QCVN 4-22:2011/BYT

QUY CHUN K THUT QUC GIA V PH GIA THC PHM - CHT NH HA

National technical regulation on Food Additive Emulsifier

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 cht nh ha 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 cht nh ha 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. Cht nh ha: l ph gia thc phm c b sung vo thc phm nhm mc ch to ra hoc duy tr dng nh tng ng nht ca hai hay nhiu thnh phn ca thc phm. 3.2. JECFA monograph 1 - Vol. 4 (JECFA monographs 1 - Combined compendium of food additive 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 ha cht ca Hip hi Ha 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. MTDI 3.6. INS (International numbering system): H thng m s quc t v ph gia thc phm.

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II. YU CU K THUT, PHNG PHP TH V LY MU 1. Yu cu k thut v phng php th i vi cc cht nh ha c quy nh ti cc ph lc ban hnh km theo Quy chun ny nh sau: 1. 1. 1. 2. 1. 3. 1. 4. 1. 5. 1. 6. 1. 7. 1. 8. 1. 9. 1. 10 . 1. 11 . 1. 12 . 1. 13 . 1. 14 . 1. 15 . 1. 16 . 1. Ph 1: Ph 2: Ph 3: Ph 4: Ph 5: Ph 6: Ph 7: Ph 8: Ph 9: Ph 10: Ph 11: Ph 12: Ph 13: Ph 14: Ph 15: Ph 16: Ph lc lc lc lc lc lc lc lc lc lc Yu cu k thut v phng php th i vi mono- v di-glycerid ca acid bo Yu cu k thut v phng php th i vi ester ca glycerol vi acid lactic v acid bo Yu cu k thut v phng php th i vi ester ca glycerol vi acid citric v acid bo Yu cu k thut v phng php th i vi ester ca glycerol vi acid diacetyl tartric v acid bo Yu cu k thut v phng php th i vi sucroglycerid Yu cu k thut v phng php th i vi ester ca polyglycerol vi cc acid bo Yu cu k thut v phng php th i vi stearyl citrat Yu cu k thut v phng php th i vi trikali ortho-phosphat Yu cu k thut v phng php th i vi cc mui amoni ca acid phosphatidic Yu cu k thut v phng php th i vi sucrose acetat isobutyrat Yu cu k thut v phng php th i vi ester glycerol ca nha cy Yu cu k thut v phng php th i vi dinatri diphosphat Yu cu k thut v phng php th i vi calci polyphosphat Yu cu k thut v phng php th i vi cc mui ca acid myristic, palmitic v stearic (Ca, Na, K, NH4) Yu cu k thut v phng php th i vi ester ca sucrose vi cc acid bo Yu cu k thut v phng php th i vi dioctyl natri sulphosucinat Yu cu k thut v phng php th i vi 2

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17 . 1. 18 . 1. 19 . 1. 20 . 1. 21 . 1. 22 . 17: Ph 18: Ph 19: Ph 20: Ph 21: Ph 22: lc stearyl tartrat Yu cu k thut v phng php th i vi sorbitan monostearat Yu cu k thut v phng php th i vi sorbitan tristearat Yu cu k thut v phng php th i vi sorbitan monolaurat Yu cu k thut v phng php th i vi sorbitan monooleat Yu cu k thut v phng php th i vi sorbitan monopalmitat

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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 cht nh ha 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.

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2. Kim tra i vi cht nh ha Vic kim tra cht lng, v sinh an ton i vi cc cht nh ha 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 cht nh ha 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.

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Ph lc 1 YU CU K THUT V PHNG PHP TH I VI MONO V DIGLYCERID CA CC ACID BO 1. Tn khc, ch s Glyceryl monostearate, glyceryl monopalmitate, glyceryl monooleate; monostearin, monopalmitin, monoolein; GMS (i vi glyceryl monostearate); INS 471 Hn hp cc ester mono- v diglyceryl mch di vi cc acid bo bo ha v khng bo ha c trong cht bo thc phm; c cha hm lng alpha-monoglycerid khng thp hn 30% v cng c th c cha cc monoglycerid ng phn; cng nh di- v triglycerid, glycerol t do, cc acid bo t do, sn phm x phng ha v nc; thng c sn xut bng cch thy phn tch glyceryl ca cc cht bo v du thc phm, nhng cng c th c sn xut bng cch este ha cc acid bo vi glycerol c hoc khng qua chng ct phn t.

2. nh ngha

Cng thc cu to

Trong OCR l ca acid bo Khi lng phn t 3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Hp th hng ngoi Acid bo Glycerol 5.2. tinh khit Nc Khng c qu 2,0 % (phng php Karl Fischer) Khng tan trong nc, tan trong ethanol, cloroform v benzen. Phi c ph hng ngoi c trng ca ester mt phn ca acid bo vi polyol. Phi c phn ng c trng ca acid bo. Phi c phn ng c trng ca glycerol. Glyceryl monostearat: 358,6 Glyceryl distearat: 625,0 Cht bo rn dng sp c mu trng hoc kem hoc dng lng snh Cht nh ho

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Ch s acid Glycerol t do X phng Ch 6. Phng php th tinh khit X phng Thm 10 g mu th vo hn hp gm 60 ml aceton v 0,15 ml dung dch xanh bromophenol (0,5%) c trung ha trc bng acid hydrocloric 0,1N hoc natri hydroxyd 0,1N. Lm m t t trn cch thy cho n khi dung dch tan hon ton, chun bng acid hydrocloric 0,1N cho n khi mu xanh bin mt. yn dung dch trong 20 pht, lm m cho n khi cn c ha tan v mu xanh ti xut hin, tip tc chun n mt mu xanh. Mi ml acid hydrocloric 0,1N tng ng vi 0,0304 g C18H33O2Na. - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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. Khng c qu 6. Khng c qu 7% Khng c qu 6%, tnh theo natri oleat. Khng c qu 2,0 mg/kg.

Ch

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Ph lc 2 YU CU K THUT V PHNG PHP TH I VI ESTER CA GLYCEROL VI ACID LACTIC V ACID BO 1. Tn khc, ch s 2. nh ngha Lactic acid esters of mono- and diglycerides; lactoglycerides INS 472b L hn hp cc ester ca glycerol vi acid lactic v acid bo trong cht bo thc phm. Sn phm thng mi c th ch r hm lng monglycerid, acid lactic, ch s acid, ch s x phng ha, hm lng acid bo t do, im ng c ca cc acid bo t do, ch s iod, hm lng glycerol t do v hm lng nc.

Cng thc cu to

Trong R1, R2 v R3 l ca acid bo, acid lactic hoc hydrogen (theo thnh phn) 3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Acid bo Acid lactic Glycerol 5.2. tinh khit Cc acid Ch 6. Phng php th tinh khit Ch - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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. 7 Ch c acid bo v acid lactic. Khng c qu 2,0 mg/kg. Khng tan trong nc lnh nhng c th phn tn trong nc nng. Phi c phn ng c trng ca acid bo. Phi c phn ng c trng ca acid lactic. Phi c phn ng c trng ca glycerol. Cht rn dng sp Cht nh ho

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Ph lc 3 YU CU K THUT V PHNG PHP TH I VI ESTER CA GLYCEROL VI ACID CITRIC V ACID BO 1. Tn khc, ch s Citric acid esters of mono- and di-glycerides, citroglycerides, CITREM; INS 472c Thu c nh qu trnh ester ho glycerol vi acid citric v acid bo thc phm hoc nh phn ng ca hn hp mono v diglycerid ca acid bo vi acid citric; ch phm bao gm cc ester hn hp ca acid citric v cc acid bo vi glycerol; c th cha lng nh cc acid bo t do, glycerol t do, acid citric t do v mono v diglycerid; c th c trung ho hon ton hoc mt phn bng NaOH hoc KOH (nh c cng b trn nhn mc).

2. nh ngha

Cng thc cu to

Trong : t nht mt trong R1, R2 hoc R3 l ca acid citric, mt l ca acid bo v R cn li c th l ca acid citric, acid bo hoc hydrogen. 3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Acid bo Acid citric Glycerol 5.2. tinh khit Tro sulfat Sn phm khng c trung ho : Khng c qu 0,5% Sn phm c trung ho hon ton hoc mt phn: Khng c qu 10% (S dng 2 g mu th, phng php I) Khng c qu 4% 8 33% (m t trong phn Phng php th) 13 50% (m t trong phn Phng php th) 37 81% (m t trong phn Phng php th) 8 Khng tan trong nc lnh, c th phn tn trong nc nng; tan trong du v m; khng tan trong ethanol lnh Phi c phn ng c trng ca acid bo . Phi c phn ng c trng ca acid citric. Phi c phn ng c trng ca glycerol. Dng du ti sp, mu trng n trng ng Cht n nh, cht nh ho, cht chng oxy ho

Glycerol t do Glycerol tng s Acid citric tng s Acid bo tng s

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Khng c qu 2,0 mg/kg.

Ch 6. Phng php th tinh khit Glycerol tng s

Cn chnh xc 2 g mu v cho vo bnh x phng ho, thm vo 50 ml dung dch KOH 0,5 M trong ethanol, v un si vi sinh hn ngc trong 30 pht. Thm chnh xc 99 ml (t buret) chloroform v 25 ml acid acetic bng vo bnh nh mc dung tch 1 lt. Chuyn ton lng dch trong bnh x phng ho vo bnh nh mc dung tch 1 lt, cho 3 ln 25 ml nc. Thm vo 500 ml nc v lc mnh trong 1 pht. Pha n th tch 1 lt bng nc, lc u v yn cho tch thnh cc lp. Dng pipet ly 50 ml dung dch acid acetic periodic TS vo trong cc cc dung tch 400 ml. Chun b 2 mu trng bng cch thm 50 ml nc vo mi cc. Dng pipet ly 50 ml dung dch mu trong nc cho vo mt trong cc cc cha 50 ml acid acetic periodic TS, lc nh cho u, y bng tm knh nhn qua c v yn trong 30 pht. Nhng khng c qu 1,5 gi. Thm 20 ml dung dch KI 15%, lc nh cho u v yn t nht 1 pht nhng khng c qu 5 pht. Khng c di nh sng trc tip hoc nh sng trng. B sung 100 ml nc v chun bng dung dch natri thiosulfat 0,1 N. S dng dng my khuy lm u dung dch. Tip tc chun cho n khi mt mu iod nu trong dung dch nc. Thm 2 ml tinh bt TS v tip tc chun cho n khi mt mu iod trong lp chloroform c tch ra trong qu trnh chun v mt mu xanh ca tinh bt iodo trong dung dch nc. Kt qu: Glycerol tng s (%): Trong : B: Lng dung dch natri thiosulfat dng chun mu trng cha 50 ml nc S: Lng dung dch natri thiosulfat dng chun mu th N: Nng dung dch natri thiosulfat 0,1 N W: Khi lng mu th c ly bng pipet em chun v c tnh theo cng thc: Trong : a: Khi lng mu (g) em x phng ha

Acid citric tng s

Nguyn tc: Mu c x phng ho bng dung dch KOH trong alcol v cc acid bo c tch ra bng trch ly. Acid citric c chuyn thnh cc dn sut trimethylsilyl (TMS) v c 9

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phn tch bng sc k kh lng. X phng ho: Cn chnh xc 1 g mu cho vo bnh y trn, thm 25 ml dung dch KOH 0,5M trong ethanol v un si vi sinh hn ngc trong 30 pht. Acid ho hn hp bng HCl v cho bc hi bng my c quay hoc bng phng php thch hp khc. Trch ly: Chuyn ton lng mu trong bnh vo mt bnh gn, s dng khng qu 50 ml nc v trch ly bng 3 ln 50 ml heptan, loi b phn dch chit heptan tch ra. Chuyn lp dung dch nc vo bnh nh mc dung tch 100 ml, trung ho, pha ti th tch 100 ml bng nc v lc u. Dn xut ho: Dng pipet ly 1 ml dung dch pha c trn, 1 ml dung dch acid tartric (1 mg/ml trong nc) cho vo bnh y trn c th y np dung tch 10 ml v c n kh. Thm 1 ml pyridin, 0,2 ml dung dch trimethyl-chlorosilan (TMCS), 0,4 ml hexamethyl-disilazan (HMDS), 0,1 ml n-methyl-ntrimethylsilyl-trifluoroacetamid (MSTFA). y np cht v lc cn thn c dung dch tan hon ton. Gia nhit bnh trong t sy 600C trong 1 gi. Sc k kh: Bt k sc k kh thch hp c th c dng vi detector ion ho ngn la v ct (thu tinh, chiu di 1,8 m v ng knh trong 2 mm), c nhi bng DC-200 10% trn chromosorb Q (80/100 mesh). Cc iu kin cho sc k kh: nhit l ct 1650C, nhit bung tim 2400C, nhit detector 2400C; tc dng kh mang nitrogen 24 ml/pht. Cch tin hnh: Bm 5 l mu cc dn xut TMS. Thi gian lu i vi acid tartric khong 12 pht v thi gian lu tng i ca acid citric/acid tartric khong2,3. Lp li cch tin hnh c m t trn trong phn Dn xut ho v Sc k kh s dng 1 ml dung dch i chng acid citric (3 mg/ml trong nc) thay th 1 ml dung dch mu th. Tnh kt qu: o din tch mi pic bng phng php thch hp. Acid citric tng (%) = Trong : ACS: Din tch pic ca acid citric (dung dch mu) ATS: Din tch pic ca acid tartric (dung dch mu) ATR: Din tch pic ca acid tartric (dung dch i chng) ACR: Din tch pic ca acid citric (dung dch i chng) WCR: Khi lng (g) ca acid citric trong 1 ml dung dch i chng 10

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W: Khi lng (g) ca mu ester glycerol vi acid citric v acid bo. Cn chnh xc 5 g mu cho vo bnh y trn dung tch 250 ml, thm vo 50 ml dung dch KOH 1 N trong ethanol, v un si vi ng sinh hn ngc trn ni cch thy trong 1 gi. Chuyn ton lng dung dch trong bnh x phng ho vo phu chit dung tch 1.000 ml, cho 3 ln 25 ml nc v thm 5 git da cam methyl TS. Thm cn thn dung dch HCl m c cho n khi mu ca dung dch chuyn sang mu r rt v lc u tch cc acid bo. Trch ly cc acid bo tch c bng ba ln 100 ml dung mi diethyl ether. Ho chung cc phn tch c v ra mi ln bng 50 ml dung dch NaCl 10% cho n khi dung dch NaCl c ra tr nn trung tnh. Lm kh dung dch ether bng Na 2SO4 khan. Sau c bc hi trn cch thy ether, thm 10 pht na trn cch thy v cn phn cn.

Acid bo tng s

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Ph lc 4 YU CU K THUT V PHNG PHP TH I VI ESTER CA GLYCEROL VI ACID DIACETYL TARTRIC V ACID BO 1. Tn khc, ch s Diacetyltartric acid esters of mono- and diglycerides, DATEM; INS 472e Tartric, acetic and fatty acid esters of glycerol, mixed; Mixed acetic and tartric acid esters of mono and diglycerides of fatty acids; INS 472f Sn phm gm hn hp cc ester ca glycerol vi acid mono, diacetyltartric v cc acid bo thc phm. N c tng hp bng phn ng gia anhydrid diacetyltartric v mono, diglycerid ca acid bo c mt acid acetic, hoc bng cch tng tc gia anhydrid acetic v mono, diglycerid ca acid bo c mt acid tartric. Do trao i nhm acyl trong ni phn t hoc gia cc phn t, c hai phng php sn xut u to ra cc thnh phn chnh nh nhau, vic to ra cc thnh phn chnh ph thuc vo t l tng i ca cc nguyn liu c bn, vo nhit v thi gian phn ng. Sn phm ny c th cha mt lng nh glycerol t do, cc acid bo t do, acid tartric v acid acetic t do. Sn phm thng mi c th c quy nh cc ch tiu k thut r hn nh ch s acid, hm lng acid tartric tng s, hm lng acid acetic t do, ch s x phng ho, ch s iod, hm lng acid bo t do, im ng c ca acid bo t do. Cng thc cu to Cc thnh phn chnh l:

2. nh ngha

Trong : 1) Mt hoc hai nhm R l ca acid bo 2) Nhm R khc l ca hoc - acid diacetylat tartric - acid monoacetylat tartric - acid tartric - acid acetic - hydrogen 3. Cm quan 4. Chc nng 5. Yu cu k thut 12 T dng lng, bt nho n dng rn ging sp (vy hoc bt) Cht nh ho

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5.1. nh tnh tan 1, 2- diol Acid bo Acid acetic Acid tartric Glycerol 5.2. tinh khit Acid Tro sulfat Acid bo t do Acid acetic tng s Acid tartric tng s Glycerol tng s Glycerol t do Ch 6. Phng php th 6.1. nh tnh 1, 2- diol Cho vi git ch acetat TS vo dung dch c 500 mg mu trong 10 ml methanol. Cht kt ta vn, mu trng khng tan c hnh thnh. Khng pht hin c cc loi acid khc ngoi acid acetic, acid tartric v acid bo. Khng c qu 0,5% c xc nh 800 25C Khng c qu 3% tnh theo acid oleic Khng c nh hn 8% v khng c qu 32% sau khi thu phn (m t trong phn Phng php th) Khng c nh hn 10% v khng c qu 40% sau khi x phng ho (m t trong phn Phng php th) Khng c nh hn 11% v khng c qu 28% sau khi x phng ho (m t trong phn Phng php th) Khng c qu 2,0% Khng c qu 2,0 mg/kg. Phn tn trong nc nng v nc lnh, tan trong methanol v ethanol. Phi c phn ng c trng ca 1,2-diol. Phi c phn ng c trng ca acid bo Phi c phn ng c trng ca acid acetic Phi c phn ng c trng ca acid tartric Phi c phn ng c trng ca glycerol

6.2. tinh khit

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Acid acetic tng s Dng c: Lp rp b dng c chng ct Hortvet Sellier ci tin nh hnh v sau, s dng ng Sellier c kch thc bn trong ln (khong 38-x 203 mm) v by chng ct dung tch ln. Cch tin hnh: Cn chnh xc 4 g mu v cho vo ng bn trong ca b dng c v lp ng vo bnh ngoi c cha khong 300 ml nc nng va un si. Thm 10 ml acid perchloric 4 N [35 ml (60 g) acid perchloric 70% trong 100 ml nc] vo mu th, v ni ng bn trong vi sinh hn lm mt bng nc qua by chng ct. Chng ct bng cch gia nhit bnh pha ngoi sao cho thu c 100 ml dung dch ct trong khong 20 - 25 pht. Tp hp cc phn dch ct c 100 ml v thm phenolphthalein TS vo mi phn, chun bng dung dch NaOH 0,5 N. Tip tc chng ct cho n khi phn 100 ml dung dch ct c cn khng qu 0,5 ml dung dch NaOH 0,5N trung ho. (Ch : khng c chng ct n cn kh). Tnh khi lng (mg) acid bay hi trong mu theo cng thc V x e, trong V l tng th tch (ml) ca dung dch NaOH 0,5N dng chun v e l h s ng lng 30,03.

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Acid tartric tng s ng chun: Chuyn 100 mg acid tartric tinh khit c cn chnh xc vo bnh nh mc dung tch 100 ml, ho tan acid tartric trong 90 ml nc v thm nc n th tch 100 ml, lc u. Ly cc phn 3, 4, 5 v 6 ml cho vo cc cuvet ring bit ph hp (kch thc 19 x 150 mm), thm lng nc thch hp c dung dch th tch 10 ml. Cho vo mi cuvet 4 ml dung dch natri metavanadat 5% va mi pha, 1 ml acid acetic. (Ch : s dng cc dung dch ny trong vng 10 pht sau khi mu tng ln). Chun b mu trng ging nh cch trn, s dng 10 ml nc thay th dung dch acid tartric. t thit b v tr bng 0 i vi mu trng v sau o hp th ca 4 dung dch acid tartric ti bc sng 520 nm vi my quang ph thch hp hoc my so mu quang in c trang b knh lc 520 nm. T s liu thu c, xy dng ng chun trn h trc to vung gc vi trc tung biu din hp th, trc honh biu din lng acid tartric tng ng (mg). Chun b mu th: Cn chnh xc 4 g mu cho vo bnh tam gic dung tch 250 ml, thm vo 80 ml dung dch KOH 0,5N v 0,5 ml phenolphthalein TS. Ni bnh vi mt sinh hn khng kh c chiu di t nht 65 cm, gia nhit hn hp trn tm nhit trong 2,5 gi. Thm vo hn hp nng khong 10% acid phosphoric cho n khi hn hp c phn ng acid r vi giy ch th Congo. Ni li bnh vi sinh hn khng kh v gia nhit cho n khi acid bo c ho lng v trong. Lm ngui v sau chuyn hn hp vo mt bnh gn dung tch 250 ml vi s h tr ca cc phn nh nc v chloroform. Chit cc acid bo c gii phng ra bng 3 ln 25 ml nc ct lin tip v cho phn nc ra vo bnh gn c cha lp nc. Chuyn ton b hn hp trong bnh gn u vo cc dung tch 250 ml, gia nhit trn ni cch thy loi b cc vt ca chloroform, lc qua giy lc mn c ra acid vo bnh nh mc dung tch 500 ml, v cui cng pha ti th tch 500 ml bng nc ct (Dung dch I). Dng pipet ly 25 ml dung dch ny cho vo bnh nh mc dung tch 100 ml v pha ti th tch 100 ml bng nc (Dung dch II). Gi phn cn li ca Dung dch I xc nh glycerol tng s. Cch tin hnh: Cho 10 ml Dung dch II c chun b trong phn Chun b mu th vo cuvet 19 x 150 mm v tip tc lm theo hng dn trong phn ng chun, bt u vi cho vo mi cuvet 4 ml dung dch metavanadat 5%. T ng chun xc nh khi lng (mg) acid tartric trong ln pha long cui cng, sau nhn vi 20 v chia kt qu cho khi lng mu gc thu c % acid tartric. Chuyn 5 ml Dung dch I chun b trong phn xc nh 15

Glycerol tng s

QCVN 4-22: 2011/BYT

Acid tartric tng s vo trong bnh bnh tm gic c np thu tinh dung tch 250 ml hoc bnh iod. Thm vo bnh 15 ml acid acetic bng v 25 ml dung dch acid periodic, c chun b bng cch ho tan 2,7 g acid periodic (H 5IO6) trong 50 ml nc, thm 950 ml acid acetic bng v lc mnh; bo qun dung dch ny trnh nh sng. Lc u hn hp trong 1 hoc 2 pht, yn trong 15 pht, thm 15 ml dung dch KI 15% v 15 ml nc, lc xoy, yn trong 1 pht v sau chun iod c gii phng ra bng dung dch natri thiosulfat 0,1N, dng tinh bt TS l cht ch th. Thc hin chun mu trng s dng nc thay th mu th. Th tch thc l s ml dung dch natri thiosulfat 0,1N dng chun glycerol v acid tartric trong mu th c 5 ml Dung dch I. T % xc nh c trong Phng php th Acid tartric tnh th tch dung dch natri thiosulfat 0,1N dng chun acid tartric. Chnh lch gia th tch thc v th tch tnh ton c dng i vi acid tartric l s ml ca dung dch natri thiosulfat 0,1N tiu th bi glycerol trong mu. Mt ml dung dch natri thiosulfat 0,1N tng ng vi 2,303 mg glycerol v 7,505 mg acid tartric.

Dng c chng ct Hortvet-Sellier ci tin Ch - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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.

16

QCVN 4-22: 2011/BYT

Ph lc 5 YU CU K THUT V PHNG PHP TH I VI SUCROGLYCERID 1. Tn khc, ch s 2. nh ngha INS 474 Sucroglycerid thu c nh phn ng gia sucrose vi cht bo hoc du n c hoc khng c mt ca dung mi. Sn phm gm c hn hp mono- v diester ca sucrose v acid bo kt hp vi mono-, div triglycerid t cht bo hoc du. Ch cc dung mi sau y c th c s dng trong sn xut: dimethyl formamid, cyclohexan, isobutanol, isopropanol v ethyl acetat. Khng mi, dng mm, dng c, dng bt c mu trng n trng nht, hoc dng gel cng. Cht nh ho

3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Acid bo ng 5.2. tinh khit Tro sulfat Ch s acid Sucrose t do Dimethylformamid Cyclohexan v isobutanol Ethyl acetat v isopropanol Ch 5.3. Hm lng 6. Phng php th 6.1. nh tnh

Khng tan trong nc lnh, tan trong ethanol. Phi c phn ng c trng ca acid bo. Phi c phn ng c trng ca ng.

Khng c qu 2% Th 2 g mu (Phng php I) Khng c qu 6 Khng c qu 5% (m t trong phn Phng php th) Khng c qu 1 mg/kg (m t trong phn Phng php th) Khng c qu 10 mg/kg, dng n cht hoc hp cht (m t trong phn Phng php th) Khng c qu 350 mg/kg, dng n cht hoc hp cht Khng c qu 2,0 mg/kg. Khng c nh hn 40% v khng c ln hn 60% ester sucrose

17

QCVN 4-22: 2011/BYT

Acid bo Cho 1 ml ethanol vo 0,1 g mu, lm m v ho tan, thm 5 ml dung dch H2SO4 long TS, gia nhit trong ni cch thy trong 30 pht v lm ngui. Cht c mu trng vng hoc du c hnh thnh, v c ho tan khi thm 3 ml diethyl ether. Cho vo 2 ml lp dung dch nc tch ra t cht c hoc du trong php th acid bo, 1 ml anthron TS vo trong ng th, cho chy cn thn vo thnh ng th; b mt ranh gii gia hai lp chuyn thnh mu xanh da tri hoc xanh l cy.

ng

6.2. tinh khit

18

QCVN 4-22: 2011/BYT

Sucrose t do Xc nh bng sc k kh lng (xem Quyn 4) Ho cht, thuc th: - Dung dch ni chun: Dung dch cholesterol trong chloroform 5 mg/ml hoc tetracosan trong chloroform 10 mg/ml - Pyridin (lm kh c qua ry phn t) - N, O- Bis-(trimethylsilyl)-acetamid (BSA) - Trimethylchlorosilan (TMCS) Cch tin hnh: Cn chnh xc 20 50 mg mu cho vo bnh silyl ha, thm 1 ml dung dch ni chun, 1 ml pyridin, 0,5 ml N, O-bis (trimethylsilyl) acetamid (BSA) v 0,5 ml trimethylchlorosilan (TMCS). y np bnh v gia nhit 700C trong 30 pht. Bm 1l vo thit b sc k kh lng. Cc iu kin sc k: Ct: - Chiu di 0,3 m - ng knh trong 4 mm - Cht liu: thu tinh - c nhi bng dexil Kh mang: nitrogen Tc dng: 40 ml/pht Detector: FID Chng trnh nhit : gi 160 0C trong 1 pht sau tng n 160 3750C vi tc 150C/pht. o cc din tch pic ca sucrose v ni chun. H s p ng (RF) c tnh t qu trnh chy sc k kh lng vi cc dung dch chun sucrose cha ni chun. Kt qu: mg ni chun x din tch pic sucrose RF = ----------------------------------------------------Din tch pic ni chun x mg sucrose v mg ni chun x din tch pic sucrose x 100 % sucrose t do = --------------------------------------------------------RF x din tch ni chun x mg mu

19

QCVN 4-22: 2011/BYT

Dimethylformamid Xc nh bng sc k kh lng Ho cht, thuc th: - Dimethyl formamid - Dimethylamin hydrochlorid - Methanol - Ethanol - HCl - NaOH Dung dch chun: Chun b dung dch gc dimethylaminhydroclorid trong ethanol 4,47 mg/ml (tng ng vi 4,0 mg/ml dimethyl formamid) v chun b mt lot cc dung dch chun tng ng vi 4; 2 v 1 g/ml dimethylformamid bng cch pha long dung dch gc vi dung dch NaOH 0,1% trong ethanol. Chun b mu: B dng c thu phn c trnh by trong phn Ph lc. Cn chnh xc 40 g mu cho vo bnh y trn dung tch 1.000 ml. Thm 500 ml dung dch NaOH 5% trong methanol, v gn bnh vo b dng c thu phn. t bnh ct bnh tam gic cha 10 ml dung dch HCl 1% trong methanol vo b dng c thu phn. Gia nhit bnh y trn v chng ct vi sinh hn ngc c nc lm mt trong 1 gi, sau ct ly 50 ml dch ct c th dng li. C dch ct c cho n kh trong cch thy si. Ho tan phn cn bng mt lng nh ethanol, thm 2,5 ml dung dch NaOH 5% trong ethanol v pha long n th tch 25 ml bng ethanol chun b mt dung dch mu. Cch tin hnh: Bm 2 l dung dch mu vo thit b sc k kh lng vi cc iu kin di y. ng chun: Chun b ng chun bng cch bm 2 l cc dung dch chun vo thit b sc k kh. Cc iu kin sc k: Ct: - Di: 2m - ng knh trong: 2 mm - Cht liu: thu tinh - Ct nhi: 10% amin 220 v KOH 10% trn nn chromosorb W yu 80/100 c ra acid yu - Luyn ct: Gia nhit ti 130 0C qua m vi tc dng nitrogen 5 ml/pht Kh mang: nitrogen Tc dng: 17 ml/pht Detector: FID 20

QCVN 4-22: 2011/BYT

Cyclohexan v isobutanol Xc nh bng sc k kh lng (xem Quyn 4) s dng cc iu kin sau: Ho cht, thuc th: - Dimethylformamid (tinh khit dng cho GLC) - Cyclohexan (dng cho quang ph UV) - Isobutanol (dng cho phn tch) Dung dch chun: Chun b dung dch gc cyclohexan v isobutanol trong dimethylformamid bng cch dng pipet ly 130 l cyclohexan v 125 l isobutanol cho vo dimethylformamid v lm y n th tch 10 ml. T dung dch gc trn pha mt lot cc dung dch chun pha long c 5, 10 v 20 mg/kg cyclohexan v isobutanol. V ng p ng bng cch bm 5 l cc dung dch chun pha long vo thit b sc k kh vi cc iu kin sau. Chun b mu th: Cn 5 g mu chnh xc n 10 mg cho vo bnh c np y thu tinh, thm vo 5 g dimethylformamid v lm m ho tan. Lm ngui v bm 5 l vo thit b sc k kh vi cc iu kin sau. Ct: - Di: 3m - ng knh trong: 4,5 mm - Cht liu: thp khng r - Cht nhi: 20% carbowax 20 M trn Chromosorb G 60/80 Kh mang: heli (p sut 1,6 bar) Detector: ion ho ngn la Nhit : - Bung bm mu: 1300C - Ct: 1300C - Detector: 2000C Xc nh nng cyclohexan v isobutanol trong dung dch mu (50%) bng cch so snh vi cc dung dch chun v nhn nng vi 2 chuyn cc kt qu cho ph hp vi sucroglycerid gc.

21

QCVN 4-22: 2011/BYT

Isopropanol v ethyl acetat Xc nh bng sc k kh (xem Quyn 4) c bung ly mu khng gian hi s dng cc iu kin sau: Ho cht, thuc th: - Isopropanol - Ethyl acetat Dung dch chun: Ly 1 g mi loi isopropanol v ethyl acetat vo mt bnh nh mc v thm nc cho n khi t th tch 100 ml, v pha long dung dch ny c cc dung dch c nng 0,02 0,4 g/100 ml. Nu cn, chun b cc dung dch chun cha ti 7 g/100 ml isopropanol v ethyl acetat. Cch tin hnh: Cn 1 g chnh xc n 0,1 g bt mu cho vo l mu. Thm 5 l nc vo l mu v np kn nhanh bng septum. l mu vo thit b sc k kh t trc chng trnh v bt u phn tch theo cc iu kin cp di y. ng chun: Ly 1 g bt ester ca sucrose vi cc acid bo, khng cha dung mi hoc cc dung mi tn d bit, cho vo l mu, thm 5 l dung dch chun v y kn bng septum. l mu vo thit b sc k kh t trc chng trnh v bt u phn tch theo cc iu kin cp di y v v c ng chun ca mi dung mi. Ct: - Di: 30 m - ng knh trong: 0,53 mm - Cht liu: mao qun silica - Fim mng: 100% methyl polysiloxan - Luyn ct: Gia nhit ti 60 0C trong 2 3 gi vi tc kh nitrogen khong 10 ml/pht. Kh mang: nitrogen Tc dng: 5 ml/pht Detector: ion ho ngn la Nhit : - Bung bm mu: 1100C - Ct: 400C. - Detector: 1100C Bung khng gian hi: - Lng mu: 1 g 0,1 g + 5 l - Nhit gia nhit mu: 800C - Thi gian gia nhit mu: 40 pht - Nhit syringe: 850C 22

QCVN 4-22: 2011/BYT

Ch - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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.

23

QCVN 4-22: 2011/BYT

6.2. nh lng Xc nh bng sc k lng cao p (xem Quyn 4) s dng cc iu kin sau: Chun b mu: Cn chnh xc 250 mg mu cho vo bnh nh mc 50 ml. Pha ti th tch 50 ml bng tetrahydrofuran v lc u. Lc qua mng lc 0,5 m. Cch tin hnh: Bm 100 l mu vo thit b sc k lng cao p n nh trc. Cc iu kin sc k: Ct: Styren-divinylbenzen copolymer i vi sc k thm thu qua gel (TSK-GEL G2000 (Supelco) hoc tng ng) Pha ng: tetrahydrofuran dng cho HPLC ui kh Tc dng: 0,7 ml/pht Detector: RI (ch s khc x) Nhit : - Ct: 380C. - Detector: 380C Ghi sc k trong khong 90 pht. Tnh % ester sucrose trong mu: % ester sucrose = 100 A/T Trong : A: Tng cc din tch pic i vi 3 thnh phn chnh, mono-, di- v tri-ester, ra gii ln lt khong 65, 68 v 73 pht. T: Tng din tch tt c cc pic ra gii trong khong 90 pht Ph lc:

24

QCVN 4-22: 2011/BYT

25

QCVN 4-22: 2011/BYT

Ph lc 6 YU CU K THUT V PHNG PHP TH I VI ESTER CA POLYGLYCEROL VI CC ACID BO 1. Tn khc, ch s 2. nh ngha Polyglycerol fatty acid esters, glyceryl fatty acid esters; INS 475 L cc ester mt phn hn hp c to thnh bng phn ng glycerol polymer ha vi m v du n, hoc vi acid bo; c th cha mt lng nh mono-, di- v triglyceride, glycerol t do v polyglycerol, acid bo t do v mui natri ca cc acid bo; polymer ha c th thay i v c ch ra bng mt con s (th d l tri-, 3), l s trung bnh nhm (gc) glycerol tnh cho 1 phn t polyglycerol. Mt polyglycerol nht nh bao gm mt s phn b cc mnh phn t c trng cho polymer ha danh nh ca n. Bng cch thay i t l cng nh bn cht cc cht bo hoc acid bo phn ng vi polyglycerol, ngi ta thu c mt nhm ln v a dng cc sn phm ester ny; Cc sn phm ester thng mi cn c c trng vi 1 s ch tiu khc nh ch s x phng ha, im ng c ca acid bo t do, ch s iod, ch s hydroxyl v hm lng tro.

Cng thc cu to

Trong gi tr trung bnh ca n khong l 3 v mi R 1, R2, R3 c th l ca acid bo hoc hydro. 3. Cm quan Dng lng nh du ti rt nht, c mu vng sng n h phch; cht do hoc nha c mu vng nht n mu nu; cht rn hoc cht sp c mu vng nht n mu nu. Cht nh ho

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

T rt a nc ti rt a du, nhng l mt loi cht c xu hng phn tn trong nc v ha tan trong dung mi hu c v cc loi du. Phi c phn ng c trng ca acid bo. Phi c phn ng c trng ca glycerol v polyglycerol.

Acid bo Glycerol v polyglycerol 5.2. tinh khit Acid

Khng pht hin c acid no ngoi acid bo.

26

QCVN 4-22: 2011/BYT

Polyglycerol Phn polyglycerol gm c khng nh hn 70% di-, tri-, tetraglycerol v phi c khng qu 10% polyglycerol bng hoc cao hn heptaglycerol. Khng c qu 2,0 mg/kg.

Ch 6. Phng php th 6.1. nh tnh Glycerol v polyglycerol

Chm 5 20 l lp dung dch nc thu c trong php th acid bo Php th nh tnh cc nhm chc dc theo cc chm i chng glycerol trn giy Whatman s 3 v trin khai sc k i xung trong 36 gi vi h dung mi isopropanol: nc (90:10 ). Vt glycerol di chuyn c 40 cm v tip sau l cc polyglycerol c pht hin ln lt pha di sau glycerol khi giy c phun hoc permanganat trong aceton hoc bc nitrat trong amoniac.

6.2. tinh khit Ch - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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.

27

QCVN 4-22: 2011/BYT

Ph lc 7 YU CU K THUT V PHNG PHP TH I VI STEARYL CITRAT 1. Tn khc, ch s INS 484 ADI = 0 - 50 mg/kg th trng, c thit lp ti k hp ln th 17 ca JECFA (1973). c tng hp bng cch ester ha acid citric vi cn stearylic thng mi, c th cha n-octadecanol v n 50% n-hexadecanol v t c cc yu cu di y. Ch phm thng mi c th c phn loi theo ch s x phng ha; hm lng v thnh phn ca cn stearylic; ch s iod; ch s acid v hm lng acid citric. Thnh phn gn ng

2. nh ngha

Cng thc cu to

Trong R1, R2 v R3 ln lt c th l C18H37 (stearyl), C16H33 (palmityl) hoc H 3. M t 4. Chc nng 5. Tnh cht 5.1. nh tnh tan Cn stearylic Citrat 5.2. tinh khit Cc acid khc v ru khc Cht khng tan trong cloroform Ch 6. Phng php th 6.1. nh tnh Khng c c cc acid khc ngoi acid citric v cc ru khc ngoi cc ru c mt trong ch phm cn stearylic thng mi. Khng c qu 0,5% (m t trong phn Phng php th). Khng c qu 2,0 mg/kg. Khng tan trong nc v trong ethanol lnh, tan trong ethanol nng. Phi c phn ng c trng ca cn stearylic. Phi c phn ng c trng ca citrat. Hp cht nhn, mu kem. Cht to phc kim loi, cht nh ha.

28

QCVN 4-22: 2011/BYT

Cn stearylic Thy phn 2 g mu th bng cch un hi lu trong 1 gi vi 50 ml dung dch natri hydroxyd (TS) thu c dung dch nc. Lm mt v chit vi ether du ha. Ly lp ether du ha v cho bay hi trn a s hoc thy tinh. Phn khng bay hi phi c khong nng chy t 43 o n 58o. Ly 5 ml dung dch nc thu c trong php th cn stearylic, thm 1 ml dung dch calci clorid (TS) v 3 git dung dch xanh da tri bromothymol (TS), acid ha nh bng dung dch acid hydrocloric long (TS). Thm dung dch natri hydroxyd (TS) n khi dung dch chuyn thnh mu xanh da tri sng, sau un si dung dch trong 3 pht, lc nh trong khi un. Trong dung dch xut hin kt ta trng, kt ta ny khng tan trong dung dch natri hydroxyd (TS) nhng tan trong dung dch acid acetic (TS). Ly 10 ml dung dch nc thu c trong php th cn stearylic, thm 1 ml dung dch thy ngn (II) sulfat (TS). un si hn hp v thm vi git dung dch kali permanganat (TS). Trong dung dch xut hin kt ta trng l mui thy ngn ca acid aceton dicarboxylic.

Citrat

6.2. tinh khit Cht khng tan trong cloroform Ha tan khong 50g (cn chnh xc n mg) mu trong 400 ml cloroform. Lc dung dch qua phu lc thy tinh xp s 3 cn b (chnh xc n mg). Gi phu lc cn m, ra phn khng tan bng cloroform, sau sy kh ti 100 o. - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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.

Ch

29

QCVN 4-22: 2011/BYT

Ph lc 8 YU CU K THUT V PHNG PHP TH I VI TRIKALI ortho-PHOSPHAT 1. Tn khc, ch s Tripotassium phosphate; Tribasic potassium phosphate; Potassium phosphate. INS 340iii MTDI=70mg/kg th trng tnh theo Phospho t cc ngun thc phm

2. nh ngha Tn ha hc Ch s C.A.S. Cng thc ha hc Khi lng phn t 3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan pH Kali Phosphat ortho-phosphat 5.2. tinh khit
o Gim khi lng khi nung Dng khan: Khng c qu 3% (120 C trong 2 gi, sau 0 800 C trong 30 pht) Dng hydrat: Khng c qu 23% (120 oC trong 2 gi, sau 800oC trong 30 pht)

Trikali phosphat; Trikali orthophosphat; Trikali monophosphat 7778-53-2 Dng khan: K3PO4 Dng hydrat: K3PO4.xH2O 212,27 (dng khan) Dng tinh th hoc ht khng mu hoc mu trng, khng mi; dng hydrat gm monohydrat v trihydrat m, cht nh ha, cht to phc kim loi

D tan trong nc, khng tan trong ethanol. 11,5 - 12,5 (dung dch 1/100). Phi c phn ng c trng ca kali. Phi c phn ng c trng ca phosphat. Phi c phn ng c trng ca ortho-phosphat.

Cc cht khng tan trong nc Fluorid Arsen Ch

Khng c qu 0,2%. Khng c qu 10,0 mg/kg. Khng c qu 3,0 mg/kg (th theo hng dn ti JECFA monograph 1 - Vol.4 - phng php II). Khng c qu 4,0 mg/kg. 30

QCVN 4-22: 2011/BYT

5.3. Hm lng K3PO4 6. Phng php th 6.1. nh tnh Kali Thm 1 th tch dung dch natri hydrogen tartrat bo ha v 1 th tch dung dch ethanol vo dung dch mu th (1/100) v lc. Trong dung dch phi xut hin kt ta tinh th trng. Thm 1ml dung dch acid nitric v 5ml dung dch nhm molybdat (TS) vo 5ml dung dch mu th (1/100) v un nng. Trong dung dch phi xut hin kt qu vng nht. Ha tan 0,1g mu th trong 10ml nc ct, acid ha nh dung dch bng acid acetic long (TS) v thm 1ml bc nitrat (TS). Trong dung dch phi xut hin kt ta mu vng Khng c thp hn 97,0% sau khi nung

Phosphat

ortho-phosphat

6.2. tinh khit

31

QCVN 4-22: 2011/BYT

Fluorid Cn 5 g mu th cho vo mt bnh cu ct 250 ml, thm 25 ml nc, 50 ml acid percloric, 5 git dung dch bc nitrat (1 trong 2) v vi vin bi thy tinh. Ni bnh cu vi 1 sinh hn c gn nhit k v mt ng mao qun sc kh, u nhit k v ng mao qun phi ngp trong dung dch cha trong bnh. Gn mt phu nh git cha nc hoc b sinh hi nc vo ng mao qun. t bnh trn tm aming c l sao cho 1/3 y bnh tip xc vi ngn la. Tin hnh ct n khi nhit trong bnh t 135 o. Thm nc t phu vo bnh hoc cho hi nc qua bnh duy tr nhit trong bnh lun t 135o-140o. Tip tc ct n khi thu c 225-240 ml dch ct sau pha long dch ct thu c vi nc n 250 ml, lc u. Ly 50 ml dung dch ny cho vo mt ng Nessler 100 ml. Trong mt ng Nessler tng t khc ly 50 ml lm mu chng. Thm vo mi ng 0,1 ml dung dch natri alizarinsulfonat (nng 1/4000) lc v 1 ml dung dch hydroxylamin (nng 1/4000) mi pha, lc u. Thm tng git dung dch natri hydroxyd 0,05 N vo ng cha dch ct, va thm va khuy, n khi mu ng cha dch ct ging vi mu ng chng l mu hng nht. Sau thm vo mi ng ng chnh xc 1 ml dung dch acid hydrocloric 0,1 N v lc u. S dng buret chia vch 0,05 ml thm t t va dung dch thori nitrat (nng 1/4000) vo ng cha dch ct sau khi lc u mu ca dung dch chuyn v mu hng nht. Ghi th tch dung dch thori nitrat (nng 1/4000) s dng, thm chnh xc lng dung dch thori nitrat (nng 1/4000) nh vy vo ng chng, lc u. Dng buret thm dung dch natri fluorid (TS) (10 g F / ml) vo ng chng cho mu hai ng ging nhau sau khi pha long v cng th tch. Lc u, yn cho bt kh thot ht trc khi so mu. Kim tra im tng ng bng cch thm 1-2 git dung dch natri fluorid (TS) vo ng chng. S thay i mu r rt s xy ra. Ghi li th tch dung dch natri fluorid (TS) s dng. Th tch dung dch natri fluorid (TS) s dng khng c qu 1 ml. - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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.

Ch

32

QCVN 4-22: 2011/BYT

6.3. nh lng Cn mu th (chnh xc n mg), khi lng tng ng vi 8g K3PO4 khan, ha tan vo trong 40 ml nc ct trong cc 400ml. Thm 100ml dung dch acid hydrocloric 1N. Sc khng kh kh carbon dioxyd vo dung dch, ch sao cho bt kh nh, thi gian sc 30 pht ui ht carbon dioxyd, y nh cc trnh mt dung dch trong khi sc. Ra np v thnh cc bng nc ct v cho in cc ca pH k thch hp vo dung dch. Chun dung dch bng dung dch natri hydroxyd 1N n im un ti pH~4, sau tnh th tch (A) dung dch acid hydrocloric 1N s dng. Bo v dung dch trnh hp th carbon dioxyd t khng kh v tip tc chun vi dung dch natri hydroxyd 1N n im un ti pH~8,8. Tnh th tch (B) dung dch natri hydroxyd 1N tiu tn trong chun ny. Nu (A) 2(B), mi ml natri hydroxyd 1N trong th tch (B) tng ng vi 212,3mg K3PO4. Nu (A) < 2(B), mi ml natri hydroxyd 1N trong hiu th tch (A)-(B) tng ng vi 212,3mg K3PO4. Ph lc 9 YU CU K THUT V PHNG PHP TH I VI CC MUI AMONI CA ACID PHOSPHATIDIC 1. Tn khc, ch s Ammonium phosphatides, Emulsifier YN, Mixed ammonium salts of phosphorylated glycerides; INS 442 ADI = 0 -30 mg/kg th trng Ch phm ny ch yu gm hn hp cc hp cht amoni ca cc acid phosphatidic bt ngun t cht bo n c (thng l du ht ci hydrogen ha mt phn). L mt mono- hoc diglycerid c th c gn phosphor. Tuy nhin, hai ester phosphor c th lin kt vi nhau dng phosphatidyl phosphatid. Ch phm c sn xut bng cch phn hy cht bo bng glyceryl, sau phosphoryl ha bng phosphor pentoxyd, v trung tnh ha bng amoniac. Ch phm thng mi c th c quy nh thm nhng chi tit c th nh hm lng nc, tp cht khng tan trong hexan, cht v c khng tan trong hexan, gi tr pH v hm lng triglycerid.

2. nh ngha

Cng thc cu to (thnh phn gn ng)

Trong R c th l mt gc mono- hoc di-glycerid 3. Cm quan Cht na rn trn nhn. 33

QCVN 4-22: 2011/BYT

4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Phosphat Acid bo Glyceryl 5.2. tinh khit Ch 5.3. Hm lng Khng c qu 2,0 mg/kg. Hm lng phosphor khng c nh hn 3,0% v khng ln hn 3,4% theo khi lng; hm lng nitrogen amoni khng c nh hn 1,2% v khng ln hn 1,5%. Khng tan trong nc, tan mt phn trong ethanol v aceton; tan trong cht bo. Phi c phn ng c trng ca phosphat. Phi c phn ng c trng ca acid bo. Phi c phn ng c trng ca glyceryl. Cht nh ha

6. Phng php th 6.1. nh tnh Phosphat Nung 1 g mu th vi 2 g natri carbonat khan. ngui v ha tan cn trong 5 ml nc v 5 ml acid nitric. Thm 5 ml amoni molybdat (TS) v un ti si. Xut hin kt ta mu vng. un hi lu 1 g ch phm th trong 1 gi vi 25 ml kali hydroxyd 0,5 N trong ethanol. Amoniac bay ra t pha u ca ng sinh hn hi lu, nhn ra bng mi ca n v phn ng trn giy qu m. Lm lnh cn ti 0 oC, ta x phng kali to thnh.

Acid bo

6.2. tinh khit Ch - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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. Phng php quang ph hp th Thuc th v dung dch lm vic Acid sulfuric: t trng tng i 1,84 Acid nitric: t trng tng i 1,42 Acid perchloric: 60%, t trng tng i 1,54 Dung dch vanadat-molybdat: Ha tan ring trong nc 20 g amoni molybdat v 1 g amoni vanadat. Trn hai dung dch, thm 140 ml acid nitric c v pha long ti 1000 ml bng 34

6.3. nh lng Phosphor

QCVN 4-22: 2011/BYT

nc. Lc u. Dung dch phosphat chun: Dung dch gc: Ha tan 3,8346 g kali dihydro phosphat, lm kh 110C, vo trong nc v pha long ti 1000 ml; 1 ml dung dch ny = 2,0 mg P 2O5. Dung dch chun lm vic phosphat: Chuyn 50,0 ml dung dch phosphat chun (1ml = 2,0 mg P 2O5) vo bnh nh mc 500 ml v pha long ti vch bng nc v lc u. 1 ml dung dch chun lm vic phosphat = 0,2 mg P2O5. Tin hnh Cn chnh xc 1,5 ti 1,6 g mu th vo mt bt thy tinh nh v cho vo trong bnh Kjeldahl 300-ml cha 5 ml acid sulfuric v 10 ml acid nitric. un nng bnh, u tin un nh v lc lin tc, sau un mnh hn trn ngn la trn. Thm nhiu ln ti cc thi im khc nhau mi ln mt lng acid nitric, lm ngui bnh trc khi thm, v tip tc un ti khi thu c dch phn hy trong v c mu vng kim. ngui, thm 5 ml acid percloric 60% v tip tc oxy ha ti khi to thnh khi acid trng trong bnh. Li lm ngui v thm 5 ml nc v tip tc un ti khi ht khi trng bay ln. ngui, pha long cn thn bng nc, li ngui v chuyn ton lng sang bnh nh mc 500-ml. Pha long ti vch bng nc v lc u (Dung dch th). Lm mt mu trng phn hy ng theo cch nh trn nhng khng cho mu th v s dng cng th tch acid nh dng v c ha t mu th (Dung dch trng phn hy). Cho vo cc bnh nh mc 100-ml ring, thm t buret: (a) 25,0 ml dung dch chun lm vic phosphat (= 5,0 mg P2O5), (b) 30,0 ml dung dch chun lm vic phosphat (= 6,0 mg P2O5), (c) 25 ml dung dch th cha t 5 n 6 mg P2O5 Cho vo mi bnh cha phosphor chun, (a) v (b), mt lng dung dch trng phn hy c cng th tch nh dung dch cho vo (c), b i vi vt phosphor c th c do thuc th phn hy acid m n c th c trong dung dch th. Cho vo mi bnh 25 ml thuc th vanadat-molybdat, lc u, pha long bng nc ti gn 100 ml, lc u, iu chnh nhit ca dung dch ti 20C, pha long ti vch bng nc v lc u. Sau 10 pht, o hp th ca c dung dch 6 mg P 2O5 v dung dch th, mu trng l dung dch cha 5 mg cht chun. Dng cc o 1 cm v o bc sng 420 nm, hoc knh lc Ilford 604 nu dng my so mu quang in. Tnh ton
A t 0,873 % phosphor = 5 + A 6mg m

Trong : At = hiu hp th gia chun 5 mg v dung dch th A6 mg = hiu hp th gia chun 6 mg v 5 mg 35

QCVN 4-22: 2011/BYT

m = khi lng mu th (g) Thit b ct ko hi nc Xc nh nitrogen amoni Thit b gm mt bnh 2-L y nt cao su, lp xuyn qua nt c mt ng thy tinh khong 7,5 cm, c lp sao cho u di ca ng gn st y bnh, v mt on ng hnh ch L ngn hn lp sao cho ng cha ngang khong 6 mm bn di b mt ca nt, hot ng nh ng ra ca lung hi nc. Bnh c thm nc ct acid ha nh bng acid sulfuric long (TS) ti khong 2/3 th tch bnh v vi mnh thy tinh xp trnh tro khi hn hp trong bnh si mnh. Bnh ny dng lm ngun cung cp hi nc. C th lp mt vi vo bnh nu mun d dng b sung nc vo bnh gia cc ln xc nh. ng ra ca hi c ni qua mt by ngng ti u vo ca u ct hi, ni vo mt bnh cu y trn c ngn 1-L c bnh c B34. u ct phi sao cho ng u vo ca hi gn chm y bnh 1-L v u ra phi ni vi hai by trnh bn, mt gn nh ca bnh 1-L v ci kia gn nh ca mt sinh hn thng ng, ni c B19, ni vi u ct. Sinh hn thng phi lp vi ng u ra ko di, chm ti y ca mt bnh nn 500-ml. Thuc th Dung dch acid boric 2% (kl/tt) trong nc Dung dch natri hydroxyd 40% (kl/tt) trong nc Acid hydrocloric 0,02 N Hn hp ch th: Trn 5,0 ml dung dch xanh lc bromocresol 0,1% (kl/tt) trong alcol v 2,0 ml dung dch methyl 0,1% (kl/tt) trong alcol v pha long ti 30 ml bng alcol 95%. Dch silicon 200/50 MS Tin hnh Lp v cho hi nc i qua thit b. Cn chnh xc khong 0,2 g mu th mui amoni ca acid phosphatidic trung tnh vo trong mt l thy tinh nh (ng knh khong 18 mm, su khong 12 mm). Chuyn l v mu th cn vo bnh ct v thm khong 250 ml nc ct. Ni u ct v by chng bn vi bnh ct v sinh hn ng, lp sinh hn sao cho u ra ngp di b mt ca 10 ml acid boric 2% v 1 ml hn hp ch th ng trong bnh nn 500-ml. Thm vo bnh ct qua phu gn bng mt ng cao su gn vo ng dn hi nc vo, 75 ml natri hydroxyd 40%, v ra vo trong bng nc ct. B phu v ni ng vo ca hi vi ngun cung cp hi. (C th cho natri hydroxyd vo bnh qua phu c kha, lp vi bnh ct nu mun v ra pha trong bng nc ct. Nu lm nh vy phi duy tr mt lp cht lng trn phu trong sut qu trnh thm v ct). Ct ko mnh bng hi nc hn hp trong bnh ct v thu ly 200 ml dch ct vo trong acid boric. Trong qu trnh ct khuy nh bnh ct nu cn, trnh mu th b lng ng xung quanh b mt pha trn ca bnh. Khi thu lng dch ct theo yu cu, h thp bnh hng, ngng cung cp hi nc v ra pha trong ca sinh hn v bn ngoi u pha di vi mt lng nh nc ct, gp dch ra vo bnh 36

QCVN 4-22: 2011/BYT

hng. Chun hn hp trong bnh hng bng acid hydrocloric 0,02 N. Lm t nht mt mu trng tin hnh theo cng cch nhng khng c mu th. Trong qu trnh ct c th c bt xut hin ca hn hp trong bnh ct. Nu c, thm 2 git dch silicon vo bnh ct ti thi im cho mu th; v mt lng tng t c cho vo mu trng. Tnh ton Mi ml HCl 0,02 N = 0,2802 mg nitrogen. Vt V o 28,02 % Nitrogen = mt Trong Vt = th tch dung dch HCl 0,02 N dng chun mu th (ml) Vo = th tch dung dch HCl 0,02 N dng chun mu trng (ml) mt = khi lng mu th (mg). Ph lc 10 YU CU K THUT V PHNG PHP TH I VI SUCROSE ACETAT ISOBUTYRAT 1. Tn khc, ch s 2. nh ngha SAIB INS 444 Hn hp c to thnh bng cch este ha sucrose thc phm vi anhydrid acetic v anhydrid isobutyric v chng ct. Hn hp bao gm tt c cc este vi t l phn t ca acetat v isobutyrat l khong 2:6 Sucrose diacetat hexaisobutyrat 137204-24-1; 27216-37-1; 126-13-6 C40H62O19 i vi sucrose diacetat hexaisobutyrat

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

Trong R1 = -CH2OCOCH3 R2 = -CH2OCOCH(CH3)2, hoc -OCOCH(CH3)2 Khi lng phn t 3. Cm quan 832 856 C40H62O19 = 846,9 Dng lng mu vng nht, khng cn v c mi nh 37

QCVN 4-22: 2011/BYT

4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Ch s khc x Khi lng ring Hp th hng ngoi 5.2. tinh khit Ch s acid Ch s x phng ha Triacetin Ch 5.3. Hm lng C40H62O19 6. Phng php th 6.1. tinh khit Triacetin S dng phng php sc k kh nh sau: Dng c, thit b: My sc k kh c trang b detector ion ha ngn la Ct: thp khng r, di 1,5 m, ng knh trong l 3,2 mm. Chun b mu: Pha long mu th bng cch cho thm mt th tch carbon disulfid tng ng. iu kin: Pha tnh: SE-30, 3% Pha rn: Chromosorb AW-DMCS, 80-100 mesh Kh mang: Heli Tc kh: 20 ml/min Nhit - Ct: Chng trnh nhit : ngay sau khi bm mu, nhit ct tng t 1000 (ln n 3000C, tc tng 100C/pht. - Nhit bung bm: 3000C Th tch bm mu: 1 l - Th theo JECFA monograph 1 - Vol.4. 38 Khng c qu 0,2 (s dng 50 g mu th). Gia 524 v 540 (s dng 1 g mu th) Khng c qu 0,1%. Khng c qu 2 mg/kg. Khng c nh hn 98,8% v khng c qu 101,9%. Khng tan trong nc, tan trong hu ht dung mi hu c. n (40, D): 1,4492 1,4504 d (25, 25): 1,141 1,151 Ph hng ngoi ca mu th phn tn trong kali bromid tng ng vi ph hng ngoi chun trong Ph lc. Cht nh ha, cht iu chnh m c, cht to c i vi ung khng cn

Ch

QCVN 4-22: 2011/BYT

- Xc nh bng k thut quang ph 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. 6.2. nh lng S dng ch s x phng ha, tnh % C 40H62O19 theo cng thc:

Trong SV = ch s x phng ha Ph hng ngoi Sucrose acetat isobutyrat

39

QCVN 4-22: 2011/BYT

Ph lc 11 YU CU K THUT V PHNG PHP TH I VI ESTER CA GLYCEROL VI NHA CY 1. Tn khc, ch s 2. nh ngha Ester gum; INS 445 Ester phc hp ca triglycerol v diglycerol vi cc acid t nha g thng thu c nh qu trnh trch ly gc cy thng lu nm bng dung mi sau c tinh ch trong h dung mi lng-lng. Loi tr khi cc yu cu k thut ny l cc dn xut ca gum nha, dch tit ra t cy thng sng, nha du thng cao v sn phm ph ca qu trnh sn xut bt giy. Sn phm cui cng gm c 90% acid nha v 10% cht trung tnh (cht khng acid). Phn acid nha l mt phc hp ca cc ng phn acid monocarboxylic diterpenoid c cng thc thc nghim in hnh l C20H30O2, trong thnh phn chnh l acid abietic. Glycerol ester ca nha g thng c tinh ch bng phng php bc hi nc hoc chng ct ngc dng. 8050-30-4 Cht rn mu vng n mu h phch nht Thnh phn base trong ko cao su, cht nh ho, cht n nh, cht n, cht iu chnh hng liu trong ung

M s C.A.S. 3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Hp th hng ngoi Alcol nha v glycerol nha 5.2. tinh khit Khng c mt nha du cao (Lu hunh) T trng

Khng tan trong nc, tan trong aceton Ph hng ngoi ca lp mng mu c t trn tm kali bromid tng ng vi ph hng ngoi in hnh di y Phi c phn ng c trng ca alcol nha v glycerol nha.

t yu cu (m t trong phn phng php th). d (20, 25) : Khng c nh hn 0,935 khi xc nh trong dung dch 50% trong d-limonen (97%, im si 175,5 176,00C, d (20, 4) : 0,84) 82 900C (m tr trong phn Phng php th) 39 15 45 (m tr trong phn Phng php th ) Khng c qu 2,0 mg/kg. 40

im nng chy Ch s acid Ch s hydroxyl Ch

QCVN 4-22: 2011/BYT

6. Phng php th 6.1. nh tnh Alcol nha v glycerol nha Cc nhm ester phc tp trong ester ca glycerol nha g thng c kh bng phn ng vi mt hydrid kim loi (natri bis (2-methoxy-ethoxy) nhm dihydrid) trong dung dch toluen to thnh mt hn hp ca alcol nha v glycerol nha. Cht kh d sau c thu phn bng dung dch acid to thnh hai pha. Sc k kh ca pha toluen to ra mt sc k ca alcol nha cu thnh ester nha g thng v c th phn bit vi cc ester gum v ester nha du cao. Nha g c th phn bit c vi nha du cao da vo t l ca abietyl v dehydroabietyl alcol: trong nha g abietyl chim u th, trong khi nha du cao, dehydroabietyl chim u th. Nha g c th phn bit c vi gum rosin da vo t l ca isopimaryl v palustryl alcol: trong nha g isopimaryl chim u th, trong khi gum rosin, palustryl chim u th. Sc k ca pha trung tnh trong nc trn ct khc kim tra s xut hin ca glycerol. Thit b, dng c: - Sc k kh c trang b detector ion ho ngn la - Ct sc k I: Column I: DB-1 methyl silicon (ngoi quan v lin kt cho), ct mao qun di 15 m, ng knh trong 0,53 mm, dy 1,5 m, di nhit 60 - 300/3200C (V d: J & W Scientific Inc,. Cat. No. 125-1012). C ng dn bm hi nhanh, trc tip. - Ct sc k II: : DB-Wax polyethyleneglycol (ngoi quan v lin kt cho), ct mao qun di 15 m, ng knh trong 0,53 mm, dy 1,0 m, di nhit 20 - 230 0C (V d: J & W Scientific Inc,. Cat. No. 125-7012). - B ghi: 0 1V - Syringe: 1 l - Pipet: 3,0 ml; 5,0 ml - Bnh tam gic: 25 ml - L: 17 ml - Cn phn tch: cn chnh xc n 0,001 g - ng li tm c vch nh mc: 15 ml - My li tm: c th t tc 3.200 vng/pht - Thanh khuy: c bao bng Teflon, kch thc 1 inch - My khuy t Ho cht, thuc th: - Toluen tinh khit - Natri Vitride TM [ natri bis (2-methoxyethoxy) nhm dihydrid, thng xp x 70% trong toluen (khong 3,5 mol/l)] (Tp on ho cht Fluka ., Hauppage, NY, M). Ly 10 ml cho vo bnh nh mc dung tch 100 ml. Pha 41

QCVN 4-22: 2011/BYT

n th tch 100 ml bng toluen v lc u. - Dung dch thu phn: Thm t t 50 ml dung dch H 2SO4 m c, tinh khit vo 200 ml nc ct ng thi khuy trong chu nc . Lm ngui ti nhit phng. - Dung dch phenolphthalein: 1% trong ethanol. - Dung dch NaOH: Ho tan 16 g NaOH trong 70 80 ml nc ct v lm ngui ti nhit phng. Pha ti 100 ml bng nc ct v lc u. trong bnh polyethylen. - 1, 4- Butandiol: 99% - Glycerol: 99% - Dung dch ni chun: Cn 0,1 g 1, 4- butandiol cho vo bnh nh mc dung tch 100 ml. Pha ti th tch 100 ml bng nc ct v lc u. - Dung dch glycerol: Cn 0,1 g 1, 4- butandiol v 0,1 g glycerol cho vo bnh nh mc dung tch 100 ml. Pha ti th tch 100 ml bng nc ct v lc u. iu kin chy sc k kh: I (alcol nha) Nhit : - Ct: 1900C, ng nhit - u vo: 2500C - Detector: 2500C Tc dng: - Kh mang (He): 30 ml/pht ti 63 psi - H2: 30 ml/pht - Khng kh: 240 ml/pht II (glycerol) Nhit : - Ct: t theo chng trnh, 120 200 0C vi tc 60C/pht - u vo: 2500C - Detector: 2500C Tc dng: - Kh mang (He): 30 ml/pht ti 63 psi - H2: 30 ml/pht - Khng kh: 240 ml/pht Cch tin hnh I (i vi nha): Cn 250 300 mg mu cho vo bnh tam gic dung tch 25 ml c thanh khuy t. Dng pipet ly 5,0 ml toluen cho vo bnh v khuy t cho n khi mu c ho tan. Ly 5,0 ml Natri VitrideTM cho vo bnh, y np bnh v khuy trong 30 pht. M np ra ng thi khuy, dng pipet ly 3,0 ml dung dch thu phn cho vo bnh. Tip tc khuy trong 3 pht. Chuyn hn hp trong bnh vo ng li tm, 42

QCVN 4-22: 2011/BYT

y np v lc u. Cho thot kh v li tm vi tc 2.800 3.200 vng/pht trong 5 pht. Bm 0,5 l lp trn vo thit b sc k kh vi cc iu kin k thut t v ghi sc k . So snh vi cc sc k c biu th pha di xc nhn th t thi gian lu ca cc alcol nha. Cch tin hnh II (i vi glycerol) Dung pipet hoc syringe hypodermic, loi b lp toluen v mt phn dung dch nc v li khong 2 ml dung dch nc trong ng li tm. Thm 1 git dung dch phenolphthalein v trung ho bng dung dch NaOH. Cc mui nhm s kt ta. Dng pipet ly 5 ml dung dch ni chun cho vo ng, pha n 15 ml bng nc ct, y np, lc v li tm vi tc 2.800 3.200 vng/pht trong 5 pht. Bm 1 l dch trong trn b mt vo thit b sc k kh vi cc iu kin k thut t v ghi sc k . Bm 1 l dung dch glycerol v ghi sc k . o cc thi gian lu ca bt k cc pic so vi 1,4- butandiol. So snh cc thi gian lu vi thi gian lu ca glycerol. 6.2. tinh khit Khng c nha du cao (Lu hunh) Khi cc hp cht hu c c cha lu hunh c gia nhit vi s c mt ca natri format, lu hunh chuyn ho thnh hydrogen sulfid c pht hin bng cch s dng giy ch acetat. Php th dng tnh cho thy vic s dng nha du cao thay cho nha g. Thit b, dng c: - ng th: loi tiu chun, kch thc 10 x 70 mm, lm bng thu tinh chu nhit. - n Bunsen: Thng dng c nh dng ngn la nh Ho cht, thuc th: - Dung dch natri format: Ho tan 20 g natri format tinh khit (NaOOCH) trong 100 ml nc ct. - Giy th ch acetat: dng thng mi c sn ca hu ht cc nh cung cp ho cht Cch tin hnh: Cn 40 50 mg mu cho vo ng th v thm 1 -2 git dung dch natri format 20% (khi lng/th tch). di giy th ch acetat trn ming ng th. Gia nhit ng th bng ngn la cho n khi khi c hnh thnh v tip xc vi giy th. Tip tc gia nhit trong 2 5 pht. Khng c c m en ca ch sulfid hnh thnh, nu c m en th chng t s c mt ca cc hp cht cha lu hunh. (Gii hn pht hin: 50 mg/kg lu hunh).

43

QCVN 4-22: 2011/BYT

im nng chy im nng chy c nh ngha l nhit ti mt a mu c gi bn trong mt chic nhn nm ngang gim xung mt khong 25,4 mm di trng lng ca mt cu thp khi mu c gia nhit vi tc qui nh trong chu cch thy hoc cch glycerol. Thit b: Thit b c minh ho trong Hnh 1 v Hnh 2 gm c cc b phn c m t trong cc phn di y. Chic nhn: c lm bng ng thau ph hp vi kch thc trnh by trong Hnh 1a. Nu cn, vng c th c gn bng cch hn hoc cch khc thun tin vi mt dy ng thau loi 13 B&S (ng knh 0,06 0,08 inch hoc 1,52 2,03 mm) c trnh by Hnh 2a. Qu cu: thng dng qu cu c lm bng thp c ng knh 3/8 inch (9,53 mm), khi lng 3,45 3,55 g. Hng dn chnh tm qu cu: Hng dn chnh tm qu cu c lm ng thau v c hnh dng, kch thc chung c minh ha trong Hnh 1c, c th c s dng nu mun. Dng c cha: S dng bnh thu tinh chu nhit nh cc Griffin dng ln dung tch 800 ml, ng knh khng c nh hn 3,34 inch (8,5 cm) v chiu su tnh t y n phn loe ra khng c nh hn 5 inch (12,7 cm). Dng c gi chic nhn v nhit k: Bt k dng c hu hiu no ph hp vi cc yu cu sau dng gi vng v nhit k u c th c s dng: (1) Vng nhn c th c gi v tr nm ngang; (2) khi s dng thit b m t trong Hnh 1d, y ca vng trn nm trn v cch tm nm ngang pha di 1 inch (25,4 mm), b mt y ca tm nm ngang cch t nht 0,5 inch (13 mm) v khng c qu 0,75 inch (18 mm) so vi y ca dng c cha bn trn, chiu su ca dch lng trong dng c cha khng c nh hn 4,0 inch (10,2 cm); (3) khi s dng thit b a ra Hnh 1e, y ca vng trn cch 1 inch (25,4 mm) so vi y ca dng c cha pha trn, vi im di cng ca thanh ngh t trn y ca dng c cha v chiu su ca dch lng trong dng c cha khng c nh hn 4,0 inch (10,2 cm) nh m t Hnh 1 a, b v c; (4) trong c hai dng c, nhit k c treo ln sao cho khong cch t y ca bu nhit k n y ca chic nhn khong 0,5 inch (13 mm) nhng khng c chm vo nhn. Nhit k thu ngn: Ph thuc vo im nng chy ca mu, s dng hoc nhit k ASTM 15C (-2 n 80 0C) i vi loi c im nng chy thp hoc nhit k ASTM 16C (30 - 200 0C) i vi loi c im nng chy cao . 44

QCVN 4-22: 2011/BYT

My khuy: S dng my khuy thch hp vi tc 500 700 vng/pht. m bo phn tn nhit u trong mi trng nng, hng ca trc quay nn y dch lng ln pha trn. (Xem Hnh 2d ). Chun b mu: Ly mu i din gm cc mnh va mi b v c b mt cha b oxy ho. No lp trn b mt mu thu cc mnh v ngay trc khi s dng, trnh bao gm cc bt mn hoc bi. Lng mu ly thng t nht gp i lng cn cho s chic nhn yu cu nhng khng c nh hn 40 g. Ngay sau lm nng chy mu trong dng c cha sch, s dng t sy, tm gia nhit hoc cch ct hoc cch du ngn qu nhit cc b. Trnh hnh thnh bt kh trong mu nng chy, khng c gia nhit trn nhit cn thit mu d dng m khng bao gm bt kh. Thi gian tnh t lc bt u gia nhit cho n lc mu khng qu 15 pht. Ngay trc khi y cc chic nhn, gia nhit mu trc n nhit ti mu c rt. Trong khi rt, cc vng trn c gi trn tm ng thau. Rt mu vo cc vng trn tha cho ngui. Lm ngui t nht 30 pht v sau ct b phn mu tha bng dao h nng hoc dao bay. S dng dng c cha sch v mt mu mi nu php th c lp li. Cch tin hnh: Mu c im nng chy trn 800C: Rt glycerol vo bnh thu tinh ti su khng c nh hn 4 inch (10,2 cm) v khng c qu 4,25 inch (10,8 cm). Nhit ban u ca chu l 320C. i vi nha thng (gm c nha g), glycerol cn c lm ngui n nhit khng c nh hn 450C di im nng chy d on trc nhng khng c thp hn 350C. V tr trc ca trc khuy gn thnh sau ca dng c cha, vi cc li dao khng chm vo thnh v phn di ca cnh dao cch nh ca vng 0,75 in (18 mm). Tr khi c hng dn chnh tm cu c s dng, lm lm nh trung tm ca mu bng cch nhn qu cu hoc mt que trn c h nng dng cho mu cng, vo mu ti thi im ny . Treo vng trn c cha cc mu trong chu glycerol b mt di ca vng trn y cch b mt ca tm ngang thp hn l 1,0 inch (25,4 mm) (xem Hnh 1d), t nht 0,5 inch (13 mm) v khng qu 0,75 inch (18 mm) trn so vi y ca bnh thu tinh, hoc 1,0 inch (25,4 mm) trn so vi y ca dng c cha (xem Hnh 2e). t qu cu trong glycerol nhng khng c t ln trn mu th. Treo nhit k ASTM 16C trong glycerol sao cho y bu ca nhit k cch y chic nhn khong 0,5 inch (13 mm) nhng khng c chm vo chic nhn. Duy tr nhit ban u ca glycerol trong 15 pht v sau dng kp thch hp t qu cu vo trung tm b mt trn ca mu trong chic nhn. Bt u khuy v tip tc khuy vi tc 500 45

QCVN 4-22: 2011/BYT

700 vng/pht cho n khi hon tt vic xc nh. Gia nhit theo tc tng nhit ca glycerol 5 0C/pht, trnh nh hng ca cn bng cch s dng cc tm chn nu cn thit. [Ch : Tc tng nhit s khng i v s khng c tnh theo gi tr trung bnh trong ton b qu trnh th. Loi b tt c cc php th trong tc tng nhit vt qu 0.5C cho bt k qu trnh no sau 3 pht u]. Ghi li im nng chy, chnh l nhit trn nhit k ti mu chm vo tm nm ngang pha di (xem Hnh 1d) hoc y ca dng c cha (xem Hnh 2e). Khng hiu chnh thn nhit k. Mu c im nng chy di 80 0C: thc hin theo cch tin hnh trn, ngoi tr s dng nhit k ASTM 15C v s dng nc va un si c lm mt ti 5 0C to mi trng nhit. i vi nha thng (gm c nha g), s dng nc c lm ngui n nhit khng c nh hn 450C di im nng chy d on trc nhng khng c thp hn 50C. Ch s hydroxyl Ch s hydroxyl c nh ngha l s mg KOH dng trung ho carbamat sulfonyl c kh nng kt hp vi cc nhm hydroxyl trong 1 g mu. Cc nhm hydroxyl trong hp cht hu c phn ng nhanh vi p toluensulfonyl isocyanat d (p - TSI) trong dung mi tr to thnh carbamat sulfonyl. Sau phn ng ca thuc th d vi nc to thnh p toluensulfonamid, carbamat sulfonyl c tnh acid c chun bng trc quan hoc thit b chun bng dung dch KOH trong methanol, pht trin im tng ng bng o th hoc bng ch th mu. Cc hiu chnh c thc hin i vi mu trng v s c mt ca bt k cc hp cht c tnh acid. Thit b, dng c: - Thit b chun t ng: s dng thang o pH 0 14, t tc 4 (khong 2 ml/pht) v mt buret 20 ml. - in cc thu tinh - in cc calomel - Bnh tam gic: dung tch 250 ml c khp ni thu tinh hnh trn - Sinh hn Allihn: di 300 mm c khp ni thu tinh hnh trn khp vi bnh tam gic. Ho cht, thuc th: - Acid benzoic, t tiu chun cht chun gc - Methanol, tinh khit, dng khan - Toluen, tinh khit, sy kh qua ry phn t 3 - KOH 0,1 N pha trong methanol: Ho tan 7 g KOH trong 1 lt methanol. Chun ho bng cch cn chnh xc n 0,0001 g, khong 46

QCVN 4-22: 2011/BYT

0,12 g acid benzoic vo cc dung tch 150 ml. Cho thanh khuy t v 80 ml hn hp toluen/methanol theo t l 1/1. Khuy u dung dch trong khi chun vi dung dch KOH trong methanol. Dung dch KOH cn phi cho xung di b mt ca dung dch, cch xa in cc. Tnh ton nng dung dich KOH trong methanol da theo phng trnh tnh ton c biu th di y. - Tetrahydrofuran (THF): sy qua m v cho qua ry phn t 3 - p-Toluenesulfonyl isocyanat (p-TSI) - Dung dch p-Toluenesulfonyl isocyanat: khong 0,22 M hoc 4,5 mEq/20 ml dung dch. Dng pipet ly 15 ml pToluenesulfonyl isocyanat (p-TSI) cho vo 500 ml tetrahydrofuran (THF) kh v lc u. Ch : V isocyanat c bn cht rt c nn khi chun b v s dng dung dch isocyanat phi c tin hnh trong t ht. Cn phi eo gng tay khi cm vo isocyanat v dung dch ca isocyanat trnh tip xc vi da. - Dung dch ch th bromocresol ta: cho 0,1 g bromocresol ta vo 18,5 ml dung dch NaOH 0,1 N v pha ti th tch 250 ml bng nc ct. Cch tin hnh: Cn 1 - 1,5 g mu chnh xc n 0,0001 g cho vo bnh tam gic kh dung tch 250 ml c ni vi khp ni thu tinh hnh trn. Dng pipet ly 10 ml dung dch THF kh cho vo bnh v ho tan mu. Dng pipet ly 20 ml dung dch p TSI cho vo bnh v lc u. Cho vi ht bt, ni vi sinh hn v bnh trn tm gia nhit. Gia nhit n khi si vi sinh hn ngc trong 10 pht. Trong qu trnh chng ct b sung thm nc thch hp qua sinh hn: Mu trng Mu th Chun vi ch th mu 2 2 Chun bng o th 1 1 Ly bnh v sinh hn ra khi tm gia nhit v ngui ti nhit phng. Ra sinh hn bng 5 ml THF. Tho sinh hn ra khi bnh v chuyn ton b phn dch trong bnh vo cc dung tch 150 ml vi s tr gip ca 50 ml THF. Cho thanh khuy t v nhng cc in cc vo hoc thm 20 git dung dch ch th. Va khuy va chun bng thit b chun o th hoc chun vi ch th mu trc quan vi dung dch KOH trong methanol. Dung dch KOH 47

QCVN 4-22: 2011/BYT

cn phi cho xung di b mt ca dung dch cch xa in cc. Xc nh th tch dung dch KOH dng chun mu th v chun mu trng c thc hin qua tt c cc bc tin hnh trn. Tnh kt qu: Nng ca dung dch KOH trong methanol (N):

Trong : W: Khi lng acid benzoic 122,1: ng lng ca acid benzoic KOH: S ml dung dch KOH trong methanol dng chun Ch s hydroxyl trong mu:

Trong : A: S ml dung dch KOH dng chun mu th B: S ml dung dch KOH dng chun mu trng N: Nng dung dch KOH trong alcol 56,1: S mg KOH/mEq AV: Ch s acid Asen Chun b dung dch mu nh sau: Cn chnh xc 1 g mu v cho vo bnh Kjeldahl, t u mt ng ca bnh trong qu bu bc hi c gn vi my ht nc, thm 5 ml dung dch H2SO4 v 4 ml dung dch hydrogen peroxyd 30% v ph mu trn ngn la nh. Tip tc thm 2 ml peroxyd trong 2 ln, phn ng lng cn gia hai ln thm cho n khi tt c cht hu c b ph hu, khi ca acid H 2SO4 bc ra mnh v dung dch tr ln khng mu. Duy tr cc iu kin xy ho ti tt c cc thi im trong qu trnh ph mu bng cch b sung peroxyd mi khi hn hp chuyn mu nu hoc ti mu. (Lng peroxyd cn ph hon ton mu s thay i nhng trong mt s trng hp mc ti a c th l 200 ml, iu ny ph thuc vo trng thi t nhin ca mu). Lm ngui, ch thm 10 ml nc, li cho bc hi n khi to thnh khi mnh v lm ngui. Chuyn dung dch ti bnh to arsin, ra bnh Kjeldahl v qu bu bng nc, thm nc ra vo bnh v pha ti 35 ml bng nc. Dung dch ny phi p ng c cc yu cu ca Gii hn th arsenic. Cn chnh xc 5 g mu v cho vo a s hoc bt s, un trn tm gia nhit cho n khi than ho hon ton, sau gia nhit trong l nung 480 0C 8 gi hoc qua m, ngui. Thm cn thn 5 ml acid nitric, lm bay hi n kh 48

Ch

QCVN 4-22: 2011/BYT

trn tm gia nhit, sau nung trong l nung 480 0C trong 15 pht, v lm ngui. Trch ly tro trong 10 ml nc trong 2 ln, lc mi phn dch chit vo mt bnh gn. Lc qua bt k cht khng tan trn dng c lc bng 6 ml dung dch amoni citrat TS, 2 ml hydroxylamin hydrochlorid TS v 5 ml nc, thm nc ra c lc vo bnh gn. Tip tc theo hng dn trong phn Cch tin hnh trong th Gii hn, bt u vi Thm 2 git phenol TS .... vo bnh gn, s dng 10 g ion ch (Pb) trong mu i chng. Ph hng ngoi Glycerol ca nha g thng

Sc k kh ca cc alcol nha thng

49

QCVN 4-22: 2011/BYT

Thit b, dng c - im nng chy

Hnh 1

Hnh 2

50

QCVN 4-22: 2011/BYT

Ph lc 12 YU CU K THUT V PHNG PHP TH I VI DINATRI DIPHOSPHAT 1. Tn khc, ch s Disodium pyrophosphate; Disodium dihydrogen diphosphate; acid sodium pyrophosphate. INS 450i MTDI=70mg/kg th trng tnh cho Phospho t cc ngun thc phm

2. nh ngha Tn ha hc Ch s C.A.S. Cng thc ha hc Khi lng phn t 3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan pH Natri Phosphat 5.2. tinh khit Gim khi lng khi sy kh Cc cht khng tan trong nc Fluorid Arsen Ch 5.3. Hm lng Na2H2P2O7 Khng c qu 0,5% (105oC trong 4 gi) Khng c qu 1%. Khng c qu 10,0 mg/kg Khng c qu 3,0 mg/kg Khng c qu 4,0 mg/kg Dng khan, hemihydrat v monohydrat: khng thp hn 97,0% tnh theo ch phm khan. Dng dodecahydrat: khng thp hn 92,0% tnh theo ch phm sau khi nung. Tan trong nc. 3,7-5,0 (dung dch 1/100). Phi c phn ng c trng ca natri. Phi c phn ng c trng ca phosphat. Disodium dihydrogen diphosphate; Disodium dihydrogen pyrophosphate 7758-16-9 Dng khan: Na2H2P2O7 221,94 Dng tinh th hoc ht mu trng. Cht nh ha, m, cht to phc kim loi, cht to xp

6. Phng php th 51

QCVN 4-22: 2011/BYT

6.1. tinh khit Cc cht khng tan trong nc Ha tan 10 g mu th trong 100 ml nc nng. Lc qua mt chn lc cn b. Ra cn khng tan trn chn lc bng nc nng. Sy kh ti (105oC trong 2 gi). ngui v cn. Cn 400mg (chnh xc n mg) mu th c sy kh trc ti (105oC trong 4 gi), ha tan trong 100 ml nc trong cc 400 ml. Chnh pH dung dch v 3,8 bng dung dch acid hydrocloric long (TS) hoc dung dch natri hydroxyd (TS), dng pH k xc nh pH dung dch. Sau thm 50 ml dung dch km sulfat (125 g ZnSO 4.7H2O ha tan trong nc, pha long n 1000ml, lc v chnh pH v 3,8), yn trong khong 2 pht. Chun acid c gii phng bng dung dch natri hydroxyd 0,1 N n khi pH dung dch li tr v 3,8. Sau mi ln thm natri hydroxyd gn n im tng ng, cn mt thi gian nu c kt ta km hydroxyd th kt ta c th tan ht. Mi ml natri hydroxyd 0,1 N tng ng vi 11,10mg Na2H2P2O7.

6.2. nh lng

52

QCVN 4-22: 2011/BYT

Ph lc 13 YU CU K THUT V PHNG PHP TH I VI CALCI POLYPHOSPHAT 1. Tn khc, ch s INS 452iv MTDI=70mg/kg th trng tnh cho phospho t cc ngun thc phm Hn hp d th ca cc mui calci ca acid polyphosphoric c cng thc chung l Hn+2PnOn+1. Tinh th hoc bt khng mi, khng mu. Cht nh ha, Cht gi m, cht to phc kim loi, cht to kt cu.

2. nh ngha 3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Phosphat Calci 5.2. tinh khit

Thng khng tan hon ton trong nc, tan trong mi trng acid. Phi c phn ng c trng ca phosphat. Phi c phn ng c trng ca calci (Dch th chun b nh trong phn th phosphat)

o Gim khi lng khi nung Khng c qu 2%. Sy ti 105 trong 4 g sau nung ti 550o trong 30 pht.

Phosphat vng Fluorid Arsen Ch 5.3. Hm lng P2O5 6. Phng php th 6.1. nh tnh Phosphat

Khng c qu 8% tnh theo hm lng P2O5. Khng c qu 10,0 mg/kg. Khng c qu 3,0 mg/kg. Khng c qu 4,0 mg/kg. Khng c thp hn 50,0% v khng c qu 71,0% P2O5 tnh theo ch phm sau khi nung.

Cn 0,5g mu, trn vi 10ml acid nitric v 50ml nc. un si trong 30 pht v lm mt. Dung dch ny c s dng lm dch th.

6.2. tinh khit Arsen - Th theo hng dn ti JECFA monograph 1 - Vol.4 phng php II,. - Ha tan 1 g mu th trong 15 ml dung dch acid hydrocloric long (TS), thm 20 ml nc, dung dch ny c s dng lm dch th). 53

QCVN 4-22: 2011/BYT

Ch - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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 khong 300 mg mu th, chnh xc n mg, ha tan trong 15 ml acid nitric v 30 ml nc ct. un si trong khong 30 pht, pha long vi nc n 100 ml. un nng n 60oC, thm lng d dung dch amoni molybdat (TS) v un 50oC trong 30 pht. Lc v ra kt ta bng acid nitric long (dung dch 1/36), tip theo bng dung dch kali nitrat (1/100) n khi dch ra khng cn acid vi qu tm. Ha tan kt ta trong 50 ml dung dch natri hydroxyd 1N, thm dung dch phenolphtalein (TS) v chun lng natri hydroxyd d bng acid sulfuric 1N. Mi ml dung dch natri hydroxyd 1 N tng ng vi 3,086mg P2O5.

6.3. nh lng

54

QCVN 4-22: 2011/BYT

Ph lc 14 YU CU K THUT V PHNG PHP TH I VI CC MUI CA CC ACID BO 1. Tn khc, ch s Salts of fatty acids INS 470 ADI khng gii hn Cc sn phm ny cha mui calci, kali v natri ca cc acid myristic, oleic, palmitic, stearic hoc hn hp cc acid ny thu c t m v du n. Sn phm thng mi cn c phn loi da trn: - Ch s x phng ha; - im ng c ca cc acid bo thu c t mu th; - Ch s iod; - Cn cn li sau khi nung bao gm c hm lng cc cation; - m. Tinh th rn hoc na rn, bng, mu trng hoc vng nht, hoc bt mu trng hoc trng vng. Cht chng ng vn, Cht nh ha.

2. nh ngha

3. Cm quan

4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Cation Thnh phn acid bo 5.2. tinh khit Cc acid bo t do Cc cht khng x phng ha Ch 5.3. Hm lng 6. Phng php th 6.1. nh tnh

Cc mui kali v natri tan trong nc v ethanol, cc mui calci khng tan trong nc, ethanol v ether. Phi c phn ng c trng ca cation. Phi c phn ng c trng ca thnh phn acid bo

Khng c qu 3% Khng c qu 2 %. Khng c qu 2,0 mg/kg. Khng thp hn 95 % tng cc mui acid bo, tnh theo khi lng kh.

55

QCVN 4-22: 2011/BYT

Cation un nng 1 g mu th vi hn hp gm 25 ml nc v 5 ml acid hydrocloric. Cc acid bo to thnh lp ni trn b mt thnh mt lp rn hoc du, tan trong hexan. Sau khi lm ngui, gn ly lp nc v cho bay hi n kh. Ha tan cht rn cn li trong nc v th cc cation thch hp. S dng phng php nh lng nh tnh ring cc acid bo c trong thnh phn ca mu. Cc acid bo chim lng ln nht phi ph hp vi cc acid c a ra trn nhn sn phm.

Thnh phn acid bo

6.2. tinh khit Cc acid bo t do Xc nh cc acid bo t do theo hng dn trong phn Phng php xc nh Acid bo t do. Tnh hm lng acid bo t do, s dng h s tng ng (e) tng ng vi 1/10 khi lng phn t ca mui.

56

QCVN 4-22: 2011/BYT

Cc cht khng x phng ha Cc cht khng x phng ha l ton b lng sn phm c mt trong cc cht bo sau khi c x phng ha vi cc hydroxyd kim loi kim v chit bng mt dung mi nht nh, vn khng bay hi trong cc iu kin xc nh ca php th. Cc cht ny bao gm cc lipid c ngun gc t nhin nh cc sterol, cc ru no mch di hn, cc phm mu v cc hydrocarbon cng nh bt k mt tp cht khng bay hi no 1030C c mt trong sn phm. Cn khong 5 g mu th (chnh xc n 0,01g) c trn u vo bnh cu y trn 250 ml. Thm 50 ml dung dch kali hydroxyd ~ 0,5 N v thm mt t bt. Gn bnh vo sinh hn hi lu, un si nh trong 1 gi. Ngng un. Thm 100 ml nc ct qua u sinh hn v lc. Sau khi lm mt, chuyn dung dch vo phu tch. Trng ra bnh v bt vi ln bng diethyl ether (dng khong 100 ml) v dch ra vo phu tch. ng nt v lc mnh trong 1 pht, u n x p sut bng cch lt ngc phu tch v m kha vi. yn phu tch cho n khi 2 pha han ton tch lp. Sau rt ly cng trit cng tt dung dch x phng vo phu tch th 2. Chit dung dch x phng trong ethanol thm hai ln na, mi ln vi 100 ml diethyl ether theo cch trn. Gp 3 dch chit ether vo 1 phu tch c sn 40 ml nc. Nh nhng quay trn phu tch cha dch chit v 40 ml nc. Nu lc qu mnh trong giai on ny, nh tng c th hnh thnh. hn hp tch lp hon ton v rt b lp nc di. Ra lp ether hai ln na vi 40 ml nc, lc u v loi b lp nc di sau khi hn hp phn lp. Vi mi dung dch ra rt ra n khi ch cn khong 2 ml, sau quay trn phu tch quanh trc, i vi pht, rt b phn tch ra, ng kha vi khi ether bt u chy qua l ca kha vi. Ra dung dch ether nhiu ln bng 40 ml dung dch kali hydroxyd, 40 ml nc, ra thm mt ln bng 40 ml dung dch kali hydroxyd, sau ra t nht 2 ln na bng 40 ml nc. Lin tc ra bng nc cho n khi dung dch ht mu hng khi th bng ch th l dung dch phenophthalein. Chuyn ton lng tng t mt dung dch ether qua cung ca phu tch vo bnh cu kh v cn b khi lng chnh xc n 0,0001 g. Lm bay hi dung mi bng cch ct trong b cch thy. Thm 5 ml aceton v loi b hon ton dung mi bay hi bng cch thi khng kh nh, nghing bnh cu khi quay trong b cch thy. Sy kh phn cn cn li nhit 1032 C trong 15 pht, t bnh cu theo phng gn nh nm ngang. Lm ngui trong bnh ht m v cn (chnh xc n 0,0001 g). Lp li qu trnh sy trong mi 15 pht lin tip cho n khi chnh 57

QCVN 4-22: 2011/BYT

Ch - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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.

58

QCVN 4-22: 2011/BYT

6.2. nh lng Nguyn tc: X phng ha cc mui v ester ha cc acid bo bng methanol, c mt bo trifluorid, methanol kim. Tin hnh sc k lng kh cc ester methyl ca acid bo. Phn A - Chun b cc ester methyl ca acid bo Thit b - Bnh y trn c nhm 50 v 100 ml. - Sinh hn hi lu, chiu di hiu dng 20 to 30 cm, c khp ni nhm tng thch vi bnh cu. - Cc phu tch 250 ml. - ng ng vo dn kh nitrogen. - ng nghim c nt thy tinh nhm. - Pipet chia , c th tch t nht l 10 ml, c gn qu bp cao su, hoc pipet t ng. Thuc th - Heptan, dng cho sc k (Ch 2 v 4) - Du nh c ct li (nhit si t 40-60 ), ch s brom < 1, khng c cn, hoc hexan (Ch 2) - Natri sulfat khan - Dung dch Natri hydroxyd trong methanol ~ 0.5 N: Ha tan 2g natri hydroxyd trong 100 ml methanol cha khng qu 0.5% (kl/kl) nc. Nu gi dung dch tng i lu c th to thnh mt lng nh kt ta mu trng ca natri. iu ny khng nh hng n kt qu iu ch methyl ester. - Dung dch bo trifluorid trong methanol, 12 n 25% (kl/kl) . Ch phm thng mi sn c l dung dch 14 v 50% (ch 2) Cnh bo: Bo trifluorid c. V vy th nghim vin khng nn pha dung dch bo trifluorid trong methanol t methanol v boron trifluorid. (ch 3). - Dung dch bo ha natri clorid trong nc. - Methyl , dung dch 1 g/l trong ethanol 60% (tt/tt) - Nitrogen, c hm lng oxy < 5 mg/kg Tin hnh Do bo trifluorid c, cc th nghim sau tt nht nn tin hnh trong t ht. Cc dng c thy tinh phi c ra bng nc ngay sau khi s dng. Sy kh mu nhit 105 C n khi lng khng i, c 2 gi th kim tra khi lng 1 ln. Cn khang 350mg (chnh xc n mg) mu sy kh. C th ly lng mu ln hoc nh hn 350 mg, tuy nhin, th tch ca bnh cu v lng thuc th s dng phi ph hp vi s liu trong bng sau: Khi lng NaOH 0.5 Dung dch BF3 Th tch bnh mu N trong methanol (ml) (mg) (ml) (ml) 59

QCVN 4-22: 2011/BYT

60

QCVN 4-22: 2011/BYT

Ph lc 15 YU CU K THUT V PHNG PHP TH I VI ESTER CA SUCROSE VI CC ACID BO 1. Tn khc, ch s 2. nh ngha Sucrose fatty acid esters; INS 473 Mono, di, tri ester ca sucrose vi cc acid bo thc phm, c iu ch t sucrose v methyl, ethyl ester ca cc acid bo thc phm bng cch ester ho vi s c mt ca cht xc tc, hoc tch t sucroglycerid. Ch cc dung mi sau c th c s dng sn xut: dimethylformamid, dimethyl sulfoxid, ethyl acetat, isopropanol, propylen glycol, isobutanol v methyl ethyl keton. Dng gel cng, dng cht c mm hoc dng bt c mu trng n trng hi xm Cht nh ho

3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Acid bo ng 5.2. tinh khit Tro sulfat Ch s acid Sucrose t do Dimethylformamid Dimethyl sulfoxid Ethyl acetat, isopropanol v propylen glycol Isobutanol Methanol

t tan trong nc, tan trong ethanol. Phi c phn ng c trng ca acid bo. Phi c phn ng c trng ca ng.

Khng c qu 2% Th 1 g mu (Phng php I) Khng c qu 6 Khng c qu 5% (m t trong phn Phng php th) Khng c qu 1,0 mg/kg (m t trong phn Phng php th) Khng c qu 2,0 mg/kg (m t trong phn Phng php th) Khng c qu 350,0 mg/kg, dng n cht hoc hp cht (m t trong phn Phng php th) Khng c qu 10,0 mg/kg (m t trong phn Phng php th) Khng c qu 10,0 mg/kg (m t trong phn Phng php th)

61

QCVN 4-22: 2011/BYT

Methyl ethyl keton Ch 5.3. Hm lng 6. Phng php th 6.1. nh tnh Acid bo Cho 1 ml ethanol vo 0,1 g mu, lm m v ho tan, thm 5 ml dung dch H2SO4 long TS, gia nhit trong cch thy trong 30 pht v lm ngui. Cht c hoc du mu trng vng c hnh thnh, khng c mi ca acid isobutyric v c ho tan khi thm 3 ml diethyl ether. S dng lp dung dch nc tch khi t diethyl ether cho Php th ng. Cho vo 2 ml lp dung dch nc tch khi diethyl ether trong php th acid bo, 1 ml anthron TS vo trong ng th, cho cn thn theo thnh ng nghim; b mt ranh gii gia hai lp chuyn thnh mu xanh da tri hoc xanh l cy. Khng c qu 10,0 mg/kg (m t trong phn Phng php th Khng c qu 2,0 mg/kg. Khng c nh hn 80,0% ester sucrose

ng

6.2. tinh khit

62

QCVN 4-22: 2011/BYT

Sucrose t do Xc nh bng sc k kh lng (Quyn 4) Dung dch chun: Chun b dung dch gc cha 5 mg/ml sucrose trong N, Ndimethylformamid. Chun b mt lot cc dung dch chun cha 0,5; 1,25 v 2,5 mg/ml sucrose bng cch pha long dung dch gc vi pyridin. Dung dch ni chun: Cn chnh xc 0,25 g octacosan cho vo bnh nh mc dung tch 50 ml, thm vo 25 ml tetrahydrofuran ho tan octacosan, v nh mc n th tch 50 ml bng pyridin. Cc iu kin sc k: Ct: 2% Dexsil 300GC trn Uniport HP 80/100 mesh (t phn cc, 2,1 m x 3.2 mm ng knh trong) hoc tng ng Kh mang: nitrogen Detector: FID Nhit : - Bung bm mu: 2800C - Ct: gi 1600C trong 1 pht sau tng t 160 n 3000C vi tc 150C/pht, gi trong 60 pht 3000C. - Detector: 3200C Thi gian lu ln lt ca sucrose v octacosan o c vi cc iu kin trn xp x 8,2 v 9,8 pht. Cch tin hnh: Cn chnh xc 20 50 mg mu cho vo ng li tm, thm 1 ml dung dch ni chun, 1 ml pyridin, 0,4 ml N, O-bis (trimethylsilyl) acetamid (BSA) v 0,2 ml trimethylchlorosilan (TMCS). Sau khi y np ng, lc v yn trong 5 pht nhit phng. Bm 1 l vo thit b sc k kh lng. ng chun: Chun b dung dch chun silylat theo cch tin hnh nh nhau s dng 1 ml mi loi dung dch chun thay th mu v pyridin. V ng chun trn th lng sucrose (mg) trong 1 ml dung dch chun (biu th trn trc X) so vi t l din tch pic ca sucrose/ni chun (biu th trn trc Y). o din tch pic i vi sucrose v ni chun. Tnh t l din tch cc pic ca chng, v tnh c hm lng sucrose trong mu da vo ng chun. Lng sucrose pht hin c (mg) % sucrose = ----------------------------------------------------- x 100 Khi lng mu (mg)

63

QCVN 4-22: 2011/BYT

Dimethylformamid Xc nh bng sc k kh lng Dung dch chun: Chun b dung dch gc cha 1 mg/ml dimethylformamid trong tetrahydrofuran. Chun b mt lot cc dung dch chun cha 0,05; 0,1 v 0,2 g/ml dimethylformamid bng cch pha long dung dch gc vi tetrahydrofuran. Cc iu kin sc k: Ct: polyethylen glycol (di 30 m x 0,32 mm ng knh trong vi dy lp phim mng 0,5 m) Kh mang: heli p sut: 150 kPa (p sut khng i) Detector: detector nitrogen phosphor (NPD) [(tn khc: Detector nhit ion ngn la (FTD)] Nhit : - Bung bm mu: 1800C - Ct: gi 400C trong 2 pht sau tng t 40 n 160 0C vi tc 200C/pht, gi trong 2 pht 1600C. - Detector: 3250C Phng php tim mu: Tim khng chia dng 1,0 l mu bng bm t ng, sau bt u lm sch sau 1 pht. Thi gian lu ca dimethylformamid o c vi cc iu kin trn xp x 6,4 pht. Cch tin hnh: Cn chnh xc 2 g mu cho vo bnh nh mc dung tch 20 ml, thm vo 10 ml tetrahydrofuran ho tan mu, thm tetrahydrofuran vo bnh n th tch 20 ml v lc u dung dch. Bm 1,0 l dung dch mu vo thit b sc k. ng chun: Chun b hng ngy bng cch bm 1,0 l mi dung dch chun vo thit b sc k. Tnh nng dimethylformamid (CDFA) theo cng thc sau: Trong : C: Nng dimethylformamid pht hin c (g/ml) W: Khi lng mu (g) Lu : Detector nitrogen phosphor khng nhy vi cc thnh phn khng cha N hoc P. V th, ct mao qun c th tc vi cc hp cht bay hi thp, mc d ng nn ca sc k n nh. Do , ct phi c luyn thng xuyn. Luyn qua m (dng kh mang theo hng ngc 1800C ) c thc hin sau khong mi t 15 mu.

64

QCVN 4-22: 2011/BYT

Dimethyl sulfoxid Xc nh bng sc k kh lng Dung dch chun: Chun b dung dch gc 0,25 mg/ml dimethyl sulfoxid trong tetrahydrofuran. Chun b mt lot cc dung dch chun cha 0,5; 1 v 5 g/ml dimethyl sulfoxid bng cch pha long dung dch gc vi tetrahydrofuran. Cc iu kin sc k: Ct: 10% PEG 20M v 3% KOH trn Gas Chrom Z (chiu di 2 m x 3 mm ng knh trong). Tng nhit l ti 1800C vi tc 100C/pht v n nh 24 48 gi vi iu kin tc dng kh nitrogen 30 40 ml/pht luyn ct. Kh mang: nitrogen Tc dng: 50 ml/pht Detector: detector trc quang ngn la (s dng b lc lu hunh 394 nm) Nhit : - Bung bm mu: 2100C - Ct: 1600C. Thi gian lu ca dimethyl sulfoxid o c vi cc iu kin trn xp x 3,4 pht. Cch tin hnh: Cn chnh xc 5 g mu cho vo bnh nh mc dung tch 25 ml, thm vo 10 ml tetrahydrofuran ho tan mu, thm tetrahydrofuran vo bnh n th tch 20 ml v lc u dung dch. Bm 3 l dung dch mu vo thit b sc k. ng chun: Chun b hng ngy bng cch bm 3 l mi dung dch chun vo thit b sc k. Tnh nng dimethyl sulfoxid (CDMSO) theo cng thc sau:

Trong : C: Nng dimethyl sulfoxid pht hin c (g/ml) W: Khi lng mu (g)

65

QCVN 4-22: 2011/BYT

Propylen glycol Xc nh bng sc k kh lng Dung dch ni chun: Chun b dung dch 500 g/ml ca ethylen glycol trong pyridin Dung dch chun: Chun b dung dch 50 g/ml ca propylen glycol trong pyridin Cc iu kin sc k: Ct: Polydimethylsiloxan (chiu di 30 m x 0,32 mm ng knh trong vi dy lp phim mng 0,25 m) Kh mang: heli Tc dng: 1,5 ml/pht (dng khng i) Detector: FID Nhit : - Bung bm mu: 2300C - Ct: gi 600C trong 5 pht sau tng t 60 n 250 0C vi tc 200C/pht, gi trong 5 pht 2500C. - Detector: 2500C Thi gian lu ln lt ca dn xut ethylen glycol v dn xut propylen glycol xp x 7,7 pht v 7,9 pht. Cch tin hnh: Cn chnh xc 1 g mu cho vo bnh nh mc dung tch 10 ml, thm vo 100 l dung dch ni chun. Ho tan v nh mc n th tch 10 ml bng pyridin. Ly 0,5 ml dung dch mu cho vo ng li tm, v thm 0,25 ml 1,1,1,3,3,3 hexamethyldisilazan (HMDS) v 0,1 ml trimethylchlorosilan (TMCS). Sau khi y cht np ng, lc mnh, yn trong 30 pht nhit phng, sau li tm. Bm 1 l dung dch ni trn mt ng sau li tm vo thit b sc k. ng chun: Tin hnh sc k nh trn, trong dng 0,05; 0,2; 0,5 v 1ml dung dch chun thay th cho dung dch mu th. Tnh nng propylen glycol (CPG) theo cng thc sau:

Trong : C: Nng propylen glycol pht hin c (g/ml) W: Khi lng mu (g) Lu : Nht thit phi lm sch cng bm mu v luyn ct 3000C sau mi t 20 mu phn tch, bi ct b nhim bn.

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QCVN 4-22: 2011/BYT

Methanol, isopropanol, isobutanol, ethyl acetat v methyl ethyl keton Xc nh bng sc k kh c bung ly mu khng gian hi. Dung dch chun: Chun b dung dch chun A cha 4.000 mg/l mi loi methanol, isopropanol, isobutanol, ethyl acetat v methyl ethyl keton bng cch cn chnh xc 0,2 g mi dung mi v cho vo bnh nh mc dung tch 50 ml cha khong 20 ml nc, sau thm nc vo n th tch 50 ml. Chun b cc dung dch cha 2.000 mg/l (dung dch chun B) v 1.000 mg/l (dung dch chun C) bng cch pha long dung dch chun A. Cch tin hnh: Cn chnh xc 4 g mu cho vo 4 l (mi l 1 g mu). Cho 5 l nc vo l th nht, cho ln lt vo l th hai, th ba, th bn dung dch chun A, B v C; y nhanh septum. (Nng ca mi dung mi sau khi thm 5 l dung dch chun A, B, C vo 1 g mu tng ng ln lt bng 20, 10 v 5 mg/kg mu). l mu trong bung ly mu khng gian hi v phn tch vi cc iu kin sau: Ct: 100% Polydimethylsiloxan (chiu di 30 m x 0,53 mm ng knh trong vi dy mng phim mng 1,5 m), v d: DB 1 c sn xut bi Cng ty TNHH J & W) Kh mang: nitrogen Tc dng: 3,5 ml/pht Detector: FID Nhit : - Bung bm mu: 1100C - Ct: 400C. - Detector: 1100C Bung ly mu khng gian hi: - Nhit cch nhit mu: 800C - Thi k cch nhit mu: 40 pht - Nhit syringe: 850C - Bm mu dng kh: 1,0 ml Kt qu: th biu din mi tng quan gia lng thm vo vi din tch pic i vi mi dung mi s dng cc kt qu phn tch. Mi tng quan phi tuyn tnh (R 2 > 0,99). Ngoi suy v xc nh x phn b chn, w i, v tnh nng dung mi Ci (mg/kg) trong mu: Trong : wi: Phn b chn ca ng thng tng quan s dng phng php thm chun (g) W: Khi lng mu (g) 67

QCVN 4-22: 2011/BYT

Ch - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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. Xc nh bng HPLC s dng cc iu kin sau: Cch tin hnh: Cn chnh xc 250 mg mu v cho vo bnh nh mc dung tch 50 ml. Pha ti th tch 50 ml bng tetrahydrofuran v lc u. Lc qua mng lc 0,5 m. Bm 100 l mu vo thit b sc k n nh trc. Cc iu kin sc k: Ct: Styren-divinylbenzen copolymer i vi sc k thm thu qua gel (TSK-GEL G2000 (Tosoh) hoc tng ng) Pha ng: tetrahydrofuran dng cho HPLC bi kh Tc dng: 0,7 ml/pht Detector: RI Nhit : - Ct: 380C. - Detector: 380C Ghi sc k trong khong 90 pht. Tnh % ester sucrose trong mu: % ester sucrose = 100 A/T Trong : A: Tng cc din tch pic i vi 3 thnh phn chnh, mono-, di- v tri-ester, ra gii ln lt khong 65, 68 v 73 pht. T: Tng din tch tt c cc pic ra gii trong khong 90 pht

6.3. nh lng

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QCVN 4-22: 2011/BYT

Ph lc 16 YU CU K THUT V PHNG PHP TH I VI DIOTYL NATRI SULFOSUCINAT 1. Tn khc, ch s DSS, natri docusat, INS 480 ADI = 0 - 0,1 mg/kg th trng

2. nh ngha Tn ha hc Natri1,4-bis-(2-ethylhexyl)-sulfosuccinat; Mui natri ca acid ester sulfo-1,4-bis-(2-ethylhexyl) butanedioic; dioctyl natri sulfosuccinat 577-11-7 C20H37NaO7S

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 Ph hng ngoi

444,56 Cht rn dng do, ging sp, mu trng c mi c trng ging octanol, nhng khng c mi ca cc dung mi khc. Cht nh ha, cht lm m

t tan trong nc, d tan trong ethanol v glycerol Ph hng ngoi ca mu phn tn trong kali bromid tng ng vi ph ca mu chun i chiu (Chun i chiu USP dioctyl sulfosuccinate c cng ty United States Pharmacopoeia, 12601 Twinbrook Parkway, Rockville, Md 20852, USA) 5.8 - 6.9 (dung dch 1/100)

pH 5.2. tinh khit Gim khi lng khi lm kh trong ca dung dch

Khng c qu 2 % (105C trong 2 gi) Ha tan 25 g mu th vo 94 ml ethanol. Dung dch khng c vn c sau 24 gi.

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QCVN 4-22: 2011/BYT

Tro sulfat Trong khong t 15,5 % n 16,2 %. (th theo hng dn ti JECFA monograph 1 - Vol.4; Phng php I, cn 1 g mu) Khng c qu 0,4 %. Khng c qu 2,0 mg/kg. Khng thp hn 98,5 % tnh theo khi lng kh.

Bis-(2-ethylhexyl)-maleat Ch 5.3. Hm lng 6. Phng php th 6.1. tinh khit

70

QCVN 4-22: 2011/BYT

Bis-(2-ethylhexyl)-maleat Thuc th - Cht in gii h tr: Ha tan 21,2 g lithi perchlorat (LiClO4) khan vo 175 ml nc trong cc 250 ml. iu chnh pH ca dung dch ny n 3,0 bng cch thm tng git acid acetic bng (khong 1 n 2 git l ), dng mt my o pH ph hp. Chuyn ton lng dung dch vo bnh nh mc 200 ml, pha long n vch bng nc, v trn u. - Dung dch chun: Chuyn 100 - 110 mg (cn chnh xc) bis-(2-ethyl-hexyl)-maleat, (bis-(2-ethylhexyl)-maleat vi tinh khit ph hp c tn khc l OT-35 ca Cng ty American Cyanamid, Pine Chemicals Department, Pearl River, New York 10965, USA), vo bnh 100 ml. Ghi li khi lng chnh xc, chnh xc n 0,1 mg, Wa. Pha long n vch bng nc, trn u. - Dung dch mu th gc: Chuyn 12,5 g mu th, c cn chnh xc, vo cc 150 ml. Ghi li khi lng chnh xc, Ws, chnh xc n 10 mg. Thm 80-90 ml isopropanol, khuy bng a thy tinh n khi mu tan hon ton. Chuyn ton lng dung dch ny vo bnh nh mc 250 ml bng isopropanol. Pha long n vch bng isopropanol, trn u. - Dung dch th A: Dng pipet ly 50.0 ml "Dung dch mu th gc" v 20,0 ml "Dung dch in gii h tr " vo bnh nh mc 100 ml. Pha long vi isopropanol n khong di vch nh mc 15 mm, y nt, lc u v trong 2 pht. Pha long tip bng isopropanol n vch, y nt, lc u. Phi thu c dung dch tuyt i trong. - Dung dch th B: Dng pipet ly 50,0 ml "Dung dch mu th gc", 10,0 ml "Dung dch chun", v 20,0 ml Dung dch in gii h tr" vo bnh nh mc 100 ml, v tip tc qui trnh nh miu t trong phn Dung dch th A - Mu trng: Dng pipet ly 20,0 ml "Dung dch in gii h tr" vo bnh nh mc 100-ml, pha long n vch bng isopropanol, trn u. Tin hnh Ra bnh cc ph H-cell vi ln bng tng lng nh Dung dch th A, sau y mt na bnh bng dung dch ny, t mt mu giy thm ln ming bnh v cho dng kh nitrogen chy t t qua dung dch trong 15 pht (Ch : Trc ht phi bo ha nitrogen bng cch cho chy qua bnh sc ph hp c cha isopropanol). Sau 15 pht, hng dng nitrogen ln b mt dung dch v ly giy ra khi bnh. Vi mt cc ph k ph hp c hiu chun trc (nh cc ph k Metrohm Polacord E-261, hoc tng ng t mt th phn cc -1,3V. iu chnh nhy dng ti thang thp nht (nhy nht) m ti cc dao dng dng vn nm trong thang. Ghi li cc ph ti nhy ny, qut th trong khong t -0,9 n -1,5 V, dng in cc so snh l in cc calomel bo ha. Ghi li dao ng trung bnh A ti th -1,3 V, v dao ng 71

QCVN 4-22: 2011/BYT

Ch - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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. Cc dung dch - Dung dch mu: Chuyn khong 3,8 g mu th, trc c sy kh 105C trong 2 gi v c cn chnh xc, vo bnh nh mc 500 ml, ha tan trong cloroform. Pha long n vch bng dung mi trn, trn u. - Dung dch tetra-n-butylamoni iodid: Chuyn 1,250 g tetran-butylamoni iodid vo bnh nh mc 500 ml, pha long n vch bng nc, trn u. - Dung dch mui: Ha tan 100 g natri sulfat khan v 10 g natri carbonat vo mt lng nc va c c th tch cui cng l 1000 ml. Tin hnh Dng pipet ly 10,0 ml "Dung dch mu" vo bnh nh mc 250 ml, thm vo 40 ml cloroform, 50 ml "Dung dch mui", 10 git dunh dch xanh bromophenol (TS). Chun bng "Dung dch tetra-n-butylamoni iodid" n khi bt u xut hin mu xanh trong lp cloroform sau khi lc mnh dung dch. Tnh hm lng (%) C 20H37NaO7S theo cng thc:

6.2. nh lng

trong V = th tch (ml) ca dung dch tetra-n-butylamoni iodid dng chun . 444,6 = khi lng phn t ca dioctyl natri sulfosuccinat W = khi lng (g) ca mu th 369,4 = khi lng phn t ca tetra-n-butylamoni iodid.

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QCVN 4-22: 2011/BYT

Ph lc 17 YU CU K THUT V PHNG PHP TH I VI STEARYL TARTRAT 1. Tn khc, ch s 2. nh ngha Stearyl palmityl tartrat; INS 483 Sn phm ca qu trnh ester ho acid tartric vi cn stearylic thng mi, v c bn gm cn stearylic v palmitylic; gm thnh phn chnh l diester, vi lng nh monoester v cc nguyn liu ban u khng thay i. Cc thnh phn chnh l distearyl tartrat, dipalmityl tartrat v stearylpalmityl tartrat. Distearyl tartrat: C40H78O6 Dipalmityl tartrat: C36H70O6 Stearylpalmityl tartrat: C38H74O6

Tn ho hc Cng thc ho hc

Cng thc cu to

Khi lng phn t

Trong R l (CH2)17CH3 hoc (CH2)15CH3 Distearyl tartrat: 655,06 Dipalmityl tartrat: 598,95 Stearylpalmityl tartrat: 627 Cht nhn c mu kem Cht nh ha, cht x l bt

3. Cm quan 4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan Nhit nng chy Ch s hydroxyl Tartrat 5.2. tinh khit Tro sulfat Acid tartric tng s Cht khng x phng ha

Khng tan trong nc, khng tan trong ethanol lnh, tan trong ethanol nng. 67 770C 200 220 Phi c phn ng c trng ca tartrat.

Khng c qu 0,5% (Th trn 2 g mu, phng php I ) Khng c nh hn 18% v khng c qu 35% (m t trong phn Phng php th) Khng c nh hn 77% v khng c qu 83% (m t trong phn Phng php th)

73

QCVN 4-22: 2011/BYT

Ch s acid Ch 5.3. Hm lng 6. Phng php th 6.1. tinh khit Khng c qu 6 (m t trong phn Phng php th) Khng c qu 2,0 mg/kg. Khng c nh hn 90% hm lng ester tng s tng ng vi ch s ester trong khong 163 180.

74

QCVN 4-22: 2011/BYT

Acid tartric tng s ng chun: Cn 100 mg acid tartric tinh khit cho vo bnh nh mc dung tch 100 ml, ho tan trong khong 90 ml nc, thm nc vo cho n khi t th tch 100 ml v lc u. Chuyn cc phn tch bit 3, 4, 5, 6 ml vo trong cc cuvet ph hp 19 x 150 mm, thm lng nc va n 10 ml. Thm vo 4 ml dung dch natri metavanadat 5% va chun b v 1 ml acid acetic vo mi cuvet. (Ch : s dng cc dung dch ny trong 10 pht sau khi xut hin mu). Chun b mu trng theo cch tng t, s dng 10 ml nc thay th dung dch acid tartric. t trn thit b hp th bng 0 i vi mu trng, sau o hp th ca 4 dung dch acid tartric bc sng 520 nm vi my quang ph thch hp hoc my so mu quang in c trang b knh lc 520 nm. T cc kt qu thu c, v ng chun trn h to vung gc vi trc tung biu din hp th v trc honh biu din hm lng acid tartric (mg). Chun b mu th: Cn chnh xc 4 g mu cho vo bnh tam gic dung tch 250 ml v thm vo 80 ml dung dch KOH 0,5N v 0,5 ml phenolphthalein TS. Ni bnh vi mt sinh hn khng kh c chiu di t nht 65 cm v gia nhit hn hp trn tm bn nng trong 2,5 gi. Thm vo hn hp nng acid phosphoric 10% cho n khi c phn ng acid r rt pht hin c bng giy th congo. Li kt ni sinh hn kh v gia nhit cho n khi acid bo ho lng v trong. Lm ngui v chuyn hn hp vo mt phu chit dung tch 250 ml bng cc phn nh nc v chloroform. Chit tch cc acid bo c gii phng ra bng 3 ln lin tip 25 ml nc v cho phn nc ra vo phu chit cha lp nc. Chuyn cc thnh phn c trong phu chit th nht vo cc 250 ml, gia nhit trn chu hi nc loi cc vt chloroform, lc qua giy lc mn vo bnh nh mc dung tch 500 ml v cui cng pha ti th tch 500 ml bng nc (Dung dch I). Dng pipet ly 25 ml dung dch ny cho vo bnh nh mc dung tch 100 ml v pha ti th tch 100 ml bng nc (Dung dch II). Cch tin hnh: Chuyn 10 ml Dung dch II c chun b trong phn Chun b mu th vo cuvet 19 x 150. Thm vo 4 ml dung dch natri metavanadat 5% va chun b v 1 ml acid acetic. (Ch : s dng cc dung dch ny trong 10 pht sau khi xut hin mu). Chun b mu trng theo cch tng t, s dng 10 ml nc thay th dung dch acid tartric. t trn thit b o hp th bng 0 i vi mu trng, sau o hp th ca 4 dung dch acid tartric bc sng 520 nm vi my quang ph thch hp hoc my so mu quang in c trang b knh lc 520 nm. T ng chun xc nh hm lng acid tartric (mg) trong dung dch pha cui cng, nhn hm lng acid tartric xc nh c vi 20 v chia cho trng lng mu ban u 75

QCVN 4-22: 2011/BYT

Cht khng x phng ha Cch tin hnh ny xc nh nhng cht ha tan trong cht bo m khng c x phng ho bi kim nhng c ha tan trong dung mi cht bo thng thng. Cch tin hnh: Cn chnh xc 5 g mu cho vo bnh dung tch 250 ml, thm dung dch c 2 g KOH trong 40 ml alcol v un si nh c sinh hn ngc trong 1 gi. Chuyn ton b hn hp cha trong bnh vo ng ong c np y thu tinh (chiu cao khong 30 cm, ng knh 3,5 cm v c vch nh mc 40, 80 v 130 ml). Ra bnh bng alcol ph hp n th tch 40 ml trong ng ong, thm nc m, sau bng nc lnh cho n khi tng th tch t 80 ml. Cui cng ra bnh bng vi ml ether du ha, cho phn dung mi ra vo ng ong, lm mt hn hp trong ng ong n nhit phng v thm 50 ml ether du ha. y np, lc ng ong mnh t nht trong 1 pht, hai lp tch ra trong. Ht lp bn trn hon ton nu c th m khng ht bt k mt cht dch no ca lp di, ly phn ether cho vo trong mt phu tch dung tch 500 ml. Lp li qu trnh trch ly v ht dch t nht 6 ln bng cc phn 50 ml ether du ha, lc mnh mi ln. Ra hn hp phn trch ly bng cc phn 25 ml alcol 10% v lc mnh cho n khi nc ra c trung ho s dng cht ch th phenolphthalein, v loi b phn nc ra. Chuyn phn ether tch c vo cc bit khi lng v ra phu tch bng 10 ml ether, sau cho nc ra vo cc. C ether trong cch thy n kh v lm kh phn cn n khi lng khng i, tt nht 75 80 0C p sut chn khng khng qu 200 mm Hg, hoc 100 0C p sut thng trong 30 pht. Lm mt trong bnh ht m v cn thu c khi lng cha hiu chnh ca cht khng x phng. Xc nh hm lng acid bo trong cn bng cch sau: Ho tan cn trong 50 ml alcol m (cha phenolphthalein TS v trc c trung ho bng NaOH ti mu hng nht), chun bng dung dch NaOH 0,02N cho n khi xut hin mu hng nht. Mi ml dung dch NaOH 0,02N tng ng vi 5,659 mg acid bo, tnh theo acid oleic. Hiu s khi lng cha hiu chnh ca cht khng x phng v khi lng acid bo tnh c t php chun trn l khi lng cht khng x phng ha trong mu th.

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Ch s acid Cn chnh xc 1 g mu v ho tan trong 20 ml ethanol 95% nng, trc c trung ho c s dng 0,5 ml cht ch th phenolphthalein TS. Lm ngui dung dch sau trung ho bng cch chun vi dung dch KOH 0,01N trong ethanol. 0,561 x th tch dung dch KOH dng chun (ml) Ch s acid = -----------------------------------------------------------------------Khi lng mu (g) Gi li dung dch c trung ho cho phn nh lng. - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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. Thm chnh xc 50 ml dung dch KOH 0,1N trong ethanol vo dung dch c trung ho trn t phn xc nh ch s acid v un si u trong 2 pht. Lm ngui dung dch v chun d KOH bng B ml dung dch HCl 0,1N. Thc hin xc nh vi mu trng ( bng A ml dung dch HCl 0,1N). 5,611 x (A B) Ch s ester = ----------------------------------------Khi lng mu (g)

Ch

6.2. nh lng

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Ph lc 18 YU CU K THUT V PHNG PHP TH I VI SORBITAL MONOSTEARAT 1. Tn khc, ch s 2. nh ngha INS 491 ADI=0-25mg/kg th trng Hn hp bao gm cc ester mt phn ca sorbitol v cc dn cht mono- v dianhydrid ca n vi acid stearic thc phm. 1338-41-6 Gm acid palmitic este ha vi polyol dn xut t sorbitol bao gm cc loi sau:

Ch s C.A.S Cng thc cu to

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

1,4-Sorbitan

Isosorbid

Ht hay mnh c mu kem nht n vng nu hoc dng sp rn, c mi nh c trng. Cht nh ha

Ti nhit cao hn im chy, tan trong toluen, dioxan, carbon tetraclorid, ether, methanol, ethanolv anilin; khng tan trong ether du ha v aceton, khng tan trong nc lnh nhng c th phn tn trong nc m.Ti nhit trn 50oC tan dng sng m trong du khong v ethyl acetat. 50-52oC. Ph hng ngoi ca mu th c trng cho este mt phn ca acid bo vi polyol

Khong nhit ng c Hp th hng ngoi 5.2. tinh khit Nc Ch s acid Ch s x phng ha

Khng c qu 1,5% (phng php Karl Fischer). Khng c thp hn 5 v khng c qu 10. Khng c thp hn 147 v khng c qu 157. 78

QCVN 4-22: 2011/BYT

Ch s Hydroxyl Ch 5.3. Hm lng Khng c thp hn 235 v khng c qu 260. Khng c qu 2,0 mg/kg. X phng ha 100g mu phi thu c khong 31,5g polyol v 73g acid bo. Hm lng polyol khng c thp hn 95% trong hn hp sorbitol, 1,4-sorbitan v isosorbid.

6. Phng php th 6.1. tinh khit Ch - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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. Tin hnh theo hng dn trong chuyn lun xc nh hm lng sorbitan ester (JECFA monograph 1 - Vol.4 ).

6.2. nh lng

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Ph lc 19 YU CU K THUT V PHNG PHP TH I VI SORBITAN TRISTEARAT 1. Tn khc, ch s 2. nh ngha INS 492 ADI=0-25mg/kg th trng Hn hp bao gm cc ester mt phn ca sorbitol v cc dn cht mono- v dianhydrid ca n vi acid stearic thc phm. 26658-19-5 Gm acid stearic este ha vi polyol dn xut t sorbitol bao gm cc loi sau:

Ch s C.A.S Cng thc cu to

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

1,4-Sorbitan

Isosorbid

Ht hay mnh c mu kem nht n vng nu hoc dng sp rn Cht nh ha

Kh tan trong toluen, ether, carbon tetraclorid, ethyl acetat; c th phn tn trong ether du ho, du khong, du thc vt, aceton v dioxan; khng tan trong nc, methanol v ethanol (th theo hng dn ti JECFA monograph 1 Vol.4 ). 47-50oC.

Khong nhit ng c 5.2. tinh khit Nc Tro sulfat Ch s acid Ch s x phng ha Ch s Hydroxyl

Khng c qu 1,5% (phng php Karl Fischer). Khng c qu 0,5%. Khng c qu 15. Khng c thp hn 176 v khng c qu 188. Khng c thp hn 66 v khng c qu 80. 80

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Ch 5.3. Hm lng Khng c qu 2,0 mg/kg. X phng ha 100g mu phi thu c khng thp hn 14g v khng c qu 21g polyol v khng thp hn 85g v khng c qu 92g acid bo. Hm lng polyol khng c thp hn 95% trong hn hp sorbitol, 1,4-sorbitan v isosorbid.

6. Phng php th 6.1. tinh khit Ch - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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 25g mu th, chnh xc n mg, chuyn vo bnh cu y trn 500 ml, thm 250 ml cn v 7,5g kali hydroxyd v trn u. Lp bnh cu vi sinh hn thch hp, un hi lu hn hp trong t 1-2 gi sau chuyn hn hp sang cc 800 ml, trng sch bnh bng 100 ml nc ct v gp dch ra vo cc. Cho bay hi ht cn trn b cch thu, thnh thong thm nc thay th lng cn bay hi. Dng qu trnh ui cn khi khng cn mi cn bay ra. Hiu chnh th tch cui cng cho 250 ml bng nc nng. Trung ho dung dch x phng vi acid sulfuric long (1/2), thm 10% d v un nng, khuy u cho n khi acid bo tch lp. Chuyn lp acid bo sang phu chit 500 ml, ra 3 hoc 4 ln, mi ln vi 20 ml nc nng loi ht cc polyol, gp dch ra vi pha nc cha polyol thu c t qu trnh x phng ho. Chit pha nc ny 3 ln 20 ml ether du ho/ln, ly lp ether du ho gp vi phn acid bo, cho bay hi ti kh trong mt a xc nh b, ngui v cn. Trung ho dung dch cha polyol bng dung dch kali hydroxyd 1/10 ti pH = 7, s dng pH k thch hp. Cho bay hi dung dch ny ti gn cn, tch phn polyol khi cc mui bng cch chit vi ln vi cn nng. Cho bay hi dch chit cn trn b cch thu ti kh trong mt a xc nh b, ngui v cn. Trnh kh qu trong qu trnh un nng. Tin hnh nh lng mt mu khc (25g) theo hng dn chuyn lun xc nh hm lng ester sorbitan xc nh % ester sorbitan.

6.2. nh lng

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Ph lc 20 YU CU K THUT V PHNG PHP TH I VI SORBITAN MONOLAURAT 1. Tn khc, ch s 2. nh ngha INS 493 ADI=0-25mg/kg th trng Hn hp bao gm cc ester mt phn ca sorbitol v cc dn cht mono- v dianhydrid ca n vi acid lauric thc phm. 1338-39-2 Gm acid lauric este ha vi polyol dn xut t sorbitol bao gm cc loi sau:

Ch s C.A.S Cng thc cu to

Sorbitol 3. Cm quan

1,4-Sorbitan

Isosorbid

Dng lng nht snh c mu h phch, dng ht hay mnh c mu kem nht n vng nu hoc dng sp rn, c mi nh c trng. Cht nh ha, cht n nh

4. Chc nng 5. Yu cu k thut 5.1. nh tnh tan 5.2. tinh khit Nc Tro sulfat Ch s acid Ch s x phng ha Ch s Hydroxyl Ch

C th phn tn c trong nc nng v nc lnh.

Khng c qu 2,0% (phng php Karl Fischer). Khng c qu 0,5%. Khng c qu 7. Khng c thp hn 155 v khng c qu 170. Khng c thp hn 330 v khng c qu 358. Khng c qu 2,0 mg/kg.

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5.3. Hm lng X phng ha 100g mu phi thu c khng thp hn 36g v khng c qu 49g polyol, khng thp hn 56g v khng c qu 68g acid bo. Hm lng polyol khng c thp hn 95% trong hn hp sorbitol, 1,4sorbitan v isosorbid.

6. Phng php th 6.1. tinh khit Ch - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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 25g mu th, chnh xc n mg, chuyn vo bnh cu y trn 500 ml, thm 250 ml cn v 7,5g kali hydroxyd v trn u. Lp bnh cu vi sinh hn thch hp, un hi lu hn hp trong t 1-2 gi sau chuyn hn hp sang cc 800 ml, trng sch bnh bng 100 ml nc ct v gp dch ra vo cc. Cho bay hi ht cn trn b cch thu, thnh thong thm nc thay th lng cn bay hi. Dng qu trnh ui cn khi khng cn mi cn bay ra. Hiu chnh th tch cui cng cho 250 ml bng nc nng. Trung ho dung dch x phng vi acid sulfuric long (1/2), thm 10% d v un nng, khuy u cho n khi acid bo tch lp. Chuyn lp acid bo sang phu chit 500 ml, ra 3 hoc 4 ln, mi ln vi 20 ml nc nng loi ht cc polyol, gp dch ra vi pha nc cha polyol thu c t qu trnh x phng ho. Chit pha nc ny 3 ln 20 ml ether du ho/ln, ly lp ether du ho gp vi phn acid bo, cho bay hi ti kh trong mt a xc nh b, ngui v cn. Trung ho dung dch cha polyol bng dung dch kali hydroxyd 1/10 ti pH = 7, s dng pH k thch hp. Cho bay hi dung dch ny ti gn cn, tch phn polyol khi cc mui bng cch chit vi ln vi cn nng. Cho bay hi dch chit cn trn b cch thu ti kh trong mt a xc nh b, ngui v cn. Trnh kh qu trong qu trnh un nng.

6.2. nh lng

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Ph lc 21 YU CU K THUT V PHNG PHP TH I VI SORBITAN MONOOLEAT 1. Tn khc, ch s 2. nh ngha INS 494 ADI=0-25mg/kg th trng Hn hp bao gm cc ester mt phn ca sorbitol v cc dn cht mono- v dianhydrid ca n vi acid oleic thc phm (R). Thnh phn ln nht l 1,4-Sorbitan monooleat, thnh phn t hn l isosorbid monooleat, sorbitan dioleat v sorbitan trioleat. 1338-43-8 Gm acid oleic este ha vi polyol dn xut t sorbitol bao gm cc loi sau:

Ch s C.A.S Cng thc cu to

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

1,4-Sorbitan

Isosorbid

Dng st snh mu h phch, dng ht hay mnh c mu kem nht n vng nu hoc dng sp rn, c mi nh . Cht nh ha, cht n nh

Ti nhit cao hn im chy, tan trong ethanol, ether, ethylacetat, anilin, toluen, dioxan, ether du ha, carbon tetraclorid; khng tan trong nc lnh nhng c th phn tn trong nc m. Phn acid oleic thu c bng cch x phng ho sorbitan monooleat trong qu trnh nh lng c ch s iod t 80 n 100.

Ch s iod

5.2. tinh khit Nc Tro sulfat Ch s acid Ch s x phng ha Khng c qu 2% (phng php Karl Fischer). Khng c qu 0,5%. Khng c qu 8. Khng c thp hn 145 v khng c qu 160. 84

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Ch s Hydroxyl Ch 5.3. Hm lng Khng c thp hn 193 v khng c qu 210. Khng c qu 2,0 mg/kg X phng ha 100g mu phi thu c khng thp hn 28g v khng c qu 32g polyol, khng thp hn 73g v khng c qu 77g acid bo. Hm lng polyol khng c thp hn 95% trong hn hp sorbitol, 1,4-sorbitan v isosorbid.

6. Phng php th 6.1. tinh khit Ch - Th theo JECFA monograph 1 - Vol.4. - Xc nh bng k thut quang ph 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 25g mu th, chnh xc n mg, chuyn vo bnh cu y trn 500 ml, thm 250 ml cn v 7,5g kali hydroxyd v trn u. Lp bnh cu vi sinh hn thch hp, un hi lu hn hp trong t 1-2 gi sau chuyn hn hp sang cc 800 ml, trng sch bnh bng 100 ml nc ct v gp dch ra vo cc. Cho bay hi ht cn trn b cch thu, thnh thong thm nc thay th lng cn bay hi. Dng qu trnh ui cn khi khng cn mi cn bay ra. Hiu chnh th tch cui cng cho 250 ml bng nc nng. Trung ho dung dch x phng vi acid sulfuric long (1/2), thm 10% d v un nng, khuy u cho n khi acid bo tch lp. Chuyn lp acid bo sang phu chit 500 ml, ra 3 hoc 4 ln, mi ln vi 20 ml nc nng loi ht cc polyol, gp dch ra vi pha nc cha polyol thu c t qu trnh x phng ho. Chit pha nc ny 3 ln 20 ml ether du ho/ln, ly lp ether du ho gp vi phn acid bo, cho bay hi ti kh trong mt a xc nh b, ngui v cn. Trung ho dung dch cha polyol bng dung dch kali hydroxyd 1/10 ti pH = 7, s dng pH k thch hp. Cho bay hi dung dch ny ti gn cn, tch phn polyol khi cc mui bng cch chit vi ln vi cn nng. Cho bay hi dch chit cn trn b cch thu ti kh trong mt a xc nh b, ngui v cn. Trnh kh qu trong qu trnh un nng. Tin hnh nh lng mt mu khc (25g) theo hng dn chuyn lun xc nh hm lng sorbitan ester xc nh % sorbitan ester.

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