• N g h i e n CLFU - K y thuat<br />
<br />
4.6 mm; 5 urn), mixture of acetonitrile and 10 preparations and, especially in the study on<br />
mM phosphate buffer with pH adjusted to 2.5 desloratadine stability.<br />
containing 30 mM TEA (25:75, v/v) as eluent in Tai lieu t h a m k h a o<br />
isocratic mode, flow rate of 1.2 ml/min., and 1. D. d. Rao, et al., Journal of Pharmaceutical and<br />
detection wavelength of 278 nm. The method Biomedical Analysis, 51 (2010), 736-742.<br />
enables a reliable statement concerning 2. Razid, et al., Dhaka Univ. J. Pharm. Sci,<br />
quantitative determination of desloratadine in (2006)5,1-4.<br />
commercially available pharmaceutical 3. Bo Y Te, Dupe Diin Viet Nam IV (2010), PL-122.<br />
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D i n h l i r ( m g d o n g t h d i m o t so a c i d b e o<br />
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o m e g a - 3 , o m e g a - 6 t r o n g d a u t h y c v a t<br />
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b a n g k y t h u a t s i c k y k h i k h d i p h 6 ( G C - M S )<br />
<br />
Nguyln Thanh Thong , Pham Thi Thanh Ha , Tran Cao Son<br />
1 1 2<br />
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Trudng dai hoc Duqc Ha Noi<br />
1<br />
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Vien Kiem nghiem An toan Ve sinh Thuc pham Qudc gia<br />
2<br />
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Dat van de so cao nhu 30-1. Ty le toi uu dupe cho la 4-1<br />
Cung vdi su phat trien cua kinh te, ty le mlc hoSc thlp hon . m<br />
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benh tim mach ngay cang cao va xuit hien d Cac acid beo omega-3, omega-6 c6 nhiiu<br />
nhiiu Ida tuoi. Chi dp dinh dudng lipid khdng trong cac loai diu thuc vat nhu diu d$u tuong,<br />
hpp li la mpt trong nhung nguyen nhan hang diu dua, diu ep, dau bip, dau mam lua mi, diu<br />
diu cua can benh nay. Cac acid beo omega-3 hat cai... Viec chiit xuit cac loai diu thuc vat<br />
co tac dung lam giam cholesterol va triglycerid kha don gian nen d i dang thu dupe ham lupng<br />
mau, giam nguy ca dau tim, giam nguy co dot Idn acid beo omega-3, omega-6 vdi chi phi thlp.<br />
quy do thieu mau cue bp. Ngoai ra, nhdm acid Tren thi gidi chua cd nhieu nghien cuu v i ham<br />
beo omega-3 con cin thilt cho phat trien hoan lupng acid bdo omega trong dau thuc vat l | 5 , 6<br />
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thien chdc nang nhin cua mit, giam nguy ca dai Hien nay, d Viet Nam cung chua co nghien cuu<br />
thao dudng, giam mdc dp nang va so con hen ve ham lupng cac loai acid beo omega 3,<br />
phi quan, giam trieu chdng viem khdp dang omega 6 trong diu thuc vat. Phuang phap phan<br />
thlp, ching trim cam... Mpt so nghien cdu- bao tich acid beo cung da dupe nhiiu tac gia nghien<br />
cao co thi co hieu dng chong ung thu cua acid cuu trong cac d l i tupng nhu diu gle, vien nang<br />
beo omega-3 (dac biet la ung thu vu, dai trang diu ca " . Tuy nhien d Viet Nam hien chua c6<br />
[1 4]<br />
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<br />
va ung thu tuyln tiln liet) . Tuy nhien, cac acid<br />
m<br />
phuang phap chinh thuc de phan tich acid beo<br />
beo nay nlu su dung vdi lilu lupng qua Idn c6 trong diu thuc vat. Trong cac phuang phap cd<br />
thi lam tang nguy co dot quy xuit huyit, tang the ung dung de phan tich acid beo thi phuong<br />
xac suit cua mpt s i benh. FDA khuyln cao phap sic ky khoi pho cho kit qua tach tot nhat,<br />
ring tong lupng acid beo Omega-3 tu chi dp dong thdi cd dp tin cay cao , [8]<br />
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an khong nen vupt qua 3 gam mii ngay, trong Do dd, chung toi nghien cuu phuang ph£p<br />
dd cd khong qua 2 gam mii ngay la tu cac chat djnh lupng mdt so acid beo omega-3, omega-6<br />
bo sung dinh dudng. Ngoai ra, ty le giua trong diu thuc v$t bing ky thuat sic ky khi khoi<br />
omega-6 va omega-3 su dung rat quan trong. pho (GC-MS) vdi cac muc tieu sau:<br />
Mpt ti le omega-6 qua cao va omega-3 qua thlp - Xay dung phuang phap djnh lupng dong<br />
diu co anh hudng khong t i t cho sue khoe. Chi thdi mpt so acid beo omega-3 va omega-6 trong<br />
dd 3n phuang Tay hien dai thudng c6 ty le cua diu thuc vat bing ky thuat sic ky khi khoi pho.<br />
acid omega-6 va omega-3 vupt qua 10-1, mdt - Thim djnh phuong phap da xay dung.<br />
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32 TAP CHI DUQC HQC - 10/2010 (SO 414 NAM 50)<br />
N g h i e n CLFU - Ky thuat<br />
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- Crng dung phuang phap de xac djnh ham - Dung dich natri methylat 5%/methanol: Can<br />
lugng cac acid beo trong mdt s i loai dau thuc vat. 5g natri methylat, djnh muc trong binh dinh muc<br />
D6i tux^ng va phu/cng p h a p n g h i e n 100ml bing methanol. Bao quan trong tu lanh.<br />
Dieu kien sac ky<br />
CLPU<br />
Ddi tugng nghien cuu Chuong trinh nhiet dp: Nhiet dp diu 100°C<br />
giu trong 1 phut; t£ng 20°C/ phut len 165°C, sau<br />
Cac loai dau thuc vat c6 nguon goc khac<br />
dd tang 5°C/1 phut len 260°C, giu trong 16,75<br />
nhau dugc lay tren dia ban Ha Npi: dau dau<br />
phut. Tong thdi gian phan tich 40 phut.<br />
tuang, dau me, dau oliu...<br />
- Nhiet dp buong bom miu: 250°C, bam<br />
Thiet bi<br />
khdng chia ddng , thdi gian khdng chia ddng la<br />
- May sic ky khi khoi pho GC-MS model<br />
1 phut. Thi tich tiem 1 ul. ,<br />
Polaris Q cua hang Thermo hoSc tuong duang.<br />
- Nhiet dp transferline: 260°C.<br />
- Cot sac ki khi DB5-MS (60 m x 0,25 mm x<br />
0,25 urn). - Khi mang: Heli, tic dp 1 ml/ phut<br />
- Cac thiet bj can thiet khac... - Nhiet dp buong ion: 200°C.<br />
Hoa chat, thuoc thu - Thdi gian clt dung mdi: 7 phut.<br />
Chuan hdn hgp 37 FAMEs (dang methylester - Chi dp chay FULL SCAN dai tu 50-250 m/z.<br />
cua acid bdo) vdi nong dp tong 10 mg/ml trong Ket qua<br />
methylen clorid, dupe cung cap bdi Supelco. Phuang phap d i n xuit hda<br />
Chuan ndi methyl nonadecanoat cua Sigma- O i phan tich dugc bing sic ky khi, miu<br />
Aldrich. phan tich cin phai dugc chuyln thanh dang hai<br />
Natri clorid, Natri hydroxyd, methanol, n- trong dilu kien thuc nghiem. Dilu nay khdng<br />
hexan, boron trifluorid 50%/methanol cua Merck. thuc hien dugc vdi cac acid beo, dac biet la cac<br />
Chuan bi cac dung djch acid bdo nhieu carbon. Vi vay, cin chuyln cac<br />
- Dung dich chuin noi: Can chinh xac 0,1g chit bdo va acid bdo trong diu thuc vat thanh<br />
methyl nonadecanoat hda tan trong n-hexan din xuit de bay hoi d l cd t h i phan tich dugc<br />
vua du 50ml, thu dupe dung dich chuan ndi cd bing GC/MS. Ben canh dd, din xuit hda cdn<br />
nong dp 2000 p.g/ml. cho phdp chuyen cac acid beo v l dang khdng<br />
- Dung dich natri hydroxyd 0,5M pha trong phan cue nen han chi dugc kha n£ng hip phu<br />
methanol: Can 2g NaOH djnh muc trong binh trong qua trinh sic ky khi do tuang tac cua<br />
djnh muc 100 ml bing methanol. Bao quan nhung nhdm phan cue gay ra.<br />
trong tu lanh. Khao sat mdt so phuang phap din xuat hda va<br />
- Dung dich boron trifluorid 7%/methanol: Hut so sanh kit qua thu dupe khi phan tich cung mdt<br />
3,5ml dung djch BF vao binh djnh muc 25 ml.<br />
3 miu: lua chpn phuong phap din xuit vdi natri<br />
Oinh muc den vach bing n-hexan. Bao quan hydroxyd/methanol va borontrifluorid/ methanol vi<br />
trong tu lanh. phuang phap nay cd hieu suit din xuit cao nhit.<br />
Bang 1: So Sclnri kit qui giua mot sd phuong phip din xuit<br />
Dan xuat bang Natri Dan xuat bang NaOH/MeOH Din xuat bang BFi/MeOH<br />
Acid beo methylat/MeOH (mg/100gf va BFi/MeOH (mg/IOOg/ 111<br />
(mg/100gf ' 0<br />
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Acid linoleic 25,694 30,060 12,00<br />
Acid arachidonic 0,004 0,006 0,005<br />
Acid y-linolenic 0,124 0,174 0,062<br />
EPA 0,075 0,107 0,071<br />
Tinh phu hgp cua he thing sic ky Tinh phu hgp cua he thong sic ki khi<br />
Cac acid bdo phan tich sau khi dugc din dugc khao sat bing each bom 6 l i n cung 1<br />
xuit hda dugc xac djnh bing sic ky khi khoi dung djch chuan hdn hgp cac acid da dugc<br />
pho (GC-MS) va so sanh vdi pho chuin trong methyl ester hda (FAMEs) vao may sic ki<br />
thu vien pho chuin Mainlib. khi.<br />
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TAP CHI DUQC HQC - 10/2010 (SO 414 NAM 50) 33<br />
• Nghien c i m - K y thuat<br />
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Bang 2: Kit qua khao sat do lap lai thdi gian co dp lap lai cua thdi gian luu (RSDu - Ky thuat<br />
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Bang 5: Khao sat tinh dung cua phwong tuyet dli cua Z < 2. Nhu vay, phuang phap<br />
score<br />
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phap ddi voi acid linoleic (C18:2n6) xay dung da dat yeu ciu v l tinh dung.<br />
Lan Ket qua thu Ket qua cdng Dp lap lai cua phuang phap<br />
phan tich dwcrc (g/100g) bo(g/100g) Phan tich 6 lin khac nhau cung 1 miu<br />
Lan 1 3,81 4,04 -1,211 diu thuc vat. Danh gia kit qua thong qua dp<br />
Lan 2 4,10 4,04 0,316 lech chuin tuang doi (RSD%) cua ham<br />
So sanh ket qua phan tich vdi ket qua trong lupng cac chit phan tich dupe qua 6 lin<br />
mlu chuan, ket qua thu dime deu co gia trj phan tich do.<br />
Bang 6: Do lap lai cua phwong phap<br />
Lwcmg chat phan tich duoc (g/100g)<br />
Lan Acid omega-3 Acid omega-6<br />
khao sat Acid Acid Acid Acid Acid<br />
Eicosa-penta-enoic Docosa-hexa-enoic linoleic arachidonic dihomo-y-linolenic<br />
1 0,78 1,90 36,13 2,99 0,0161<br />
2 0,74 1,82 35,81 2,92 0,0170<br />
3 0,75 1,96 36,16 3,09 0,0164<br />
4 0,74 1,87 36,35 2,99 0,0172<br />
5 0,73 1,85 • 36,40 2,97 0,0164<br />
6 0,73 1,93 35,81 2,95 0,0161<br />
TB 0,75 1,89 36,11 2,98 0,0165<br />
SD 0,018 0,05 0,25 0,058 0,00046<br />
RSD% 2,45 2,75 0,69 1,94 2,80<br />
Ket qua cho thay gia trj do lech chuan tuong ddi diu nho hon 2,8%. Nhu vay quy trinh da xay dung<br />
RSD% cua cac chat phan tich trong 6 lan phan tich la phii hpp va dam bao cho viec phan tich.<br />
Lfng dung phuang phap d i phan tich mdt sd miu diu thuc vat<br />
Bang 7: Kit qua phan tich mot sd miu diu thirc vat<br />
Ham lwcmg cac chat phan tich dwoc (g/100g)<br />
Acid linoleic Acid arachidonic EPA Acid dihomo-gamma-linolenic DHA<br />
1 36,11 0,0024 0,010 0,017<br />
2 7,41 2,98 0,75 0,194 1,89<br />
3 6,16 0,085 0,075<br />
4 14,63<br />
5 21,78<br />
6 36,35<br />
7 48,51<br />
Ban luan lupng va deu dat yeu cau. Vdi ket qua nay,<br />
Kit qua thu dupe cho thiy phuang phap din chung toi d i xuit ung dung phuong phap nay<br />
xuit-cac acid beo sang dang methylester bing tai cac phong thi nghiem de phan tich cac acid<br />
NaOH/Methanol va BF /Methanol co hieu suit beo omega-3, omega-6 noi tren.<br />
3<br />
<br />
din xuit cao nhit. Phuang phap da xay dung co kha nang<br />
Phuong phap GC-MS xay dung da dinh tinh phan tach tot tdi 32 acid beo khac nhau trong<br />
va dinh lupng dupe dong thdi mpt so acid beo hin hpp chuin 37 acid beo. Vi vay, chung toi d l<br />
omega-3, omega-6 trong diu thuc vat. Phuong nghj tilp tuc hoan thien quy trinh phan tich va<br />
phap nay da dupe thim djnh tren mpt s i chi thim dinh d l dinh lupng dupe ding thdi cac<br />
tieu nhu tinh chpn lpc, khoang tuyin tinh, dp lap acid beo con lai trong hin hop chuin 37 acid<br />
lai, dp dung, gidi han phat hien, gidi han dinh beo.<br />
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TAP CHI DUQC HQC - 10/2010 (SO 414 NAM 50) 35<br />
• N g h i e n CLFU - Ky thuat<br />
<br />
RT: 7.48 - 39 86<br />
100 NL:<br />
4 55E6<br />
TIC F: MS<br />
Std 1 0_20_<br />
80 30ppm<br />
70<br />
60<br />
<br />
<br />
<br />
40 1 1 48 21 .40 |<br />
30 i 20.03<br />
14.70 > 30.90<br />
20 8 66 13.12 30.21 ! p<br />
o 38 51<br />
O ( o I<br />
1 8 34 24 37.45<br />
- 8 i q 5 i<br />
o : ,~J ^~~} ff, X Jl.„ JI_J J M 131.68^ JL 37.02.I [<br />
10 15 20 25 30 35<br />
Time (min)<br />
Sic dd phan f/ch hon hop chuin 37 FAMEs<br />
Summary Khanh, Trinh Van Lau, Xac djnh cac acid beo trong<br />
dau mang hat gac bang phuong phap sac ky khi<br />
Fatty ac/ds from vegetable oil is<br />
(GC/FID), Kiem nghiem thuoc, (2005) 3(9), tr. 21-25.<br />
transesterified into fatty acid methyl esters<br />
4. Nguyln Tudng Vy, Tran Tu An, Oang Due<br />
(FAMEs) by using sodium hydroxid in methanol Khanh, Trjnh Van Lau, Xac dinh thanh phan cac acid<br />
and borontrifluoride in methanol. FAMEs then beo dau gac bang sic ky khi khoi pho (GC/MS),<br />
was extracted by hexan and determined by gas Duoc hoc, (2007) 3, tr. 28-30.<br />
chromatography coupled with mass spectrometry 5. Bruno J., Nicholas L. S., William D. J.,<br />
(GC-MS). Separation of FAMEs using a 60m x Determination of the fatty-acid composition of<br />
0,25mm fused silica capillary column coated with soybean oil by high-pressure liquid chromatography,<br />
a 0.25 pm film of poly-(5% diphenyl/ 95% (1982) Talanta, 29. pp. 54 -56.<br />
dimethyl siloxane). Simultaneous determination 6. Chen S. H., Chen K. C, Lien H. M.,<br />
of several omega-3 and omega-6 fatty acids was Determination of fatty acids in vegetable oil by<br />
performed using full scan MS (50-250 m/z). The reversed-phase liquid chromatography with<br />
method was validated, showing good linearity (R fluorescence detection, Journal of Chromatography A<br />
(1999), 849, pp. 357-369.<br />
> 0.999) in a range of 0.5 to 20 pg/ml, high<br />
7. Hibbeln, J.R.. Healthy intakes ofw3 and 106<br />
trueness and precision (RSD
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