic anion exchanger car t r idge column (360 mg): A Provisional Translation from the Japanese Original 29/100 polyethylene column of 8-9 mm in inner diameter packed wi th 360 mg of t r imethylamino acrylamide copolymer s i lane-bonded s i l ica gel , or a column equivalent to the specif ied one in separa t ion capabi l i ty. Water : Water produced for l iquid chromatography. Sodium sul fa te (anhydrous) : Sodium sulfa te (anhydrous)(specia l grade) . I f the reagent i s found to include any substance that may interfere wi th analys is of the target composi t ional substances f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Phosphate buffer solut ion (pH 2.5) : Dissolve monopotass ium dihydrogen monophosphate (1 .36 g) in water to make a 800-ml solut ion, and add phosphor ic ac id to adjus t the pH to 2.5 . Add water to the adjus ted solut ion to make a 1 ,000-ml solut ion. 3 . Reference s tandard Quinoxal ine-2-carboxyl ic ac id: This product conta ins not less than 99% of quinoxal ine-2-carboxyl ic ac id , and i ts mel t ing point i s 208°C. 4 . Procedure a . Extract ion Weigh 5.00 g of the tes t sample , previously ground. . For muscle , remove the fa t layer as much as poss ible before gr inding. Then, add 100 of methanol /0 .3% metaphosphor ic ac id solut ion (3:7) and homogenize the mixture . Fi l ter the homogenized sample by suct ion in to a ro tary vacuum evaporator through a f i l ter paper covered with a 2-mm thick layer of d ia tomaceous ear th . Wash the res idue on the surface of the f i l ter paper wi th 10 ml of methanol /0 .3% metaphosphor ic ac id solut ion (3:7) and f i l ter the washings , in to the rotary vacuum evaporator. Concentra te the mixture to 30 ml a t 45°C or lower and add 0 .1 ml of phosphor ic ac id. b . Clean-up i . Octadecyls i lane-bonded s i l ica gel column chromatography Add 5 ml of methanol to an octadecyls i lane-bonded s i l ica gel car t r idge column (500 mg) fol lowed by 10 ml of water and discard the eff luent . Pour the solut ion obta ined by the ext rac t ion descr ibed in 4-a in to the column fol lowed by 20 ml of phosphate buffer solut ion (pH 2.5) and discard the eff luent . Pour 10 ml of methanol in to the column and col lec t the eluate in a ro tary vacuum evaporator, and then remove the methanol a t 40°C or lower. Dissolve the res idue in 5 ml of water. Provisional Translation from the Japanese Original 30/100 i i . St rongly bas ic anion exchanger column chromatography Pour 5 ml of water into a s t rongly bas ic anion exchanger car t r idge column (360 mg) and discard the eff luent . Pour the solut ion obtained in 4-b- i in to the column fol lowed by 2 ml of water, 10 ml of e thanol and 5 ml of water, and discard the eff luent . Pour 5 ml of 0 .1 mol / l hydrochlor ic ac id in to the column and col lect the e luate in a 50-ml tes t tube . Add 3 ml of 3 mol / l hydrochlor ic acid and 15 ml of e thyl aceta te and shake the mixture v igorously for f ive minutes us ing a shaker, and then leave i t to s tand. Transfer the e thyl aceta te layer in to a 100-ml conical f lask . Add 15 ml of e thyl aceta te to the aqueous layer in the tes t tube and repeat the above procedure , and then add the e thyl aceta te layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occas ional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator and remove the e thyl acetate at 40°C or lower. Dissolve the res idue in 1 .0 ml of acetoni t r i le /phosphate buffer solut ion (pH 2.5) (1 :4) , which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard. Test ing condi t ions Column packing: Octadecyls i lane-bonded s i l ica gel (5 µm in par t ic le s ize) . Column: A s ta in less tube (4 .0-6 .0 mm in inner d iameter, 150 mm in length) . Column temperature : 40°C Detector : Operate wi th an absorpt ion wavelength of 245 nm. Mobile phase: Use acetoni t r i le /phosphate buffer solut ion (pH 2.5) (1 :4) . Adjust the f low ra te so that quinoxal ine-2-carboxyl ic ac id f lows out in approximately 10 minutes . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the same condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. Provisional Translation from the Japanese Original 31/100 c . Conf i rmat ion tes ts Perform l iquid chromatography/mass spect rometry under the same condi t ions descr ibed in 5-a , but the mobi le phase should be acetoni t r i l /water (1 :4) . Test resul ts obta ined must be the same as those obta ined for the reference s tandard. Determine the quant i ty us ing e i ther the peak height or peak area method, i f required. (8) Coumaphos analyt ica l method 1 . Apparatus A gas chromatograph wi th an a lkal i f lame ionizat ion detector (GC-FID), a f lame photometr ic detector (GC-FPD, in terference f i l ter for phosphorus determinat ion, wavelength: 526 nm), or a h ighly-sensi t ive n i t rogen phosphorus detector (GC-NPD), and a gas chromatograph/mass spectrometer (GC/MS or GC/MS/MS). 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Reagents des ignated as “specia l grade” in th is sect ion must meet the requirements for “specia l grade” specif ied in the Japan Indust r ia l Standards for the reagents . Acetoni t r i le : Use a ro tary vacuum evaporator on 300 mL of acetoni t r i le to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Acetone: Use a ro tary vacuum evaporator on 300 mL of acetone to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Sodium chlor ide: Sodium chlor ide (specia l grade) . I f the reagent i s found to include any substance that may in ter fere wi th the analysis of substances to be analyzed f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Si l ica gel for column chromatography (63-200 µm in par t ic le s ize) : Heat s i l ica gel made for column chromatography (63-200 µm in par t ic le s ize) a t 130°C for 12 hours or longer. Cool down to room temperature in a des iccator. Provisional Translation from the Japanese Original 32/100 Dia tomaceous ear th : Diatomaceous ear th for chemical analys is . Ethyl acetate : Use a ro tary vacuum evaporator on 300 mL of e thyl aceta te to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion in to a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11g . n-Hexane: Use a ro tary vacuum evaporator on 300 mL of n -hexane to evaporate unt i l 5 mL is lef t . For analys is , in ject 5 μL of the solut ion in to a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11g . Water : Dis t i l led water. I f the dis t i l led water i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Sodium sul fa te (anhydrous) : Sodium sulfa te (anhydrous)(specia l grade) . I f the reagent i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tural chemicals , wash wi th a solvent such as n -hexane before use . 3 . Reference s tandard Coumaphos: This product contains not less than 98% of coumaphos , and i t s mel t ing point is 95°C. 4 . Procedure a . Extract ion i . Cereal gra ins , legumes/pulses and seeds Weigh 10.0 g of the tes t sample , previously ground so as to pass through a s tandard mesh s ieve (420 µm). Add 20 ml of water to the obtained sample and leave i t to s tand for two hours . Then, add 100 ml of acetone and homogenize the mixture for three minutes . Fi l ter the homogenized sample by suct ion in to a ro tary vacuum evaporator through a f i l ter paper covered wi th a 1-cm thick layer of d ia tomaceous ear th . Col lec t the res idue on the surface of the f i l ter paper and add 50 ml of acetone, and then homogenize the mixture for three minutes . Repeat the above procedure and combine the f i l t ra te in to the rotary vacuum evaporator, and then concentra te the mixture to approximate ly 30 ml a t 40°C or lower. Provisional Translation from the Japanese Original 33/100 Transfer the concentra ted solut ion to a 300-ml separa t ing funnel a l ready conta ining 100 ml of sa tura ted sodium chlor ide solut ion. Wash the eggplant-shaped f lask of the above rotary vacuum evaporator wi th 100 ml of e thyl aceta te/n-hexane (1:4) and add the washings to the separa t ing funnel above. Shake the mixture v igorously for f ive minutes us ing a shaker and leave i t to s tand, and then t ransfer e thyl aceta te and n-hexane layers to a 300-ml conical f lask . Add 50 ml of e thyl acetate /n-hexane (1:4) to the aqueous layer and repeat the above procedure , and then add e thyl acetate and n-hexane layers to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes with occasional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of n-hexane and wash twice the res idue on the surface of the f i l ter paper wi th the washings . Add the washings in to the rotary vacuum evaporator and remove the ethyl acetate and n-hexane a t 40°C or lower. Add 30 ml of n-hexane to the residue and t ransfer the mixture to a 100-ml separa t ing funnel , and then add 30 ml of n-hexane-satura ted acetoni t r i le . Shake the mixture v igorously for f ive minutes us ing a shaker and leave i t to s tand, and then t ransfer the acetoni t r i le layer to the ro tary vacuum evaporator. Add 30 ml of n-hexane-satura ted acetoni t r i le to the n-hexane layer and repeat the above procedure twice , and then combine the acetoni t r i le layer in to the rotary vacuum evaporator. Remove the acetoni t r i le a t 40°C or lower and dissolve the res idue in 5-ml of acetone/n-hexane (1:1) . i i . Frui t , vegetables , teas and hops In case of f ru i t and vegetables , weigh accurate ly 1 kg of the test sample and add an appropr iate amount of water, i f required. Homogenize the mixture and mesure out a sample equiva lent to 20 .0 g . In case of teas , weigh 5 .00 g of the tes t sample and add 20 ml of water, and then leave i t to s tand for two hours . In case of hops , weigh 5 .00 g of the tes t sample , previously ground, and add 20 ml of water, and then leave i t to s tand for two hours . Then, add 100 ml of acetone and homogenize the mixture for three minutes . Fi l ter the homogenized sample by suct ion in to a ro tary vacuum evaporator through a f i l ter paper covered wi th a 1-cm thick Provisional Translation from the Japanese Original 34/100 layer of d ia tomaceous ear th . Col lec t the res idue on the surface of the f i l ter paper and add 50 ml of acetone and homogenize the mixture for three minutes . Repeat the above procedure and combine the f i l t ra te in to the rotary vacuum evaporator, and then concentra te the mixture to approximate ly 30ml a t 40℃ or lower. Transfer the concentra ted solut ion to a 300-ml separa t ing funnel a l ready conta ining 100 ml of sa tura ted sodium chlor ide solut ion. Wash the eggplant-shaped f lask of the above rotary vacuum evaporator wi th 100 ml of e thyl aceta te/n-hexane (1:4) and add the washings to the separa t ing funnel above. Shake the mixture v igorously for f ive minutes us ing a shaker and leave the i t to s tand, and then t ransfer the e thyl acetate and n-hexane layers to a 300-ml conical f lask . Add 50 ml of e thyl aceta te/n-hexane (1:4) to the aqueous layer and repeat the above procedure twice, and then combine the e thyl aceta te /n-hexane lyers to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occas ional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of n-hexane and wash twice the res idue on the surface of the f i l ter paper. Add the washings in to the rotary vacuum evaporator and remove the ethyl aceta te and n-hexane a t 40°C or lower. Dissolve the res idue in 5 ml of acetone/n-hexane (1 :1) . i i i . Foods except those l i s ted in i and i i above Obtains the extracts according to the methods descr ibed in i or i i . b . Clean-up Add 5 g of s i l ica gel for column chromatography (63-200 µm in par t ic le s ize) suspended in acetone/n-hexane (1 :1) into a chromatograph tube (15 mm in inner d iameter, 300 mm in length) and add over approximate ly 5 g of sodium sulfa te (anhydrous) into the column. Pour acetone/n-hexane (1 :1) into the column unt i l only a smal l amount remains on the packing of the column and pour the solut ion obta ined by the ext ract ion descr ibed in 4-a in to the column. Pour 100 ml of acetone/n-hexane (1:1) in to the column and col lec t the e luate in to a ro tary vacuum evaporator, and then remove the acetone and n-hexane at 40°C or lower. Dissolve the res idue in acetone to make exact ly 5 ml of solut ion, which is used as the sample solut ion. 5 . Determinat ion Provisional Translation from the Japanese Original 35/100 a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard. Tes t ing condi t ions 1 Column: A s i l ica te g lass capi l lary column (0 .53 mm in inner d iameter, 10-30 m in length) coated wi th methyl s i l icone for gas chromatography to a th ickness of 1 .5 µm Column temperature : Hold the column temperature a t 80°C for one minute , fo l lowed by an increase of 8°C every minute unt i l reaching 250°C, and hold for f ive minutes . In le t temperature : 230°C Detector : Operate at 280°C Gas f low ra te : Use hel ium as the carr ier gas . Adjust the f low ra te to the opt imal condi t ion. Adjus t the f lows of a i r and hydrogen to the opt imal condi t ions . Tes t ing condi t ions 2 Column: A s i l ica te g lass capi l lary column (0 .32 mm in inner d iameter, 10-30 m in length) coated wi th 50% tr i f luoro propyl methyl s i l icone for gas chromatography to a thickness of 0 .25 µm Column temperature : Hold the column temperature a t 70°C for one minute , fo l lowed by an increase of 25°C every minute unt i l reaching 125°C, af ter which increase the temperature by 10°C every minute unt i l reaching 235°C, and hold for 12 minutes . In le t temperature : 230°C Detector : Operate at 280°C Gas f low ra te : Use hel ium as the carr ier gas . Adjust the f low ra te opt imal condi t ion. Adjus t the f lows of a ir and hydrogen to the opt imal condi t ions . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the same condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. c . Conf i rmat ion tes ts Perform gas chromatography/mass spect rometry under the same condi t ions descr ibed in 5-a . Tes t resul ts obtained must be the same as those obta ined for the reference s tandard. Determine the quant i ty Provisional Translation from the Japanese Original 36/100 us ing e i ther the peak height or peak area method, i f required. (9) Chloramphenicol analyt ica l method 1 . Apparatus A l iquid chromatograph/mass spect rometer (LC/MS or LC/MS/MS) 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solut ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Acetoni t r i le : Acetoni tr i le produced for l iquid chromatography. Divinylbenzene-N -v inylpyrrol id ine copolymer car t r idge column (60 mg): A polyethylene column of 12-13 mm in inner d iameter packed wi th 60 mg of d ivinylbenzen-N -vinylpyrrol idone copolymer, or a column equivalent to the speci f ied one in separa t ion capabi l i ty. Divinylbenzene-N -vinylpyrrol id ine copolymer car t r idge column (200 mg): A polyethylene column of 12-13 mm in inner d iameter packed wi th 200 mg of d ivinylbenzen-N -v inylpyrrol idone copolymer, or a column equivalent to the speci f ied one in separa t ion capabi l i ty. Water : Water produced for l iquid chromatography. 3 . Reference s tandard Chloramphenicol : This product conta ins not less than 99% of chloramphenicol , and i t s decomposi t ion point i s 208°C. 4 . Procedure a . Extract ion i . Honey Weigh 5 .00 g of the tes t sample , previously homogenized, and dissolve i t in 20 ml of water. i i . Royal je l ly Weigh 1.00 g of the tes t sample , previously homogenized, add 60 ml of methanol /1% metaphosphor ic ac id (3 :2) , and homogenize the mixture again . Fi l ter the homogenized sample by suct ion in to a ro tary vacuum evaporator through a f i l ter paper covered wi th a 2-mm thick layer of d ia tomaceous ear th . Wash the residue on the f i l ter paper wi th 15 ml of methanol /1% metaphosphor ic ac id (3:2) and f i l ter the washings by suct ion. Combine the f i l t ra tes in to the ro tary vacuum evaporator and concentra te the mixture to 2 ml a t 45ºC or lower. i i i . Foods other than those given in sect ions i and i i Provisional Translation from the Japanese Original 37/100 Weigh 5 .00 g of the tes t sample , previously ground. For muscle , remove the fa t layer as much as poss ible before gr inding. Add 100 ml of methanol /1% metaphosphoric ac id (3 :2) and homogenize the mixture again . Fi l ter the homogenized sample by suct ion in to a ro tary vacuum evaporator through a f i l ter paper covered wi th a 2-mm thick layer of d ia tomaceous ear th . Wash the residue on the f i l ter paper wi th 10 ml of methanol /1% metaphosphor ic ac id (3:2) and f i l ter the washings by suct ion. Combine the f i l t ra tes in to the rotary vacuum evaporator and cocentra te the mixture to 30 ml a t 45ºC or lower. b . Clean-up i . Honey Pour 5 ml of methanol and 5 ml of water in to a d ivinylbenzene-N v inylpyrrol id ine copolymer car t r idge column (60 mg) and discard the eff luent . Pour the solut ion obta ined by the ext rac t ion descr ibed in 4-a in to the column fol lowed by 5 ml of 20% (vol) methanol , and discard the eff luent . Pour 6 ml of 60% (vol) methanol and col lect the e luate in to a ro tary vacuum evaporator and remove the methanol and water a t 45°C or lower. Dissolve the res idue in 1 .0 ml of acetoni t r i le /water (3 :7) , which is used as the sample solut ion. i i . Royal je l ly Pour 10 ml of methanol and 10 ml of water into a d ivinylbenzene-N v inylpyrrol id ine copolymer car t r idge column (200 mg) and discard the eff luent . Pour the solut ion obta ined by the ext ract ion descr ibed in 4-a in to the column fol lowed by 4 ml of water and 4 ml of 5% (vol) methanol and discard the eff luent . Pour 10 ml of 60% (vol) methanol and col lect the e luate into a ro tary vacuum evaporator, and remove the methanol and water a t 45°C or lower. Dissolve the res idue in 1 .0 ml of acetoni t r i le /water (3 :7) , which is used as the sample solut ion. i i i . Foods other than those given in sect ions i and i i Pour 5 ml of methanol and 5 ml of water in to a d ivinylbenzene-N v inylpyrrol id ine copolymer car t r idge column (60 mg) and discard the eff luent . Pour the solut ion obta ined by the ext rac t ion descr ibed in 4-a in to the column fol lowed by 10 ml of water, and discard the eff luent . Pour 10 ml of methanol and col lec t the eluate in to a ro tary vacuum evaporator and remove the methanol a t 40°C or lower. Provisional Translation from the Japanese Original 38/100 Dissolve the res idue in 1 .0 ml of acetoni t r i le /water (3 :7) , which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fo l lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard. Test ing condi t ions Column packing: Octadecyls i lanized s i l ica gel (2–5 µm in par t ic le s ize) . Column: A s ta inless s tee l tube (2 .0–6.0 mm in inner d iameter, 100–250 mm in length) . Column temperature : 40°C Mobile phase: Use acetoni t r i le /10 mmol/ l ammonium formate (3 :7) . Adjus t the f low ra te so that chloramphenicol f lows out in approximate ly 5 minutes . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the same condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. (10) Chlorpromazine analyt ical method 1 . Apparatus A l iquid chromatograph/ tandem mass spect rometer (LC/MS/MS). 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Acetoni t r i le : Acetoni tr i le produced for l iquid chromatography. St rongly ac idic ca t ion exchanger car t r idge column (500 mg): A polyethylene column of 8-9 mm in inner d iameter packed wi th 500 mg of benzenesulfonyl propyl s i lane-bonded s i l ica gel , or a column equivalent to the specif ied one in separat ion capabi l i ty. Water : Water produced for l iquid chromatography. Methanol : Methanol produced for l iquid chromatography. 3 . Reference s tandard Chlorpromazine hydrochlor ide: This product contains not less than 98% of chlorpromazine hydrochlor ide , and i t s mel t ing point i s Provisional Translation from the Japanese Original 39/100 194-196°C. 4 . Procedure a . Extract ion Weigh 5 .00 g of the tes t sample , previously ground, and homogenize i t wi th 25 ml of e thyl aceta te and 1 ml of 4 mol / l potass ium carbonate solut ion. Centr i fuge the mixture a t 3 ,000 rpm for f ive minutes And t ransfer the e thyl acetate layer in to a ro tary vacuum evaporator. Add 25 ml of e thyl acetate to the res idue and repeat the above procedure and t ransfer the ethyl aceta te layer in to the rotary vacuum evaporator, and then remove the e thyl acetate a t 40°C or lower. Add 30 ml each of acetoni t r i le and acetoni t r i le-sa tura ted n-hexane to the res idue and shake the mixture v igorously for f ive minutes us ing a shaker and leave i t to s tand, and then t ransfer the acetoni t r i le layer to a 100-ml separa t ing funnel . Add 30 ml of acetoni t r i le-sa tura ted n-hexane and repeat the above procedure . Col lec t the acetoni t r i le layer in to the rotary vacuum evaporator and remove the acetoni t r i le a t 40°C or lower. Dissolve the res idue in 10 ml of methanol /1 .2% metaphosphor ic ac id solut ion (2:3) and f i l ter the mixture through a cot ton plug. b . Clean-up Pour 3 ml of methanol fo l lowed by 3 ml of water in to a s t rongly ac idic cat ion exchanger car t r idge column (500 mg) and discard the eff luent . Pour the solut ion obta ined by the ext rac t ion descr ibed in 4-a in to the column fol lowed by 5 ml of water and discard the eff luent . Pour 15 ml of methanol /0 .1 mol / l dipotass ium hydrogen or thophosphate solut ion (9:1) to the column and col lec t the eluate in to a ro tary vacuum evaporator, and then remove the water and methanol a t 40°C or lower. Dissolve the res idue in 1 .0 ml of methanol , which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard. Test ing condi t ions Column packing: Octadecyls i lane-bonded s i l ica gel (2-5 µm in par t ic le s ize) . Provisional Translation from the Japanese Original 40/100 Column: A s ta in less tube (2 .0-6 .0 mm in inner d iameter, 100-250 mm in length) . Column temperature : 40°C Mobile phase: Use acetoni t r i le / formic acid/water (500:1:500) . Adjus t the f low ra te so that chlorpromazine f lows out in approximate ly 15 minutes . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the same condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. (11) Diethyls t i lbes t rol analyt ica l method 1 . Apparatus A l iquid chromatograph/ tandem mass spect rometer (LC/MS/MS) 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Reagents des ignated as “specia l grade” in th is sect ion must meet the requirements for “specia l grade” specif ied in the Japan Indust r ia l Standards for the reagents . Acetoni t r i le : Acetoni tr i le produced for l iquid chromatography. Glucuronidase solut ion: Should conta in 100,000 uni ts /ml of beta-D glucuronidase ext racted f rom Helix pomatia and ref ined. Dichloromethane: Dichloromethane (specia l grade) . Weakly bas ic anion exchanger car t r idge column (500 mg): A polyethylene column of 8-9 mm in inner d iameter packed wi th 500 mg of d ie thylaminopropyl weakly bas ic anion exchanger produced for column chromatography, or a column equivalent to the speci f ied one in separat ion capabi l i ty. Water : Water produced for l iquid chromatography. Sodium sulfa te (anhydrous) : Sodium sulfa te (anhydrous)(specia l grade) . I f the reagent i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tural chemicals , wash wi th a solvent such as d ichloromethane before use . . 3 . Reference s tandard Die thyls t i lbes t ro l : This product conta ins not less than 99% of d ie thyls t i lbes t rol , and i t s decomposi t ion point i s 208°C. 4 . Procedure Provisional Translation from the Japanese Original 41/100 a . Extract ion Weigh 5 .00 g of the tes t sample , previously ground, and homogenize i t wi th 11 ml of 0 .04 mol / l sodium aceta te solut ion, and then add acet ic ac id to adjus t the pH to 4 .25-4.75. Add 100 µl of g lucuronidase solut ion to the mixture and leave i t to s tand for 14 hours a t 37°C. Add 16 ml of acetoni t r i le and shake the mixture vigorously for f ive minutes us ing a shaker. Centr i fuge the mixture a t 3 ,000 rpm for f ive minutes and t ransfer the supernatant in to a ro tary vacuum evaporator. Add 16 ml of acetoni t r i le to the res idue and repeat the above procedure twice and t ransfer the supernatant in to the rotary vacuum evaporator, and then concentra te the solut ion to approximate ly 3 ml a t 50°C or lower. Add 50 ml of d ichloromethane and 200 ml of 5% sodium chlor ide solut ion to the concentra ted solut ion , and then shake i t v igorously for f ive minutes us ing a shaker. Col lect the d ichloromethane layer and dehydrate wi th sodium sulfa te (anhydrous) . Add 50 ml of d ichloromethane to the aqueous layer and repeat the above procedure twice, and then combine the dichloromethane layer. Add an adequate amount of sodium sulfa te (anhydrous) and leave i t to s tand for 15 minutes wi th occas ional shaking. Fi l ter the content into a ro tary vacuum evaporator and remove the dichloromethane at 40°C or lower. Dissolve the res idue is in 4 ml of n-hexane/benzene (3:1) . b . Clean-up Pour 10 ml of n-hexane/benzene (3 :1) in to a weakly basic anion exchanger car t r idge column (500 mg) and discard the eff luent . Pour the solut ion obta ined by the ext rac t ion descr ibed in 4-a in to the column fol lowed by 2 ml of n-hexane/benzene (3:1) and 4 ml of d ichloromethane, and discard the eff luent . Pour 8 ml of d ichloro methane/methanol (9 :1) in to the column and col lec t the e luate in to a ro tary vacuum evaporator, and then remove the d ichloromethane and methanol a t 40°C or lower. Dissolve the res idue in 0 .5 ml of acetoni t r i le /water (1 :1) , which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard. Test ing condi t ions Provisional Translation from the Japanese Original 42/100 Column packing: Octadecyls i lane-bonded s i l ica gel (2-5 µm in par t ic le s ize) . Column: A s ta in less tube (2 .0-6 .0 mm in inner d iameter, 100-250 mm in length) . Column temperature : 40°C Mobile phase: Use acetoni t r i le /0 .002 mol / l ammonium aceta te (1:1) . Adjust the f low ra te so that d ie thyls t i lbes t rol f lows out in approximate ly seven minutes . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the same condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. (12) Analyt ica l method for d imetr idazole , metronidazole and ronidazole 1 . Apparatus A l iquid chromatograph/ tandem mass spect rometer (LC/MS/MS) 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Acetoni t r i le : Acetoni tr i le produced for l iquid chromatography. Methanol : Methanol produced for l iquid chromatography. Water : Water produced for l iquid chromatography. 3 . Reference s tandard Dimetr idazole : This product contains not less than 99% of d imetr idazole , and i t s mel t ing point i s 138-139°C. Metronidazole : This product conta ins not less than 99% of metronidazole , and i t s mel t ing point i s 158-160°C. Ronidazole: This product conta ins not less than 99% of ronidazole , and i t s mel t ing point i s 167-169°C. 4 . Procedure Weigh 5 .00 g of the tes t sample , previously ground, and homogenize i t wi th 20 ml of acetoni t r i le . Centr i fuge the mixture a t 3 ,000 rpm for f ive minutes and t ransfer the acetoni t r i le layer to a 100-ml separat ing funnel . Add 20 ml of n-hexane and shake the mixture vigorously, and then leave i t to s tand. Transfer the acetoni t r i le layer in to a rotary vacuum evaporator and add 5 ml of n-propanol , and then remove the acetoni t r i le and n-propanol a t 40°C or lower. Dissolve the res idue in Provisional Translation from the Japanese Original 43/100 1 .0 ml of methanol and f i l te r the mixture through a membrane f i l ter wi th a pore s ize of 0 .2 µm, which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard. Test ing condi t ions Column packing: Octadecyls i lane-bonded s i l ica gel (2-5 µm in par t ic le s ize) . Column: A s ta in less tube (2 .0-6 .0 mm in inner d iameter, 100-250 mm in length) . Column temperature : 40°C Mobile phase: Use acetoni t r i l /water (1 :9) . Adjust the f low ra te so that d imetr idazole , metronidazole and ronidazole f low out in 4-10 minutes . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the same condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. (13) Daminozide analyt ica l method 1 . Apparatus A gas chromatograph wi th an a lkal i f lame ionizat ion detector (GC-FID) or h ighly-sensi t ive n i t rogen phosphorus detector (GC-NPD), a gas chromatograph/mass spectrometer (GC/MS or GC/MS/MS), and s team dis t i l la t ion apparatus . The steam dis t i l la t ion apparatus should be made of g lass and roughly as shown in the fo l lowing f igure: A: 1 ,000-ml round bot tom f lask ( for s team generat ion) B: 1 ,000-ml round bot tom f lask ( for d is t i l la t ion) C: Dis t i l la t ion t rap D: Cool ing tube E: 100-ml conical f lask 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those Provisional Translation from the Japanese Original 44/100 l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Reagents des ignated as “specia l grade” in th is sect ion must meet the requirements for “specia l grade” specif ied in the Japan Indust r ia l Standards for the reagents . Acetone: Use a ro tary vacuum evaporator on 300 mL of acetone to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Basic a lumina car t r idge column (1 ,710 mg): A polyethylene column of 8-9 mm in inner d iameter packed wi th 1 ,710 mg of bas ic a lumina, or a column equivalent to the speci f ied one in separa t ion capabi l i ty. Si l icone for defoaming: Si l icone produced for defoaming. ο -Ni t robenzaldehyde: ο -Ni t robenzaldehyde (specia l grade) . 1% ο -Ni t robenzaldehyde-methanol so lu t ion : Disso lve ο -n i t robenzaldehyde (100 mg) in 10 ml of methanol . Prepare immediate ly before use . Phenolphthale in reagent : Dissolve phenolphthalein (1 g) in 100 ml of e thanol . n-Hexane: Use a ro tary vacuum evaporator on 300 mL of n -hexane to evaporate unt i l 5 mL is lef t . For analys is , in ject 5 μL of the solut ion in to a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11g . Water : Dis t i l led water. I f the dis t i l led water i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Methanol : Use a ro tary vacuum evaporator on 300 mL of methanol to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Phosphate buffer solut ion (pH 5) : Dissolve monopotassium dihydrogen monophosphate (13.15 g) and dipotass ium hydrogen orthophosphate (0 .59 g) in water to make a 100-ml solut ion. 3 . Reference s tandard Dimethylhydrazine: This product conta ins not less than 97% of Provisional Translation from the Japanese Original 45/100 1 ,1-dimethylhydrazine , and i t s boi l ing point i s 62-64°C. 4 . Procedure a . Extract ion i . Cereal gra ins , legumes/pulses , seeds , f ru i t , vegetables , matcha and hops In case cereal gra ins , legumes/pulses and seeds , weigh 5 .0 g of test samples so as to pass through a s tandard mesh s ieve (420 µm). In case f ru i t and vegetables , weigh accurate ly 1 kg of the tes t sample and add an appropr ia te amount of water to the sample , i f required. Homogenize the mix ture and measure out a sample equivalent to 10.0 g . In case of matcha, weigh 5 .0 g of the tes t sample . In case of hops , weigh 5 .0 g of the tes t sample , previously ground. Then, add 80 ml of water to the obta ined sample and shake i t v igorously for 30 minutes , and then f i l ter the mixture by suct ion through a glass f iber f i l ter. Col lect the res idue on the f i l ter and add 40 ml of water, and then shake the mixture for f ive minutes . Repeat the above procedure and combine the f i l t ra te to a 1 ,000-ml round bot tom f lask ( for d is t i l la t ion) . i i . Teas except matcha Weigh 6 .0 g of the tes t sample and soak i t in 360 ml of water a t 100°C, and then leave i t to s tand a t room temperature for f ive minutes . Transfer 120 ml of the cooled f i l t ra te in to a 1 ,000-ml round bot tom f lask ( for d is t i l la t ion) . i i i . Foods except those l i s ted in i and i i above Obtain the extracts according to the methods descr ibed in i . b . Dis t i l la t ion Add 60 g of sodium hydroxide (65 g for vegetables and f rui t ) b i t by bi t in to the round bot tom f lask descr ibed above wi th the f lask water cool ing. Add 1-2 drops of defoaming s i l icone to the d issolved solut ion and a t tach the f lask to a d is t i l la t ion apparatus immediately. Separa te ly, a t tach a 100-ml conical f lask containing 5 ml of phosphate buffer solut ion (pH 5) and one drop of phenolphthalein reagent to a s team dis t i l le r and heat a 1 ,000-ml round bot tom f lask ( for s team generat ion). Dis t i l l the solut ion unt i l the dis t i l la te comes to 45 ml and conf i rm that the dis t i l la te remains color less . Adjust the heater so as to complete the d is t i l la t ion in about 15 minutes . Provisional Translation from the Japanese Original 46/100 c . Der ivat iza t ion Add 1 ml of 1% ο -n i t robenzaldehyde-methanol solut ion to the above dis t i l la te and shake the mixture , and then leave i t to s tand for two hours at 30°C. Add 50 ml of n-hexane and shake the mixture for f ive minutes , and then leave i t to s tand. Col lect the n-hexane layer and f i l ter in a 200-ml eggplant-shaped f lask through a l iquid phase separa t ion f i l ter paper. Add 50 ml of n-hexane to the aqueous layer and repeat the above procedure , and then combine the n-hexane layer to the eggplant-shaped f lask above. Wash the res idue on the f i l ter wi th 10 ml of n-hexane and combine the washings to the eggplant -shaped f lask above, and then remove the n-hexane a t 40°C or lower. Dissolve the res idue in 5 ml of acetone/n-hexane (1:19) . d . Clean-up Pour 10 ml of acetone/n-hexane (1 :19) in to a bas ic a lumina car t r idge column (1 ,710 mg) and discard the eff luent . Pour the solut ion obtained in 4-c in to the column fol lowed by 10 ml of acetone/ n-hexane (1:19) and col lect the e luate in to a ro tary vacuum evaporator, and then remove the acetone and n-hexane a t 40°C or lower. Dissolve the res idue in acetone to make exact ly 5 ml of solut ion, which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard under the procedure descr ibed in 4-c . Test ing condi t ions Column: A s i l ica te g lass capi l lary column (0 .25 mm in inner d iameter, 10-30 m in length) coated wi th 5% phenyl methyl s i l icone for gas chromatography to a th ickness of 0 .25 µm Column temperature : Hold the column temperature a t 60°C for two minutes , fo l lowed by an increase of 10°C every minute unt i l reaching 280°C, and hold for f ive minutes . In le t temperature : 280°C Detector : Operate at 280°C. Gas f low ra te : Use hel ium as the carr ier gas . Adjust the f low ra te so that dimethylhydrazine der ivat ives f low out in approximate ly 13 minutes . Adjus t the f lows of a i r and hydrogen to the opt imal Provisional Translation from the Japanese Original 47/100 condi t ions . b . Quant i ta t ive tes ts Determine the content of d imethylhydrazine f rom the tes t resul ts obtained under the same condi t ions descr ibed in 5-a us ing e i ther the peak height or peak area method. Fur thermore , de termine the content of daminozide using the fo l lowing formula: Daminozide content (ppm) = 2 .67 x d imethylhydrazine content (ppm) c . Conf i rmat ion tes ts Perform gas chromatography/mass spect rometry under the same condi t ions descr ibed in 5-a . Test resul ts obta ined for the reference s tandard must be the same as those obta ined under the procedure descr ibed in 4-c . Determine the quant i ty us ing e i ther the peak height or peak area method, i f required. d . Cal ibra t ion curve Assay a solut ion of d imethylhydrazine (1 ml) combined wi th 5 ml of phosphate buffer solut ion (pH 5) and 40 ml of water wi th the same procedure descr ibed in 4-c . (14) Ni t rofurans analyt ica l method Analyse 3-amino-2-oxazol id inone, 1-aminohydantoin , 3-amino-5 morphol inomethyl-2-oxazol id inone and semicarbazide . 1 . Apparatus A l iquid chromatograph/ tandem mass spect rometer (LC/MS/MS) 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Reagents des ignated as “specia l grade” in th is sect ion must meet the requirements for “specia l grade” specif ied in the Japan Indust r ia l Standards for the reagents . Acetoni t r i le : Acetoni tr i le produced for l iquid chromatography. Porous diatomaceous ear th column ( to hold 20 ml of solut ion): A polyethylene column of 20-30 mm in inner d iameter packed wi th granular porous dia tomaceous ear th produced for column chromatography that can hold 20 ml of solut ion, or a column equivalent to the specif ied one in separat ion capabi l i ty. ο -Ni t robenzaldehyde: o -Ni t robenzaldehyde (specia l grade) . Water : Water produced for l iquid chromatography. Provisional Translation from the Japanese Original 48/100 3 . Reference s tandard 3-amino-2-oxazol id inone: This product conta ins not less than 99% of 3-amino-2-oxazol id inone, and i t s decomposi t ion point is 65-67°C. 1-aminohydantoin hydrochlor ide: This product conta ins not less than 90% of 1-aminohydantoin hydrochloride , and i t s decomposi t ion point i s 201-205°C. 3-amino-5-morphol inomethyl-2-oxazol id inone: This product conta ins not less than 99% of 3-amino-5-morphol inomethyl-2-oxazol id inone and i t s decomposi t ion point i s 115-120°C. Semicarbazide hydrochlor ide: This product conta ins not less than 99% of semicarbazide hydrochlor ide , and i t s decomposi t ion point i s 175-177°C. 4 . Procedure a . Extract ion Weigh 5 .00 g of the tes t sample , previously ground, and homogenize i t wi th 70 ml of 0 .1 mol / l hydrochlor ic acid . Centr i fuge the mixture at 2 ,500 rpm for f ive minutes and col lect the supernatant , and then add 0 .1 mol / l hydrochlor ic ac id to make 100 ml of solut ion. b . Der ivat iza t ion Transfer 10 ml of the solut ion obta ined by the ext rac t ion descr ibed in 4-a and add 0 .4 ml of 0 .05 mol / l ο -n i t robenzaldehyde-dimethyl sul foxide solut ion, and then leave i t to s tand for 16 hours a t 37°C. Add 5 ml of 0 .1 mol / l d ipotass ium hydrogen or thophosphate solut ion to the solut ion and add approximately 0 .8 ml of a 1 mol / l sodium hydroxide solut ion to adjus t the pH to 7-8 . When some res idue remains in the solut ion, centr i fuge a t 2 ,500 rpm for f ive minutes and col lec t the supernatant . b . Clean-up Pour the solut ion obtained by the der ivat iza t ion descr ibed in 4-b in to a porous dia tomaceous ear th column ( to hold 20 ml of solut ion) . Leave the column to s tand for f ive minutes and add 100 ml of e thyl aceta te .Collec t the eluate in to a ro tary vacuum evaporator and remove the e thyl aceta te a t 40°C or lower. Dissolve the res idue in 1 .0 ml of acetoni t r i le /water (1 :1) , which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts Provisional Translation from the Japanese Original 49/100 obtained must be the same as those obta ined for the reference s tandards under the procedure descr ibed in 4-b and 4-c. . Test ing condi t ions Column packing: Octadecyls i lane-bonded s i l ica gel (2-5 µm in par t ic le s ize) . Column: A s ta in less tube (2 .0-6 .0 mm in inner d iameter, 100-250 mm in length) . Column temperature : 40°C Mobile phase: Use a mixture of acetoni t r i le and 0 .1% acet ic ac id solut ion. Create a concentra t ion gradient of 1 :4 to 4:1 acetoni tr i le / 0 .1% acet ic acid solut ion in 15 minutes . Adjust the f low ra te so that 3-amino-2-oxazol id inone f lows out in approximate ly 12 minutes . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the same condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. (15) Propham analyt ica l method 1 . Apparatus A gas chromatograph/mass spectrometer (GC/MS or GC/MS/MS) 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Reagents des ignated as “specia l grade” in th is sect ion must meet the requirements for “specia l grade” specif ied in the Japan Indust r ia l Standards for the reagents . Acetoni t r i le : Use a ro tary vacuum evaporator on 300 mL of acetoni t r i le to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Acetone: Use a ro tary vacuum evaporator on 300 mL of acetone to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Sodium chlor ide: Sodium chlor ide (specia l grade) . I f the reagent i s Provisional Translation from the Japanese Original 50/100 found to include any substance that may in ter fere wi th the analysis of substances to be analyzed f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Octadecylsi lane-bonded s i l ica gel car t r idge column (1 ,000 mg): A polyethylene column of 12-13 mm in inner d iameter packed wi th 1 ,000 mg of octadecyls i lane-bonded si l ica gel , or a column equivalent to the specif ied one in separat ion capabi l i ty. Graphi te carbon and aminopropyls i lane-bonded si l ica gel laminated car t r idge column (500 mg/ 500 mg): A polyethylene column of 12-13 mm in inner d iameter packed with 500 mg each of graphi te carbon and aminopropyls i lane-bonded si l ica gel , or a column equivalent to the speci f ied one in separa t ion capabi l i ty. Diatomaceous ear th : Diatomaceous ear th for chemical analys is . Toluene: Use a ro tary vacuum evaporator on 300 mL of toluene to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . n-Hexane: Use a ro tary vacuum evaporator on 300 mL of n -hexane to evaporate unt i l 5 mL is lef t . For analys is , in ject 5 μL of the solut ion in to a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11g . Water : Dis t i l led water. I f the dis t i l led water i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Sodium sulfa te (anhydrous) : Sodium sulfa te (anhydrous)(specia l grade) . I f the reagent i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tural chemicals , wash wi th a solvent such as n -hexane before use . 0 .5 mol / l phosphate buffer solut ion (pH 7.0) : Dissolve 136.85 g of secondary sodium phosphate (dodecahydrate) and 17.92 g of sodium phosphate monobasic (d ihydrate) in water to make a 1 ,000-ml solut ion. 3 . Reference s tandard Propham: This product conta ins not less than 99% of propham, and i t s Provisional Translation from the Japanese Original 51/100 mel t ing point i s 87°C. 4 . Procedure a . Extract ion i . Cereal gra ins , legumes/pulses and seeds Weigh 10.0 g of tes t samples , previously ground so as to pass through a s tandard mesh s ieve (420 µm). Add 20 ml of water to the obtained sample and leave i t to s tand for 15 minutes . Then, add 50 ml of acetoni t r i le and homogenize the mixture for three minutes . Fi l ter the homogenized sample by suct ion through a f i l ter paper covered wi th a 1-cm thick layer of d ia tomaceous ear th . Col lec t the res idue on the surface of the f i l ter paper and add 20 ml of acetoni t r i le , and then homogenize the mixture for three minutes. Repeat the above procedure and combine the f i l t ra te , and then add acetoni t r i le to make a 100-ml solut ion. Transfer 20 ml of the above solut ion to a 100-ml separa t ing funnel and add 10 g of sodium chlor ide and 20 ml of 0 .5 mol / l phosphate buffer solut ion (pH 7.0) . Shake the mixture v igorously for 10 minutes us ing a shaker and leave i t to s tand, and then col lec t the acetoni t r i le layer. Pour 10 ml of acetoni t r i le in to an octadecyls i lane-bonded s i l ica gel car t r idge column (1 ,000 mg) and discard the eff luent . Pour the above acetoni t r i le layer in to the column and col lec t the e luate in a 50-ml conical f lask Pour 2 ml of acetoni t r i le in to the column and combine the e luate to the f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occas ional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 10 ml of acetoni t r i le and wash twice the res idue on the surface of the f i l ter paper wi th . the washings . Add the washings in to the rotary vacuum evaporator and remove the acetoni t r i le a t 40°C or lower. Dissolve the res idue in 2 ml of to luene/acetoni t r i l (1 :3) . i i . Frui t , vegetables , teas and hops In case of f ru i t and vegetables , weigh accurate ly 1 kg of the test sample and add an appropr ia te amount of water to the sample , i f required. Homogenize the mix ture and measure out a sample equivalent to 20.0 g . In case of teas and hops, weigh 5 .00 g of the tes t sample , previously Provisional Translation from the Japanese Original 52/100 ground, and add 20 ml of water, and then leave i t to s tand for 15 minutes . Then, add 50 ml of acetoni t r i le and homogenize the mixture for three minutes . Fi l ter the homogenized sample by suct ion through a f i l ter paper covered wi th a 1-cm thick layer of d ia tomaceous ear th . Col lec t the res idue on the surface of the f i l ter paper and add 20 ml of acetoni t r i le , and then homogenize the mixture for three minutes. Repeat the above procedure and combine the f i l t ra te , and then add acetoni t r i le to make a 100-ml solut ion. Transfer 20 ml of the above solut ion to a 100-ml separa t ing funnel and add 10 g of sodium chlor ide and 20 ml of 0 .5 mol / l phosphate buffer solut ion (pH 7.0) . Shake the mixture v igorously for 10 minutes us ing a shaker and leave i t to s tand, and then col lec t the acetoni t r i le layer in a 50-ml conical f lask . Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occasional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 10 ml of acetoni t r i le and wash twice the res idue on the surface of the f i l ter paper wi th the washings . Add the washings in to the rotary vacuum evaporator and remove the acetoni t r i le a t 40°C or lower. Dissolve the res idue in 2 ml of to luene/acetoni t r i l (1 :3) . i i i . Foods except those l i s ted in i and i i above Obtain the extracts according to the methods descr ibed in i or i i . b . Clean-up Pour 10 ml of to luene/acetoni t r i le (1 :3) in to a graphi te carbon and aminopropyls i lane-bonded s i l ica gel laminated car t r idge column (500 mg/500 mg) and discard the eff luent . Pour the solut ion obta ined by the ext rac t ion descr ibed in 4-a in to the column fol lowed by 20 ml of to luene/acetoni t r i le (1 :3) . Col lect the e luate in to a ro tary vacuum evaporator and remove the to luene and acetoni t r i le a t 40°C or lower. Dissolve the res idue in acetone/n-hexane (1:1) to make exact ly 2 ml (1 ml for cereal gra ins , legumes/pulses , seeds , teas and hops) of solut ion, which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference Provisional Translation from the Japanese Original 53/100 s tandard. Test ing condi t ions Column: A s i l ica te g lass capi l lary column (0 .25 mm in inner d iameter, 30 m in length) coated wi th 5% phenyl methyl s i l icone for gas chromatography to a th ickness of 0 .25 µm Column temperature : Hold the column temperature a t 50°C for one minute , fo l lowed by an increase of 25°C every minute unt i l reaching 125°C, af ter which increase the temperature by 10°C every minute unt i l reaching 300°C, and hold for 6 .5 minutes . In le t temperature : 250°C Detector : Operate at 280°C Gas f low ra te : Use hel ium as the carr ier gas . Adjust the f low ra te to the opt imal condi t ions. In jec t ion method: Spl i t less in jec t ion method. Measured mass s ize (m/z) : 93, 137 and 179 b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the same condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. (16) Malachi te green analyt ica l method Analyze malachi te green and leucomalachi te green. 1 . Apparatus A l iquid chromatograph/mass spect rometer (LC/MS or LC/MS/MS) 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Tolut ions , Etc. , Par t I I Food Addit ives . Reagents des ignated as “specia l grade” in th is sect ion must meet the requirements for “specia l grade” specif ied in the Japan Indust r ia l Standards for the reagents . Acetoni t r i le : Acetoni tr i le produced for l iquid chromatography. Ammonium formate : Ammonium formate (specia l grade) . St rongly ac idic ca t ion exchanger car t r idge column (500 mg): A polyethylene column of 8-9 mm in inner d iameter packed wi th 500 mg of benzenesulfonyl propyls i lanized s i l ica gel , or a column equivalent to the specif ied one in separa t ion capabi l i ty. Ci t r ic acid–phosphate buffer (pH 3.0) Solut ion 1: Dissolve 63.0g of c i t r ic ac id in water to make 1 ,000 mL of Provisional Translation from the Japanese Original 54/100 so lu t ion. Solut ion 2: Dissolve 215 g of d isodium phosphate in water to make 1 ,000 mL of solut ion. Mix Solut ion 1 and Solut ion 2 and adjus t the pH to 3 .0 . Dichloromethane: Dichloromethane (specia l grade) . Water : Water produced for l iquid chromatography. Sodium sulfa te (anhydrous) : Sodium sulfa te (anhydrous)(specia l grade) . When i t contains substances that may interfere wi th the analys is , wash i t wi th e thyl aceta te before us ing. 3 . Reference s tandard Malachi te Green Oxala te : This product conta ins not less than 99% of malachi te green oxala te , and i t s decomposi t ion point i s 164ºC. Leucomalachi te Green: This product conta ins not less than 99% of leucomalachi te green, and i t s decomposi t ion point i s 103ºC. 4 . Procedure a . Extract ion Weigh 5 .00 g of the tes t sample , previously ground, and homogenize i t wi th 10 mL of c i t r ic ac id-phosphate buffer (pH 3.0) . Add 15 mL of acetoni t r i le and shake the mixture vigorously for f ive minutes us ing a shaker. Centr i fuge the mixture a t 3 ,000 rpm for f ive minutes and col lec t the acetoni t r i le-water layer. Add 15mL of acetoni t r i le to the res idue and repeat the above procedure , and then combine the acetoni t r i le layer wi th the acetoni t r i le-water layer. Add 5 mL of n -hexane to the combined layers and shake the mixture v igorously for f ive minutes us ing a shaker and leave i t to s tand, and then col lect the acetoni t r i le-water layer. Add 5 mL of n -hexane to the col lec ted layer and repeat the above procedure to col lec t the acetoni t r i le-water layer. Add 50 mL of 20% sodium chlor ide and 10 mL of d ichloromethane to the col lec ted layer and shake i t v igorously for f ive minutes us ing the shaker and leave i t to s tand, and then col lec t the acetoni t r i le d ichloromethane layer. Add an adequate amount of sodium sulfa te (anhydrous) and leave i t to s tand for 15 minutes wi th occas ional shaking, and then f i l ter the mixture . b . Clean-up Pour 5 mL of acetoni t r i le into a s t rongly ac idic ca t ion exchanger Provisional Translation from the Japanese Original 55/100 car t r idge column (500 mg) and discard the eff luent . Pour the solut ion obtained by the ext ract ion descr ibed in 4–a in to the column fol lowed by 5 mL of acetoni t r i le and discard the eff luent . Pour 10 mL of acetoni t r i le /ammonia water (9 :1) and col lect the e luate in to a ro tary vacuum evaporator, and then remove the acetoni t r i le and ammonia water a t 40ºC or lower. Dissolve the res idue in 1 .0 mL of acetoni t r i le , which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandards . Test ing condi t ions Column packing: Octadecyls i lane-bonded s i l ica gel (2-5µm in par t ic le s ize) . Column: A s ta in less tube (2 .0-6 .0 mm in inner d iameter, 100-250 mm in length) . Column temperature : 40°C Mobile phase: Use a mixture of acetoni t r i le and 0 .01 mol /L ammonium formate . Create a concentra t ion gradient of 1 :9 to 1:0 acetoni t r i le /0 .01 mol /L ammonium formate in 20 minutes , and mainta in a ra t io of 1 :0 for 10 minutes . Adjust the f low ra te so that malachi te green f lows out in approximate ly 10 minutes . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the same condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. 6 . Regardless of the provis ions in 5 , ingredients of agr icul tura l chemicals and other chemical substances l i s ted in the f i rs t column of the table in (1) must not be conta ined in foods a t levels exceeding the l imi ts s t ipula ted in the th i rd column of the same table according to the food categor ies shown in the second column of the same table . In associa t ion wi th th is regula t ion, foods l i s ted in the “foods” column in the table in (2) below shal l be tested us ing the par t l i s ted in the “samples” column in the table as a sample . In addi t ion, in the substances used as ingredients of agr icul tural chemicals and other chemical substances l i s ted in the f i rs t Provisional Translation from the Japanese Original 56/100 column of the table in (1) below, which are s t ipula ted to be “Not detected” in the th i rd column of the same table in the foods l i s ted in the second column of the same table , no ingredient of agr icul tura l chemicals or o ther chemical substances shal l be detected when these foods are tes ted us ing the tes t methods s t ipulated in (3) to (10) below. (1) The maximum res idue l imi ts of substances used as ingredients of agr icul tural chemicals in foods (MRLs Lis t ) (2) Samples Food Sample mater ia l Bar ley and buckwheat Threshed seeds Wheat and rye Husked seeds Rice (brown r ice) Husked seeds Corn (maize) Seeds wi th the husk, the s i lk and the cores removed Other cereal gra ins Threshed seeds Peas , beans (dry)* , broad beans and soybeans (dry) Seeds wi thout the pods Peanuts , dry With the shel ls removed Other legumes/pulses Seeds wi thout the pods Apricot , mume plum, cherry, Japanese plum ( including prune) and nectar ine With the peduncles and the seeds removed Peach With the skins and the seeds removed Orange ( including navel orange) , grapefrui t , c i t rus natsudaidai (whole) , l ime and lemon Whole f rui t Ci t rus natsudaidai (pulp) and unshu orange (pulp) With the peels removed Ci t rus natsudaidai , peels With the calyxes removed Other c i t rus f rui ts Whole f rui t Pear, Japanese pear, quince and apple With the blossom scars , the cores and the peduncles removed Loquat With the peduncles , the skins and the seeds removed Avocado and mango With the seeds removed Provisional Translation from the Japanese Original 57/100 Kiwifrui t With the skins removed Guava With the calyxes removed Date With the calyxes and the seeds removed Pineapple With the tops removed Pass ion f rui t and papaya Whole f ru i t Banana With the pedicels removed Strawberry, cranberry, huckleberry, b lackberry and blueberry With the calyxes removed Raspberry Whole f rui t Other berr ies With the calyxes removed Japanese pers immon With the calyxes and the seeds removed Watermelon, makuwauri melon and melons With the r inds removed Grape With the peduncles removed Other f rui ts Edible por t ions Turnip ( roots) and Japanese radish ( roots , including radish) With the dir t l ight ly r insed off wi th water Turnip ( leaves) , watercress , kale , Japanese radish ( leaves , including radish) and brussels sprouts With the decayed leaves removed Caul i f lower and broccol i With the leaves removed Cabbage and Chinese cabbage A sample consis t ing of one por t ion f rom each of four heads , wi th each head equal ly cut into four por t ions , wi thout the decayed outer leaves and the cores . Kyona and komatsuna ( Japanese mustard spinach) With the roots and the decayed leaves removed Horseradish Roots wi th the di r t l ight ly r insed off wi th water Qing-geng-cai and other cruci ferous vegetables Edible por t ions Sweet potato , konjac , taro , pota to, yam and other pota toes With the dir t l ight ly r insed off wi th water Provisional Translation from the Japanese Original 58/100 Pumpkin ( including squash) , cucumber ( including gherkin) and or iental pickl ing melon (vegetable) With the vines removed Other cucurbi taceous vegetables Edible por t ions Art ichoke, endive and chicory With the decayed leaves removed Burdock and sa ls i fy A sample th inly s l iced then ground wi th a meat gr inder, the leaves having been removed and the d i r t having been l ight ly r insed off with water Shungiku With the roots and the decayed leaves removed Let tuce ( including cos le t tuce and leaf le t tuce) With the decayed outer leaves and the cores removed Other composi te vegetables Edible por t ions Shi i take mushroom, but ton mushroom and other mushrooms Edible por t ions Celery, pars ley and mitsuba With the roots and the decayed leaves removed Carrot and parsnip With the dir t l ight ly r insed off Other umbel l i ferous vegetables Edible por t ions Tomato, egg plant and pimiento (sweet pepper) With the calyxes removed Other solanceous vegetables Edible por t ions Asparagus Stems Onion, gar l ic , welsh ( including leek) and mul t ip lying onion With the outer skins and the root hai r removed Nira and other l i l iaceous vegetables Edible por t ions Green soybeans , k idney beans (wi th pods , immature) and peas (wi th pods , immature) With the pedicels removed Okra With the calyxes removed Sugarcane With the husks removed Ginger With the leaves removed, and wi th the di r t l ight ly r insed off wi th water Provisional Translation from the Japanese Original 59/100 Sugar beet With the dir t l ight ly r insed off wi th water Spinach With red roots lef t on , and wi th the root hai r and the decayed leaves removed Bamboo shoots and other vegetables Edible por t ions Sesame seeds , rapeseeds , sunf lower seeds , saff lower seeds , cot ton seed and other oi l seeds Seeds only Almond, g inkgo nut , ches tnut , walnut , pecan and other nuts With the husks removed Cacao beans and coffee beans Beans wi thout the pods Tea Tea leaves Hop Dried f lowers Other spices and other herbs Edible por t ions (3) 2 ,4 ,5-T analyt ica l method Perform according to 5 (3) . (4) Amitrol analyt ica l method Perform according to 5 (5) . (5) Analyt ica l method for a ldr in , endr in and die ldr in 1 . Apparatus A gas chromatograph wi th an e lec t ron capture detector (GC-ECD) and a gas chromatograph/mass spectrometer (GC/MS or GC/MS/MS) 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Reagents des ignated as “specia l grade” in th is sect ion must meet the requirements for “specia l grade” specif ied in the Japan Indust r ia l Standards for the reagents . Acetoni t r i le : Use a ro tary vacuum evaporator on 300 mL of acetoni t r i le to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Acetone: Use a ro tary vacuum evaporator on 300 mL of acetone to Provisional Translation from the Japanese Original 60/100 concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Ether : Use a ro tary vacuum evaporator on 300 mL of d ie thyl e ther to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Sodium chlor ide: Sodium chlor ide (specia l grade) . I f the reagent i s found to include any substance that may in ter fere wi th the analysis of substances to be analyzed f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Synthet ic magnesium s i l ica te (Flor is i l ) for column chromatography: Heat f lor is i l (150-250 μm in par t ic le s ize) a t 130°C for 12 hours or longer. Cool down to room temperature in a des iccator. Diatomaceous ear th : Diatomaceous ear th for chemical analys is . n-Hexane: Use a ro tary vacuum evaporator on 300 mL of n -hexane to evaporate unt i l 5 mL is lef t . For analys is , in ject 5 μL of the solut ion in to a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11g . Water : Dis t i l led water. I f the dis t i l led water i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Sodium sul fa te (anhydrous) : Sodium sulfa te (anhydrous)(specia l grade) . I f the reagent i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tural chemicals , wash wi th a solvent such as n -hexane before use . 3 . Reference s tandard Aldr in : This product conta ins not less than 97% of a ldr in , and i t s mel t ing point i s 102-104°C. Endr in: This product conta ins not less than 98% of endr in , and i t s decomposi t ion point i s 200°C. Dieldr in : This product conta ins not less than 98% of die ldr in , and i t s mel t ing point i s 177-179°C. Provisional Translation from the Japanese Original 61/100 4 . Procedure a . Extract ion i . Cereal gra ins , legumes/pulses and seeds Weigh 10.0 g of tes t samples , previously ground so as to pass through a s tandard mesh s ieve (420 µm). Add 20 ml of water to the obtained sample and leave i t to s tand for two hours . Then, add 100 ml of acetone and homogenize the mixture for three minutes . Fi l ter the homogenized sample by suct ion in to a ro tary vacuum evaporator through a f i l ter paper covered wi th a 1-cm thick layer of d ia tomaceous ear th . Col lec t the res idue on the surface of the f i l ter paper and add 50 ml of acetone, and then homogenize the mixture for three minutes . Repeat the above procedure and combine the f i l t ra te in to the rotary vacuum evaporator, and then concentra te the mixture to approximate ly 30 ml a t 40°C or lower. Transfer the concentra ted solut ion to a 300-ml separa t ing funnel a l ready conta ining 100 ml of 10% sodium chlor ide solut ion. Wash the eggplant-shaped f lask of the above rotary vacuum evaporator wi th 100 ml of n-hexane and add the washings to the separa t ing funnel above. Shake the mixture v igorously for f ive minutes using a shaker and leave i t to s tand, and then t ransfer the n-hexane layer to a 300-ml conical f lask . Add 50 ml of n-hexane to the aqueous layer and repeat the above procedure , and then add the n-hexane layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes with occasional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of n-hexane and wash twice the res idue on the surface of the f i l ter paper wi th the washings . Add the washing in to the ro tary vacuum evaporator and remove the n-hexane at 40°C or lower. Add 20 ml of n-hexane to the residue and t ransfer the mixture to a 100-ml separa t ing funnel , and then add 40 ml of n-hexane-satura ted acetoni t r i le . Shake the mixture v igorously for f ive minutes us ing a shaker and leave i t to s tand, and then t ransfer the acetoni t r i le layer in to the ro tary vacuum evaporator. Add 40 ml of n-hexane-satura ted acetoni t r i le to the n-hexane layer and repeat the above procedure twice , and then combine the acetoni t r i le layer in to the rotary vacuum evaporator. Remove the acetoni t r i le a t 40°C or lower and Provisional Translation from the Japanese Original 62/100 d issolve the res idue in n-hexane to make exact ly 5 ml of solut ion. i i . Frui t , vegetables , matcha In case of f ru i t and vegetables , weigh accurate ly 1 kg of the test sample and add an appropr ia te amount of water to the sample , i f required. Homogenize the mix ture and measure out a sample equivalent to 20.0 g . In case of matcha, weigh 5 .00 g of the tes t sample and add 20 ml of water, and then leave i t to s tand for two hours . Then, add 100 ml of acetone and homogenize the mixture for three minutes . Fi l ter the homogenized sample by suct ion in to a ro tary vacuum evaporator through a f i l ter paper covered wi th a 1-cm thick layer of d ia tomaceous ear th . Col lec t the res idue on the surface of the f i l ter paper and add 50 ml of acetone, and then homogenize the mixture for three minutes . Repeat the above procedure and combine the f i l t ra te in to the rotary vacuum evaporator, and then concentra te the mixture to approximate ly 30 ml a t 40°C or lower. Transfer the concentra ted solut ion to a 300-ml separa t ing funnel a l ready conta ining 100 ml of 10% sodium chlor ide solut ion. Wash the eggplant-shaped f lask of the above rotary vacuum evaporator wi th 100 ml of n-hexane and add the washings to the separa t ing funnel above. Shake the mixture v igorously for f ive minutes using a shaker and leave i t to s tand, and then t ransfer the n-hexane layer to a 300-ml conical f lask . Add 50 ml of n-hexane to the aqueous layer and repeat the above procedure twice , and then combine the n-hexane layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes with occasional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of n-hexane and wash twice the res idue on the surface of the f i l ter paper. Add the washings into the ro tary vacuum evaporator and remove the n-hexane a t 40°C or lower. Dissolve the res idue in n-hexane to make exact ly 10 ml of solut ion. i i i . Teas ( l imi ted to unfermented tea) o ther than matcha Weigh 9 .00 g of the tes t sample and soak i t in 540 ml of water a t 100°C, and the leave i t to s tand a t room temperature for f ive minutes . Transfer 360 ml of the cooled f i l t ra te in to a 500-ml conical f lask . Add 100 ml of acetone and 2 ml of sa tura ted lead aceta te Provisional Translation from the Japanese Original 63/100 so lut ion to the above f lask and leave i t to s tand for one hour a t room temperature . Fi l ter the mixtue by suct ion in to a 1 ,000-ml separa t ing funnel through a f i l ter paper covered wi th a 1-cm th ick layer of d ia tomaceous ear th . Wash the above conical f lask with 50 ml of acetone and wash the res idue on the surface of the f i l ter paper wi th the washings . Add the washings in to the above separa t ing funnel and a lso add 30 g of sodium chlor ide and 100 ml of n-hexane. Shake the mixture v igorously for f ive minutes and leave i t to s tand, and then t ransfer the n-hexane layer to a 300-ml conical f lask . Add 100 ml of n-hexane to the aqueous layer and repeat the above procedure, and then add the n-hexane layer to the conical f lask above. Add an adequate amount of asodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occas ional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask 20 ml of n-hexane and wash twice the res idue on the surface of the f i l ter paper wi th the washings . Add the washings in to the rotary vacuum evaporator and remove the n-hexane a t 40°C or lower. Dissolve the res idue in n-hexane to make exact ly 5 ml of solut ion. b . Clean-up Add 10 g of f lor is i l for column chromatography suspended in n-hexane in to a chromatograph tube (15 mm in inner d iameter, 300 mm in length) and add over approximate ly 5 g of sodium sul fa te (anhydrous) in to the column. Spi l l out the n-hexane unt i l only a smal l amount remains on the packing of the column and pour 2 ml of the solut ion obta ined by the ext ract ion descr ibed in 4-a in to the column. Pour 200 ml of e ther /n-hexane (3 :17) in to the column and col lect the e luate in to a ro tary vacuum evaporator, and then remove the e ther and n-hexane a t 40°C or lower. Dissolve the res idue in n-hexane to make exact ly 2 ml of solut ion, which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard. Tes t ing condi t ions 1 Column: A s i l ica te g lass capi l lary column (0 .25 mm in inner d iameter, 10-30 m in length) coated wi th methyl s i l icone for gas Provisional Translation from the Japanese Original 64/100 chromatography to a th ickness of 0 .25 µm. Column temperature : Hold the column temperature a t 50°C for one minute , fo l lowed by an increase of 25°C every minute unt i l reaching 175°C, af ter which increase the temperature by 10°C every minute unt i l reaching 300°C, and hold for f ive minutes . In le t temperature : 230°C Detector : Operate at 300°C Gas f low ra te : Use hel ium as the carr ier gas . Adjust the f low ra te so that a ldr in f lows out in approximately 10 minutes . Tes t ing condi t ions 2 Column: A s i l ica te g lass capi l lary column (0 .25 mm in inner d iameter, 10-30 m in length) coated wi th 14% cyanopropylphenyl-methyl s i l icone for gas chromatography to a th ickness of 0 .25 µm. . Column temperature : Hold the column temperature a t 80°C for two minutes , fo l lowed by an increase of 30°C every minute unt i l reaching 190°C, af ter which increase the temperature by 3 .6°C every minute unt i l reaching 250°C, and hold for e ight minutes . In le t temperature : 230°C Detector : Operate at 300°C Gas f low ra te : Use hel ium as the carr ier gas . Adjust the f low ra te so that a ldr in f lows out in approximately 10 minutes . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. c . Conf i rmat ion tes ts Perform gas chromatography/mass spect rometry under the condi t ions descr ibed in 5-a . Test resul ts obta ined must be the same as those obtained for the reference s tandard. Determine the quant i ty using e i ther the peak height or peak area method, i f required. (6) Captafol analyt ica l method Perform according to 5 (6) . (7) Quinoxal ine-2-carboxyl ic ac id analyt ical method Performed according to 5 (7) . (8) Cyhexat in analyt ica l method Performed according to 5 (4) . Provisional Translation from the Japanese Original 65/100 (9) Daminozide analyt ica l method Performed according to 5 (13) . (10) Analyt ica l method for t r iazophos and para thion 1 . Apparatus A gas chromatograph wi th an a lkal i f lame ionizat ion detector (GC-FID), a f lame photometr ic detector (GC-FPD, in terference f i l ter for phosphorus determinat ion, wavelength: 526 nm), or a h ighly-sensi t ive n i t rogen phosphorus detector (GC-NPD), and a gas chromatograph/mass spectrometer (GC/MS or GC/MS/MS) 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Reagents des ignated as “specia l grade” in th is sect ion must meet the requirements for “specia l grade” specif ied in the Japan Indust r ia l Standards for the reagents . Acetoni t r i le : Use a ro tary vacuum evaporator on 300 mL of acetoni t r i le to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Acetone: Use a ro tary vacuum evaporator on 300 mL of acetone to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion into a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11 g . Sodium chlor ide: Sodium chlor ide (specia l grade) . I f the reagent i s found to include any substance that may in ter fere wi th the analysis of substances to be analyzed f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Si l ica gel for column chromatography (63-200 µm in par t ic le s ize) : Heat s i l ica gel made for column chromatography (63-200 µm in par t ic le s ize) a t 130°C for 12 hours or longer. Cool down to room temperature in a des iccator. Diatomaceous ear th : Diatomaceous ear th for chemical analys is . Ethyl acetate : Use a rotary vacuum evaporator on 300 mL of e thyl aceta te to concentra te to the point of dryness . Dissolve the res idue in 5 mL of n -hexane. For analys is , in ject 5 μL of the solut ion in to a Provisional Translation from the Japanese Original 66/100 GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as high as or lower than the peaks of γ -BHC at 2 x 10 - 11g . n-Hexane: Use a ro tary vacuum evaporator on 300 mL of n -hexane to evaporate unt i l 5 mL is lef t . For analys is , in ject 5 μL of the solut ion in to a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11g . Water : Dis t i l led water. I f the dis t i l led water i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . Sodium sulfa te (anhydrous) : Sodium sulfa te (anhydrous)(specia l grade) . I f the reagent i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tural chemicals , wash wi th a solvent such as n -hexane before use . 3 . Reference s tandard Tr iazophos: This product conta ins not less than 98% of t r iazophos, and i t s mel t ing point i s 0-5°C. Para thion: This product conta ins not less than 97% of para thion, and i t s boi l ing point i s 375°C. 4 . Procedure a . Extract ion i . Cereal gra ins and legumes/pulses Weigh 10.0 g of tes t sample , previously ground so as to pass through a s tandard mesh s ieve (420 µm). Add 20 ml of water to the obta ined sample and leave i t to s tand for two hours . Then, add 100 ml of acetone and homogenize the mixture for three minutes . Fi l ter the homogenized sample by suct ion in to a ro tary vacuum evaporator through a f i l ter paper covered wi th a 1-cm thick layer of d ia tomaceous ear th . Col lec t the res idue on the surface of the f i l ter paper and add 50 ml of acetone, and then homogenize the mixture for three minutes . Repeat the above procedure and combine the f i l t ra te in to the rotary vacuum evaporator, and then concentra te the mixtue to approximate ly 30ml a t 40℃ or lower. Transfer the concentra ted solut ion to a 300-ml separa t ing funnel a l ready conta ining 100 ml of sa tura ted sodium chlor ide solut ion. Provisional Translation from the Japanese Original 67/100 Wash the eggplant-shaped f lask of the above rotary vacuum evaporator wi th 100 ml of e thyl aceta te/n-hexane (1:4) and add the washings to the separa t ing funnel above. Shake the mixture v igorously for f ive minutes us ing a shaker and leave i t to s tand, and then t ransfer the layers of e thyl aceta te and n-hexane to a 300-ml conical f lask . Add 50 ml of e thyl aceta te/n-hexane (1:4) to the aqueous layer and repeat the above procedure , and then add the layers of e thyl aceta te and n-hexane to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occas ional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of n-hexane and wash twice the res idue on the surface of the f i l ter paper wi th the washings . Add the washing in to the rotary vacuum evaporator and remove the e thyl aceta te and n-hexane at 40°C or lower. Add 30 ml of n-hexane the res idue and t ransfer the mixture to a 100-ml separa t ing funnel , and then add 30 ml of n-hexane-satura ted acetoni t r i le . Shake the mixture v igorously for f ive minutes us ing a shaker and leave i t to s tand, and then t ransfer the acetoni t r i le layer in to the ro tary vacuum evaporator. Add 30 ml of n-hexane-satura ted acetoni t r i le to the n-hexane layer and repeat the above procedure twice , and then combine the acetoni t r i le layer in to the rotary vacuum evaporator above. Remove the acetoni t r i le a t 40°C or lower and dissolve the res idue in 5 ml of acetone/n-hexane (1 :1) . i i . Frui t and vegetables Weigh accurate ly 1 kg of the tes t sample and add an appropr ia te amount of water to the sample , i f required. Homogenize the mixture and measure out a sample equivalent to 20.0 g . Then, add 100 ml of acetone and homogenize the mixture for three minutes . Fi l ter the homogenized sample by suct ion in to a ro tary vacuum evaporator using f i l ter paper covered wi th a 1-cm thick layer of d ia tomaceous ear th . Col lec t the res idue on the surface of the f i l ter paper and add 50 ml of acetone, and then homogenize the mixture for three minutes . Repeat the above procedure and combine the f i l t ra te in to the rotary vacuum evaporator, and then concentra te the mixture to approximate ly 30 ml a t 40°C or lower. Transfer the concentra ted solut ion to a 300-ml separa t ing funnel Provisional Translation from the Japanese Original 68/100 a l ready conta ining 100 ml of sa tura ted sodium chlor ide solut ion. Wash the eggplant-shaped f lask of the above rotary vacuum evaporator wi th 100 ml of e thyl aceta te/n-hexane (1:4) and add the washing to the separa t ing funnel above. Shake the mixture v igorously for f ive minutes us ing a shaker and leave i t to s tand, and then t ransfer the layers of e thyl aceta te and n-hexane to a 300-ml conical f lask . Add 50 ml of e thyl aceta te/n-hexane (1:4) to the aqueous layer and repeat the above procedure twice, and then combine the e thyl aceta te and n-hexane layer to the conical f lask above. Add an adequate amount of sodium sulfa te (anhydrous) to the f lask and leave i t to s tand for 15 minutes wi th occasional shaking. Fi l ter the content of the f lask in to a ro tary vacuum evaporator. Wash the f lask wi th 20 ml of n-hexane and wash twice the res idue on the surface of the f i l ter paper. Add the washings in to the rotary vacuum evaporator and remove the ethyl aceta te and n-hexane a t 40°C or lower. Dissolve the res idue in 5 ml of acetone/n-hexane (1 :1) . b . Clean-up Add 5 g of s i l ica gel for column chromatography (63-200 µm in par t ic le s ize) suspended in acetone/n-hexane (1 :1) into a chromatograph tube (15 mm in inner d iameter, 300 mm in length) and add over approximately 5 g of sodium sulfa te (anhydrous) to the column. Pour acetone and n-hexane (1:1) in to the column unt i l only a smal l amount remains on the packing of the column. Pour the solut ion obtained by the ext rac t ion descr ibed in 4-a in to the column fol lowed by 100 ml of acetone/n-hexane (1:1) and col lect the e luate into a ro tary vacuum evaporator, and then remove the acetone and n-hexane a t 40°C or lower. Dissolve the res idue in acetone to make exact ly 5 ml of solut ion, which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard. Tes t ing condi t ions 1 Column: A s i l ica te g lass capi l lary column (0 .53 mm in inner d iameter, 10-30 m in length) coated wi th methyl s i l icone for gas chromatography to a th ickness of 1 .5 µm. Provisional Translation from the Japanese Original 69/100 Column temperature : Hold the column temperature a t 80°C for one minute , fo l lowed by an increase of 8°C every minute unt i l reaching 250°C, and hold for f ive minutes . In le t temperature : 230°C Detector : Operate at 280°C Gas f low ra te : Use hel ium as the carr ier gas. Adjust the f low ra te to the opt imal condi t ion. Adjus t the f lows of a i r and hydrogen to the opt imal condi t ions . Tes t ing condi t ions 2 Column: A s i l ica te g lass capi l lary column (0 .32 mm in inner d iameter, 10-30 m in length) coated wi th 50% tr i f luoro propyl methyl s i l icone for gas chromatography to a thickness of 0 .25 µm. Column temperature : Hold the column temperature a t 70°C for one minute , fo l lowed by an increase of 25°C every minute unt i l reaching 125°C, af ter which increase the temperature by 10°C every minute unt i l reaching 235°C, and hold for 12 minutes . In le t temperature : 230°C Detector : Operate at 280°C Gas f low ra te : Use hel ium as the carr ier gas . Adjust the f low ra te to the opt imal condi t ion. Adjus t the f lows of a i r and hydrogen to the opt imal condi t ions . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. c . Conf i rmat ion tes ts Perform gas chromatography/mass spect rometry under the condi t ions descr ibed in 5-a . Test resul ts obta ined must be the same as those obtained for the reference s tandard. Determine the quant i ty using e i ther the peak height or peak area method, i f required. 7 . In addi t ion to the substances s t ipula ted under 6 , ingredients of agr icul tural chemicals and other chemical substances l i s ted in the f i rs t column of the table in (1) must not be conta ined in foods a t levels exceeding the l imi ts s t ipula ted in the th i rd column of the same table according to the food categor ies shown in the second column of the same table . Concerning th is , foods l i s ted in the “foods” column in the table in Provisional Translation from the Japanese Original 70/100 (2) shal l be tes ted using the part l i s ted in the “samples” column in the table as a sample . In addi t ion, in the substances used as ingredients of agr icul tural chemicals and other chemical substances l i s ted in the f i rs t column of the table in (1) , which are s t ipulated to be “Not detected” in the th i rd column of the same table in the foods l i s ted in the second column of the same table , no ingredient of agricul tura l chemicals or o ther chemical substances shal l be detected when these foods are tes ted us ing the tes t methods s t ipula ted in (3) to (8) .” (1) The maximum res idue l imi ts of substances used as ingredients of agr icul tural chemicals in foods (Provis ional MRLs Lis t ) (2) Samples Food Sample mater ia l Bar ley and buckwheat Threshed seeds Wheat and rye Husked seeds Rice (brown r ice) Husked seeds Corn (maize) Seeds wi th the husk, the s i lk and the cores removed Other cereal gra ins Threshed seeds Peas , beans (dry)* , broad beans and soybeans (dry) Seeds wi thout the pods Peanuts , dry With the shel ls removed Other legumes/pulses Seeds wi thout the pods Apricot , mume plum, cherry, Japanese plum ( including prune) and nectar ine With the peduncles and the seeds removed Peach With the skins and the seeds removed Orange ( including navel orange) , grapefrui t , c i t rus natsudaidai (whole) , l ime and lemon Whole f rui t Ci t rus natsudaidai (pulp) and unshu orange (pulp) With the peels removed Ci t rus natsudaidai , peels With the calyxes removed Other c i t rus f rui ts Whole f rui t Provisional Translation from the Japanese Original 71/100 Pear, Japanese pear, quince and apple With the blossom scars , the cores and the peduncles removed Loquat With the peduncles , the skins and the seeds removed Avocado and mango With the seeds removed Kiwifrui t With the skins removed Guava With the calyxes removed Date With the calyxes and the seeds removed Pineapple With the tops removed Pass ion f rui t and papaya Whole f ru i t Banana With the pedicels removed Strawberry, cranberry, huckleberry, b lackberry and blueberry With the calyxes removed Raspberry Whole f rui t Other berr ies With the calyxes removed Japanese pers immon With the calyxes and the seeds removed Watermelon, makuwauri melon and melons With the r inds removed Grape With the peduncles removed Other f rui ts Edible por t ions Turnip ( roots) and Japanese radish ( roots , including radish) With the dir t l ight ly r insed off wi th water Turnip ( leaves) , watercress , kale , Japanese radish ( leaves , including radish) and brussels sprouts With the decayed leaves removed Caul i f lower and broccol i With the leaves removed Cabbage and Chinese cabbage A sample consis t ing of one por t ion f rom each of four heads , wi th each head equal ly cut into four por t ions , wi thout the decayed outer leaves and the cores Kyona and komatsuna ( Japanese mustard spinach) With the roots and the decayed leaves removed Provisional Translation from the Japanese Original 72/100 Horseradish Roots wi th the di r t l ight ly r insed off wi th water Qing-geng-cai and other cruci ferous vegetables Edible por t ions Sweet potato , konjac , taro , pota to, yam and other pota toes With the dir t l ight ly r insed off wi th water Pumpkin ( including squash) , cucumber ( including gherkin) and or iental pickl ing melon (vegetable) With the vines removed Other cucurbi taceous vegetables Edible por t ions Art ichoke, endive and chicory With the decayed leaves removed Burdock and sa ls i fy A sample th inly s l iced then ground wi th a meat gr inder, the leaves having been removed and the d i r t having been l ight ly r insed off with water Shungiku With the roots and the decayed leaves removed Let tuce ( including cos le t tuce and leaf le t tuce) With the decayed outer leaves and the cores removed Other composi te vegetables Edible por t ions Shi i take mushroom, but ton mushroom and other mushrooms Edible por t ions Celery, pars ley and mitsuba With the roots and the decayed leaves removed Carrot and parsnip With the dir t l ight ly r insed off Other umbel l i ferous vegetables Edible por t ions Tomato, egg plant and pimiento (sweet pepper) With the calyxes removed Other solanceous vegetables Edible por t ions Asparagus Stems Onion, gar l ic , welsh ( including leek) and mul t ip lying onion With the outer skins and the root hai r removed Nira and other l i l iaceous vegetables Edible por t ions Provisional Translation from the Japanese Original 73/100 Green soybeans , k idney beans (wi th pods , immature) and peas (wi th pods , immature) With the pedicels removed Okra With the calyxes removed Sugarcane With the husks removed Ginger With the leaves removed, and wi th the di r t l ight ly r insed off wi th water Sugar beet With the dir t l ight ly r insed off wi th water Spinach With red roots lef t on , and wi th the root hai r and the decayed leaves removed Bamboo shoots and other vegetables Edible por t ions Sesame seeds , rapeseeds , sunf lower seeds , saff lower seeds , cot ton seed and other oi l seeds Seeds only Almond, g inkgo nut , ches tnut , walnut , pecan and other nuts With the husks removed Cacao beans and coffee beans Beans wi thout the pods Tea Tea leaves Hop Dried f lowers Other spices and other herbs Edible por t ions (3) Analyt ica l method for a ldr in , endr in and die ldr in Perform according to 6 (5) . (4) Clenbuterol analyt ica l method 1 . Apparatus A l iquid chromatograph/ tandem mass spect rometer (LC/MS/MS) 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Acetoni t r i le : Acetoni tr i le produced for l iquid chromatography. Provisional Translation from the Japanese Original 74/100 St rongly ac idic cat ion exchanger car t r idge column (500 mg): A polyethylene column of 8-9 mm in inner diameter packed wi th 500 mg of benzenesulfoni l propyl s i lane-bonded s i l ica gel , or a column equivalent to the specif ied one separa t ion capabi l i ty. Water : Water produced for l iquid chromatography. Methanol : Methanol produced for l iquid chromatography. 3 . Reference s tandard Clenbuterol hydrochlor ide: This product conta ins not less than 98% of c lenbuterol hydrochlor ide , and i t s mel t ing point i s 174-176°C. 4 . Procedure a . Extract ion Weigh 5 .00 g of the tes t sample , previously ground, and homogenize i t wi th 25 ml of e thyl aceta te and 1 ml of 4 mol / l potass ium carbonate solut ion. Centr i fuge the mixture a t 3 ,000 rpm for f ive minutes and col lec t the e thyl acetate layer in to a ro tary vacuum evaporator. Add 25 ml of e thyl acetate to the res idue and repeat the above procedure and combine the ethyl aceta te layer, and then remove the e thyl acetate a t 40°C or lower. Add 30 ml each of acetoni t r i le and acetoni t r i le sa tura ted n-hexane to the res idue and shake the mixture v igorously for f ive minutes us ing a shaker, and then leave i t to s tand. Transfer the acetoni t r i le layer to a 100-ml separa t ing funnel and add 30 ml of acetoni t r i le-sa tura ted n-hexane. Repeat the above procedure and col lec t the acetoni t r i le layer in to the rotary vacuum evaporator, and then remove the acetoni t r i le a t 40°C or lower. Dissolve the res idue in 10 ml of methanol /1 .2% metaphosphor ic ac id solut ion (2:3) and f i l ter the mixture through a cot ton plug. b . Clean-up Pour 3 ml of methanol fo l lowed by 3 ml of water in to a s t rongly ac idic cat ion exchanger car t r idge column (500 mg) and discard the eff luent . Pour the solut ion obta ined by the ext rac t ion descr ibed in 4-a in to the column fol lowed by 5 ml of water and discard the eff luent . Pour 15 ml of methanol /0 .1 mol / l d ipotass ium hydrogen or tho phosphate solut ion (9 :1) in to the column and col lec t the e luate in to a ro tary vacuum evaporator, and then remove the water and methanol a t 40°C or lower. Dissolve the res idue in 1 .0 ml of methanol , which is used as the sample solut ion. 5 . Determinat ion Provisional Translation from the Japanese Original 75/100 a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard. Test ing condi t ions Column packing: Octadecyls i lane-bonded s i l ica gel (2-5 µm in par t ic le s ize) . Column: A s ta in less tube (2 .0-6 .0 mm in inner d iameter, 100-250 mm in length) . Column temperature : 40°C Mobile phase: Use acetoni t r i le / formic acid/water (500:1:500) . Adjus t the f low ra te so that c lenbuterol f lows out in approximate ly 10 minutes . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. (5) Dexamethasone analyt ica l method 1 . Apparatus A l iquid chromatograph/ tandem mass spect rometer (LC/MS/MS) 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Acetoni t r i le : Acetoni tr i le produced for l iquid chromatography. Octadecylsi lane-bonded s i l ica gel car t r idge column (360 mg): A polyethylene column of 8-9 mm in inner d iameter packed wi th 360 mg of octadecyls i lane-bonded s i l ica gel , or a column equivalent to the speci f ied one in separa t ion capabi l i ty. Synthet ic magnesium s i l icate (Flor is i l ) for column chromatography: Heat f lor is i l (150-250 μm in par t ic le s ize) a t 130°C for 12 hours or longer. Cool down to room temperature in a des iccator. Water : Water produced for l iquid chromatography. Methanol : Methanol produced for l iquid chromatography. Phosphate buffer solut ion (pH 5.0) : Solut ion 1: Dissolve 27.2 g of monopotass ium dihydrogen mono phosphate in water to make a 1 ,000 ml of so lut ion. Provisional Translation from the Japanese Original 76/100 Solut ion 2: Dissolve 3.48 g of d ipotass ium hydrogen or thophosphate in water to make a 100 ml of solut ion. Mix Solut ion 1 and Solut ion 2 and adjus t the pH to 5 .0 . 3 . Reference s tandard Dexamethasone: This product conta ins not less than 99% of dexamethasone, and i t s mel t ing point i s 262-264°C. 4 . Procedure a . Extract ion Weigh 5 .00 g of the tes t sample , previously ground, and homogenize i t wi th 30 ml of 95% acetoni t r i le solut ion. Centr i fuge the mixture a t 2 ,500 rpm for f ive minutes and col lect the acetoni t r i le layer. Add 30 ml of 95% acetoni t r i le solut ion to the res idue and repeat the above procedure , and then col lec t the acetoni t r i le layer. b . Clean-up i . F lor is i l for column chromatography Add 8 g of f lor is i l for column chromatography suspended in acetoni t r i le in to a chromatograph tube (15 mm in inner d iameter, 300 mm in length) . Spi l l out the acetoni t r i le unt i l only a smal l amount remains on the packing of the column and pour 100 ml of acetoni t r i le , and then discard the eff luent . Pour the solut ion obtained by the ext ract ion descr ibed in 4-a into the column fol lowed by 30 ml of acetoni t r i le and then col lect the e luate into a 300-ml separa t ing funnel . Add 50 ml of n-hexane the funnel and shake i t v igorously us ing a shaker for three minutes , and then leave i t to s tand. Col lect the acetoni t r i le layer in to a ro tary vacuum evaporator and remove the acetoni t r i le a t 40°C or lower. Dissolve the res idue in 4 ml of phosphate buffer solut ion (pH 5.0) and add 6 ml of water. i i . Octadecyls i lane-bonded s i l ica gel column chromatography Pour 10 ml of methanol , 10 ml of water and 2 ml of phosphate buffer solut ion (pH 5.0) in to the oc tadecyls i lane-bonded s i l ica gel car t r idge column (360 mg) in that order, and discard the eff luent . Pour the solut ion obta ined by chromatography descr ibed in 4-b- i in to the column fol lowed by 5 ml of phosphate buffer solut ion (pH 5.0) and 10 ml of 25% methanol solu t ion, .and then discard the eff luent . Pour 10 ml of 60% acetoni t r i le solut ion into the column and col lect the e luate in to a ro tary vacuum evaporator, and then remove the acetoni t r i le and water a t 40°C or lower. Dissolve the Provisional Translation from the Japanese Original 77/100 res idue in 0 .5 ml of 10% acetoni t r i le solut ion, which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard. Test ing condi t ions Column packing: Octadecyls i lane-bonded s i l ica gel (2-5 µm in par t ic le s ize) . Column: A s ta in less tube (2 .0-6 .0 mm in inner d iameter, 100-250 mm in length) . Column temperature : 40°C Mobile phase: Use acetoni t r i le / formic acid/water (1 ,200:1:800) . Adjus t the f low ra te so that dexamethasone f lows out in approximate ly 7-10 minutes . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. (6) Analyt ica l method for t r iazophos and para thion Perform according to 6 (10) . (7) Analyt ica l method for a lpha- t renbolone and beta- t renbolone 1 . Apparatus A high-performance l iquid chromatograph wi th an ul t raviole t spect rophotometr ic detector (HPLC-UV) and a l iquid chromatograph/ mass spectrometer (LC/MS or LC/MS/MS) 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Reagents des ignated as “specia l grade” in th is sect ion must meet the requirements for “specia l grade” specif ied in the Japan Indust r ia l Standards for the reagents . Weakly bas ic anion exchanger car t r idge column (500 mg): A polyethylene column of 8-9 mm in inner d iameter packed wi th 500 mg of d ie thylaminopropyl weakly bas ic anion exchanger produced for Provisional Translation from the Japanese Original 78/100 column chromatography, or a column equivalent to the speci f ied one in separat ion capabi l i ty. Hydroxypropyl dextran for column chromatography: Dextran (25-100 µm in par t ic le s ize) chemical ly bonded wi th a hydroxypropyl group produced for column chromatography. Dichloromethane: Dichloromethane (specia l grade) . 3 . Reference s tandard Alpha- t renbolone: This product conta ins not less than 94% of a lpha- t renbolone, and i t s mel t ing point i s 110°C. Beta- t renbolone: This product conta ins not less than 99% of beta- t renbolone, and i ts mel t ing point i s 186°C. 4 . Procedure a . Extract ion Weigh 5 .00 g out of tes t sample , previously ground. For muscle , remove the fa t layer as much as poss ible before gr inding. Add 20 ml of acetoni t r i le /methanol (4 :1) and homogenize the mixture again . Centr i fuge a t 2 ,600 rpm for f ive minutes and col lec t the supernatant in a 100-ml separa t ing funnel . Add 20 ml of acetoni t r i le /methanol (4 :1) to the precipi ta te and centr i fuge the mixture under the above condi t ions , and then combine the acetoni t r i le and methanol layer to the separa t ing funnel above. Add 20 ml of water-sa tura ted n-hexane to the funnel and shake i t v igorously for f ive minutes , and then leave i t to s tand. Transfer the acetoni t r i le and methanol layer to a 200-ml separa t ing funnel . Add 40 ml each of 5% sodium sulfa te solu t ion and dichloromethane to the funnel and shake i t v igorously for f ive minutes and leave i t to s tand, and then t ransfer the dichloromethane layer in to a ro tary vacuum evaporator. Add 15 ml of d ichloromethane to the aqueous layer and repeat the above procedure and combine the dichloromethane layer in to the rotary vacuum evaporator, and then remove the dichloromethane a t 40°C or lower. Dissolve the res idue in 1 ml of n-hexane/benzene (3:1) . b . Clean-up i . Weakly bas ic anion exchanger column chromatography Pour 6 ml of n-hexane/benzene (3:1) in to a weakly bas ic anion exchanger car t r idge column (500 mg) and discard the eff luent . Pour the the solut ion obta ined by the ext rac t ion descr ibed in 4-a into the column fol lowed by 2 ml of n-hexane/benzene (3:1) and discard the Provisional Translation from the Japanese Original 79/100 e ff luent . Pour 3 ml of d ichloromethane/methanol (9 :1) in to the column and col lect the e luate in to a ro tary vacuum evaporator, and then evaporate the solvent to near dryness under n i t rogen a t 40°C or lower. Dissolve the res idue in 0 .5 ml of benzene/methanol (17:3) . i i . Hydroxypropyl dextran for column chromatography Add hydroxypropyl dextran for column chromatography suspended in benzene/methanol (17:3) , previously lef t to s tand for 12 hours , in to a polyethylene column (6 mm in inner d iameter) so as to obtain a 120-mm long layer of dextran . Spi l l out the benzene/methanol (17:3) unt i l only a smal l amount remains on the packing of the column. Pour the solut ion obtained by chromatography descr ibed in 4-b- i in to th is column fol lowed by 10 ml of benzene/methanol (17:3) . Discard 2 .0 ml of the f i rs t eff luent and col lec t the next e luate in to a ro tary vacuum evaporator, and then evaporate the solvent to near dryness under ni t rogen a t 40°C or lower. Dissolve the res idue in 0 .5 ml of acetoni t r i le /water (1 :1) , which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perfom qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard. Test ing condi t ions Column packing: Octadecyls i lane-bonded s i l ica gel (5 µm in par t ic le s ize) . Column: A s ta in less tube (4 .0-6 .0 mm in inner d iameter, 150 mm in length) . Column temperature : 40°C Detector : Operate wi th an absorpt ion wavelength of 340 nm. Mobile phase: Use acetoni t r i le /water (5 :6) . Adjust the f low ra te so that a lpha- trenbolone f lows out in approximate ly 10 minutes . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. c . Conf i rmat ion tes ts Perform l iquid chromatography/mass spect rometry under the Provisional Translation from the Japanese Original 80/100 condi t ions descr ibed in 5-a . Tes t resul ts obtained must be the same as those obta ined for the reference s tandard. Determine the quant i ty us ing e i ther the peak height or peak area method, i f required. (8) Ethylene dibromide analyt ica l method 1 . Apparatus A gas chromatograph wi th an e lec t ron capture detector (GC-ECD), a gas chromatograph/mass spect rometer (GC/MS or GC/MS/MS) and Dean Stark dis t i l la t ion apparatus The Dean Stark dis t i l la t ion apparatus i s roughly as shown in the fol lowing f igure: A: Dis t i l la t ion f lask B: Cool ing tube C: Dis t i l la t ion t rap 2 . Reagents and tes t solut ions In addi t ion to the reagents and tes t solu t ions l i s ted below, use those l i s ted in Sect ion C Reagents /Test Solut ions , Etc . , Par t I I Food addi t ives . Synthet ic magnesium s i l icate (Flor is i l ) for column chromatography: Heat f lor is i l (150-250 μm in par t ic le s ize) a t 130°C for 12 hours or longer. Cool down to room temperature in a des iccator. Si l icone for defoaming: Si l icone produced for defoaming. n-Hexane: Use a ro tary vacuum evaporator on 300 mL of n -hexane to evaporate unt i l 5 mL is lef t . For analys is , in ject 5 μL of the solut ion in to a GC-ECD. Peaks other than that of n -hexane in the resul t ing chromatogram should be as h igh as or lower than the peaks of γ -BHC at 2 x 10 - 11g . Water : Dis t i l led water. I f the dis t i l led water i s found to include any substance that may in terfere wi th analys is of the target composi t ional substances f rom agr icul tura l chemicals , wash wi th a solvent such as n -hexane before use . 3 . Reference s tandard Ethylene dibromide: This product conta ins not less than 99% of Provisional Translation from the Japanese Original 81/100 e thylene dibromide, and i t s boi l ing point i s 131.5°C. 4 . Procedure a . Extract ion Homogenize about 1 kg of a test sample and t ransfer 100 g of the homogenized sample in to a 1 ,000-ml dis t i l la t ion f lask , and then add 200 ml of water and 10 ml of n-hexane to the f lask . Add a few drops of defoaming s i l icone and boi l ing s tones to the mixture and a t tach the f lask to the Dean Stark dis t i l lat ion apparatus to be heated and ref luxed for one hour. After cool ing, remove most of the water in the dis t i l la t ion t rap and f i l ter the n-hexane layer in to a 10-ml measur ing f lask through a l iquid phase separa t ion f i l ter paper. Wash the t rap wi th a smal l amount of n-hexane and f i l ter the washings through the f i l ter paper above. Combine the f i l t ra te wi th the n-hexane layer to make exact ly a 10-ml solut ion. b . Clean-up Transfer the solut ion obtained by the ext rac t ion descr ibed in 4-a to a 10-ml tes t tube with a g lass s topper. Add approximate ly 1 g of f lor is i l for column chromatography to the tes t tube and shake i t v igorously, and then leave i t to s tand a t room temperature for about 15 minutes . Col lec t about 5 ml of supernatant in another tes t tube wi th a g lass s topper, which is used as the sample solut ion. 5 . Determinat ion a . Qual i ta t ive tes ts Perform qual i ta t ive tes ts under the fol lowing condi t ions . Test resul ts obtained must be the same as those obta ined for the reference s tandard. Test ing condi t ions Column: A s i l ica te g lass capi l lary column (0 .32 mm in inner d iameter, length in 30 m) coated wi th 6% cyanopropylphenyl-methyl s i l icone for gas chromatography to a thickness of 1 .8 µm. Column temperature : Hold the column temperature a t 50°C for two minute , fo l lowed by an increase of 5°C every minute unt i l reaching 110°C, af ter which increase the temperature by 30°C every minute unt i l reaching 260°C, and hold for f ive minutes . In le t temperature : 250°C Detector : Operate at 300°C Gas f low ra te : Use hel ium as the carr ier gas . Adjust the f low ra te so Provisional Translation from the Japanese Original 82/100 tha t e thylene dibromide f lows out in approximate ly e ight minutes . b . Quant i ta t ive tes ts Determine the quant i ty f rom the tes t resul ts obta ined under the condi t ions descr ibed in 5-a using e i ther the peak height or peak area method. c . Conf i rmat ion tes ts Perform gas chromatography/mass spect rometry under the condi t ions descr ibed in 5-a . Test resul ts obta ined must be the same as those obtained for the reference s tandard. Determine the quant i ty using e i ther the peak height or peak area method, i f required. 8 . In cases where a substance used as an ingredient of an agr icul tura l chemical (excluding substances determined by the Minis ter of Heal th , Labour and Welfare as having no potent ia l to cause damage to human heal th under Paragraph 3 of Art ic le 11 of the Law), for which no composi t ional s tandards are s t ipula ted in 5 to 7 , i s the same as the substance natura l ly contained in the food, the amount of the substance conta ined in the food shal l not exceed the amount normal ly conta ined in the food. This , however, does not apply to foods conta ining substances tha t may cause damage to human heal th a t the amount normal ly conta ined. 9 . In cases where substances used as ingredients of agr icul tura l chemicals and other chemical substances that are l i s ted in the f i rs t column of the fo l lowing table are conta ined in a food, the amount of the substances shal l not exceed the amount s t ipulated in the th i rd column of the same table according to the food categories shown in the second column of the same table . The maximum res idue l imi ts of substances used as ingredients of agr icul tural chemicals in foods (excluding foods l i s ted in the second column of the table in 6 (1) and the second column of the table in 7 (1)) : Column 1 Column 2 Column 3 AZINPHOS-METHYL Other spices , dr ied 0 .5 ppm ACEPHATE Other spices , dr ied (except the f rui t of Sansho ( Japanese pepper)) 0 .2 ppm Provisional Translation from the Japanese Original 83/100 AMITRAZ Cot tonseed oi l (except ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the Japanese Agricul tura l Standards (JAS) for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm ALDICARB Peanut o i l ( l imi ted to ref ined peanut oi l and peanut sa lad oi l that meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .01 ppm Cot tonseed oi l ( l imi ted to ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .01 ppm IPRODIONE Other spices , dr ied ( l imi ted to seeds) 0 .05 ppm Other spices , dr ied ( l imi ted to roots or rhizomes) 0 .1 ppm IMIDACLOPRID Wheat f lour (except whole gra in) 0 .03 ppm Wheat bran 0 .3 ppm ETHION Other spices , dr ied ( l imi ted to f rui ts ) 5 ppm Other spices , dr ied ( l imi ted to seeds) 3 ppm Other spices , dr ied ( l imi ted to roots or rhizomes) 0 .3 ppm ETHEPHON Rais in 5 ppm Fig , dr ied 10 ppm Provisional Translation from the Japanese Original 84/100 ENDOSULFAN Cot tonseed oi l (except ref ined cot tonseed oi l and cot tonseed sa lad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .5 ppm Other spices , dr ied ( l imi ted to f rui ts ) 5 ppm Other spices , dr ied ( l imi ted to seeds) 1 ppm Other spices , dr ied ( l imi ted to roots or rhizomes) 0 .5 ppm CARBARYL Rice bran 170 ppm Mil led r ice 1 ppm Wheat f lour (except whole gra in) 0 .2 ppm Wheat germ 1 ppm Wheat bran 2 ppm Corn oi l (except edible corn oi l that meets the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .1 ppm Soybean oi l (except edible soybean oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .2 ppm Tomato ju ice 3 ppm Tomato paste 10 ppm Edible o l ive oi l ( l imi ted to v i rg in o i l ) 25 ppm Sunf lower oi l (except edible sunf lower oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm Provisional Translation from the Japanese Original 85/100 QUINTOZENE Other spices , dr ied ( l imi ted to f rui ts ) 0 .02 ppm Other spices , dr ied ( l imi ted to seeds) 0 .1 ppm Other spices , dr ied ( l imi ted to roots or rhizomes) 2 ppm GLYPHOSATE Cot tonseed oi l ( l imi ted to ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm Cot tonseed oi l (except ref ined cot tonseed oi l and cot tonseed sa lad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm GLUFOSINATE Sunflower oi l (except edible sunf lower oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm Rapeseed oi l (except ref ined rapeseed oi l and rapeseed salad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm KRESOXIM-METHYL Rais in 2 ppm Edible o l ive oi l ( l imi ted to v i rg in o l ive o i l ) 0 .7 ppm CLETHODIM Soybean oi l ( l imi ted to edible soybean oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .5 ppm Provisional Translation from the Japanese Original 86/100 Soybean oi l (except edible soybean oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 1 ppm Sunf lower oi l (except edible sunf lower oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .1 ppm Cot tonseed oi l ( l imi ted to ref ined cot tonseed oi l and cot ton seed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .5 ppm Cot tonseed oi l (except ref ined cot tonseed oi l and cot ton seed sa lad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .5 ppm Rapeseed oi l ( l imi ted to ref ined rapeseed oi l and rapeseed salad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .5 ppm Rapeseed oi l (except ref ined rapeseed oi l and rapeseed salad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .5 ppm CHLORDANE Mil led r ice 0 .02 ppm Provisional Translation from the Japanese Original 87/100 Soybean oi l ( l imi ted to edible soybean oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .02 ppm Soybean oi l (except edible soybean oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm Cot tonseed oi l (except ref ined cot tonseed oi l and cot tonseed sa lad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm Linseed oi l (except ref ined l inseed oi l ) 0 .05 ppm CHLORPYRIFOS Wheat f lour (except whole gra in) 0 .1 ppm Corn oi l ( l imi ted to edible corn oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .2 ppm Soybean oi l ( l imi ted to edible soybean oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm Cot tonseed oi l ( l imi ted to ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm Provisional Translation from the Japanese Original 88/100 Cot tonseed oi l (except ref ined cot tonseed oi l and cot tonseed sa lad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm Other spices , dr ied ( l imi ted to f rui ts ) 1 ppm Other spices , dr ied ( l imi ted to seeds) 5 ppm Other spices , dr ied ( l imi ted to roots or rhizomes) 1 ppm CHLORPYRIFOS-METHYL Wheat f lour (except whole gra in) 2 ppm Wheat bran 20 ppm Other spices , dr ied ( l imi ted to f rui ts ) 0 .3 ppm Other spices , dr ied ( l imi ted to seeds) 1 ppm Other spices , dr ied ( l imi ted to roots or rhizomes) 5 ppm CHLORMEQUAT Wheat f lour ( l imi ted to whole gra in) 5 ppm Wheat f lour (except whole gra in) 2 ppm Wheat bran 10 ppm Rye f lour ( l imi ted to whole gra in) 4 ppm Rye f lour (except whole gra in) 3 ppm Rye bran 10 ppm Rapeseed oi l (except ref ined rapeseed oi l and rapeseed salad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .1 ppm FENBUTATIN OXIDE Plum, dr ied 10 ppm Rais in 20 ppm Provisional Translation from the Japanese Original 89/100 DICHLORVOS and NALED (as to ta l ) Wheat f lour ( l imi ted to whole gra in) 2 ppm Wheat f lour (except whole gra in) 1 ppm Wheat bran 10 ppm Wheat germ 10 ppm Other spices , dr ied 0 .1 ppm DIQUAT Mil led r ice 0 .2 ppm Wheat f lour ( l imi ted to whole gra in) 2 ppm Wheat f lour (except whole gra in) 0 .5 ppm Wheat bran 5 ppm Vegetable oi l (except ref ined vegetable oi l ) 0 .05 ppm DICOFOL Plum, dr ied 3 ppm Cot tonseed oi l ( l imi ted to ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .5 ppm Cot tonseed oi l (except ref ined cot tonseed oi l and cot tonseed sa lad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .5 ppm Other spices , dr ied ( l imi ted to f rui ts ) 0 .1 ppm Other spices , dr ied ( l imi ted to seeds) 0 .05 ppm Other spices , dr ied ( l imi ted to roots or rhizomes) 0 .1 ppm DISULFOTON Other spices , dr ied 0 .05 ppm Provisional Translation from the Japanese Original 90/100 CYHALOTHRIN Cot tonseed oi l ( l imi ted to ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .02 ppm Cot tonseed oi l (except ref ined cot tonseed oi l and cot tonseed sa lad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .02 ppm DIPHENYLAMINE Apple ju ice 0 .5 ppm CYPRODINIL Wheat bran 2 ppm Plum, dr ied 5 ppm Rais in 5 ppm CYPERMETHRIN Other spices , dr ied ( l imi ted to f rui ts ) 0 .1 ppm Other spices , dr ied ( l imi ted to roots or rhizomes) 0 .2 ppm Vegetable o i l ( l imi ted to ref ined vegetable o i l ) 0 .5 ppm DIMETHIPIN Cot tonseed oi l ( l imi ted to ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .1 ppm Cot tonseed oi l (except ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .1 ppm DIMETHOATE Edible o l ive o i l (except v i rg in oi l ) 0 .05 ppm Other spices , dr ied ( l imi ted to f rui ts ) 0 .5 ppm Provisional Translation from the Japanese Original 91/100 Other spices , dr ied ( l imi ted to seeds) 5 ppm Other spices , dr ied ( l imi ted to roots or rhizomes) 0 .1 ppm BROMIDE Wheat f lour ( l imi ted to whole gra in) 50 ppm Peach, dr ied 50 ppm Plum, dr ied 20 ppm Rais in 100 ppm Date , dr ied 100 ppm Fig , dr ied 250 ppm Frui ts (dr ied f rui ts except peach, p lum, grape, date and f ig) 30 ppm Other herbs, dr ied 400 ppm SPINOSAD Wheat bran 2 ppm Rais in 1 ppm Cot tonseed oi l ( l imi ted to ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .01 ppm Cot tonseed oi l (except ref ined cot tonseed oi l and cot tonseed sa lad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .01 ppm DIAZINON Plum, dr ied 2 ppm Other spices , dr ied ( l imi ted to f rui ts ) 0 .1 ppm Other spices , dr ied ( l imi ted to seeds) 5 ppm Other spices , dr ied ( l imi ted to roots or rhizomes) 0 .5 ppm Provisional Translation from the Japanese Original 92/100 THIODICARB and METHOMYL Wheat f lour (except whole gra in) 0 .03 ppm Wheat germ 2 ppm Wheat bran 3 ppm Corn oi l ( l imi ted to edible corn oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .02 ppm Soybean oi l ( l imi ted to edible soybean oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .2 ppm Soybean oi l (except edible soybean oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .2 ppm Cot tonseed oi l ( l imi ted to ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .04 ppm TEBUCONAZOLE Rais in 3 ppm TEBUFENOZIDE Rais in 2 ppm DELTAMETHRIN and TRALOMETHRIN (as to ta l ) Wheat f lour ( l imi ted to whole gra in) 2 ppm Wheat f lour (except whole gra in) 0 .3 ppm Wheat bran 5 ppm TERBUFOS Rapeseed oi l (except ref ined rapeseed oi l and rapeseed salad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm Provisional Translation from the Japanese Original 93/100 PARAQUAT Sunflower oi l ( l imi ted to edible sunf lower oi l that meets the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm Sunf lower oi l (except edible sunf lower oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm Cot tonseed oi l ( l imi ted to ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm PARATHION Other spices , dr ied ( l imi ted to f rui ts ) 0 .2 ppm Other spices , dr ied ( l imi ted to seeds) 0 .1 ppm Other spices , dr ied ( l imi ted to roots or rhizomes) 0 .2 ppm PARATHION-METHYL Rais in 1 ppm Other spices , dr ied ( l imi ted to f rui ts ) 5 ppm Other spices , dr ied ( l imi ted to seeds) 5 ppm Other spices , dr ied ( l imi ted to roots or rhizomes) 3 ppm BIORESMETHRIN Wheat f lour ( l imi ted to whole gra in) 1 ppm Wheat f lour (except whole gra in) 1 ppm Wheat germ 3 ppm Wheat bran 5 ppm BIFENTHRIN Wheat f lour ( l imi ted to whole gra in) 0 .5 ppm Wheat f lour (except whole gra in) 0 .2 ppm Provisional Translation from the Japanese Original 94/100 Wheat bran 2 ppm PIPERONYL BUTOXIDE Wheat f lour ( l imi ted to whole gra in) 30 ppm Wheat f lour (except whole gra in) 10 ppm Wheat germ 90 ppm Wheat bran 80 ppm Corn oi l (except edible corn oi l that meets the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 80 ppm Tomato ju ice 0 .3 ppm Juice of c i t rus f rui ts 0 .05 ppm Frui ts , dr ied 0 .2 ppm PYRIPROXYFEN Cot tonseed oi l ( l imi ted to ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .01 ppm Cot tonseed oi l (except ref ined cot tonseed oi l and cot tonseed sa lad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .01 ppm PIRIMICARB Other spices , dr ied ( l imi ted to seeds) 5 ppm PIRIMIPHOS-METHYL Wheat bran 15 ppm Other spices , dr ied ( l imi ted to f rui ts ) 0 .5 ppm Other spices , dr ied ( l imi ted to seeds) 3 ppm PYRETHRINS Frui ts , dr ied 0 .2 ppm VINCLOZOLIN Other spices , dr ied 0 .05 ppm Provisional Translation from the Japanese Original 95/100 FAMOXADONE Wheat bran 0 .2 ppm Rais in 5 ppm FENAMIPHOS Peanut o i l (except ref ined peanut oi l and peanut sa lad oi l that meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm Cot tonseed oi l (except ref ined cot tonseed oi l and cot tonseed sa lad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm FENITROTHION Wheat f lour ( l imi ted to whole gra in) 5 ppm Wheat bran 20 ppm Other spices , dr ied ( l imi ted to f rui ts ) 1 ppm Other spices , dr ied ( l imi ted to seeds) 7 ppm Other spices , dr ied ( l imi ted to roots or rhizomes) 0 .1 ppm FENTHION Edible o l ive oi l ( l imi ted to v i rg in o i l ) 1 ppm PHENTHOATE Other spices , dr ied ( l imi ted to seeds) 7 ppm FENVALERATE Wheat f lour ( l imi ted to whole gra in) 2 ppm Wheat f lour (except whole gra in) 0 .2 ppm Wheat bran 5 ppm Cot tonseed oi l ( l imi ted to ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .1 ppm Provisional Translation from the Japanese Original 96/100 Cot tonseed oi l (except ref ined cot tonseed oi l and cot tonseed sa lad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .1 ppm FENPROPATHRIN Cot tonseed oi l (except ref ined cot tonseed oi l and cot tonseed sa lad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 3 ppm FLUDIOXONIL Nira , dr ied 50 ppm Basi l , dr ied 50 ppm FLUSILAZOLE Rais in 1 ppm FLUTOLANIL Rice bran 10 ppm Mil led r ice 1 ppm PROCHLORAZ Wheat bran 7 ppm Sunf lower oi l ( l imi ted to edible sunf lower oi l that meets the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 1 ppm PROCYMIDONE Sunflower oi l ( l imi ted to edible sunf lower oi l that meets the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .5 ppm PROPARGITE Corn f lour 0 .2 ppm Corn oi l ( l imi ted to edible corn oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .5 ppm Provisional Translation from the Japanese Original 97/100 Corn oi l (except edible corn oi l that meets the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .7 ppm Peanut o i l ( l imi ted to ref ined peanut oi l and peanut sa lad oi l that meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .3 ppm Peanut o i l (except ref ined peanut oi l and peanut sa lad oi l that meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .3 ppm Orange ju ice 0 .3 ppm Apple ju ice 0 .2 ppm Grape ju ice 1 ppm Rais in 12 ppm Cot tonseed oi l ( l imi ted to ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .2 ppm PROFENOFOS Cot tonseed oi l ( l imi ted to ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm HEPTACHLOR Soybean oi l ( l imi ted to edible soybean oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .02 ppm Provisional Translation from the Japanese Original 98/100 Soybean oi l (except edible soybean oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .5 ppm PERMETHRIN Wheat f lour ( l imi ted to whole gra in) 2 ppm Wheat f lour (except whole gra in) 0 .5 ppm Wheat germ 2 ppm Wheat bran 5 ppm Soybean oi l (except edible soybean oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .1 ppm Sunf lower oi l ( l imi ted to edible sunf lower oi l that meets the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 1 ppm Sunf lower oi l (except edible sunf lower oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 1 ppm Cot tonseed oi l ( l imi ted to ref ined cot tonseed oi l and cot tonseed sa lad oi l tha t meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .1 ppm Other spices , dr ied 0 .05 ppm PENCONAZOLE Rais in 0 .5 ppm PHOSALONE Other spices , dr ied ( l imi ted to f rui ts ) 2 ppm Other spices , dr ied ( l imi ted to seeds) 2 ppm Provisional Translation from the Japanese Original 99/100 Other spices , dr ied ( l imi ted to roots or rhizomes) 3 ppm PHORATE Peanut o i l ( l imi ted to ref ined peanut oi l and peanut sa lad oi l that meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm Peanut o i l (except ref ined peanut oi l and peanut sa lad oi l that meet the JAS for Edible Vegetable Fats and Oils , and other edible o i ls that meet s tandards equivalent to or s t r ic ter than JAS) 0 .05 ppm Other spices , dr ied ( l imi ted to f rui ts ) 0 .1 ppm Other spices , dr ied ( l imi ted to seeds) 0 .5 ppm Other spices , dr ied ( l imi ted to roots or rhizomes) 0 .1 ppm MALATHION Tomato ju ice 0 .01 ppm Other spices , dr ied ( l imi ted to f rui ts ) 1 ppm Other spices , dr ied ( l imi ted to seeds) 2 ppm Other spices , dr ied ( l imi ted to roots or rhizomes) 0 .5 ppm METHAMIDOPHOS Other spices , dr ied (except the f rui t of Sansho ( Japanese pepper)) 0 .1 ppm METALAXYL and MEFENOXAM Other spices , dr ied ( l imi ted to seeds) 5 ppm METHIDATHION Edible o l ive oi l ( l imi ted to v i rg in o i l ) 2 ppm Cot tonseed oi l (except ref ined cot tonseed oi l and cot tonseed sa lad oi l that meet the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 2 ppm Provisional Translation from the Japanese Original 100/100 METHOXYFENOZIDE Plum, dr ied 2 ppm Rais in 3 ppm METHOPRENE Wheat f lour ( l imi ted to whole gra in) 5 ppm Wheat f lour (except whole gra in) 2 ppm Wheat bran 10 ppm Corn oi l ( l imi ted to edible corn oi l that meets the JAS for Edible Vegetable Fats and Oi ls , and other edible o i ls tha t meet s tandards equivalent to or s t r ic ter than JAS) 0 .2 ppm HYDROGEN PHOSPHIDE Vegetables , dr ied 0 .01 ppm Frui ts , dr ied 0 .01 ppm 10. In addi t ion to the provis ions of 6 or 9 , when food products are manufactured or processed us ing foods for which composi t ional s tandards are speci f ied in 6 through 9 , the foods used must comply wi th the s tandards given in 6 through 9 .