XAY D I / N G
PHl/CfNG PH
A P PHAT H I E N CHAT M A U M E T A N I LYELLOW CO MAT
T R A I P H E PTRONG SON MOI VA SON MONG TAY
NGHIEM THI ANH MINH, VU TUNG LAM
Trir&ng Dai hgc Duftc Hd Noi LE DINH CHI, LE THI HUONG HOA
Vien Kiem nghiem thudc Trung irong
1. Dat van de
Metanil yellow la mot chat mau tdng hgp, cdn cd ten khac la "Tropaeolin G" (Cdng thiic phan tii:
CjgHj^NjOjSNa, khdi lugng phan tu: 375,39; ma sd mau: CI 13065). Metanil yellow dugc su dung rdng rai trong cdng nghiep thudc da, giay, det. Da cd nhiiu tai lieu tren the gidi va khu vuc de cap din tac hai cua metanil yellow tdi siic khde cua con ngudi khi diing dudng udng hoac ngam qua da. Theo Hiep dinh hda hgp quan ly my pham ciia ASEAN nam 2003, metanil yellow la mdt trong nhieu chat mau bi cam sii dung trong my phim. Tuy nhien, hien nay d Viet Nam viec kiem tra cac chit mau bi cim su dung trong my phim cdn nhieu han che. De gdp phan tien tdi thiet lap he thdng quy trinh phan tich chuan de phat hien cac chit bi cam su dung trong my pham, trong nghien ciiu nay chiing tdi tiln hanh xay dimg phuang phap phat hien metanil yellow cd mat trai phep trong doi tugng miu my pham la cac loai son mdi va son mdng tay, ddng thdi ap dung phuang phap xay dimg dugc de kiem tra su cd mat cua metanil yellow trong mdt sd mau my pham dang luu hanh tren thi trudng.
\_J
\ //Hinh 1. Cong thuc cdu tao ciia Metanil yellow 2. Thvc nghiem
2.1. Hda chat, thiet bf, dung cu 2.1.1. Hoa chdt
Methanol tinh khilt HPLC (Merck, Diic).
Triethylamin tinh khiet phan tich (Merck, Diic).
Acid acetic 30% tinh khilt phan tich (Merck, Diic).
- n-Hexan tinh khiet phan tich (Merck, Diic).
Acid phosphoric tinh khiet phan tich (Merck, Diic).
N, N dimethylformamid (DMF) tinh khiet phan tich (Merck, Diic).
Nudc cit dugc loc qua mang milipore 0,45 ^im.
- Chit chuan Metanil yellow (Merck, Diic).
2.1.2. Thiet bi, dungcu
- May sac ky ldng Shimadzu dugc trang bi detector PDA.
May do pH 691 Metrohm.
Bd loc dung mdi, loc mau vdi mang Igc 0,45 iim.
Can phan tich Mettler Toledo cd do chinh xac 0,1 mg.
- Cac dung cu thiiy tinh chinh xac: binh dinh miic, pipet chinh xac, dng dong...
2.2. Boi twang vd phiromg phdp nghien ciru 2.2.1. Phuang phdp nghien cuu
Phuang phap phat hien metanil yellow cd mat trai phep trong my pham dugc xay dung dua tren ca sd ky thuat sic ky ldng hieu nang cao (HPLC). Su cd mat ciia metanil yellow trong mau my pham dugc danh gia ca ve mat dinh tinh va dinh lugng.
- Dinh tinh:
+ Dua vdo thdi gian luu: so sanh thdi gian luu cua pic chinh tren sac ky dd cua dung dich thu vdi thdi gian luu cua pic chinh tren sac ky dd cua dung dich chuan.
+ Dua vdo pho hdp thu UV-VIS thu duac nhd detector PDA: so sanh phd hap thu UV-VIS lay d dinh pic nghi la metanil yellow tren sac ky dd cua dung dich thii vdi phd UV-VIS liy d dinh pic metanil yellow tren sac ky do cua dung dich chuan, danh gia he sd tuang ddng (similarity index) giiia 2 phd.
- Dinh luang:
Khi da khang dinh su cd mat cua metanil yellow trong mau my pham, ham lugng metanil yellow trong mau thu dugc xac dinh dua vao dien tich pic hay chieu cao pic metanil yellow thu dugc tren sac ky dd cua dung dich chuan va dung dich thu, ndng do cua metanil yellow trong dung dich chuan va lugng mau thii dem phan tich.
T^p chi KIEM NGHIfM THU6C - So 2.2011; Tap 9,(32)
2.2.2. Doi tuang nghien cuu
Phuang phap nghien ciiu dugc xay dimg va ap dung dl kiem tra phat hien metanil yellow trong 6 mau son mdi va 8 mau son mdng tay.
3. Ket qua va ban ludn
3.1. Xay dimg phuang phdp phan tich 3.1.1. Quy trinh xic ly mdu my phdm
a) Lua chpn dung moi chiet metanil yellow khdi nen mdu my phdm
Sau qua trinh khao sat so bd, chiing tdi nhan thay hdn hgp DMF va acid phosphoric (95:5) la dung mdi chiet metanil yellow cd nhieu uu diem hon ca:
- Cho phep chiet dugc metanil yellow ra khdi nen mau son mdi va son mdng tay mdt each chgn Igc, it lan tap chat.
- Quy trinh xu ly mau don gian, de thuc hien.
Vi vay, trong nghien ciiu nay chiing tdi su dung dung mdi chiet mau la hdn hgp DMF va acid phosphoric (95:5).
b) Quy trinh xir ly mdu my phdm
Can chinh xac mdt lugng khoang 2,0 g mau my pham vao cdc cd md 25 ml, dimg dGa thiiy tinh phan tan mau deu trong cdc, them khoang 5 ml dung mdi chiet mau, khuay ki, chuyen dung dich thu dugc vao binh gan. Them khoang 20 ml n-hexan vao bmh gan.
Trang lai cdc dung miu bang dung mdi chiit miu 2 lan, mdi lan khoang 5 ml rdi chuyen vao binh gan, lac ky trong 5 phiit, de lang cho phan ldp, gan lay phan dung mdi chiet miu (ldp dudi). Neu ldp n-hexan cdn lai trong binh gan van cdn mau, them 4 ml dung mdi chiet mau vao binh gan de chiet lan 2, lac, de lang, gan lay phan dung mdi chiet mau. Gdp djch chiet, them dung mdi chiit mau viia du 20,0 ml, lac deu. Lgc qua giay Igc, rdi Igc lai qua mang lgc 0,45 |j.m de tiem sac ky.
3.1.2. Quy trinh phdn tich metanil yellow bdng HPLC Qua trinh phan tich dugc tien hanh tren he thdng may sac ky ldng hieu nang cao dugc trang bi detector PDA vdi cac thdng sd cua he thdng sac ky nhu sau:
• Pha tinh: cdt Apollo C18 (250 x 4,6 mm; kich ca hat 5 [xm) hoac tuang duang.
• Detector PDA dat trong dai budc sdng tu 200- 900 nm. Budc sdng lay sac ky dd: 424 nm.
• Pha ddng: methanol - dem pH 6,2 (55:45).
• Tdc do ddng: 1,0 ml/phiit.
• The tich tiem miu: 20 jil.
• Nhiet do cdt: nhiet do phdng.
Cach chuan bi dung dich dem pH 6,2: chuan bi dung dich triethylamin 3 mM trong nudc, chinh pH vl 6,2 bing acid acetic 30 %. Lgc qua mang loc 0,45 H-m.
- Dimg dich chuin:
+ D I danh gia khoang tuyen tinh, chiing tdi chuin bi mot day dung dich chuan cd ndng do metanil yellow chinh xac khoang 0,4; 1,0; 2,0; 4,0; 8,0; va 20,0 ng/ml trong hdn hgp DMF - acid phosphoric (95:5), lgc qua mang lgc 0,45 |j.m.
-I- D I kiim tra sir cd mat cua metanil yellow trong miu my phim, chiing tdi chuin bi mdt dung dich chuin cd ndng do metanil yellow chinh xac khoang 0,4^g/ml trong hdn hgp DMF acid phosphoric (95:5), lgc qua mang lgc 0,45 p,m.
Dung dich thu: su dung dung dich thu dugc sau khi xu ly mau theo quy trinh da trinh bay d muc 3.1.l.b.
3.2. Tham ^nh phirong phdp 3.2.1. Dp ddc hieu
Su dimg cac dieu kien phan tich da trinh bay d muc 3.1.2, chiing tdi da tien hanh phan tich cac miu sau:
MIU tring: miu my phim khdng chiia metanil yellow dugc xu: ly theo quy trinh d muc 3.1.1 .b.
Mau tu tao: mau my pham khdng chiia metanil yellow dugc dung lam mau trang cd trdn them mot lugng chfnh xac metanil yellow chuan va xii ly theo quy trinh d muc 3.1.1 .b.
Dung dich chuin metanil yellow: pha mot dung dich chuin metanil yellow trong dung mdi chiit miu de dugc dung dich cd ndng do khoang 20 p,g/ml.
Ket qua:
- Ddi vdi dung dich chuan, tren sic ky dd chi xuat hien mdt pic ro rang cua metanil yellow.
- Ddi vdi miu tring, tren sic ky dd khdng co pic nao xuat hien tuang iing vdi thdi gian luu cua metanil yellow.
Ddi vdi mau tu tao, sic ky dd cho ta mot pic tuomg ling vdi pic cua metanil yellow tren sic ky do cua dung dich chuan ve thdi gian luu va do tuong thich khi so sanh phd UV-VIS (he sd tuong ddng ~ 1).
ddng thdi dugc tach rieng khdi cac pic khac.
Sac ky dd ciia dung dich chuin, miu tring va mau tu tao, ciing kit qua so sanh phd UV-VIS giiia pi' metanil yellow tren sac ky dd ciia dung dich chuin va pic cd thdi gian luu tuang iing tren sic ky dd cua raa"
tu tao duac the hien d hinh 2.
10 Tap chi KIEM NGHIEM THU6C - So 2.2011; Tap 9.(32)
IL ii
ISO 17.B
Minutes
Thai gian (phiit) g
0 0 Z.A s o T S 10O 138 ISO I T S
Minutes
Thai gian (phut) C D
Hinh 2. Kit qud ddnh gid dp ddc hieu A: SKD dung dich chudn metanil yellow; B: SKD mdu my phdm khdng chua metanil yellow; C: SKD mdu tu tao; D: Kit qud so sdnh pho UV-VIS ciia pic metanil yellow
tren SKD A vdpic co ciing thdi gian lim tren SKD C.
3.2.2. Khoang tuyin tinh
De ddnh gia khoang tuyen tinh, chiing tdi chuan bi mdt day dung dich chuan cd ndng do metanil yellow chinh xac khoang 0,4; 1,0; 2,0; 4,0; 8,0; va 20,0 Hg/ml trong hdn hgp DMF - H3P04(95:5), lgc qua mang lgc 0,45 |im. Cac dung djch chuin nay dugc lan lugt tiem vao he thdng HPLC sii dung cac dieu kien
sic ky d muc 3.1.2. Ket qua dap iing tren sac ky dd (thdng qua dien tich pic) cho thiy phuang phap phan tich da xay dung cho dap iing tuyen tinh tdt vdi metanil yellow trong khoang ndng do phan tich tu 0,4 den 20,0 ng/ml vdi he sd tuong quan tuyln tinh r = 0,9999 (Bang 1).
Bdng 1. Kit qud khao sdt khoang tuyen tinh cua metanil yellow
TT 1 2 3 4 5 6
Phuong trinh hoi qui Hf so tuang quan
Nong dp (Hg/ml)
0,4 1 2 4 8 20
Dien tich pic (mAU.s)
25935 66750 129224 256215 489277 1189123 y = 59116 x+10639
r = 0,9999
1400000
sr 1200000 59116x +10639
5 10 15 N6ng do metanil yellow (mcg/ml)
,T?pchi KltM NGHI$M THU6C - S6 2.2011; Tap 9.(32)
20
11
3.2.3. Dp lap lai vd dp diing
Viec danh gia do lap lai - do diing dugc thuc hien ddng thdi tren 2 miu tu tao, mdt miu dua tren nen son mdi khdng chiia metanil yellow, mdt miu tren nin son mdng tay khdng chiia metanil yellow. Mot lugng chinh xac metanil yellow chuin se dugc dua vao cac nin mau my pham ke tren. Mdi miu tu tao se dugc tien hanh xu ly theo quy trinh d muc 3.1.1 .b va phan tich theo diiu kien d sic ky d muc 3.1.2 thanh 6 lan ddc lap. Do lap lai cua ca phuang phap se dugc danh gia qua kit qua ham lugng metanil yellow thu dugc sau 6 lan phan tich ddc lap miu tu tao. Do diing cua phuang phap dugc danh gia bing each so sanh lugng metanil yellow tim lai dugc trong mau tu tao so vdi lugng metanil yellow cho vao ban dau (bang 2 va bang 3).
Bdng 2. Ket qud khao sdt dp diing vd dp lap lai cua phuang phdp tren mdu tu tao co nen son moi khong chira metanil yellow
TT
1 2 3 4 5 6
Khdi luong my pham nen (g)
2,0164 2,0072 2,0010 2,0112 2,0340 2,0150
Lirong metanil yellow them vdo (fig)
8,20 8,20 8,20 8,20 8,20 8,20
Luong metanil yellow dm thay (pg)
7,80 8,04 8,07 8,06 7,95 8,05 RSD (%) sau 6 lan phan tich dgc lap
Ty le thu hoi trung binh (%)
Ty le thu hoi (%)
95,1 98,0 98,4 98,3 97,0 98,2 1,3%
97,5%
3. Kit qud khao sdt dp dung vd dp lap lai cua phuang phdp tren mdu tu tao co nen san mong tay khdng chira metanil yellow
TT
1 2 3 4 5 6
Khoi luong my pham nin (g)
2,0424 2,0200 2,0210 2,0184 2,0206 2,0294
Luong metanil yellow them vdo (l^g)
8,00 8,00 8,00 8,00 8,00 8,00
Luong metanil yellow dm thdy (ng)
6,84 7,04 6,78 6,87 6,58 6,92 RSD (%) sau 6 lan phan tich dgc lap
Ty le thu hoi trung binh (%)
Tylethuh6i(%)
85,5 88,0 84,8 85,9 85,6 86,5 2,3%
86,0%
Ket qua thuc nghiem tren hai miu tu tao cho thay, phuang phap cd do lap lai tdt, the hien qua gia tri RSD thap sau 6 lan phan tich dgc lap (1,3 % vdi miu tu tao tren nen son mdi va 2,3 % vdi miu tu tao tren nen son mdng tay). Ve do diing, mau tu tao tren nin son mdi cho ty le thu hdi tdt han tu 95,1 % din 98,4 %. Ty le thu hdi tren miu tu tao cd nin son mdng tay dao ddng trong khoang tu 84,8 % din 88,0 %.
Nhu vay, tren ca hai loai nen miu my pham, vdi lugng
metanil yellow cho vao nen miu rit nhd (chi xap xi 8 |ig), phuang phap phan tich xay dimg da the hiSn do diing chip nhan dugc, phii hgp vdi muc dich ciia phuang phap la phat hien metanil yellow cd mat trai phep trong son mdi va son mdng tay.
3.2.4. Gidi han phdt hien - Gidi han dinh luang Gidi han phat hien va gidi han dinh lugng ciia phuang phap dugc xac dinh qua thuc nghi?m each phan tich cac miu tu tao trong nin miu my ]
12 Tap chi KIEM NGHIEM THUOC - So 2.2011; TSp 9.(32)
CO ndng dg metanil yellow chuan dugc them vao giam din. Kit qua thuc nghiem cho thiy:
Vdi ndng do metanil yellow trong dung dich chuin tiem sic ky la 0,4 fig/ml, pic metanil yellow tren sic ky dd cao hon 10 lin so vdi dudng nin. Nhu vay phuang phap cd gidi han dinh lugng cho metanil yellow la 0,4 jig/ml.
Vdi ndng do metanil yellow trong dung dich chuin tiem sac ky la 0,12 \ig/ml, pic metanil yellow tren sic ky dd cao han 3 lin so vdi dudng nin. Nhu vay phuang phap cd gidi han phat hien cho metanil yellow la 0,12 ng/ml.
3.2.5. Ap dung phuang phdp di Mim tra mptsd mdu my phdm mua tren thi trudng
Phuang phap phat hien metanil yellow da dugc chiing tdi ap dung dl kiem tra 6 miu son mdi va 8 mau son mdng tay mua tren thj trudng. Kit qua da phat hien thay mdt mau son mdng tay khdng cd nhan mac, dia chi nha san xuat cho pic cd cung thdi gian luu va phd UV-VIS (vdi he sd tucmg ddng khi so sanh phd bang 0,9913) nhu pic metanil yellow chuan.
Nhirng ket qua dinh tinh tren cho phep ket luan mau chiia metanil yellow (xem hinh 3). Tinh toan dinh lugng cho thay ham lugng metanil yellow trong mau la 28,33 ^g/g.
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1 A
1 / » \ 1
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g
A, U _ , ^'
A B ITmh 3. Sdc ky do vd kit qud so sdnh pho UV-VIS cua mdu san mong tay khong ro nguon goc co chira
Metanil yellow. A: SKD ciia mdu; B: Kit qud so sdnh pho UV- VIS cua pie co thdi gian luu tuang duang vdi metanil yellow tren SKD A vdpic metanil yellow tren SKD cua dung dich chudn.
4. Kit luSn
Phuang phap phat hien metanil yellow cd mat trai phep trong son mdi va son mdng tay dugc xay dung dua tren quy trinh chiet ldng - ldng de tach metanil yellow tu nen mau cung quy trinh phan tich metanil yellow bang HPLC pha dao dugc trang bi detector PDA. Metanil yellow dugc phat hien dinh tinh nhd so sanh thdi gian luu va phd hap thu UV-VIS cua pic nghi ngd tren sic k^ do ciia dung dich thu vdi pic metanil yellow chuan tren sac ky dd cua dung dich chuin. Ket qua tham djnh ve do dac hieu, khoang
tuyln tinh, do diing, do lap lai, gidi han dinh lugng va gidi han phat hien chiing minh phuang phap da xay dung dap iing dugc yeu cau ap dung de viec phat hien metanil yellow cd mat trai phep trong mau son mdi va son mdng tay, ddng thdi cd do diing va do lap lai tdt ve ket qua dinh lugng metanil yellow. Phuang phap da dugc ap dung de kiem tra 6 mau son mdi va 8 mau son mdng tay mua tren thi trudng, trong dd da phat hien dugc mdt mau son mdng tay chiia metanil yellow vdi ham lugng 28,33 ng/g.
Tai lieu tham khao
1. Ming-Ren Fuh, Kang-Jung Chia, Talanta, sd 56, tap 4, tr.663-671,2002.
2. Identification of prohibited colorants in cosmetic products by TLC and HPLC, phuang phap hoa hgp cua ASEAN (ma sd ACM SIN 02).
SUMMARY
A method using samplepretreatment by liquid/liquid extraction, following by HPLC analyzing in reversed-phase conditions and diode array detector was developed for determination of banned colorant metanil yellow illegally presented in lipstick and nail polish. The present of metanil yellow is identified by comparing retention time and
UV- VIS absorption spectrum of any suspected peak in chromatogram of sample preparation to that of the peak of
T^p chi KIEM NGHIfM THU6C - S6 2.2011; Tap 9.(32) 13
