thông tư 27-2010-ttbyt

B Y T _______ S: 27/2010/TT-BYT

CNG HO X HI CH NGHA VIT NAM c lp - T do - Hnh phc ____________________________ H Ni, ngy 20 thng 5 nm 2010

THNG T Ban hnh Quy chun k thut quc gia v ph gia thc phm - phm mu ____________ B TRNG B Y T Cn c Lut Tiu chun v Quy chun k thut ngy 29 thng 6 nm 2006 v Ngh nh s 127/2007/N-CP ngy 01 thng 8 nm 2007 ca Chnh ph quy nh chi tit thi hnh mt s iu ca Lut Tiu chun v Quy chun k thut; Cn c Php lnh V sinh an ton thc phm ngy 07 thng 8 nm 2003 v Ngh nh s 163/2004/N-CP ngy 07 thng 9 nm 2004 ca Chnh ph quy nh chi tit thi hnh mt s iu ca Php lnh V sinh an ton thc phm; Cn c Ngh nh s 188/2007/N-CP ngy 27 thng 12 nm 2007 ca Chnh ph quy nh chc nng, nhim v, quyn hn v c cu t chc ca B Y t; Theo ngh ca Cc trng Cc An ton v sinh thc phm, V trng V Khoa hc v o to, V trng V Php ch, QUY NH: iu 1. Ban hnh km theo Thng t ny: QCVN 4-10: 2010/BYT - Quy chun k thut quc gia v ph gia thc phm Phm mu. iu 2. Thng t ny c hiu lc t ngy 01 thng 01 nm 2011. iu 3. Cc trng Cc An ton v sinh thc phm, Th trng cc n v thuc B Y t, cc n v trc thuc B Y t; Gim c S Y t cc tnh, thnh ph trc thuc trung ng v cc t chc, c nhn c lin quan chu trch nhim thi hnh Thng t ny./. Ni nhn:- VPCP (Vn x, Cng bo, Cng TTT Chnh ph); - Cc B, c quan ngang B, c quan thuc CP; - B T php (Cc Kim tra VBQPPL); - B trng Nguyn Quc Triu ( bo co); - UBND cc tnh, thnh ph trc thuc TW; - Vin Kim st nhn dn ti cao; - To n nhn dn ti cao; - S Y t cc tnh, thnh ph trc thuc TW; - Chi cc ATTP cc tnh, thnh ph trc thuc TW; - TTYTDP cc tnh, thnh ph trc thuc TW; - Cc c quan KTNN v thc phm nhp khu; - Tng Cc Tiu chun - o lng - Cht lng; - Website B Y t; - Lu: VT, K2T, PC, ATTP.

KT. B TRNG TH TRNG TH TRNG

Trnh Qun Hun

CNG HO X HI CH NGHA VIT NAM QCVN 4-10: 2010/BYT

QUY CHUN K THUT QUC GIA V PH GIA THC PHM - PHM MU National technical regulation on Food Additives - Colours

H NI - 2010

Li ni u QCVN 4-10: 2010/BYT do Ban son tho quy chun k thut quc gia v Ph gia thc phm v cht h tr ch bin bin son, Cc An ton v sinh thc phm trnh duyt v c ban hnh theo Thng t s 27/2010/TT-BYT ngy 20 thng 5 nm 2010 ca B trng B Y t.

QUY CHUN K THUT QUC GIA V PH GIA THC PHM - PHM MU National technical regulation on Food Additives - Colours I. QUY NH CHUNG 1. Phm vi iu chnh Quy chun k thut quc gia (sau y gi tt l Quy chun) ny quy nh cc yu cu k thut v qun l v cht lng, v sinh an ton i vi cc phm mu c s dng vi mc ch lm ph gia thc phm. 2. i tng p dng Quy chun ny p dng i vi: 2.1. T chc, c nhn nhp khu, xut khu, sn xut, bun bn v s dng cc phm mu lm ph gia thc phm (sau y gi tt l t chc, c nhn). 2.2. C quan qun l nh nc c lin quan. 3. Gii thch t ng v ch vit tt: 3.1. Phm mu: l ph gia thc phm c s dng vi mc ch tng cng hoc khi phc mu cho sn phm thc phm. 3.2. JECFA monograph 1 - Vol. 4 (JECFA monographs 1 - Combined compendium of food addiditive specifications; Joint FAO/WHO expert committee on food additives; Volume 4 - Analytical methods, test procedures and laboratory solutions used by and referenced in the food additive specifications; FAO, 2006): Cc yu cu k thut i vi ph gia thc phm, Tp 4 Cc phng php phn tch, quy trnh th nghim, dung dch th nghim c s dng (hoc tham chiu) trong yu cu k thut i vi ph gia thc phm; JECFA bin son; FAO ban hnh nm 2006. 3.3. M s C.A.S (Chemical Abstracts Service): M s ng k ho cht ca Hip hi Ho cht Hoa K. 3.4. TS (test solution): Dung dch thuc th. 3.5. ADI (Acceptable daily intake): Lng n vo hng ngy chp nhn c. 3.6. INS (International numbering system): H thng m s quc t v ph gia thc phm. II. YU CU K THUT V PHNG PHP TH V LY MU 1. Yu cu k thut v phng php th i vi phm mu c quy nh ti cc ph lc ban hnh km theo Quy chun ny nh sau : 1.1. Ph lc 1: Quy nh k thut v phng php th i vi curcumin 1.2. Ph lc 2: Quy nh k thut v phng php th i vi riboflavin 1.3. Ph lc 3: Quy nh k thut v phng php th i vi tatrazin 1.4. Ph lc 4: Quy nh k thut v phng php th i vi quinolin 1.5. Ph lc 5: Quy nh k thut v phng php th i vi sunset yellow FCF 1.6. Ph lc 6: Quy nh k thut v phng php th i vi carmin 1.7. Ph lc 7: Quy nh k thut v phng php th i vi carmoisin 1.8. Ph lc 8: Quy nh k thut v phng php th i vi amaranth 1.9. Ph lc 9: Quy nh k thut v phng php th i vi ponceau 4R 1.10. Ph lc 10: Quy nh k thut v phng php th i vi erythrosin 1.11. Ph lc 11: Quy nh k thut v phng php th i vi red 2G 1.12. Ph lc 12: Quy nh k thut v phng php th i vi allura red AC 1.13. Ph lc 13: Quy nh k thut v phng php th i vi indigotin 1.14. Ph lc 14: Quy nh k thut v phng php th i vi brilliant blue FCF 1.15. Ph lc 15: Quy nh k thut v phng php th i vi clorophyl 1.16. Ph lc 16: Quy nh k thut v phng php th i vi clorophyl phc ng 1.17. Ph lc 17: Quy nh k thut v phng php th i vi mui Natri, Kali ca Clorophyl phc ng 1.18. Ph lc 18: Quy nh k thut v phng php th i vi green S 1.19. Ph lc 19: Quy nh k thut v phng php th i vi caramen 1.20. Ph lc 20: Quy nh k thut v phng php th i vi brilliant black PN 1.21. Ph lc 21: Quy nh k thut v phng php th i vi brown HT 1.22. Ph lc 22: Quy nh k thut v phng php th i vi -caroten tng hp 1.23. Ph lc 23: Quy nh k thut v phng php th i vi caroten thc vt

1.24. Ph lc 24: Quy nh k thut v phng php th i vi cao annatto 1.25. Ph lc 25: Quy nh k thut v phng php th i vi -Apo-carotenal 1.26. Ph lc 26: Quy nh k thut v phng php th i vi este methyl (hoc Ethyl) ca acid -Apo-8'-Carotenoic 1.27. Ph lc 27: Quy nh k thut v phng php th i vi cao v nho 1.28. Ph lc 28: Quy nh k thut v phng php th i vi dioxyd titan 1.29. Ph lc 29: Quy nh k thut v phng php th i vi oxyd st 1.30. Ph lc 30: Quy nh k thut v phng php th i vi fast green FCF 2. Cc yu cu k thut quy nh trong Quy chun ny c th theo JECFA monograph 1 - Vol. 4, ngoi tr mt s php th ring c m t trong cc ph lc. Cc phng php th c hng dn trong Quy chun ny khng bt buc phi p dng, c th s dng cc phng php th khc tng ng. 3. Ly mu theo hng dn ti Thng t 16/2009/TT-BKHCN ngy 02 thng 6 nm 2009 ca B Khoa hc v Cng ngh v hng dn kim tra nh nc v cht lng hng ha lu thng trn th trng v cc quy nh khc ca php lut c lin quan. III. YU CU QUN L 1. Cng b hp quy 1.1. Cc phm mu phi c cng b ph hp vi cc quy nh ti Quy chun ny. 1.2. Phng thc, trnh t, th tc cng b hp quy c thc hin theo Quy nh v chng nhn hp chun, chng nhn hp quy v cng b hp chun, cng b hp quy ban hnh km theo Quyt nh s 24/2007/Q-BKHCN ngy 28 thng 9 nm 2007 ca B trng B Khoa hc v Cng ngh v cc quy nh ca php lut. 2. Kim tra i vi phm mu Vic kim tra cht lng, v sinh an ton i vi cc phm mu phi thc hin theo cc quy nh ca php lut. IV. TRCH NHIM CA T CHC, C NHN 1. T chc, c nhn phi cng b hp quy ph hp vi cc quy nh k thut ti Quy chun ny, ng k bn cng b hp quy ti Cc An ton v sinh thc phm v bo m cht lng, v sinh an ton theo ng ni dung cng b. 2. T chc, c nhn ch c nhp khu, xut khu, sn xut, bun bn v s dng cc phm mu sau khi hon tt ng k bn cng b hp quy v bo m cht lng, v sinh an ton, ghi nhn ph hp vi cc quy nh ca php lut. V. T CHC THC HIN 1. Giao Cc An ton v sinh thc phm ch tr, phi hp vi cc c quan chc nng c lin quan hng dn trin khai v t chc vic thc hin Quy chun ny. 2. Cn c vo yu cu qun l, Cc An ton v sinh thc phm c trch nhim kin ngh B Y t sa i, b sung Quy chun ny. 3. Trng hp hng dn ca quc t v phng php th v cc quy nh ca php lut vin dn trong Quy chun ny c sa i, b sung hoc thay th th p dng theo vn bn mi.

PH LC 1 YU CU K THUT V PHNG PHP TH I VI CURCUMIN 1. Tn khc, ch s 2. nh ngha Vng ngh, Kurkum INS: 100i ADI = 0 3 mg/kg th trng. Curcumin c sn xut bng cch chit c ngh - thn r ca cy Curcuma longa L. (Curcuma domestica Valeton) bng dung mi. thu c bt curcumin c hm lng cao, dch chit c tinh ch bng phng php kt tinh. Sn phm thu c cha cc curcumin, thnh phn mu chnh l 1,7-bis-(4-hydroxy3-methoxy-phenyl)-hepta-1,6-dien-3,5-dion (tn khc: Curcumin, Diferuloylmethan, CI Natural Yellow 3, CI (1975) 75300) v dn cht desmethoxy- v bis-desmethoxy- ca n theo t l khng c nh. Trong ch phm c th cha lng nh du v nha t nguyn liu. Ch cc dung mi sau y c th c s dng trong qu trnh chit v tinh ch curcumin: Aceton, methanol, ethanol, iso-propanol, hexan v ethyl acetat. Carbon dioxyd siu ti hn cng c th c s dng trong qu trnh chit. Cc thnh phn mu chnh: I. 1,7-Bis-(4-hydroxy-3-methoxyphenyl)-hepta-1,6-dien3,5-dion II. 1-(4-Hydroxyphenyl)-7-(4-hydroxy-3methoxyphenyl)-hepta-1,6-dien-3,5-dion III. 1,7-Bis-(4-hydroxyphenyl)-hepta-1,6-dien-3,5-dion I. 458-37-7 II. 33171-16-3 III. 33171-05-0 I. C21H20O6 II. C20H18O5 III. C19H16O4

Tn ha hc

M s C.A.S. Cng thc ha hc Cng thc cu to

Khi lng phn t 3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Cc phn ng mu Sc k bn mng 5.2. tinh khit Tn d dung mi

I. R1= R2=OCH3 II. R1= OCH3; R2=H III. R1= R2=H I. 368,39 II. 338,39 III. 308,39 Bt tinh th mu vng cam. Phm mu. Khng tan trong nc v diethyl ether; tan trong ethanol v acid acetic bng. Dung dch mu th trong ethanol c mu vng v cho hunh quang xanh nht. Thm dung dch mu th ny vo acid sulfuric c s c mu thm. t yu cu (m t trong phn Phng php th). Aceton: Hexan: Methanol: Ethanol: Khng c qu 30 mg/kg Khng c qu 25 mg/kg

Khng c qu 50 mg/kg

Isopropanol: Ethyl acetat:

Ch 5.3. Hm lng 6. Phng php th 6.1. nh tnh Cc phn ng mu

Khng c qu 2 mg/kg Khng nh hn 90% tng cc thnh phn mu. X l dung dch mu th trong nc hoc trong ethanol long vi acid hydrocloric n khi mu cam bt u xut hin. Chia dung dch sau khi x l thnh 2 phn, thm bt (hoc tinh th) acid boric vo 1 phn. Mu hng s xut hin nhanh sau khi thm, mu s d nhn bit hn khi so vi phn khng thm acid boric. Phn ng c th tin hnh bng cch nhng mt mu giy lc vo dch mu th tan trong ethanol, lm kh ti 100o, sau tm t mu giy lc bng dung dch acid boric long thm vi git acid hydrocloric. kh, s xut hin mu hng. Chm 5 L dch th (0,01 g mu th trong 1 ml ethanol 95%) trn 1 bn mng sc k (cellulose vi tinh th; 0,1 mm). Khai trin sc k trong bnh cha hn hp dung mi pha ng 3-methyl-1butanol/ethanol/nc/dung dch amoniac (4:4:2:1), dung mi khai trin trn bn mng 10-15 cm t vch xut pht. Kim tra, quan st di nh sng thng v n UV: Di nh sng thng v UV c 2 hoc 3 vt mu vng vi Rf trong khong 0,2 - 0,4. Di n UV c cc vt vi Rf khong 0,6 v 0,8 Cc vt u c hunh quang mu vng di n UV. Xc nh bng k thut hp th nguyn t thch hp cho hm lng quy nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c. Cn 0,08 g mu th (chnh xc n mg), chuyn vo bnh nh mc 200 ml v ho tan bng lc vi ethanol, nh mc n vch bng ethanol. Dng pipet ht 1 ml dung dch v cho vo bnh nh mc 100 ml, nh mc n vch bng ethanol. Xc nh hp th quang ti 425 nm, cuvet o dy 1 cm. Tnh t l tng hm lng cht mu trong mu th theo cng thc: Trong : A = hp th ca dung dch mu th W = Khi lng mu th 1607 = hp th ring ca dung dch curcumin chun trong ethanol ti 425 nm. Vic xc nh hp th quang phi tin hnh ngay khi pha xong dung dch, bi v mu ca dung dch c th nht dn theo thi gian.

Sc k bn mng

6.2 tinh khit Ch

6.3. nh lng

PH LC 2 YU CU K THUT V PHNG PHP TH I VI RIBOFLAVIN 1. Tn khc, ch s 2. nh ngha Tn ha hc M s C.A.S. Cng thc ha hc Cng thc cu to Vitamin B2, Lactoflavin INS: 101i ADI = 0 - 0,5 mg/kg th trng. Riboflavin; 3,10-dihydro-7,8-dimethyl-10-[(2S,3S,4R)-2,3,4,5tetrahydroxypentyl]benzo-[g]pteridin-2,4-dion; 7,8-dimethyl10-(1'-Dribityl) isoalloxazin. 83-88-5 C17H20N4O6

Khi lng phn t 3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Quang ph

376,37 Bt tinh th mu vng n vng cam, c mi nh. Phm mu. Rt kh tan trong nc, thc t khng tan trong cn, ether, aceton v cloroform; rt d tan trong dung dch kim long. S dng dung dch mu th trong nc, chun b trong phn nh lng, xc nh hp th quang (A) ti cc bc sng 267 nm, 375 nm, 444 nm. T s A375/A267 nm trong khong 0,31 v 0,33. T s A444/A267 nm trong khong 0,36 v 0,39. [ ]20,D: trong khong -115o v -140o. t yu cu (m t trong phn Phng php th).

Gc quay cc ring Phn ng mu 5.2. tinh khit Gim khi lng khi lm Khng c qu 1,5%. kh Tro sulfat Khng c qu 0,1%. Cc cht mu ph t yu cu (m t trong phn Phng php th). Cc amin thm bc nht Khng c qu 100 mg/kg tnh theo anilin. Ch Khng c qu 2 mg/kg. 5.3. Hm lng C17H20N4O6 Khng c thp hn 98%. 6. Phng php th 6.1 nh tnh Gc quay cc ring Sy kh mu ti 100 oC trong 4 g. Cn 50 mg v ha tan trong dung dch natri hydroxyd 0,05 N khng c carbonat v pha long n 10 ml cng bng dung dch ny. o quay quang ca dung dch trong 30 pht k t khi pha dung dch. Phn ng mu Ha tan khong 1 mg mu vo 100 ml nc. Dung dch c mu vng xanh nht khi cho nh sng truyn qua v c hunh quang xanh vng m vi nh sng phn x. Mu s b mt khi thm acid v c hoc kim v c. 6.2. tinh khit

Gim khi lng khi lm Sy ti 105 oC, trong 4 gi kh Tro sulfat - Th theo JECFA monograph 1 - vol.4 - Mu th: cn 2 g Cc cht mu ph Chun b dung dch chun khng c lumiflavin cho th nghim bng cch pha long 3 ml dung dch kali dicromat 0,1 N vi nc n 1000 ml. Cho mt t cloroform qua ct nhm oxyd loi ht ethanol. Ly 10 ml cloroform ny, thm 35 mg mu, lc trong 5 pht v lc. Mt mu ca dch lc khng c m hn mt mu ca 10 ml chun khi quan st trong dng c cha cng loi Ch Th theo hng dn ti JECFA monograph 1 - vol.4. S dng k thut hp th nguyn t thch hp vi hm lng ch nh xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong Ph lc 1: Cc phng php th chung phn cc phng php phn tch cng c 6.3. nh lng Tin hnh nh lng trong mi trng t nh sng. Phn tn 65,0 mg mu th trong 5 ml nc trong bnh nh mc 500 ml mu nu, cn m bo rng ton b mu th b tm t, ha tan trong 5 ml dung dch natri hydroxyd 2 N. Ngay khi mu tan hon ton, thm 100 ml nc v 2,5 ml acid acetic bng v pha long bng nc n 500 ml. Ly 20,0 ml dung dch ny, thm 3,5 ml dung dch natri acetat 1,4% (kl/tt) v pha long vi nc n 200 ml. o hp th quang (A) ti cc i hp th 444 nm. % Riboflavin = [(A5000) / (328W)] 1,367 trong A = hp th ca dung dch mu th ti 444 nm W = Khi lng mu th, tnh bng g.

PH LC 3 YU CU K THUT V PHNG PHP TH I VI TARTRAZIN 1. Tn khc, ch s 2. nh ngha CI Food Yellow 4, FD&C Yellow No. 5; S CI (1975): 19140 INS: 102 ADI = 0 - 7,5 mg/kg th trng. Ch yu gm Trinatri 5-hydroxy-1-(4-sulfonphenyl)-4-(4sulfonphenylazo)-H-pyrazol-3-carboxylat cng cc cht mu ph cng vi NaCl v (hoc) Na2SO4 l cc thnh phn khng mu chnh. C th chuyn thnh cht mu nhm (aluminium lake) tng ng, trong trng hp ny p dng quy nh chung i vi cc loi cht mu nhm. Trinatri 5-hydroxy-1-(4-sulfonphenyl)-4-(4-sulfonphenylazo)H-pyrazol-3-carboxylat 1934-21-0 C16H9N4Na3O9S2

Tn ha hc M s C.A.S. Cng thc ha hc Cng thc cu to

Khi lng phn t 534,37 3. Cm quan Dng bt hoc ht mu cam nht. 4. Chc nng Phm mu. 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc, t tan trong ethanol. nh tnh cc cht mu Phi c phn ng c trng ca cc cht mu 5.2. tinh khit Gim khi lng khi lm Khng c qu 15% cng vi clorid v sulfat tnh theo mui kh ti 135 oC natri. Cht khng tan trong nc Khng c qu 0,2%. Ch Khng c qu 2 mg/kg. Cc cht mu ph Khng c qu 1%. Cc cht hu c ngoi cht Khng c qu 0,5% (tng cc acid Tetrahydroxysuccinic; mu acid 4-Hydrazinbenzensulfonic; acid 4-Aminobenzensulfonic; acid 5-Oxo-1-(4-sulfophenyl)-2pyrazolin-3-carboxylic; acid 4,4'-Diazoaminodi (acid benzensulfonic). Cc amin Khng c qu 0,01% tnh theo anilin. thm bc nht khng sulfon ho Cc cht c th chit bng Khng c qu 0,2%. ether 5.3. Hm lng Khng c thp hn 85% tng cc cht mu. 6. Phng php th 6.1. tinh khit Ch - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng k thut hp th nguyn t thch hp vi hm lng ch nh xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4, Cc phng php th chung phn cc phng php phn tch cng c.

Cc cht mu ph

- Th theo hng dn ti JECFA monograph 1-Vol. 4 - S dng iu kin sau: Dung mi khai trin l dung mi s 4; chiu cao ca tuyn dung mi ~ 12 cm. Cc cht hu c ngoi cht - Th theo hng dn ti JECFA monograph 1-Vol. 4 mu - S dng k thut HPLC vi cc iu kin nh sau: HPLC ra gii gradient 2 n 100% vi tc tng c nh 2%/pht. 6.2. nh lng Tin hnh theo hng dn trong chuyn lun xc nh tng hm lng bng chun vi Titan (III) clorid (trong Ph lc 1: Cc phng php th chung) nh sau: Cn 0,6 - 0,7 g mu th; m 15 g natri hydro tartrat; Mi ml TiCl3 0,1 N tng ng vi 13,56 mg cc cht mu (D).

PH LC 4 YU CU K THUT V PHNG PHP TH I VI QUINOLIN 1. Tn khc, ch s 2. nh ngha CI food yellow 13, CI (1975) No. 47005, INS: 104 ADI = 0 10 mg/kg th trng. Ch phm thu c bng cch sulfonat ho 2-(2-quinolyl)-1,3indandion hoc hn hp cha khong hai phn ba 2-(2-quinolyl)1,3-indandion v mt phn ba 2-[2-(6-methyl-quinolyl)]1,3indandion; Ch phm cha hn hp mui natri ca cc disulfonat (ch yu), cc monosulfonat and cc trisulfonat ca cc hp cht nu trn v cc cht mu ph cng vi natri clorid v/hoc natri sulfat l cc thnh phn chnh khng to mu. Quinolin c th chuyn thnh dng mu nhm tng ng, trong trng hp ny p dng quy chun chung cho dng mu nhm ca cc phm mu. Dinatri 2-(1,3-dioxo-2-indanyl)-6,8-quinolin sulfat; dinatri 2-(2quinolyl)-indan-1,3-dion disulfonat (thnh phn chnh) 8004-72-0 (acid disulfonic khng methyl ha) C18H9NNa2O8S2 (thnh phn mu chnh) Thnh phn mu chnh


Khi lng phn t 3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc, t tan trong ethanol nh tnh cc cht mu Phi c phn ng c trng ca cc cht mu. 5.2. tinh khit Gim khi lng khi sy Khng c qu 30% cng vi clorid v sulfat tnh theo mui ti 135o natri. Cht khng tan trong nc Khng c qu 0,2%. Ch Khng c qu 2 mg/kg. Km Khng c qu 50 mg/kg. Cc cht mu ph Tng hm lng 2 cht 2-(2-quinolyl)-1,3-indandion v 2-[2-(6methylquinolyl)]-1,3-indandion Khng c qu 4 mg/kg Cc hp cht hu c ngoi Tng hm lng cc cht 2-methylquinolin, acid 2cc thnh phn methylquinolinsulfonic, acid phtalic, 2,6-dimethylquinolin, acid mu 2,6-dimethylquinolin sulfonic Khng c qu 0,5% Cc amin thn bc nht Khng c qu 0,01% tnh theo anilin. khng sulfonat ha Cht c th chit bng Khng c qu 0,2%. ether 5.3. Hm lng Khng nh hn 70% tng cc thnh phn mu. i vi ch phm quinolin vng sn xut t 2-(2-quinolyl)-1,3-indandion s c cc thnh phn sau: - Khng nh hn 80% dinatri 2-(2-quinolyl)-indan- 1,3diondisulfonat;

6 mui: R1 = SO3Na, R2 = H 8 mui: R1, R2 = SO3H Thnh phn mu chnh: 477,38 Bt hoc ht nh mu vng. Phm mu.

- Khng c qu 15% natri 2-(2-quinolyl)-indan-1,3dionmonosulfonat; - Khng c qu 7% trinatri 2-(2-quinolyl)-indan-1,3diontrisulfonat; 6. Phng php th 6.1. tinh khit Ch Th theo hng dn ti JECFA monograph 1-Vol. 4. S dng k thut hp th nguyn t thch hp vi hm lng ch nh xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4, Cc phng php th chung phn cc phng php phn tch cng c. Cc cht mu ph Th gii hn ca 2-(2-quinolyl)-1,3-indandion v 2-[2-(6methylquinolyl)]-1,3-indandion. S dng thit b v ether c cht lng theo m t trong chuyn lun xc nh cht c th chit bng ether v tin hnh chit nh sau: Ra dch chit 2 ln mi ln vi 25 ml nc ct. Cho ether bay hi dch chit cn khong 5 ml, sau cho vo t sy ti nhit 105o ui ht ether. Ho tan cn cn li trong cloroform v pha long n chnh xc 10 ml. Xc dnh hp th quang ca dung dch ny ti bc sng hp th cc i (khong 420 nm) s dng cloroform lm dung dch i chng. Gi tr hp th quang = 0,27 tng ng vi gii hn ca 2-(2-quinolyl)1,3-indandion l 4 mg/kg. 2-[2-(6-methylquinolyl)]-1,3indandion c nh gi nh 2-(2-quinolyl)-1,3-indandion. Cc hp cht hu c ngoi Cc hp cht hu c ngoi cc thnh phn mu trong quinolin cc thnh phn ch c trong ch phm sn xut t 2-(2-quinolyl)-1,3-indandion. mu Xc nh bng phng php sc k lng, vi cc iu kin nh sau: Thit b: Sc k lng phn gii cao c b iu khin gradien nng pha ng. Detector: Detector UV cho HPLC c th o hp th quang ti 254 nm. Ct: Nucleosil, 250X4 mm, 7 m. H dung mi: A: m acetat pH 4,6:nc (1:10). m acetat bao gm dung dch NaOH 1M : acid acetic 1 M : nc (5:10:35). B: (A):methanol (20:80). Nng mu 1%(m/v) trong dung mi A. iu kin gradient nng pha ng: Pht %A 0 100 15 65 20 50 25 0 36 0 42 100 Tc dng: 1 ml/pht 6.2. nh lng Tin hnh theo hng dn trong chuyn lun Xc nh tng hm lng bng phng php quang ph (JECFA monograph 1 - Vol. 4). Dung mi: m photphat pH 7 Pha long dung dch A: t 10 ml pha n 250 ml hp th ring (a): 86,35 Bc sng hp th cc i: 415 nm %B 0 35 50 100 100 0

Ring i vi ch phm quinolin sn xut t 2-(2-quinolyl)-1,3indandion xc nh t l % ca di-, mono-, trisulfonat: S dng iu kin HPLC m t trong chuyn lun xc nh cc hp cht hu c ngoi thnh phn mu, vi nng dung dch mu th trong dung mi HPLC A l 0,05% thay cho 1%. Biu th kt qu theo t l % tng cc thnh phn mu c trong ch phm.

PH LC 5 YU CU K THUT V PHNG PHP TH I VI SUNSET YELLOW FCF 1. Tn khc, ch s 2. nh ngha CI Food Yellow 3, FD&C Yellow No. 6; S CI (1975): 15985 INS: 110 ADI = 0 - 2,5 mg/kg th trng. Ch yu gm dinatri 6-hydroxy-5-(4-sulfonphenylazo)-2naphthalen-6-sulfonat cng cc cht mu ph cng vi NaCl v (hoc) Na2SO4 l cc thnh phn khng mu chnh. C th chuyn thnh cht mu nhm (aluminium lake) tng ng, trong trng hp ny p dng quy nh chung i vi cc loi cht mu nhm. dinatri 6-hydroxy-5-(4-sulfonphenylazo)-2-naphthalen-6sulfonat. 2783-94-0 C16H10N2Na2O7S2


Khi lng phn t 452,38 3. Cm quan Dng bt hoc ht mu cam. 4. Chc nng Phm mu. 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc, t tan trong ethanol. nh tnh cc cht mu Phi c phn ng c trng ca cc cht mu. 5.2. tinh khit Gim khi lng khi lm Khng c qu 15% cng vi clorid v sulfat tnh theo mui kh ti 135oC natri. Cht khng tan trong nc Khng c qu 0,2%. Ch Khng c qu 2 mg/kg. Cc cht mu ph Khng c qu 5%. Trong khng c qu 2% cc cht mu khc ngoi trinatri 2-hydroxy-1-(4-sulfonphenylazo)naphthalen-3,6-disulfonat. Cc cht hu c ngoi cht Khng c qu 0,5% tng cc acid 4-amino-1-benzensulfonic; mu Acid 3-hydroxy-2,7-naphthalendisulfonic; Acid 6-hydroxy-2-naphthalensulfonic; Acid 7-hydroxy-1,3-naphthalendisulfonic; Acid 4,4'-diazoaminodibenzensulfonic; Acid 6,6'-dioxy-2-naphthalensulfonic. Cc amin Khng c qu 0,01% tnh theo anilin. thm bc nht khng sulfon ho Cc cht c th chit bng Khng c qu 0,2%. ether 5.3. Hm lng Khng c thp hn 85% tng cc cht mu. 6. Phng php th 6.1. tinh khit Ch - Th theo hng dn ti JECFA monograph 1-Vol. 4.

- S dng k thut hp th nguyn t thch hp vi hm lng ch nh xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c. Cc cht mu ph - Th theo hng dn ti JECFA monograph 1-Vol. 4 - S dng iu kin sau: Dung mi khai trin l dung mi s 4; chiu cao ca tuyn dung mi ~ 17 cm. Cc cht hu c ngoi cht - Th theo hng dn ti JECFA monograph 1-Vol. 4 mu - S dng k thut HPLC vi cc iu kin nh sau: HPLC ra gii gradient 2 n 100% vi tc tng c nh 4%/pht, tip theo ra gii ti 100%. 6.2. nh lng Tin hnh theo hng dn trong chuyn lun xc nh tng hm lng bng chun vi Titan (III) clorid (trong JECFA monograph 1-Vol. 4) nh sau: Cn 0,5 - 0,6 g mu th; m 10 g natri citrat; Mi ml TiCl3 0,1 N tng ng vi 11,31 mg cc cht mu (D).

PH LC 6 QUY YU CU K THUT V PHNG PHP TH I VI CARMIN 1. Tn khc, ch s Cochineal carmin, CI Natural Red 4, CI (1975) No. 75470, INS 120 ADI cho c nhm carmin l 0 - 5 mg/kg th trng tnh theo amoni carminat, cc mui Ca, K, Na quy ra tng ng. Ch phm thu c bng cch chit t phm yn chi cha xc kh ca rp son ci (Dactylopius coccus Costa) vi nc; Cht mu chnh l mt phc chelat nhm ngm nc ca acid carminic, trong phc ny t l phn t gia nhm v acid carminic l 1/2. Trong ch phm thng mi, cht mu chnh kt hp vi cation amoni; calci; kali hoc natri, dng t do hay kt hp, cc cation ny lun d. Trong ch phm c th cha protein t xc cn trng nguyn liu, c th cha mui carminat v lng d t cation nhm. Phc chelat nhm ngm nc ca acid carminic (7-beta-Dglucopyranosyl-3,5,6,8-tetrahydroxy-1-methyl-9,10-dioxoanthracen-2-carboxylic acid) 1390-65-4 (carmin) 1260-17-9 (acid carminic) Acid carminic C22H20O13 Acid carminic

2. nh ngha


Phc nhm ca acid carminic

Khi lng phn t 3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan

Acid carminic: 492,39 Dng bt hoc mnh vn mu ti m Phm mu tan ca ch phm carmin ph thuc vo bn cht ca cation c mt trong ch phm. Ch phm cha ch yu cation amoni (mui

Phn ng mu 5.2. tinh khit Gim khi lng khi lm Khng c qu 20%. kh Tro ton phn Khng c qu 12%. Protein Khng c qu 25%. Cht khng tan trong dung Khng c qu 1%. dch amoniac long Ch Khng c qu 2 mg/kg. Yu cu v vi sinh vt Salmonella: m tnh. 5.3. Hm lng C22H20O13 Khng thp hn 50% C22H20O13 tnh theo ch phm lm kh. 6. Phng php th 6.1 nh tnh Phn ng mu - Kim ha nh dung dch mu th bng cch thm 1 git dung dch natri hydroxyd (hoc kali hydroxyd) 10%. Dung dch chuyn sang mu tm. - Thm mt t tinh th natri dithionit (Na2S2O4) vo cc dung dch mu th trung tnh, kim ha, acid ha. Cc dung dch ny khng b mt mu. - Lm kh lng nh mu th trn a s. ngui hon ton, nh 1-2 git thuc th acid sulfuric vo cn cn li trn a. Mu ca cn khng thay i. - Acid ha dung dch mu th trong nc bng dung dch thuc th acid hydrocloric, th tch thuc th bng 1/3 th tch dch th, cho thm ru amylic, lc u. Tch ly lp ru amylic v ra 24 ln vi ng th tch nc ct loi han ton acid hydrocloric. Pha long phn ru amylic vi ether du ho (40-60 0 ) ng th tch hoc th tch ether du ho gp i, lc vi ln vi mt t nc ct loi ht mu. Thm, nh tng git dung dch urani acetat 5%, lc u sau mi ln thm. Dung dch chuyn sang mu xanh lc ngc c trng. 6.2 tinh khit Gim khi lng khi lm - Th theo hng dn ti JECFA monograph 1-Vol. 4 kh - sy ti 135 oC trong 3 gi. Tro ton phn - Th theo hng dn ti JECFA monograph 1-Vol. 4 - chuyn lun xc nh tro, cn 1 g mu. Protein - Th theo hng dn ti JECFA monograph 1-Vol. 4 - chuyn lun xc nh Nit phi amoni X 6,25 Cht khng tan trong dung Cn 0,25 g (chnh xc n mg) mu th c lm kh, hon dch amoniac long tan trong 2,5 ml dung dch amoniac long (160 ml dung dch thuc th amoniac c, thm nc ct n 500 ml) v pha long vi nc ct n 100 ml, thu c dung dch trong. Lc dung dch ny qua phu thu tinh xp. Ra phu vi dung dch amoniac 0,1% v lm kh n khi lng khng i ti 105o. Ch Xc nh bng k thut hp th nguyn t thch hp cho hm lng quy nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c. 6.3. nh lng Cn 100 mg (chnh xc n mg) mu th, ho tan vo 30 ml dung dch acid hydrocloric 2N si, lm ngui. Chuyn ton b dung dch sang bnh nh mc 1000 ml, pha long v nh mc ti vch bng nc ct, lc u. Xc nh hp th quang ca dung dch ti bc sng hp th cc i khong 494 nm, s dng mu trng

amoni carminat) tan tt trong nc ti pH 3,0 v pH 8,5. Ch phm cha ch yu cation calci (mui calci carminat) t tan trong nc ti pH 3,0 nhng tan tt ti pH 8,5. t yu cu (m t trong phn Phng php th)

l nc ct, cuvet o c dy 1 cm. Tnh hm lng % acid carminic trong mu th theo cng thc: (100 A 100) / (1,39 W) trong : A = hp th quang ca dung dch mu th; W = Khi lng mu th,tnh bng mg; 1,39 = hp th quang ca dung dch acid carminic nng 100 mg/1000 ml; Nu hp th quang o c khng nm trong khong gi tr 0,65 - 0,75, chun b mu th khc v hiu chnh khi lng mu th thch hp.

PH LC 7 YU CU K THUT V PHNG PHP TH I VI CARMOISIN 1. Tn khc, ch s 2. nh ngha CI Food Red 3, Azorubine; CI (1975): 14720 INS: 122 ADI = 0 - 4 mg/kg th trng. Ch yu gm dinatri 4-hydroxy-3-(4-sulfon-1-naphthylazo)-1naphthalensulfonat cng cc cht mu ph cng vi NaCl v (hoc) Na2SO4 l cc thnh phn khng mu chnh. C th chuyn thnh cht mu nhm (aluminium lake) tng ng, trong trng hp ny p dng quy nh chung i vi cc loi cht mu nhm. dinatri 4-hydroxy-3-(4-sulfon-1-naphthylazo)-1naphthalensulfonat. 3567-69-9 C20H12N2Na2O7S2


Khi lng phn t 502,44 3. Cm quan Dng bt hoc ht mu . 4. Chc nng Phm mu. 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc, t tan trong ethanol. nh tnh cc cht mu Phi c phn ng c trng ca cht mu. 5.2. tinh khit Gim khi lng khi lm Khng c qu 15% cng vi clorid v sulfat tnh theo mui kh ti 135 oC natri. Cht khng tan trong nc Khng c qu 0,2%. Ch Khng c qu 2 mg/kg. Cc cht mu ph Khng c qu 1%. Cc cht hu c ngoi cht Khng c qu 0,5% tng cc acid 4-Amino-1mu naphthalensulfonic v acid 4-hydroxy-1-naphthalensulfonic. Cc amin Khng c qu 0,01% tnh theo anilin. thm bc nht khng sulfon ho Cc cht c th chit bng Khng c qu 0,2%. ether 5.3. Hm lng Khng c thp hn 85% tng cc cht mu. 6. Phng php th 6.1 tinh khit Ch - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng k thut hp th nguyn t thch hp vi hm lng ch nh xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c. Cc cht mu ph - Th theo hng dn ti JECFA monograph 1-Vol. 4 - S dng iu kin sau: Dung mi khai trin l dung mi s 4; chiu cao ca tuyn dung mi ~ 17 cm.

Cc cht hu c ngoi cht - Th theo hng dn ti JECFA monograph 1-Vol. 4 mu - S dng k thut HPLC vi cc iu kin nh sau: HPLC ra gii gradient t 2% n 100% vi tc tng c nh 2%/pht. 6.3. nh lng Tin hnh theo hng dn trong chuyn lun xc nh tng hm lng bng chun vi Titan (III) clorid (trong JECFA monograph 1-Vol. 4) nh sau: Cn 0,5 - 0,6 g mu th; m 15 g natri hydrotartrat; Mi ml TiCl3 0,1 N tng ng vi 12,56 mg cc cht mu (D).

PH LC 8 YU CU K THUT V PHNG PHP TH I VI AMARANTH 1. Tn khc, ch s 2. nh ngha CI Food Red 9, Naphtol Rot S.; CI (1975): 16185 INS: 123 ADI = 0 - 0,5 mg/kg th trng Ch yu gm trinatri 3-hydroxy-4-(4-sulfon-1-naphthylazo)-2,7naphthalendisulfonat v cc cht mu ph cng vi NaCl v (hoc) Na2SO4 l cc thnh phn khng mu chnh. C th chuyn thnh cht mu nhm (aluminium lake) tng ng, trong trng hp ny p dng quy nh chung i vi cc loi cht mu nhm. trinatri 3-hydroxy-4-(4-sulfon-1-naphthylazo)-2,7naphthalendisulfonat 915-67-3 C20H11N2Na3O10S3


Khi lng phn t 604,48 3. Cm quan Dng bt hoc ht mu nu hoc nu sm. 4. Chc nng Phm mu. 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc, t tan trong ethanol nh tnh cc cht mu Phi c phn ng c trng ca cht mu. 5.2. tinh khit Gim khi lng khi lm Khng c qu 15% cng vi clorid v sulfat tnh theo mui kh ti 135 oC natri. Cht khng tan trong nc Khng c qu 0,2%. Ch Khng c qu 2 mg/kg. Cc cht mu ph Khng c qu 3%. Cc cht hu c ngoi cht Khng c qu 0,5% tng cc acid 4-amino-1mu naphthalensulfonic; acid 3-hydroxy-2,7-naphthalendisulfonic; acid 6-hydroxy-2naphthalensulfonic; acid 7-hydroxy-1,3-naphthalendisulfonic v acid 7-hydroxy-1,3,6- naphthalen-trisulfonic. Cc amin Khng c qu 0,01% tnh theo anilin. thm bc nht khng sulfon ho Cc cht c th chit bng Khng c qu 0,2%. ether 5.3. Hm lng Khng c thp hn 85% tng cc cht mu. 6. Phng php th 6.1 tinh khit Ch - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng k thut hp th nguyn t thch hp vi hm lng ch nh xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c.

Cc cht mu ph

- Th theo hng dn ti JECFA monograph 1-Vol. 4 - S dng iu kin sau: Dung mi khai trin l dung mi s 3; chiu cao ca tuyn dung mi ~ 17 cm, sau 1 gi khai trin. Cc cht hu c ngoi cht - Th theo hng dn ti JECFA monograph 1-Vol. 4 mu - S dng k thut HPLC vi cc iu kin nh sau: HPLC ra gii gradient 2 n 100% vi tc tng c nh 4%/pht. 6.2. nh lng Tin hnh theo hng dn trong chuyn lun xc nh tng hm lng bng chun vi Titan (III) clorid (trong JECFA monograph 1-Vol. 4) nh sau: Cn 0,7 - 0,8 g mu th; m 10 g natri citrat; Mi ml TiCl3 0,1 N tng ng vi 15,11 mg cc cht mu (D).

PH LC 9 YU CU K THUT V PHNG PHP TH I VI PONCEAU 4R 1. Tn khc, ch s CI Food Red 7, Cochineal Red A ; New Cochineal; CI (1975): 16255 INS: 124 ADI = 0 - 4 mg/kg th trng. Ch yu gm trinatri d-2-hydroxy-1-(4-sulfon-1-naphthylazo)6,8-naphthalendisulfonat cng cc cht mu ph cng vi NaCl v (hoc) Na2SO4 l cc thnh phn khng mu chnh. C th chuyn thnh cht mu nhm (aluminium lake) tng ng, trong trng hp ny p dng quy nh chung i vi cc loi cht mu nhm. Trinatri-2-hydroxy-1-(4-sulfon-1-naphthylazo)-6,8naphthalendisulfonat 2611-82-7 C20H11N2Na3O10S3.1,5H2O

2. nh ngha


Khi lng phn t 631,51 3. Cm quan Dng bt hoc ht mu . 4. Chc nng Phm mu. 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc, t tan trong ethanol. nh tnh cc cht mu Phi c phn ng c trng ca cht mu. 5.2. tinh khit Gim khi lng khi lm Khng c qu 20% cng vi clorid v sulfat tnh theo mui kh ti 135 oC natri. Cht khng tan trong nc Khng c qu 0,2%. Ch Khng c qu 2 mg/kg. Cc cht mu ph Khng c qu 1%. Cc cht hu c ngoi cht Khng c qu 0,5% tng cc acid 4-amino-1mu naphthalensulfonic; acid 7-hydroxy-1,3-naphthalendisulfonic; acid 3-hydroxy-2,7-naphthalensulfonic; acid 6-hydroxy-2naphthalensulfonic v acid 7-hydroxy-1,3,6naphthalentrisulfonic. Cc amin Khng c qu 0,01% tnh theo anilin. thm bc nht khng sulfon ho Cc cht c th chit bng Khng c qu 0,2%. ether 5.3. Hm lng Khng c thp hn 85% tng cc cht mu. 6. Phng php th 6.1. tinh khit Ch - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng k thut hp th nguyn t thch hp vi hm lng ch nh xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong

JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c. Cc cht mu ph - Th theo hng dn ti JECFA monograph 1-Vol. 4 - S dng iu kin sau: Dung mi khai trin l dung mi s 3; chiu cao ca tuyn dung mi ~ 17 cm, sau 1 gi khai trin. Cc cht hu c ngoi cht - Th theo hng dn ti JECFA monograph 1-Vol. 4 mu - S dng k thut HPLC vi cc iu kin nh sau: HPLC ra gii gradient 2 n 100% vi tc tng c nh 2%/pht. 6.2. nh lng Tin hnh theo hng dn trong chuyn lun xc nh tng hm lng bng chun vi Titan (III) clorid (trong JECFA monograph 1-Vol. 4) nh sau: Cn 0,7 - 0,8 g mu th; m 10 g natri citrat; Mi ml TiCl3 0,1 N tng ng vi 15,78 mg cc cht mu (D).

PH LC 10 YU CU K THUT V PHNG PHP TH I VI ERYTHROSIN 1. Tn khc, ch s 2. nh ngha CI Food Red 14, FD&C Red No. 3; CI (1975): 45430 INS 127 ADI = 0 - 0,1 mg/kg th trng Ch yu l mui dinatri 9-(o-carboxyphenyl)-6-hydroxy2,4,5,7-tetraiodo-3-isoxanthon monohydrat cng cc cht mu ph cng vi NaCl v (hoc) Na2SO4 l cc thnh phn khng mu chnh. C th chuyn thnh cht mu nhm (aluminium lake) tng ng, trong trng hp ny p dng quy nh chung i vi cc loi cht mu nhm. Mui dinatri 9-(o-carboxyphenyl)-6-hydroxy-2,4,5,7-tetraiodo3-isoxanthon monohydrat 16423-68-0 C20H6I4Na2O5.H2O


Khi lng phn t 897,88 3. Cm quan Dng bt hoc ht mu . 4. Chc nng Phm mu. 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc v trong ethanol nh tnh cc cht mu Phi c phn ng c trng ca cht mu. 5.2. tinh khit Gim khi lng khi lm Khng c qu 13% cng vi clorid v sulfat tnh theo mui kh ti 135 oC natri. Iod v c Khng c qu 0,1% tnh theo natri iodid. Cht khng tan trong nc Khng c qu 0,2%. Km Khng c qu 50 mg/kg. Ch Khng c qu 2 mg/kg. Cc cht mu ph Khng c qu 4% (khng bao gm Fluorescein). Fluorescein Khng c qu 20 mg/kg. Cc cht hu c ngoi cht Khng c qu 0,2% Tri iodoresorcinol mu Khng c qu 0,2% acid 2-(2,4-dihydroxy-3,5-diiodobenzoyl) benzoic Cc cht c th chit bng Khng c qu 0,2% i vi dung dch mu th pH < 7. ether 5.3. Hm lng Khng c thp hn 87% tng cc cht mu. 6. Phng php th 6.1. tinh khit Iod v c Cn 1,0 g mu th vo mt cc c m 100 ml. Thm 75 ml nc ct, dng my khuy t khuy cho tan hon ton. Cho in cc c trng cho iod v mt in cc so snh vo dung dch v t my o th thang o thch hp. c th ca h (mV). Dng buret thm dung dch bc nitrat 0,001 M, u tin mi ln thm 0,5 ml, khi gn n im tng ng (nhn bit

bng s thay i in th sau mi ln thm) gim cn 0,1 ml/ln, n nh v c in th ca h sau mi ln thm (mV). Tin hnh chun n khi thm dung dch bc nitrat 0,001 M m khng lm thay i in th dung dch nhiu. V ng biu th s ph thuc gia in th dung dch (mV) vo th tch dung dch bc nitrat 0,001 M thm vo. im tng ng c xc nh l im m th tch dung dch bc nitrat 0,001 M thm vo tng ng vi dc ca ng th ln nht. Hm lng natri iodid trong mu c tnh = chun x 0,015%. Ch - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng k thut hp th nguyn t thch hp vi hm lng ch nh xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c. Cc cht mu ph - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng iu kin sau: Dung mi khai trin l dung mi s 5; chiu cao ca tuyn dung mi ~ 17 cm. Ch : Khng bn mng sc k trc tip ra ngoi snh sng mt tri Cc cht hu c ngoi cht - Th theo hng dn ti JECFA monograph 1-Vol. 4 mu - S dng k thut sc k ct vi cc iu kin hp th nh sau: Tri-iodoresorcinol: 0,079 mgL-1cm-1 ti bc sng 233 nm, trong mi trng acid. Acid 2-(2,4-dihydroxy-3,5-di-iodobenzoyl) benzoic: 0,047 mgL-1cm-1 ti bc sng 348 nm, trong mi trng kim. Fluorescein Nguyn tc: Fluorescein c tch khi mu th bng sc k lp mng (TLC) v so snh vi sc k ca Fluorescein chun ti nng tng ng vi gii hn quy nh. Dung mi: Methanol + nc + dung dch amoniac (khi lng ring ~ 0,890) (500 ml + 400 ml + 100 ml). Chun b mu: Cn 1,0 g mu th, ha tan trong khong 50 ml dung mi v pha long n 100 ml. Chun: Cn chnh xc lng Fluorescein ( tinh ch bng cch kt tinh li trong ethanol) tng ng vi 1 g x hm lng cc cht mu trong mu (xc nh trong phn nh lng). Ha tan trong nc (hoc trong nc c 10 ml dung dch amoniac c khi lng ring ~ 0,890 nu s dng acid khng c Fluorescein), pha long v nh mc n 100 ml. Tin hnh pha long theo cc bc lin tip nh sau: Ht 1ml pha long n 100 ml bng nc. Ht 1ml pha long n 100 ml bng nc. Ht 20 ml pha long n 100 ml bng dung mi. Dung mi sc k: n-Butanol + nc + dung dch amoniac (khi lng ring ~ 0,890) + ethanol (100 ml + 44 ml + 1 ml + 22,5 ml). Tin hnh: Ln lt chm cnh nhau 25 L dung dch th v dung dch chun trn cng 1 bn mng cellulose. Khai trin trong h dung mi sc k trong 16 gi. kh bn mng. Soi di n UV v so snh hunh quang ca vt th vi vt chun. Mt

hunh quang ca vt th khng c qu vt chun. Ch : khng bn mng ra nh sng mt tri trc tip. 6.2. nh lng Cn 1 g (chnh xc n mg) mu th, ha tan trong 250 ml nc, chuyn vo mt cc 500 ml sch, thm 8,0 ml acid nitric 1,5 N, khuy u. Lc qua phu lc thy tinh xp ( xp 3, ng knh 5 cm), cha sn a thy tinh v cn b. Ra cn bng acid nitric 0,5% n khi dch ra khng b c khi th vi dung dch bc nitrat (TS), sau vi 30 ml nc ct. Sy ti nhit 135 5 oC n khi lng khng i. Cn thn ph cn bng a thy tinh, ngui trong bnh ht m v cn. Hm lng tng cc cht mu

=

Khi lng cn x 107,4 Khi lng mu th

%

Xc nh Cht khng tan trong acid hydrocloric trong mu nhm erythrosin Ha cht, thuc th: Acid hydrocloric c Acid hydrocloric 0,5% (tt/tt) Dung dch amoniac long (pha long 10 ml dung dch amoniac c khi lng ring =0,890 bng nc n 100 ml). Tin hnh: Cn khong 5 g (chnh xc n mg) mu th vo cc 500 ml. Thm 250 ml nc v 60 ml acid hydrocloric c. un si ha tan nhm trong khi erythrosin chuyn v dng acid t do, khng tan trong acid. Lc qua phu lc thy tinh xp s 4 cn b. Ra cn vi mt lng nh acid hydrocloric 0,5% nng, sau vi nc ct nng. Loi b dch ra acid, ly phu thy tinh xp ra v ra phu bng dung dch amoniac long n khi dch ra khng c mu. Sy ti 135 oC n khi lng khng i. Cn phu v tnh khi lng cn cn li v tnh hm lng % so vi khi lng mu ly.

PH LC 11 YU CU K THUT V PHNG PHP TH I VI RED 2G 1. Tn khc, ch s 2. nh ngha CI Food Red 10, Azogeranine; CI (1975): 18050 INS: 128 ADI = 0 - 1 mg/kg th trng. Ch yu gm dinatri 8-acetamido-1-hydroxy-2-phenylazo-3,6naphthalendisulfonat cng cc cht mu ph cng vi NaCl v (hoc) Na2SO4 l cc thnh phn khng mu chnh. C th chuyn thnh cht mu nhm (aluminium lake) tng ng, trong trng hp ny p dng quy nh chung i vi cc loi cht mu nhm. Dinatri 8-acetamido-1-hydroxy-2-phenylazo-3,6naphthalendisulfonat 3734-67-6 C18H13N3Na2O8S2


Khi lng phn t 509,43 3. Cm quan Dng bt hoc ht mu . 4. Chc nng Phm mu. 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc, t tan trong ethanol. nh tnh cc cht mu Phi c phn ng c trng ca cht mu. 5.2. tinh khit Gim khi lng khi lm Khng c qu 20% cng vi clorid v sulfat tnh theo mui kh ti 135 oC natri. Cht khng tan trong nc Khng c qu 0,2%. Ch Khng c qu 2 mg/kg. Cc cht mu ph Khng c qu 2%. Cc cht hu c ngoi cht Khng c qu 0,3% tng cc acid 5-acetamido-4-hydroxymu 2,7-naphthalen-2,7-disulfonic v acid 5-Amino-4-hydroxy-2,7naphthalen-2,7-disulfonic. Cc amin Khng c qu 0,01% tnh theo anilin. thm bc nht khng sulfon ho Cc cht c th chit bng Khng c qu 0,2%. ether 5.3. Hm lng Khng c thp hn 80% tng cc cht mu. 6. Phng php th 6.1. tinh khit Ch - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng k thut hp th nguyn t thch hp vi hm lng ch nh xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c. Cc cht mu ph - Th theo hng dn ti JECFA monograph 1-Vol. 4 - S dng iu kin sau: Dung mi khai trin l dung mi s 4;

chiu cao ca tuyn dung mi ~ 17 cm. Cc cht hu c ngoi cht - Th theo hng dn ti JECFA monograph 1-Vol. 4 mu - S dng k thut HPLC vi cc iu kin nh sau: HPLC ra gii gradient 2 n 100% vi tc tng c nh 2%/pht. 6.2. nh lng Tin hnh theo hng dn trong chuyn lun xc nh tng hm lng bng chun vi Titan (III) clorid (trong JECFA monograph 1-Vol. 4) nh sau: Cn 0,6 - 0,7 g mu th; m 15 g natri hydrotartrat; Mi ml TiCl3 0,1 N tng ng vi 12,74 mg cc cht mu (D).

PH LC 12 YU CU K THUT V PHNG PHP TH I VI ALLURA RED AC 1. Tn khc, ch s 2. nh ngha CI Food Red 17, FD&C Red No. 40; CI (1975): 16035 INS: 129 ADI = 0 - 7 mg/kg th trng. Ch yu gm dinatri 6-hydroxy-5-(2-methoxy-5-methyl-4sulfon-phenylazo)-2-naphthalen-sulfonat cng cc cht mu ph cng vi NaCl v (hoc) Na2SO4 l cc thnh phn khng mu chnh. C th chuyn thnh cht mu nhm (aluminium lake) tng ng, trong trng hp ny p dng quy nh chung i vi cc loi cht mu nhm. Dinatri 6-hydroxy-5-(2-methoxy-5-methyl-4-sulfon-phenylazo)2-naphthalen-sulfonat 25956-17-6 C18H14N2Na2O8S2


Khi lng phn t 496,43 3. Cm quan Dng bt hoc ht mu sm. 4. Chc nng Phm mu. 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc, t tan trong ethanol. nh tnh cc cht mu Phi c phn ng c trng ca cht mu. 5.2. tinh khit Gim khi lng khi lm Khng c qu 15% cng vi clorid v sulfat tnh theo mui kh ti 135 oC natri. Cht khng tan trong nc Khng c qu 0,2%. Ch Khng c qu 2 mg/kg. Cc cht mu ph Khng c qu 3%. Cc cht hu c ngoi cht Khng c qu 0,3% Natri 6-hydroxy-2-naphtalen sulfonat; mu Khng c qu 0,2% acid 4-amino-5-methoxy-2-methylbenzen-sulfonic; Khng c qu 1,0% dinatri 6,6'-oxybis(2-naphthalensulfonat); Cc amin Khng c qu 0,01% tnh theo anilin. thm bc nht khng sulfon ho Cc cht c th chit bng Khng c qu 0,2%. ether 5.3. Hm lng Khng c thp hn 85% tng cc cht mu. 6. Phng php th 6.1 tinh khit Ch - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng k thut hp th nguyn t thch hp vi hm lng ch nh xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong

JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c. Cc cht mu ph - Th theo hng dn ti JECFA monograph 1-Vol. 4 - S dng iu kin sau: Dung mi khai trin l dung mi s 4; chiu cao ca tuyn dung mi ~ 17 cm. Cc cht hu c ngoi cht - Th theo hng dn ti JECFA monograph 1-Vol. 4 mu - S dng k thut sc k lng di cc iu kin nh sau: HPLC ra gii gradient t 0% n 18% vi tc tng c nh 1%/pht; 18 n 62% vi tc tng c nh 7%/pht, sau ra gii ti 100%, tc dng 0,6 ml/pht. 6.2. nh lng Tin hnh theo hng dn trong chuyn lun xc nh tng hm lng bng chun vi Titan (III) clorid (trong JECFA monograph 1-Vol. 4) nh sau: Cn 0,5 - 0,6 g mu th; m 15 g natri hydrotartrat; Mi ml TiCl3 0,1 N tng ng vi 12,41 mg cc cht mu (D).

PH LC 13 YU CU K THUT V PHNG PHP TH I VI INDIGOTIN 1. Tn khc, ch s Indigocarmin; CI Food Blue 1, FD&C Blue No. 2; CI (1975): 73015 INS: 132 ADI = 0 - 5 mg/kg th trng. Ch yu gm dinatri 3,3' -dioxo-[delta2,2'-biindolin]-5,5'disulfonat v dinatri 3,3'-dioxo-[delta2,2'-biindolin]-5,7'disulfonat cng cc cht mu ph cng vi NaCl v (hoc) Na2SO4 l cc thnh phn khng mu chnh. C th chuyn thnh cht mu nhm (aluminium lake) tng ng, trong trng hp ny p dng quy nh chung i vi cc loi cht mu nhm. Dinatri 3,3' -dioxo-[delta2,2'-biindolin]-5,5'-disulfonat 860-22-0 (ng phn 5,5') C16H8N2Na2O8S2

2. nh ngha


Khi lng phn t 466,36 3. Cm quan Dng bt hoc ht mu xanh lam. 4. Chc nng Phm mu. 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc, t tan trong ethanol. nh tnh cc cht mu Phi c phn ng c trng ca cht mu. 5.2. tinh khit Gim khi lng khi lm Khng c qu 15% cng vi clorid v sulfat tnh theo mui kh ti 135 oC natri. Cht khng tan trong nc Khng c qu 0,2%. Ch Khng c qu 2 mg/kg. Cc cht mu ph Khng c qu 1% (khng bao gm dinatri 3,3'-dioxo[delta2,2'-biindolin]-5,7'-disulfonat). Cc cht hu c ngoi cht Khng c qu 0,5% tng cc acid isatin-5-sulfonic; acid 5mu sulfoanthranilic v acid anthranilic Cc amin Khng c qu 0,01% tnh theo anilin. thm bc nht khng sulfon ho Cc cht c th chit bng Khng c qu 0,2%. ether 5.3. Hm lng Khng c thp hn 85% tng cc cht mu. Khng c qu 18% dinatri 3,3'-dioxo-[delta2,2'-biindolin]-5,7'disulfonat. 6. Phng php th 6.1. tinh khit Ch - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng k thut hp th nguyn t thch hp vi hm lng ch nh xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c.

Cc cht mu ph

- Th theo hng dn ti JECFA monograph 1-Vol. 4 - S dng iu kin sau: Dung mi khai trin l dung mi s 3; Chiu cao tuyn dung mi ~ 17 cm. Ch 1: ng phn 5,7' c tch ra l mt vng mu xanh ln ngay trc vt chnh mu xanh. Khng tnh vng ny vo khi xc nh cc cht mu ph. Ch 2: 15 ml dung dch natri hydro carbonat s dng trong quy trnh chung c thay th bng dung dch acid hydrocloric 0,05 N trnh s phn hy xy ra khi indigo sulfon ha trong mi trng kim. Cc cht hu c ngoi cht - Th theo hng dn ti JECFA monograph 1-Vol. 4 mu - S dng k thut sc k lng vi cc iu kin nh sau: HPLC ra gii gradient t 2 n 100%, sau tip tc ra gii ti 100%. Cc cht c th chit bng - Th theo hng dn ti JECFA monograph 1-Vol. 4 ether - Cn 2 g mu th thay cho hng dn cn 5 g mu trong phn cc phng php chung. 6.2. nh lng Tin hnh theo hng dn trong chuyn lun xc nh tng hm lng bng chun vi Titan (III) clorid (trong JECFA monograph 1-Vol. 4) nh sau: Cn 1,0 - 1,1 g mu th; m 15 g natri hydrotartrat; Mi ml TiCl3 0,1 N tng ng vi 23,32 mg cc cht mu (D). Xc nh ng phn bng sc k giy: Tin hnh theo cc iu kin hng dn cho phn xc nh hm lng cc cht mu ph. Ct vt sc k tng ng ca ng phn trn sc k , theo hng dn ct vt mu ph. Chit vo dung mi v o hp th quang ti Smax ca n. Tin hnh o mu trng song song ti cng iu kin. S dng 0,1 ml dung dch mu th lm chun, chm ln bn giy hnh ch nht 18 cm x 0,7 cm. Hm lng (%) ng phn trong mu c tnh nh sau % = [A / As] x 20% x [D / 100] Trong : A l hp th ( hiu chnh vi mu trng) ca ng phn. As l hp th ( hiu chnh vi mu trng) ca chun. D l hm lng tng cht mu trong mu. Xc nh ng phn bng HPLC: ng phn 5,7' c tch bng HPLC vi iu kin tch cc cht mu ph nu trn v nh lng.

PH LC 14 YU CU K THUT V PHNG PHP TH I VI BRILLIANT BLUE FCF 1. Tn khc, ch s 2. nh ngha CI Food Blue 2, FD&C Blue No. 1; CI (1975): 42900 INS: 133 ADI = 0 - 1 mg/kg th trng. Ch yu gm dinatri 3-[N-ethyl-N-[4-[[4-[N-ethyl-N-(3sulfonbenzyl)-amino] phenyl] (2-sulfonphenyl)methylen]-2,5cyclohexadien-1-yliden] amonimethyl] benzensulfonat v cc ng phn ca n cng cc cht mu ph cng vi NaCl v (hoc) Na2SO4 l cc thnh phn khng mu chnh. C th chuyn thnh cht mu nhm (aluminium lake) tng ng, trong trng hp ny p dng quy nh chung i vi cc loi cht mu nhm. Dinatri 3-[N-ethyl-N-[4-[[4-[N-ethyl-N-(3-sulfonbenzyl)amino] phenyl] (2-sulfonphenyl)methylen]-2,5-cyclohexadien1-yliden] amonimethyl] benzensulfonat. Dinatri I-[4-(N-ethyl-3-sulfonatobenzylamino)phenyl]-I- [4-(Nethyl-3-sulfonatobenzyliminio)cyclohexa-2,5-dienyliden]toluen-2-sulfonat (Tn ha hc khc). 3844-45-9 C37H34N2Na2O9S3

Tn ha hc


Khi lng phn t 792,86 3. Cm quan Dng bt hoc ht mu xanh lam. 4. Chc nng Phm mu. 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc, t tan trong ethanol. nh tnh cc cht mu Phi c phn ng c trng ca cht mu. 5.2. tinh khit Gim khi lng khi lm Khng c qu 15% cng vi clorid v sulfat tnh theo mui kh ti 135 oC natri. Cht khng tan trong nc Khng c qu 0,2%. Ch Khng c qu 2 mg/kg. Crom Khng c qu 50 mg/kg. Cc cht mu ph Khng c qu 6%. Cc cht hu c ngoi cht Khng c qu 1,5% tng cc acid 2-; 3- v 4mu formylbenzensulfonic. Khng c qu 0,3% acid 3-[[N-ethyl-N-(4-sulfophenyl) amino] methyl] benzensulfonic. Leuco base Khng c qu 5%. Cc amin Khng c qu 0,01% tnh theo anilin. thm bc nht khng

sulfon ho Cc cht c th chit bng Khng c qu 0,2%. ether 5.3. Hm lng Khng c thp hn 85% tng cc cht mu. 6. Phng php th 6.1. tinh khit Ch - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng k thut hp th nguyn t thch hp vi hm lng ch nh xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c. Cc cht mu ph - Th theo hng dn ti JECFA monograph 1-Vol. 4 - S dng iu kin sau: Dung mi khai trin l dung mi s 4; Khai trin dung mi trong 20 gi. Cc cht hu c ngoi cht - Th theo hng dn ti JECFA monograph 1-Vol. 4 mu - S dng k thut sc k ct vi cc iu kin hp th nh sau: acid 3-formylbenzensulfonic: 0,0495 mgL-1cm-1 ti bc sng 246 nm, trong HCl long. acid 3-[[N-ethyl-N-(4-sulfophenyl) amino] methyl] benzensulfonic: 0,078 mgL-1cm-1 ti bc sng 277 nm, trong dung dch amoniac long. Leuco base Th theo hng dn ti JECFA monograph 1-Vol. 4 - Cn 120 5 mg mu th v tin hnh theo hng dn trong chuyn lun xc nh Leuco base trong cht mu nhm Triarylmethan sulfon ha; hp th ring (a) = 0,164 mgL-1cm-1 ti bc sng ~ 630 nm; t s = 0,9706. 6.2. nh lng Tin hnh theo hng dn trong chuyn lun xc nh tng hm lng bng chun vi Titan (III) clorid (trong JECFA monograph 1-Vol. 4) nh sau: Cn 1,8 - 1,9 g mu th; m 15 g natri hydrotartrat; Mi ml TiCl3 0,1 N tng ng vi 39,65 mg cc cht mu (D).

PH LC 15 YU CU K THUT V PHNG PHP TH I VI CHLOROPHYLL 1. Tn khc, ch s Magnesium chlorophyll, Magnesium phaeophytin; Clorophyllin; C.I. Natural Green 3; C.I. (1975): 75810; INS 140 ADI "khng gii hn" . Thu c bng cch chit c, c linh lng, tm ma v cc nguyn liu thc vt khc vi dung mi; Sn phm sau khi loi dung mi dng kt hp vi magnesi, c th loi hon ton hoc 1 phn t clorophyll thu c phaeophytin tng ng; Thnh phn mu chnh l cc phaeophytin v magnesi clorophyll. Sn phm chit sau khi loi dung mi cha cc cht mu khc nh carotenoid, sp v cht bo c ngun gc t nguyn liu. Ch c s dng nhng dung mi sau chit: aceton, dicloromethan, methanol, ethanol, propan-2-ol v hexan. Cc cht mu chnh l: Phytyl (132R,17S,18S)-3-(8-ethyl-132-methoxycarbonyl2,7,12,18-tetramethyl-13'-oxo-3-vinyl-131-132-17,18tetrahydrocyclopenta [at]-prophylrin-17-yl) propionat (Pheophytin a), hoc phc magnesi (Clorophyll a). Phytyl (132R,17S,18S)-3-(8-ethyl-7-formyl-132methoxycarbonyl-2,12,18-trimethyl-13'-oxo-3-vinyl-131-13217,18-tetrahydro-cyclopenta [at]-prophylrin-17-yl) propionat, (Pheophytin b), hoc phc magnesi (Clorophyll b). Phaeophytin a, phc magnesi: 479-61-8 Phaeophytin b, phc magnesi: 519-62-0 Phc magnesi phaeophytin a (Clorophyll a): C55H72MgN4O5 Phaeophytin a: C55H74N4O5 Phc magnesi phaeophytin b (Clorophyll a): C55H70MgN4O6 Phaeophytin b: C55H72N4O6

2. nh ngha

Tn ha hc

M s C.A.S. Cng thc ha hc

Cng thc cu to

Khi lng phn t

3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan

Trong : X = CH3 i vi cc hp cht "a" X = CHO i vi cc hp cht "b". Mt Mg s chuyn dng t clorophyll thnh phaeophytin Phc magnesi phaeophytin a (Clorophyll a): 893,51 Phaeophytin a: 871,22 Phc magnesi phaeophytin (Clorophyll a): 907,49 Phaeophytin : 885,20 Cht rn dng sp c mu t xanh oliu n xanh lc thm ty thuc vo lng magnesi kt hp. Phm mu. Khng tan trong nc; tan trong cn, diethyl ether; cloroalkan, hydrocarbon v du khng bay hi.

Sc k lp mng 5.2. tinh khit Dung mi tn d Arsen Ch

t yu cu (m t trong phn phng php th) Aceton, methanol, ethanol, propan-2-ol, hexan: Khng c qu 50 mg/kg, n cht hoc kt hp. Dicloromethan: Khng c qu 10 mg/kg Khng c qu 3 mg/kg. (Th theo hng dn ti JECFA monograph 1-Vol. 4 - Phng php II). Khng c qu 5 mg/kg. (Th theo hng dn ti JECFA monograph 1-Vol. 4. S dng k thut hp th nguyn t thch hp xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c). Tng hm lng cc phaeophytin v cc phc magnesi ca n khng thp hn 10%.

5.3. Hm lng 6. Phng php th 6.1. nh tnh Sc k lp mng

Chm dung dch mu th 1/20 trong cloroform ln bn mng Silica 60C thnh 1 vch di 2 cm. Sau khi bn mng kh, khai trin bn mng trong h dung mi gm 50% hexan, 45% cloroform v 5% ethanol (thng thng cloroform vi tinh khit thuc th cha sn 2% ethanol lm n nh. 5% ethanol l khng tnh n lng ethanol c sn ny) n khi tuyn dung mi t 15 cm cch im xut pht. dung mi bay hi, sau quan st cc vt ring bit v xc nh vt tng ng vi cu t quan tm thng qua Rf v mu. Gi tr Rf gn ng v mu ca cc vt nh sau: phaeophytin a: 0,77; mu xm/nu phaeophytin b: 0,75; mu vng/nu Clorophyll a: 0,50; mu xanh lam/xanh lc Clorophyll b: 0,63; mu vng/xanh lc Ngoi ra cn quan st thy cc vt ca -caroten ti Rf 0,81 v xanthophyll ti Rf 0,47 v 0,23. Th theo hng dn ti JECFA monograph 1-Vol. 4. Xc nh bnh Sc k kh hoc bng phng php ct cun (chuyn lun Xc nh dung mi tn d) hoc phn tch khng gian hi (chuyn lun Th gii hn dung mi tn d). - Th theo hng dn ti JECFA monograph 1-Vol. 4 - Phng php II. - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng k thut hp th nguyn t thch hp xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c).

6.2. tinh khit Dung mi tn d

Arsen Ch

6.1. nh lng

Cn khong 100 mg (chnh xc n mg) mu th, ha tan trong diethyl ether n 100 ml. Ly 2 ml dung dch ny, pha long vi diethyl ether va 25 ml. hp th quang ca mu ti nng ny khi o ti bc sng 660,4 nm khng c vt qu 0,7 so vi thang o hp th. o hp th quang (A) ca dung dch ny trong cuvet 1 cm so vi mu trng l diethyl ether ti bc sng 660,4 nm; 642,0 nm; 667,2 nm; 654,4 nm. (cc bc sng ny ln lt l cc i hp th trong diethyl ether ca clorphyll a; clorophyll b; phaeophtin a v ng phaeophytin b, tng ng). Ngoi ra o thm ti bc sng 649,8 nm v 628,2 nm. Phn dung dch pha long cn li, thm vo 1 tinh th acid oxalic, ha tan v lc u. o li hp th quang ca dung dch ti cc bc sng cho. " A" l s chnh lch hp th quang ca dung dch o ti cng bc sng trc v sau khi thm acid oxalic. Tnh nng tng cht ( mol/L) theo cc phng trnh sau: Clorophyll a = 17,7 A (660,4 nm) + 7,15 A (642,0 nm) Clorophyll b = 19,4 A (642,0 nm) - 2,92 A (660,4 nm) Phaeophytin a = -4,89 A (649,8 nm) + 0,0549 A (628,2 nm) +18,7 A (667,2 nm) + 0,0575 A (654,4 nm) - clorophyll a Phaeophytin b = -71,0 A (649,8 nm) + 2,51 A (628,2 nm) - 13,5 A (667,2 nm) + 84,3 A (654,4 nm) - clorophyll b Chuyn t mol/L sang hm lng % theo phng trnh sau: % clorophyll a =a mol 0,8935 12,5 100 Khi lng mu th (mg) mol 0,9075 12,5 100 Khi lng mu th (mg) mol 0,8712 12,5 100 Khi lng mu th (mg) mol 0,8852 12,5 100 Khi lng mu th (mg)

% clorophyll b =a

% phaeophytin a =a

% phaeophytin b =a

PH LC 16 YU CU K THUT V PHNG PHP TH I VI CHLOROPHYLL PHC NG 1. Tn khc, ch s Copper chlorophyllin, copper phaeophytin C.I. Natural Green 3; C.I. (1975) No. 75810; INS: 141i ADI = 0 - 15 mg/kg th trng. Thu c khi thm mui ng hu c vo sn phm thu c khi chit c, c linh lng, tm ma v cc nguyn liu thc vt khc vi dung mi; Sn phm sau khi loi dung mi cha cc cht mu khc nh carotenoid cng nh sp v cht bo. Cht mu chnh l ng phaeophytin. Ch c s dng nhng dung mi sau chit: aceton, dicloromethan, methanol, ethanol, propan-2-ol v hexan. Cht mu chnh dng acid ca n l [Phytyl (132R, 17S, 18S)-3-(8-ethyl-132-methoxycarbonyl2,7,12,18-tetramethyl-13-oxo-3-vinyl-131, 132,17,18tetrahydroxypenta[at]-prophyrin-17-yl) propionat] ng II (ng phaeophytin a) [Phytyl (132R, 17S, 18S)-3-(8-ethyl-7-formyl-132methoxycarbonyl-2,12,18-trimethyl-13-oxo-3-vinyl-131, 132,17,18tetrahydroxypenta[at]-prophyrin-17-yl) propionat] ng II (ng phaeophytin b). 65963-40-8 ng phaeophytin a (dng acid): C55H72CuN4O5 ng phaeophytin b (dng acid): C55H70CuN4O6

2. nh ngha

Tn ha hc

M s C.A.S Cng thc ha hc Cng thc cu to

Khi lng phn t 3. Cm quan 4. Chc nng 5. Tnh cht 5.1. nh tnh tan Quang ph Sc k lp mng 5.2. tinh khit Dung mi tn d

Trong : X = CH3 i vi hp cht "a" X = CHO i vi hp cht "b" ng phaeophytin a: 932,75 ng phaeophytin b: 943,73 Cht rn dng sp c mu t xanh lc n xanh lc thm ty thuc vo ngun nguyn liu. Phm mu. Khng tan trong nc; tan trong cn, diethyl ether; cloroalkan, hydrocarbon v du khng bay hi. A (1%, 1 cm) ca mu th trong cloroform, o ti 405 nm khng c thp hn 54. t yu cu (m t trong phn phng php th).

Aceton, methanol, ethanol, propan-2-ol, hexan: Khng c qu 50 mg/kg, n cht hoc hp cht. Dicloromethan: Khng c qu 10 mg/kg. ng t do c th ion ha Khng c qu 200 mg/kg. ng tng s Khng c qu 8% tnh theo tng s ng phaeophytin.

Arsen Ch 5.3. Hm lng 6. Phng php th 5.1. nh tnh Sc k lp mng

Khng c qu 3 mg/kg. Khng c qu 5 mg/kg. Khng thp hn 10% tng s ng phaeaphytin.

Chm dung dch mu th 1/20 trong cloroform ln bn mng Silica 60C thnh 1 vch di 2 cm. Sau khi bn mng kh, khai trin bn mng trong h dung mi gm 50% hexan, 45% cloroform v 5% ethanol (thng thng cloroform vi tinh khit thuc th cha sn 2% ethanol lm n nh. 5% ethanol l cha tnh n lng ethanol c sn ny) n khi tuyn dung mi t 15 cm cch im xut pht. dung mi bay hi, sau quan st cc vt ring bit v xc nh vt tng ng vi cu t quan tm thng qua Rf v mu. Gi tr Rf gn ng v mu ca cc vt nh sau: ng phaeophytin a: 0,5; mu xanh lc ng phaeophytin b: 0,73; mu vng/xanh lc Ngoi ra cn quan st thy cc vt ca -caroten ti Rf 0,81 v xanthophyll ti Rf 0,47 v 0,23.

5.2. tinh khit Dung mi tn d - Th theo hng dn ti JECFA monograph 1-Vol. 4. - Xc nh bnh Sc k kh hoc bng phng php ct cun (chuyn lun Xc nh dung mi tn d) hoc phn tch khng gian hi (chuyn lun Th gii hn dung mi tn d). ng t do c th ion ha Cn khong 1 g (chnh xc n mg) mu th v ha tan trong 20 ml du lc, un nng nh cho tan hon ton. Thm chnh xc 200 ml nc, khuy bng my khuy t, v iu chnh ti pH 3,0 bng cch thm cn thn acid hydrocloric 0,5 N (trnh thm qu nhanh). yn hn hp trong 10 pht. Nu cn, iu chnh li ti pH 3,0 bng cch thm cn thn acid hydrocloric 0,5 N. Chuyn sang phu tch v yn 20 pht. Lc lp nc qua giy lc Whatman No. 50, b 10 ml dch lc u. Dng dung dch ny phn tch ng bng quang ph hp th nguyn t - JECFA monograph 1-Vol. 4. ng tng s Cn khong 0,1 g (chnh xc n mg) mu th, cho vo a silica, nung nhit khng qu 500 C, ti khi loi i ton b than; lm m tro bng mt hoc hai git acid sulfuric c v tro ha li. Ha tan tro bng cch un si 3 ln, mi ln vi 5 ml acid hydrocloric 10% (kl/kl), lc dch acid thu c sau mi ln thm acid qua cng mt phu giy lc nh vo bnh nh mc 100 ml. ngui, v thm nc tinh khit ti vch. Dng dung dch ny phn tch ng bng quang ph hp th nguyn t - xem JECFA monograph 1-Vol. 4. Arsen - Th theo hng dn ti JECFA monograph 1 - Vol.4 - Phng php II. Ch - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng k thut hp th nguyn t thch hp xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c. 6.3. nh lng Cn khong 100 mg (chnh xc n mg) mu th, ha tan trong diethyl ether n 100 ml. Ly 2 ml dung dch ny, pha long vi diethyl ether va 25 ml. hp th quang ca mu ti nng ny khi o ti bc sng 660,4 nm khng c vt qu 0,7 so vi thang o hp th. o hp th quang (A) ca dung dch ny trong cuvet 1 cm so vi mu trng l diethyl ether ti bc sng 667,2 nm; 654,4 nm; 649,8 nm v 628,2 nm. (hai bc sng sau l cc i hp th trong

diethyl ether ca ng phaeophtin a v ng phaeophytin b, tng ng). Tnh nng tng cht ( mol/L) theo cc phng trnh sau: ng phaeophytin a = 45,6 A (649,8 nm) - 2,75 A (628,2 nm) + 3,10 A (667,2 nm) - 35,4 A (654,4 nm) ng phaeophytin b = -8,46 A (649,8 nm) + 20,7 A (628,2 nm) 1,69 A (667,2 nm) + 5,13 A (654,4 nm) Chuyn t mol/L sang hm lng % theo phng trnh sau:

% ng phaeophytin a =

mol 0,9327 12,5 100 Khi lng mu th (mg) mol 0,9467 12,5 100 Khi lng mu th (mg)

% ng phaeophytin b =

PH LC 17 YU CU K THUT V PHNG PHP TH I VI MUI NATRI, KALI CA CLOROPHYL PHC NG 1. Tn khc, ch s Natri ng clorophyllin; kali ng clorophyllin Chlorophyllins, C.I. (1975) No. 75810; INS 141ii ADI = 0 -15 mg/kg th trng. Mui kim ca ng Clorophyllin thu c khi thm ng vo sn phm thu c khi x phng ha dch chit dung mi t c, c linh lng, tm ma v cc nguyn liu thc vt khc; qu trnh x phng ha c th loi cc nhm ester methyl v cyclophytol v c th tch i mt phn vng pentenyl; sau khi thm ng vo clorophyllin tinh ch, nhm acid b trung tnh ha to thnh dng mui ca kali v/hoc natri; sn phm thng mi c th dng dung dch nc hoc bt kh. Ch c s dng nhng dung mi sau chit: aceton, dicloromethan, methanol, ethanol, propan-2-ol v hexan. Cht mu chnh dng acid ca n l 3-(10-Carboxylato-4-ethyl-1,3,5,8-tetramethyl-9-oxo-2vinylphorbin-7-yl) propionat, phc ng (ng clorophyllin a) 3-(10-carboxylato-4-ethyl-3-formyl-1,5,8-trimethyl-9-oxo-2vinylphorbin-7-yl) propionat, phc ng (ng clorophyllin b) Ty thuc vo mc thy phn vng cyclopentenyl c th tch i vi sn phm thu c vi chc carboxyl th ba. ng clorophyllin a (dng acid): C34H32CuN4O5 ng clorophyllin b (dng acid): C34H30CuN4O6

2. nh ngha

Tn ha hc

Cng thc ha hc Cng thc cu to

Khi lng phn t

3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Quang ph ng Natri

Trong : X = CH3 i vi hp cht "a" X = CHO i vi hp cht "b" M = Kali v/hoc natri ng clorophyllin a: 640,20 ng clorophyllin b: 654,18 Mi cht c th tng ti 18 Dalton nu vng cyclopentenyl tch i. Dung dch mu l cy sm hoc bt xanh l cy sm ti xanh lam/en. Phm mu. Tan trong nc; rt kh tan trong cn thp , keton v diethyl ether; khng tan trong cloroalkan, hydrocarbon v mt s du khng bay hi. t yu cu (m t trong phn Phng php th) t yu cu (m t trong phn Phng php th) Phi c phn ng c trng ca natri.

Kali 5.2. tinh khit Phm mu kim Dung mi tn d

Phi c phn ng c trng ca kali.

t yu cu (m t trong phn Phng php th) Aceton, methanol, ethanol, propan-2-ol, hexan: Khng c qu 50 mg/kg, n cht hoc kt hp. Dicloromethan: Khng c qu 10 mg/kg ng t do c th ion ha Khng c qu 200 mg/kg.

ng tng s Arsen Ch 5.3. Hm lng 6. Phng php th 6.1 nh tnh Quang ph ng

Khng c qu 8% tnh theo ng phaeophytin tng s. Khng c qu 3 mg/kg. Khng c qu 5 mg/kg. Khng thp hn 95% ng Clorophyllin tng s sau khi sy kh ti 100 C trong 1 gi. A (1%, 1 cm) ca mu th ( sy kh 100 C trong 1 gi), o ti 405 nm trong dung dch m phosphat pH 7,5 khng c nh hn 540. Ha tan tro sulfat ca mu th (Cn 1 g mu, Phng php I) trong 10 ml acid hydrocloric long (TS) bng cch lm nng trong b cch thy. Lc nu dung dch c, v pha long vi nc ti 10 ml. S dng dung dch ny l dung dch th cho cc php th sau: Cho dung dch amoniac (TS) vo 5 ml dung dch th kim ha. Dung dch phi xut hin mu xanh lam. Cho 0,5 ml dung dch natri diethyldithiocarbamat 1/1000 vo 5 ml dung dch th. Dung dch phi xut hin ta nu. Th theo hng dn ti JECFA monograph 1-Vol. 4 - Chun b dung dch c m t trong chuyn lun nh tnh ng. Th theo hng dn ti JECFA monograph 1-Vol. 4 - Chun b dung dch c m t trong chuyn lun nh tnh ng.

Natri Kali 6.2 tinh khit Phm mu kim

Ly 5 ml dung dch mu th 0,5% trong nc vo ng nghim, thm 1 ml acid hydrocloric 1 N v 5 ml diethyl ether. Lc k v phn lp. Mu ca lp ether khng c m hn mu xanh l cy nht. Dung mi tn d Xc nh bnh Sc k kh hoc bng phng php ct cun (chuyn lun Xc nh dung mi tn d) hoc phn tch khng gian hi (chuyn lun Th gii hn dung mi tn d). (Th theo hng dn ti JECFA monograph 1-Vol. 4) ng t do c th ion ha Cn khong 1 g (chnh xc n mg) mu th v ha tan trong 20 ml du lc, un nng nh. Thm chnh xc 200 ml nc, khuy bng my khuy t, v iu chnh ti pH 3,0 bng cch thm cn thn acid hydrocloric 0,5 N (trnh thm qu nhanh). yn hn hp trong 10 pht. Nu cn, iu chnh li ti pH 3,0 bng cch thm cn thn acid hydrocloric 0,5 N. Chuyn sang phu tch v yn 20 pht. Lc lp nc qua giy lc Whatman No. 50, b 10 ml dch lc u. Dng dung dch ny phn tch ng bng quang ph hp th nguyn t (xem JECFA monograph 1-Vol. 4). ng tng s Cn khong 0,1 g (chnh xc n mg) mu th, cho vo a silica, nung nhit khng qu 500 C, ti khi loi i ton b than; lm m tro bng mt hoc hai git acid sulfuric c v tro ha li. Ha tan tro bng cch un si 3 ln, mi ln vi 5 ml acid hydrocloric 10% (kl/kl), lc dch acid thu c sau mi ln thm acid qua cng mt phu giy lc nh vo bnh nh mc 100 ml. ngui, v thm nc tinh khit ti vch. Dng dung dch ny phn tch ng bng quang ph hp th nguyn t (xem JECFA monograph

Arsen Ch

1-Vol. 4). Th theo hng dn ti JECFA monograph 1-Vol. 4- Phng php II Th theo hng dn ti JECFA monograph 1-Vol. 4. S dng k thut hp th nguyn t thch hp xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c. Cn khong 1 g (chnh xc n mg) mu th, sy kh trc 100 C trong 1 gi, sau ha tan trong 20 ml dung dch m phosphat (pH 7,5) v pha long ti 1000 ml bng nc ct. Ly 10 ml dung dch ny v pha long ti 100 ml bng dung dch m phosphat (pH 7,5). o mt quang ca dung dch ny (0,001% kl/tt) bng my quang ph thch hp, s dng cuvet o 1 cm v rng khe 0,10 mm 403-406 nm, ghi cc i trong khong . Hm lng (%) natri ng clorophyllin c tnh theo cng thc theo cng thc sau: Mt quang 104 -------------------------------------565 khi lng mu th (g) Quy c: natri ng clorophyllin tinh khit 100% c hp th ring l 565.

6.3. nh lng

PH LC 18 YU CU K THUT V PHNG PHP TH I VI GREEN S 1. Tn khc, ch s 2. nh ngha CI food green 4, food green S; CI (1975): 44090 INS: 142 ADI khng xc nh. Ch yu gm natri N-[4-][[4-(dimethylamino) phenyl] (2hydroxy-3,6 disulfo-1-naphtalenyl) methylen]-2,5cyclohexadien-1-ylidenen-N-methylmetanamini v cc cht mu ph cng vi NaCl v (hoc) Na2SO4 l cc thnh phn khng mu chnh. C th chuyn thnh cht mu nhm (aluminium lake) tng ng, trong trng hp ny p dng quy nh chung i vi cc loi cht mu nhm. Natri N-[4-[[4-(dimethylamino) phenyl](2-hydroxy-3,6 disulfo1naphtalenyl) methylen]-2,5-cyclohexadien-1-yliden]-N methylmetanamini; hoc Natri 5-[4-dimethylamino-l-(4-dimethyliminiocyclohexa2,5-dienyliden) benzyl]-6-hydroxy-7-sulfonat-naphtalen-2sulfonat. 860-22-0 C27H25N2NaO7S2

Tn ha hc


Khi lng phn t 576,63 3. Cm quan Dng bt hoc ht mu xanh lc m. 4. Chc nng Phm mu. 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc, kh tan trong ethanol. nh tnh cc cht mu Phi c phn ng c trng ca cht mu. 5.2. tinh khit Gim khi lng khi lm Khng c qu 20% cng vi clorid v sulfat tnh theo mui kh ti 135 oC natri. Cht khng tan trong nc Khng c qu 0,2%. Ch Khng c qu 2 mg/kg. Crom Khng c qu 50 mg/kg. Cc cht mu ph Khng c qu 1%. Cc cht hu c ngoi cht Khng c qu 0,1% ru 4, 4-bis (dimethylamino) mu benzhydric. Khng c qu 0,1% 4, 4-bis (dimethylamino) benzophenon, Khng c qu 0,2% acid 3-Hydroxynaphtalen-2,7-disulfonic. Cc amin Khng c qu 0,01% tnh theo anilin. thm bc nht khng sulfon ho Leuco base Khng c qu 5,0%.

Cc cht c th chit bng Khng c qu 0,2%. ether 5.3. Hm lng Khng c thp hn 80% tng cc cht mu. 6. Phng php th 6.1. tinh khit Ch - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng k thut hp th nguyn t thch hp vi hm lng ch nh xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c. Cc cht mu ph - Th theo hng dn ti JECFA monograph 1-Vol. 4 - S dng iu kin nh sau: Dung mi khai trin l dung mi s 2, chiu cao t vch xut pht ti tuyn dung mi khong 17 cm. Cc cht hu c ngoi cht - Th theo hng dn ti JECFA monograph 1-Vol. 4 mu - S dng k thut sc k lng vi iu kin nh sau: HPLC ra gii gradient 2% n 100% vi tc tng c nh 2%/pht. Leuco base - Th theo hng dn ti JECFA monograph 1-Vol. 4 - Cn chnh xc 110 5 mg mu v tin hnh theo hng dn trong chuyn lun th Leuco base trong cht mu Triarylmethan sulfon ha. hp th ring (a)=0,1725 mgL-1cm-1 ti bc sng khong 634 nm. T s = 0,9600. 6.2. nh lng Tin hnh theo hng dn trong chuyn lun xc nh tng hm lng bng chun vi Titan (III) clorid (trong JECFA monograph 1-Vol. 4) nh sau: Cn 1,4 - 1,5 g mu th; m 15 g natri hydrotartrat; Mi ml TiCl3 0,1 N tng ng vi 28,83 mg cc cht mu (D).

PH LC 19 YU CU K THUT V PHNG PHP TH I VI CARAMEN 1. Tn khc, ch s Cht mu caramel c chia thnh bn loi. Tn khc v ch s ca mi loi nh sau: Loi I: Caramel thng, caramel caustic; INS No. 150a Loi III: Caramel amoni; INS No. 150c Loi IV: Caramel amoni sulfit, INS No. 150d ADI i vi nhm I Khng xc nh; ADI i vi nhm III = 0-160 mg/kg th trng ADI i vi loi IV 0-200 mg/kg th trng (0-150 mg/kg th trng i vi dng rn) L cc hn hp phc tp, trong mt s dng t hp keo, sn xut bng cch un nng ring carbohydrat hoc cng vi s c mt ca acid, kim hoc mui loi thc phm; ty theo ha cht dng khi sn xut phn loi nh sau: Loi I: sn xut bng cch un nng ring carbohydrat hoc cng vi s c mt ca acid, kim; khng dng hp cht amoni hoc sulfit. Loi III: sn xut bng cch un nng ring carbohydrat hoc cng vi s c mt ca acid, kim; cng vi hp cht amoni; khng dng hp cht sulfit. Loi IV: sn xut bng cch un nng ring carbohydrat hoc cng vi s c mt ca acid, kim; cng vi c hp cht sulfit v hp cht amoni. Trong tt c cc trng hp, nguyn liu carbohydrat c sn dng thng phm l cc cht lm ngt dinh dng loi thc phm c cha glucose, fructose v/hoc polyme ca chng. Cc acid v kim l acid sulfuric hoc citric thc phm v natri, kali hoc calci hydroxyd hoc hn hp ca chng loi thc phm. Hp cht amoni c dng l mt hoc bt k mt cht no sau y: amoni hydroxyd, amoni carbonat v amoni hydro carbonat, amoni phosphat, amoni sulfat, amoni sulfit v amoni hydrosulfit. Dng hp cht sulfit l mt hoc bt k mt cht no sau y: acid sulfur, kali, natri v amoni sulfit v amoni hydro sulfit. C th dng tc nhn chng ni bt loi thc phm tr gip qu trnh sn xut. Cht dng rn hoc lng, mu nu sm ti en c mi ng chy. Phm mu C th trn ln vi nc Phi c phn ng c trng ca tp mu. Loi I: Khng c qu 50 % cht mu l loi lin kt bi celulose DEAE v khng qu 50% cht mu l loi lin kt bi celulose phosphoryl. Loi III: Khng c qu 50% cht mu l loi lin kt bi celulose DEAE v hn 50% cht mu l loi lin kt bi celulose phosphoryl. Loi IV: Hn 50% cht mu l loi lin kt bi celulose DEAE v t s hp th khng qu 50. Xem m t trong phn PHNG PHP TH. Ch : Gii hn arsen v ch p dng cho tt c cc nhm caramel v c tnh da trn ch phm nguyn dng: Cc gii

2. nh ngha

3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan nh tnh tp mu Phn loi

5.2. tinh khit

hn v khong khc p dng i vi mi loi ring theo ch dn, nu khng c ch dn ring, c tnh da trn ch phm rn. Hm lng cht rn Loi I: 62-77% Loi III: 53-83% Loi IV: 40-75% Cng mu Loi I: 0,01-0,12 Loi III: 0,08-0,36 Loi IV: 0,10-0,60 Nit ton phn Loi I: Khng c qu 0,1% Loi III: 1,3 -6,8% Loi IV: 0,5-7,5% Lu hunh ton phn Loi I: Khng c qu 0,3% Loi III: Khng c qu 0,3% Loi IV: 1,4-10,0% Lu hunh dioxyd Loi I: Loi III: Loi IV: Khng c qu 0,5% Nit amoniac Loi I: Loi III: Khng c qu 0,4% Loi IV: Khng c qu 2,8% 4-Methylimidazol (MEI) Loi I: Loi III: Khng c qu 300 mg/kg & khng c qu 200 mg/kg tnh theo cht mu tng ng Loi IV: Khng c qu 1000 mg/kg & khng c qu 250 mg/kg tnh theo cht mu tng ng 2-Acetyl-4-tetrahydroxy- Loi I: butylimidazol (THI) Loi III: Khng c qu 40 mg/kg & khng c qu 25 mg/kg tnh theo cht mu tng ng. Loi IV: Arsen Khng c qu 1 mg/kg Ch Khng c qu 2 mg/kg 6. Phng php th 6.1. nh tnh Phn loi/ Cht mu lin Trong ch tiu ny cht mu lin kt bi celulose DEAE c kt bi celulose DEAE nh ngha l phn trm hp th gim i ca dung dch cht mu caramel 560 nm sau khi x l vi celulose DEAE. Thuc th ring : Celulose DEAE (diethylaminoethyl) c dung lng 0,7 mE/g, nh Cellex D ca Bio-Rad hoc Celulose DEAE c dung lng cao hn hoc thp hn tng ng vi lng cht cao hn hoc thp hn. Tin hnh: Pha dung dch cht mu caramel c hp th xp x 0,5 ti 560 nm bng cch cho mt lng cht mu caramel thch hp vo bnh nh mc 100 ml, cng vi acid hydrocloric 0,025 N. Pha long ti vch bng acid hydrocloric 0,025 N v ly tm hoc lc, nu dung dch vn c. Ly 20 ml dung dch cht mu caramel, thm 200 mg celulose DEAE, trn k trong t pht, ly tm hoc lc, v thu ly lp dung dch trong pha trn. Xc nh hp th ca dung dch cht mu caramel v lp dung dch pha trn trong cng o 1 cm 560 nm, bng my quang ph thch hp, ngay trc chun ha vi mu trng l acid hydrocloric 0,025 N. Tnh phn trm cht mu lin kt bi celulose DEAE theo cng thc: (A 1 A 2 ) 10 0 A1

Trong : A1 l hp th ca dung dch cht mu caramel 560 nm A2 l hp th ca lp dung dch pha trn sau khi x l vi Celulose DEAE 560 nm. Phn loi/ Cht mu lin Trong ch tiu ny cht mu lin kt bi celulose phosphoryl kt bi celulose c nh ngha l phn trm hp th gim i ca dung dch phosphoryl cht mu caramel 560 nm sau khi x l vi celulose phosphoryl. Thuc th ring: Celulose phosphoryl dung lng 0,85 mE/g, nh Cellex P Bio-Rad hoc Celulose phosphoryl dung lng cao hn hoc thp hn tng ng vi lng cht cao hn hoc thp hn. Tin hnh: Cho khong 200-300 mg cht mu caramel vo bnh nh mc 100 ml, pha long ti vch bng acid hydrocloric 0,025 N v ly tm hoc lc, nu dung dch vn c. Ly 40 ml dung dch cht mu caramel, thm 2,0 g celulose phosphoryl, lc k trong t pht. Ly tm hoc lc, v thu ly lp dung dch trong pha trn. Xc nh hp th ca dung dch cht mu caramel v lp dung dch pha trn trong cng o 1 cm 560 nm, bng my quang ph thch hp, ngay trc chun ha vi mu trng l acid hydrocloric 0,025 N. Tnh phn trm cht mu lin kt bi celulose phosphoryl theo cng thc: (A 1 A 2 ) 10 0 A1 Trong : A1 l hp th ca dung dch cht mu caramel 560 nm A2 l hp th ca lp dung dch pha trn sau khi x l vi Celulose phosphoryl 560 nm. Phn loi/ H s hp Trong ch tiu ny t s hp th c nh ngha l hp th th ca cht mu caramel 280 nm chia cho hp th ca cht mu caramel 560 nm. Tin hnh: Cho khong 100 mg cht mu caramel vo bnh nh mc 100 ml cng vi nc, pha long ti vch bng nc, lc u v ly tm, nu dung dch vn c. Ly 5,0 ml dung dch trong ny cho vo bnh nh mc 100 ml, pha long ti vch bng nc, v lc u. Xc nh hp th ca dung dch 0,1% trong cng o 1 cm 560 nm v ca dung dch pha long 1:20 280 nm, bng my quang ph thch hp, ngay trc chun ha vi mu trng l nc. (My quang ph thch hp l my c b n sc ha cho rng di 2 nm hoc nh hn v t l nh sng lc l 0,5% hoc nh hn.) Tnh t s hp th ca cht mu caramel bng cch chia gi tr hp th ca cht mu caramel 280 nm nhn 20 (h s pha long) cho gi tr hp th 560 nm. 6.2. tinh khit Hm lng cht rn Hm lng cht rn ca cht mu Caramel c xc nh bng cch lm kh mu th vi cht mang gm c ct thch anh tinh khit qua c ry s 40 nhng khng qua c c ry s 60 c chun b bng cch ra vi acid hydrocloric, ra ht acid, sy v nung. Trn 30,0 g ct chun b c cn chnh xc vi 1,5-2,0 g cht mu caramel c cn chnh xc v lm kh ti khi lng khng i 60oC di p sut gim 50 mmHg (6,7 kPa). Ghi khi lng cui cng ca caramel v ct. Tnh % cht rn theo cng thc sau:

% cht rn =

(m 3 m 2 ) 10 0 m1

Trong : m3 = khi lng cui cng ca caramel thm ct (g) m2 = khi lng ct (g) m1 = khi lng caramel thm vo ban u (g) Tnh ton trn c s cht rn Hm lng nit ton phn, lu hunh ton phn, nit amoni, lu hunh dioxyd, 4-MEI v THI c biu th theo cht rn. Xc nh nng (Ci) ca mi tp cht theo nguyn trng; nng (CS) theo cht rn c tnh theo cng thc: Cs = Cng mu

Ci 100 % cht rn

Trong ch tiu ny cng mu c nh ngha l hp th ca dung dch 0,1% (kl/tt) ca cht mu caramel rn trong nc 610 nm trong cng o 1 cm. Tin hnh: Cho khong 100 mg cht mu caramel vo bnh nh mc 100 ml cng nc, pha long ti vch bng nc, lc u v ly tm, nu dung dch vn c. Xc nh hp th (A610) ca dung dch trong ny trong cng o 1 cm 610 nm bng my quang ph thch hp, ngay trc chun ha vi mu trng l nc. Tnh cng mu ca cht mu caramel theo cng thc sau: A610 100 Cng mu = % cht rn Xc nh % cht rn nh m t phn Hm lng cht rn. Tnh ton trn c s cht mu tng ng: Khi c thm cc gii hn i vi 4-MEI v THI c biu th theo cht mu tng ng th cc nng trc ht c tnh trn c s cht rn nh ch dn trong phn "Tnh trn c s vo cht rn", v sau biu th theo cht mu tng ng theo cng thc: CS 0,1 Mu tng ng = cng mu Trong : CS = nng trn cht rn. Nh vy hm lng c biu th bng mt sn phm c cng mu l 0,1 n v hp th. Th theo hng dn ti JECFA monograph 1-Vol. 4- Xc nh theo ch dn trong phn Xc nh Nit (Phng php Kjeldahl) s dng Phng php II Ly mt chn ln nht c th cho va l nung in, cho vo 1-3 g MgO hoc mt lng tng ng Mg(NO 3)26H2O (6,4 19,2 g), 1 g ng saccarose lm thnh bt, v 50 ml HNO3. Thm 5-10 g cht mu caramel. Cho cng mt lng thuc th vo mt chn khc lm mu trng. C trn hi nc si ti khi thnh khi nho. t chn vo l nung ngui (25 C) v un nng t t ti khi ton b khi NO2 bay ht. ngui, ha tan v trung tnh ha bng HCl (1+2,5), thm d 5 ml. Lc, un nng ti si, v thm 5 ml dung dch BaCl22H2O 10% tng git mt. C ti 100 ml, yn qua m, lc, ra,

Nit ton phn Lu hunh ton phn

Lu hunh dioxyd

nung, v cn BaSO4. Hiu chnh kt qu i vi BaSO4 thu c trong mu trng v ghi dng mg S/100 g. Cc dng c thng mi dng phn tch lu hunh ton phn nh thit b phn tch theo qui trnh t chy Leco/chun cng c th s dng v c khuyn dng vi lng mu th khong 200 mg. Thit b S dng thit b Monier-Williams ci tin (c 5GA Scientific, Inc., Bloomfield, N.J., M) xc nh acid sulfur, hoc dng c thit k nh trnh by hnh. B dng c gm mt bnh ct y trn ba c 1000 ml c ng ni thy tinh vut thon chun 24/40. Gn km mt sinh hn Allihn 30 cm hi lu mt c pha ngoi ca bnh, v u kia ca sinh hn ni vi mt ng Tygon hoc silicon inch (c un si trc vi dung dch acid hydrocloric 1/20 v ra bng nc) ti b ng hp th (c ni hnh cu 35/20 hoc tng ng). Ni c gia ca bnh vi bnh gn hnh tr 125 ml, v gn mt on ng ti ng ch U ngn cm qua mt np cao su trn c ca bnh gn. Gn mt ng vo bng thy tinh, cong di, ti gn y bnh, vo c ngoi khc ca bnh, v ni ng vo mt bnh ra-kh 250 ml mt on ng. Bnh ra kh li ni bng mt ng ti chai (bom kh) nit.

Nghin 4,5 g pyrogalol (acid pyrogalic) vi 5 ml nc trong mt ci nh, v cho khi nho sang bnh ra kh. Tip tc nghin phn cn li, v chuyn ton b sang bnh vi 2 ln 5 ml nc. Cho kh nit i t chai kh ti bnh ui ht khng kh, v sau thm vo bnh qua phu cung di dung dch c lm ngui ca 65 g kali hydroxyd trong khong 85 ml nc. Lp u bnh vo, v sc bong bng nit qua ui ht khng kh khi khng gian pha trn. Kp cht ng c hai v tr ca chai, v ni n ti ng u vo ca bnh ct. Bnh ra kh phi c chun b nh m t, vi dung dch pyrogalol mi pha trong ngy lm. Thm ln lt vo mi ng hp th hnh ch U nh sau: hai a thy tinh 8 mm chiu di khong 25 mm, 10 ml ht thy tinh 3 mm u ra, 10,0 ml dung dch hydrogen peroxyd 3%, v 1 git methyl (TS). Lp tt c cc phn ca thit b, v kim tra xem c kn khng bng cch thi nh vo trong ng gn vi c bnh gn. Khi thi, ng kha bnh gn. yn vi pht; nu mc cht lng trong ng ch U vn ngang bng, ng tt c cc khp ni v th li. Nu h thng kht th tin hnh theo ch dn di y. Tin hnh: Phn tn khong 25 g mu th, cn chnh xc, trong 300 ml nc va un si ngui, v chuyn khi nho sang bnh qua mt phu l rng dng nc. Pha long ti khong 400 ml bng nc, v kha bnh gn li. Thm 90 ml acid hydrocloric 4 N vo bnh gn, v cho acid vo bnh bng cch thi nh vo ng c bnh gn. Kha bnh gn. M kp ng c hai v tr

Nit amoniac

4-Methylimidazol

ca bnh ra kh, v bt u thi kh nit vi tc u n bt kh. un nng bnh ct bng bp o un hi lu trong khong 20 pht. Khi t c s hi lu n nh, cp in un tip hi lu trong 1,75 gi. Tt nc trong sinh hn, v tip tc un ti khi khp ni pha u vo ca ng ch U th nht xut hin hi ngng ng v hi m. B bnh gn v tt bp. Khi khp ni u trn ca sinh hn ngui, tho khp ni v trng vo ng ch U th hai, li ng ni vi khp ra ca ng ch U th nht nhng ngt khi u vo ca ng ch U th hai. Quay ng ni ti khi u h ca n gn chm u vo ca ng ch U th nht. Thm 1 git methyl vo ng ch U th nht, v chun bng natri hydroxyd 0,1 N ti khi c mu vng sng, lc nh. Sau khi chun ng ch U th nht, tho ng ni, lp n vi ng ch U th hai u ra, v chun tng t. Ghi tng lng dung dch hai ln chun l Vt (ml). Tin hnh lm mt mu trng, th tch dung dch natri hydroxyd 0,1 N l Vo. Tnh phn trm lu hunh dioxyd trong mu th theo cng thc: (V Vo ) 0,0032 SO 2 % = t 100 m Trong m l khi lng ca mu th (g) Cho 25 ml acid sulfuric 0,1 N vo bnh hng 500 ml, v lp vi thit b ct gm ng ni bu Kjeldahl v mt sinh hn c u ra ngp pha di b mt dung dch trong bnh hng. Cho khong 2 g cht mu caramel, cn chnh xc, vo trong bnh Kjeldahl c di 800 ml, v thm vo bnh 2 g magnesi oxyd (khng c carbonat), 200 ml nc, v vi vin bt. Lc bnh trn u hn hp trong bnh, v lp nhanh bnh vi thit b ct. un nng bnh ti si, thu ly khong 100 ml dch ct vo bnh hng. Ra u ra ca ng sinh hn vi vi ml nc, cho dch ra vo bnh hng, sau thm 4 hoc 5 git ch th methyl (500 mg methyl trong 100 ml alcol), v chun bng natri hydroxyd 0,1 N, ghi th tch l Vt (ml). Tin hnh lm mt mu trng, th tch dung dch natri hydroxyd 0,1 N trung tnh l Vo. Tnh phn trm nit amoniac trong mu th theo cng thc: (V Vo ) 0,0014 100 Nit amoniac = t m Trong m l khi lng ca mu th (g) Ghi ch: Cn thng tin v mt phng php tt hn. Dng cc nguyn vt liu v thuc th sau (thuc th phi l loi ACS hoc tng ng). Nguyn vt liu: Bng thy tinh loi Pyrex, ct sc k 22 300 mm v kho PTFE (nh Kimax 17800); cc polypropylen 150 ml (nh Nalge 1201); bnh y trn 250 ml (nh Pyrex 4320); phu rt bt 75 mm; tha gt (spatula) 5 cm; my ct quay chn khng, bp a kn, ni cch thy, pipet Pasteur dng mt ln; bnh nh mc 5 ml. Thuc th: Aceton; Celit 545; methylen clorid; natri hydroxyd; v tetrahydrofuran. Tin hnh: Sau khi trn k mu th cht mu caramel bng cch lc hoc

khuy, cn 10,00 g hn hp vo cc polypropylen 150 ml. Polypropylen c coi l tt hn thy tinh v b mt k nc d chuyn ton lng mu th. Thm vo 5,0 g dung dch NaOH 3,0 N v lc k m bo pH ca mu th ln hn 12. Thm 20 g Celit 545 vo cc trn, v trn u ti khi hn hp gn kh li. Qu trnh ny khong 2 ti 3 pht. Vi mu th c hm lng nc qu cao, hn hp cht mu caramel-Celit 545 c th qu t. Trong trng hp ny, trn 5,00 g hn hp cht mu caramel vi 2,5 g NaOH 3,0 N v 15 g Celit 545 v tin hnh phn tch. Mt nt bng thy tinh Pyrex c t pha di ct sc k 22 300 mm vi kha PTFE. Hn hp cht mu caramelCelit 545 c cho vo ct qua phu 75 mm. Hn hp trong ct c nhi bng cch g nh ct theo phng thng ng khong 10 cm trn mt b mt c lt. Khi nhi tt, hn hp cht mu caramel-Celit 545 phi chim xp x 250 mm pha di ct. Cn thn khi tin hnh trnh nhi lng qu hoc cht qu. Ct nhi lng qu methylen clorid s ra gii qu nhanh v khng chit hon ton. Ct nhi qu cht dn ti dung mi ra gii kh tip cn mt s vng cht nhi. iu ny cng dn n vic chit khng hon ton. M kha, dng cc rt methylen clorid vo ct. Khi dung mi ti nt bng thy tinh, ng kho v dung mi tng tc vi hn hp trong ct 5 pht. Sau m kha v ct c ra gii tip vi methylen clorid ti khi thu c 200 ml vo bnh y trn 250 ml. Ly 1,00 ml dung dch chun ni 2 MEI (50,0 mg 2 MEI/50,0 ml methylen clorid) thm vo dch ra gii thu c. 2 MEI c tch tt khi 4 MEI trong iu kin sc k kh lng dng v khng thy c trong cht mu caramel. Sau , loi dung mi khi dch ra gii bng ct quay p sut gim 45-50 kPa v bnh cu y trn c duy tr 35 oC trong ni cch thy. Cn ca dch chit c chuyn hon ton sang bnh nh mc 5 ml bng pipet Pasteur dng mt ln, bng cch ra bnh cu nhiu ln mi ln vi mt lng nh (khong 0,75 ml) tetrahydrofuran hoc aceton. C hai dung mi c dng vi kt qu nh nhau. Sau khi trn k hn hp bng cch lc ngc bnh nhiu ln, c dch chit dng cho phn tch GLC. Dch chit sau khi chun b phi phn tch cng sm cng tt, bi mu ch n nh trong vng 1 ngy sau khi chun b. Phn tch GLC c tin hnh bng thit b sc k kh vi detector ngn la hydro. Ct thy tinh 1 m 6 mm ng knh ngoi ( 4 mm ng knh trong), thm 7,5% Carbowax 20M + 2% KOH trn 90/100 mesh Anakrom ABS. Cc thng s GLC nh sau: kh mang nit 50 ml/pht; hydro 50 ml/pht; oxy 80 ml/pht; b phn tim mu 200 oC; ct ng nhit 180 o C; detector 250 oC; th tch mu 5 L. Tt c qu trnh nh lng tin hnh dng k thut chun ni. 2-Acetyl-4-tetrahydroxy- Ghi ch: Cn thng tin v mt phng php tt hn. butylimidazol (THI) THI c chuyn thnh dn xut 2,4-dinitrophenylhydrazon (THI-DNPH) ca n. Dn cht ny c tch khi thuc th d v tp carbonyl bng HPLC dng ct RP-8, xc nh bng o hp th bc sng 385 nm. Tin hnh: Cn chnh xc cht mu caramel (200-250 mg), sau ha tan trong nc (3 ml). Chuyn ton lng dung dch sang phn pha trn ca ct t hp. Bt u ra gii bng nc, v ton

b lng nc qua cc ct khong 100 ml. Sau tho ct pha trn. Ra gii ct pha di bng HCl 0,5 N. B 10,0 ml dch ra gii ban u, sau thu ly 35 ml. C dung dch ny ti kh 40 oC v 15 torr. Ha tan cn c st trong methanol khng c carbonyl (250 L) v thuc th 2,4-dinitrophenylhydrazin (250 L). Chuyn hn hp phn ng sang l c np l tm m v bo qun 5 gi nhit phng. Tim 5 L (cng c th t 1 ti 25 L) vo ct HPLC LiChrosorb RP-8 (10 m). Pha ng gm MeOH/ H3PO4 0,1 M 50/50 (tt/tt). iu chnh thnh phn pha ng nu c tnh ca ct thay i, tu thuc nh sn xut. (Rt nn s dng cht nhi LiChrosorb RP-8, 10 m, ct 250 ( 4 mm "Vertex" ca nh sn xut Knauer, Bad Homburg, F.R.G.). Vi tc pha ng 2 ml/pht v kch c ct 250 ( 4,6 mm, THI-DNPH ra gii sau khong 6,30,1 pht. Pht hin 385 nm v o chiu cao ca pic. Hm lng c tnh da vo ng chun ca THI-DNPH trong methanol. Nguyn vt liu: Thuc th - 2,4,-Dinitrophenylhydrazin hydroclorid: 2,4dinitrophenylhydrazin thng mi (5 g) cho vo acid hydrocloric c (10 ml) trong bnh nn 100 ml, v lc nh ti khi base t do (mu ) chuyn thnh hydroclorid (mu vng). Thm vo ethanol (100 ml), lc v un nng trn cch thy si ti khi tt c cht rn ha tan. Sau khi kt tinh nhit phng, hydroclorid c lc, ra bng ether, sy nhit phng v bo qun trong bnh ht m. Trong khi bo qun, hydroclorid dn chuyn thnh base t do. Sau , c th loi bng cch ra vi dimethoxyethan. Thuc th c chun b bng cch trn 0,5 g 2,4-dinitrophenylhydrazin hydroclorid trong 15 ml methanol 5% trong dimethoxyethan trong 30 pht. Bo qun trong t lnh v kim tra nh k bng HPLC. - Nha trao i cation (mnh): Dowex 50 AG x 8, H+, 100200 mesh. - Nha trao i cation (yu): Amberlite CG AG 50 I, H+, (100200 mesh). ( lng hai hoc ba ln trc khi s dng). - Methanol, khng c carbonyl t do: Methanol c chun b theo Y. Peleg v C.H. Mannheim, J. Agr. Fd. Chem, 18 (1970) 176, bng cch x l vi thuc th Girard P. - Dimethoxyethan: Nu khng tinh khit, dimethoxyethan c tinh ch bng cch ct t 2,4-dinitrophenylhydrazin vi s c mt ca acid v ct li t natri hydroxyd. Ngay trc khi s dng cho qua ct nhm trung tnh loi peroxyd. Thit b: Ct t hp: tng t nh m t trong J. Agr. Fd. Chem. 22 (1974) 110. Ct pha trn (150 ( 12,5 mm, chiu cao nhi ti a 9 cm, hoc 200 ( 10 mm, chiu cao nhi ti a 14 cm, vi mao qun u ra c 1 mm ng knh trong) nhi cht trao i cation acid yu, chiu cao cht nhi ln lt khong 50-60, hoc 80-90 mm. Ct pha di (chiu di tng 175 mm, ng knh trong 10 mm, vi u ra mao qun v kho Teflon) nhi vi cht trao i cation acid mnh chiu cao cht nhi 60 mm. Bnh ng dung mi, dng phu nh git (100 ml) kha Teflon. Tt c cc phn c ni bng khp ni thy tinh mi chun (14,5 mm). HPLC: vi ct qui nh trn v detector UV c kh nng o 385 nm. Chun ha: Ha tan THI-DNPH trong methanol tuyt i

khng c carbonyl (khong 100 mg/L; nng ca THI: 47,58 ng/L). Dung dch ny pha long 10 ln vi methanol (4,7 ng THI/L). Dung dch chun THI-DNPH n nh t nht 20 tun nu bo qun trong t lnh. Arsen Ch - Th theo hng dn ti JECFA monograph 1-Vol. 4 - Phng php II. - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng k thut hp th nguyn t thch hp cho hm lng quy nh xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c.

PH LC 20 YU CU K THUT V PHNG PHP TH I VI BRILLIANT BLACK PN 1. Tn khc, ch s 2. nh ngha CI food Black 1, Black PN; CI (1975): 28440 INS: 151 ADI = 0 - 1 mg/kg th trng. Ch yu gm tetranatri 4-acetamido-5-hydroxy-6-[7-sulfon-4(4-sulfon-phenylazo)-1-naphthylazo]-1,7-naphthalendisulfonat cng cc cht mu ph cng vi NaCl v (hoc) Na2SO4 l cc thnh phn khng mu chnh. C th chuyn thnh cht mu nhm (aluminium lake) tng ng, trong trng hp ny p dng quy nh chung i vi cc loi cht mu nhm. tetranatri 4-acetamido-5-hydroxy-6-[7-sulfon-4-(4-sulfonphenylazo)-1-naphthylazo]-1,7-naphthalen-disulfonat. 2519-30-4 C28H17N5Na4O14S4


Khi lng phn t 867,69 3. Cm quan Dng bt hoc tinh th mu en 4. Chc nng Phm mu. 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc, t tan trong ethanol nh tnh cc cht mu Phi c phn ng c trng ca cht mu. 5.2. tinh khit Gim khi lng khi lm Khng c qu 20% cng vi clorid v sulfat tnh theo mui kh ti 135 oC natri. Cht khng tan trong nc Khng c qu 0,2%. Ch Khng c qu 2 mg/kg. Cc cht mu ph Khng c qu 4%. Cc cht hu c ngoi cht Khng c qu 0,8% tng cc acid 4-Acetamido-5-hydroxymu 1,7-naphthalendisulfonic; acid 4-Amino-5-hydroxy-1,7naphthalendisulfonic; acid 8-Amino-2-naphthalensulfonic; acid Sulfanilic v acid 4,4'-Diazoamino-di(benzensulfuric) Cc amin Khng c qu 0,01% tnh theo anilin. thm bc nht khng sulfon ho Cc cht c th chit bng Khng c qu 0,2%. ether 5.3. Hm lng Khng c thp hn 80% tng cc cht mu. 6. Phng php th 6.1. tinh khit Ch - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng k thut hp th nguyn t thch hp vi hm lng ch nh xc nh. La chn c mu th v phng php

chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c. Cc cht mu ph - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng cc iu kin sau: Dung mi khai trin: Sc k (i): s 1 Sc k (ii): s 4 Chiu cao tuyn dung mi: Sc k (i): 17 cm Sc k (ii): 17 cm.) Cc cht hu c ngoi cht - Th theo hng dn ti JECFA monograph 1-Vol. 4 mu - S dng k thut sc k lng, vi cc iu kin nh sau: HPLC ra gii gradient t 2 n 100%, tc tng c nh 2%/pht (tuyn tnh). 6.2. nh lng Tin hnh theo hng dn trong chuyn lun xc nh tng hm lng bng chun vi Titan (III) clorid (trong JECFA monograph 1-Vol. 4) nh sau: Cn 0,6 - 0,7 g mu th; m 15 g natri hydrotartrat; Mi ml TiCl3 0,1 N tng ng vi 10,86 mg cc cht mu (D).

PH LC 21 YU CU K THUT V PHNG PHP TH I VI BROWN HT 1. Tn khc, ch s 2. nh ngha CI food Brown 3, Choccolat brown HT; CI (1975): 20285 INS: 155 ADI = 0 - 1,5 mg/kg th trng. Ch yu gm dinatri 4,4'- (2,4-dihydroxy-5-hydroxymethyl1,3-phenylen-bisazo) di-1-naphthalen-sulfonat cng cc cht mu ph cng vi NaCl v (hoc) Na2SO4 l cc thnh phn khng mu chnh. C th chuyn thnh cht mu nhm (aluminium lake) tng ng, trong trng hp ny p dng quy nh chung i vi cc loi cht mu nhm. Dinatri 4,4'- (2,4-dihydroxy-5-hydroxymethyl-1,3-phenylenbisazo) di-1-naphthalen-sulfonat. 4553-89-3 C27H18N4Na2O9S2


Khi lng phn t 652,57 3. Cm quan Dng bt hoc tinh th mu nu. 4. Chc nng Phm mu. 5. Yu cu k thut 5.1. nh tnh tan Tan trong nc, khng tan trong ethanol. nh tnh cc cht mu Phi c phn ng c trng ca cht mu. 5.2. tinh khit Gim khi lng khi lm Khng c qu 30% cng vi clorid v sulfat tnh theo mui kh ti 135 oC natri. Cht khng tan trong nc Khng c qu 0,2%. Ch Khng c qu 2 mg/kg. Cc cht mu ph Khng c qu 10%. Cc cht hu c ngoi cht Khng c qu 0,7% acid 4-Aminonaphthalen-1-sulfonic. mu Cc amin Khng c qu 0,01% tnh theo anilin. thm bc nht khng sulfon ho Cc cht c th chit bng Khng c qu 0,2%. ether 5.3. Hm lng Khng c thp hn 70% tng cc cht mu 6. Phng php th 6.1 tinh khit Ch - Th theo hng dn ti JECFA monograph 1-Vol. 4. - S dng k thut hp th nguyn t thch hp vi hm lng ch nh xc nh. La chn c mu th v phng php chun b mu da trn nguyn tc ca phng php m t trong JECFA monograph 1-Vol. 4 phn cc phng php phn tch cng c. Cc cht mu ph - Th theo hng dn ti JECFA monograph 1-Vol. 4.

- S dng cc iu kin sau: Pha chun theo cch sau: Pha long 1,0 ml dung dch ch phm 1% vi nc n 100 ml, lc u. Ly 0,10 ml dung dch ny vo ng nghim, thm 5,0 ml dung dch nc:aceton (1:1 tt/tt) v sau thm 14,9 ml dung dch natri hydrocarbonat 0,05 N, lc u. Xc nh hp th ca chun - As. Dung mi khai trin: S 6. Thi gian khai trin: ~ 14 gi. Cc cht hu c ngoi cht - Th theo hng dn ti JECFA monograph 1-Vol. 4 mu - S dng k thut sc k lng, vi cc iu kin nh sau: HPLC ra gii gradient t 2 n 100%, tc tng c nh 2%/pht. 6.2. nh lng Tin hnh theo hng dn trong chuyn lun xc nh tng hm lng bng quang ph (trong JECFA monograp

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