Xây dựng quy trình định lượng desloratadin bằng phương pháp HPLC với đầu dò PDA

• N g h i e n CLFU - Ky thuat<br /> <br /> K i t luan screening showed that six compounds having<br /> 1. Da tong hop dupe 5-(2'-clorobenzyliden)(1) antibacterial activity against Eschehchia coli,<br /> va 9 din chat base Mannich (2 -10). Bacillus subtilis and Staphylococcus aureus; six<br /> 2. Ket qua thu hoat tinh sinh hpc cho thay compounds possessing antifungal activity against<br /> cac chat (1, 2, 5, 6, 9,10) deu the hi§n hoat tinh Aspergilus niger and Fusahum oxysporum.<br /> khang khuan, khang nam tuong doi cao tren Keywords: rhodanine, 5-(2-clorobenzyliden)<br /> cac chung vi khuan, nam moc kiem dinh a cac rhodanine, Mannich base, antifungal activity,<br /> muc dd khac nhau, chat ngung tu 1 va chat 2 antibacterial activity.<br /> khang 5 chung VSV kiem djnh (E. coli, B. Tai lieu t h a m k h a o<br /> subtilis, S. aureus, A. niger, F. oxysporum) vdi 1. Nguyen Quang Oat, Oinh Thj Thanh Hai, Ngo<br /> MIC = 12,5 - 25ug/ml, cac chat 5, 6, 9 khang 5 Mai Anh, Imphea Rith, Tap chi Duac hpc, (2001), 11,<br /> chung VSV kiem dinh (E. coli, B. subtilis, S. tr.13-16.<br /> aureus, A. niger, F. oxysporum) vdi MIC = 12,5 - 2. Nguyen Quang Oat, Oinh Thj Thanh Hai, Vu<br /> 50ug/ml, chat 10 khang 4 chung VSV kiem djnh Tran Anh, Phung Anh Thu, Nguyln Thi Thu Ha, Tap<br /> (B. subtilis, S. aureus, A. niger, F. oxysporum) chi Duoc hpc, (2008), 1, tr.31-34.<br /> 3. Oinh Thj Thanh Hai, Nguyln Quang Oat, Le<br /> vdiMIC = 25-50ug/ml.<br /> Mai Huang, Tap chi Duoc noc,(2003), 1, tr.18-20.<br /> Ket qua thu khang te bao ung thu cho thay ca 4. Dinh Thj Thanh Hai, Tap chi Duoc hoc, (2010),<br /> 6 chat dem thu nghidm deu kh6ng the hidn hoat 5, tr.54-58.<br /> tinh khang d6ng te bao ung thu gan (Hep-G2). 5. Akerblom E.B, J.Med.Chem, (1994), 17(6),<br /> Summary pp.759-762.<br /> By the condensation of aromatic aldehydes 6. Habib N.S, Rida S.M, Badaway E.A.M,<br /> Eur.J.Med.Chem, (1997), 32, pp.759-762.<br /> with rhodanine, compound 1 was obtained.<br /> 7. Pafahl M.C, J. Of Applied Chem, (2003), 11(3),<br /> Compound 1 underwent Mannich reaction to give pp.65-68.<br /> 9 Mannich base derivatives (2 - 10). The 8. Likhitayawuid K., Angerhofer C. et al., J. Nat.<br /> structures of synthesized compounds were Prod, (1993), 56(1), 30-38.<br /> confirmed by IR, H-NMR, C-NMR and mass<br /> 1 13<br /> 9. Vlietlinck A.J, Vanden Berger D.A, Methods in<br /> spectroscopy. The results of biological activity plants biochemistry, (1991), 6,47-68.<br /> <br /> <br /> <br /> <br /> X a y d y n g q u y t r i n h d i n h l u o n g d e s l o r a t a d i n<br /> <br /> <br /> bang phuong phap HPLC voi dau do PDA<br /> -<br /> Nguyen Thi Ngpc Giau , Nguyen Ngoc T h i Tran , Nguyen Due Tuan *<br /> 1 2 1<br /> <br /> <br /> <br /> <br /> 'Khoa Dugc, Dgi hoc Y Dugc TP. Hd Chi Minh<br /> Khoa Dugc, Trudng dgi hoc Y Dugc Cdn Tho<br /> 2<br /> <br /> <br /> <br /> <br /> Dat v l n d i den thdi diem hidn tai. Tai Viet Nam, che pham<br /> Desloratadin la mdt trong cac thuoc khang vien nen bao phim desloratadin d§ dupe dua<br /> histamin dupe lua chpn dau tien trong dieu vao nghien cuu va san xuat. Tuy nhien, chua cd<br /> trj viem mui dj ung do it tac dung phu tren tim cdng ty nao trong nude san xuat che pham<br /> mach va hau nhu khong gay buon ngu. desloratadin dang siro trong khi thuoc nhap<br /> Desloratadin la chat chuyen hda cd hoat tinh khau tu nude ngoai (Siro Aerius, SP<br /> cua loratadin, vua cd tac dung khang histamin, Laboratories) cd gia thanh kha cao. Cho den<br /> khang dj ung va khang viem, thdi gian ban thai nay vln chua cd chuyen luan cua desloratadin<br /> 27 gid va la thuoc duy nhat dupe chi djnh dieu trong tat ca cac Dupe diin. Dat moi quan tam<br /> trj viem mui dj ung cho tre tu 6 thang tuoi cho din mpt hoat chit mdi cd nhiiu uu dilm nhu<br /> <br /> 28 TAP CHI DlTOC HOC - 10/2010 (SO 414 NAM 50)<br />  N g h i e n CLFU - Ky thuat<br /> <br /> desloratadin va mong muon nghien cuu mot tinh tren che pham nguyen trang, ham i m 0,21%.<br /> phuong phap dinh lupng hoat chat nay, vi vay Trang thiet bi<br /> de tai nay dupe thuc hien nhim muc dich dua He thong sic ky Idng hi$u nang cao Alliance<br /> ra mpt quy trinh djnh lupng desloratadin trong 2695, diu dd day diod quang (Waters, My). Can<br /> che pham va theo doi dd on djnh cua phan tfch Sartorius CP 224D (Sartorius, Due).<br /> desloratadin trong qua trinh xay dung cong thuc May do pH Metrohm 744 (Metrohm, Due). May<br /> bao che siro va thanh pham sau nay. sieu am Elma (Elma, Due). Tu siy Memmert<br /> WM 500 CO (Memmert, Due).<br /> Nguyen lieu va phiFong p h a p<br /> Dung mdi va hda chat<br /> Chat ddi chieu, trang thiit bj, dung mdi va<br /> Acid phosphoric 85%, kali dihydrophosphat,<br /> hda chit natri hydroxyd, acid hydrocloric, hydroperoxyd,<br /> Chat doi chieu triethylamin dat tieu chuin phan tich (Merck).<br /> Desloratadin - So 16: 169011006 - Vien Kiim Acetonitril dat tieu chuin dung trong sic ky long<br /> nghiem thuoc TP Ho Chi Minh, ham lupng 99,64% (Merck).<br /> Ddi tupng nghien ciru<br /> Bang 1: Cac chi phim nghi§n ciru<br /> Biet dwov Ham luong Dang bao che Sold Han dung Nha san xuat<br /> Aerius 5 mg Vien bao phim 9STBA78A04 05/2011<br /> • SP Laboratories<br /> Aerius 0,5 mg/ml Siro 8ETSA94002 10/2011<br /> Phuong phap nghien cuu Dung dich miu thd<br /> Dua vao ciu true desloratadin la mpt base Vien desloratadin: cac vien bao phim dupe<br /> huu co c6 tinh phan cue trung binh va tham b6c bd mang bao, can rieng bidt 20 vien, xac<br /> khao mdt so tai lieu " ', chung toi d§ ap dung ky<br /> [1 2<br /> djnh khoi lupng trung binh cua 1 vien. Nghien<br /> thuat sic ky long hidu nSng cao pha dao vdi vien thanh bdt mjn, trdn deu. Can chinh xac<br /> pha ddng bao gom acetonitril va nude acid. Sic khoang mdt lupng bdt thuoc tuong duong 10,0<br /> ky do dupe ghi tai budc s6ng hip thu cue dai mg desloratadin, cho vao binh djnh muc 100 ml.<br /> cua desloratadin. Sau khi tim dupe dieu kien Them pha ddng khoang 65 ml, sieu am khoang<br /> sic ky thich hpp se thim djnh phuong phap 5 phut. Them pha ddng vua du the tich, lie diu,<br /> bao gom khao sat tinh phu hpp cua he thong, lpc, bd 20 ml djch lpc diu. Lpc tilp qua mang<br /> tinh dac hieu, dd dung, dp l§p lai va tinh tuyin<br /> lpc millipore 0,45 urn.<br /> tinh. Quy trinh sau khi dupe thim dinh se dupe<br /> ung dung de djnh lupng mdt so che phim chua Siro desloratadin: dung buret liy 20 ml siro<br /> desloratadin c6 tren thj trudng. Oe ap dung quy (tuong duong vdi 10 mg desloratadin), cho vao<br /> trinh nay trong viec theo ddi dp on djnh cua binh djnh muc 100 ml. Them pha ddng vua du thi<br /> desloratadin trong qua trinh xay dung cong thuc tich, lie diu, lpc qua mang lpc millipore 0,45 urn.<br /> bao chi siro va khao sat d6 on djnh cua thanh Dung djch miu tring: dupe chy|n bj tuong<br /> phim sau nay, desloratadin va che phim tu miu thu nhung khdng cd chit khao sat.<br /> nghien cuu dupe khao sat dudi cac dieu kien K i t q u a va ban l u a n<br /> nhu chiiu anh sang true tilp trong 48 gid, d§t d Khao sat diiu kien phan tich<br /> 100°C trong 48 gid, thuy phan trong dung djch<br /> Trong qua trinh thuc nghiem, cac yeu to anh<br /> HCI 0,5 N trong 48 gid, thuy phan trong dung<br /> djch NaOH 0,5 N trong 48 gid va oxy hda bdi hudng din hieu qua tach va dd chpn lpc cua<br /> H 0 3% trong 24 gid. Muc dich cua chung ldi phuang phap da dupe khao sat nhu pH nude<br /> 2 2<br /> <br /> nhim chung minh ring quy trinh vin bao dam acid, ty la dung mdi, thay the nude acid bing<br /> tinh d$c hieu ngay ca khi c6 mat cua san phim dung dich dem phosphat cd pH acid, them vao<br /> phan huy tu desloratadin. triethylamin d i cai thidn he so doi xung cua pic<br /> Chuan bi mau desloratadin. Chung tdi da xac djnh dupe diiu<br /> Dung dich miu ddi chiiu: can chinh xac kidn sic ky thich hpp de tach hoan toan<br /> khoang 10,0 mg desloratadin, cho vao binh djnh desloratadin (ke ca trong trudng hpp cd san<br /> muc 100 ml. Them pha dpng khoang 65 ml, sieu pham phan huy) vdi thdi gian phan tich ngin va<br /> am khoang 5 phut. Them pha ddng vua du the h$ so doi xung cua pic desloratadin dat yeu ciu,<br /> tich, lie diu, lpc qua mang lpc millipore 0,45 urn nim trong khoang 0,8 - 1,5 .<br /> <br /> TAP CHI DUQC HQC - 10/2010 (SO 414 NAM 50) 29<br /> • N g h i e n CLFU - Ky thuat<br /> <br /> <br /> <br /> <br /> Hinh 1: Sic ky dd dung djch miu ddi chiiu (a), dung dich miu thu- - vien desloratadin (b), dung<br /> dich miu thu - siro desloratadin (c), desloratadin duoc chiiu anh sing true tiip trong 48 gid (d),<br /> desloratadin duoc dat d 100°C trong 48 gid (e), desloratadin bj thuy phin trong HCI 0,5N trong 48 gid<br /> ( f ) , desloratadin bi thuy phin trong NaOH 0,5N trong 48 gid (g), desloratadin bj oxy hda bdi H 0 3% 2 2<br /> <br /> <br /> trong 24 gid (h), miu tring (i), siro desloratadin duoc chiiu inh sing true tiip trong 48 gid (J), siro<br /> desloratadin duoc dit d 100°C trong 48 gid (k). Dieu kien sac ky: cdt Ascentic RP C8 (250 x 4,6 mm;<br /> 5 pm), pha ddng: ACN - dem phosphat 10 mM pH 2,5 cd TEA 30 mM (25:75, tt/tt), tdc dd ddng: 1,2<br /> ml/phut, budc sdng phit hien 278 nm, thi tich tiem miu 20 ul, nhiet dd cdt: nhidt dd phdng.<br /> Nhin xef: Trong cac mlu desloratadin khao giam It, dac biet mau bj oxy hoa bing H 0 3%<br /> 2 2<br /> <br /> <br /> sat, mau de a nhiet do 100 C trong 48 gid, trong 24 gid co ham lugng giam dang ke. Mau<br /> miu dugc thuy phan trong HCI 0,5 N trong 48 siro desloratadin d l d nhiet dp 100°C trong 48<br /> gid c6 ham lugng thay doi khong dang ke, miu gid co ham lugng giam nhiiu, trong khi dudi<br /> dugc thuy phan trong NaOH 0,5 N trong 48 gid dilu kien anh sang true tilp trong 48 gid thi<br /> va dugc chieu anh sang true tilp trong 48 gid giam It.<br /> Tham djnh phuong phap<br /> Khao sat tinh phu hop cua he thong<br /> Bang 2: Kit qua khio sat tinh phu hgp cua he thdng cua phuong phip (n=6)<br /> Mau Gia trj thong ke to (phut) S k' As Ntk<br /> Xib 5,686 2.916.832 3,9 1,3 8.229<br /> Doi chieu SD 0,006 11.038 0,0 0,0 104<br /> RSD 0,10% 0,38% 0,13% 0,51% 1,27%<br /> Xib 5,720 2.957.629 3,9 1,3 8.524<br /> Vien desloratadin SD 0,003 13.303 0,0 0,0 60<br /> RSD 0,05% 0,45% 0,06% 0,29% 0,64%<br /> Xib 5,731 2.858.242 3,9 1,3 8.725<br /> Siro desloratadin SD 0,006 4.278 0,0 0,0 94<br /> RSD 0,11% 0,15% 0,14% 0,69% 1,08%<br /> sdNd7a<br /> fa: thdi gian Iwu, S: dien tich dinh, W: he so dung lugng, A : h$ so doi xung,<br /> s b k. ly thuyet bieu kien<br /> <br /> <br /> 30 TAP CHI DU'QC HQC - 10/2010 (SO 414 NAM 50)<br /> • N g h i e n CLFU - Ky thuat<br /> <br /> Nhan xet: He s6 d*6i xung co gia trj trung desloratadin khong co cac thanh phin khac<br /> binh khoang 1,3. RSD cua cac thong s6 sac ky trong tit ca cac miu.<br /> cho cac lan tiem lap lai cua mau 6*6i chieu va Nhu vay quy trinh dinh lupng desloratadin<br /> mlu thi> diu nho han 2%. Vay quy trinh dinh dat tinh chpn lpc cua mpt quy trinh dinh lupng<br /> luqng desloratadin dat tinh phu hpp cua he hoat chit trong chi phim va co the dupe ung<br /> thing l | 3<br /> dung d l theo doi dp on dinh cua desloratadin<br /> Tinh dac hieu trong qua trinh xay dung cong thuc bao chi siro<br /> Ket qua khao sat tinh dac hieu cho thiy: miu va thanh phim sau nay.<br /> tring (hinh 1i) khong co pic trung vdi pic Dp dung, dp lap lai va khoang tuyen tinh<br /> desloratadin va thdi gian luu cua pic chinh trong Bang 3: Kit qua khao sat do dung, do lap lai<br /> sic ky d l cua miu thu (hinh 1b, 1c) va miu va khoang tuyin tinh<br /> khao sat (hinh 1d, 1e, 1f, 1g, 1h, 1j, 1k) tuong Desloratadin<br /> duong vdi thdi gian luu cua pic desloratadin Phuong trinh hdi qui y = 29769x; R = 0,9997<br /> 2<br /> <br /> <br /> trong sic ky cua cua miu doi chiiu (hinh 1a).<br /> Khoang tuyen tinh (|ig/mi) 25- 150<br /> Khi them cung lupng chit doi chiiu desloratadin<br /> vao miu thu, chiiu cao va dien tich dinh cua pic 06 lap lai (n=6)<br /> desloratadin tang len glp doi so vdi trudc khi Vien desloratadin RSD = 1,39%<br /> them. Cac miu siro de d nhiet dp 100°C trong Siro desloratadin RSD = 0,42%<br /> 48 gid, miu desloratadin bi oxy hoa bdi H 0 2 2<br /> Op dung (n=9)<br /> 3%, miu desloratadin bj thuy phan trong NaOH Vien desloratadin RTB = 98,68 %<br /> 0,5 N co xuit hien pic la va pic desloratadin tach Siro desloratadin RTB = 97,96%<br /> hoan toan cac pic nay. Pho UV-Vis tai thdi gian RTB: ty le hoi phuc trung binh<br /> luu cua pic desloratadin trong miu thu va miu Kit qua thing ke cho thiy quy trinh phan<br /> khao sat giong pho UV-Vis tai thdi gian luu cua tich co khoang tuyln tinh rpng vdi he so tuong<br /> pic desloratadin trong miu dli chiiu. Su dung quan cao, ty le hoi phuc nim trong khoang 97%<br /> chuc nang kiim tra dp tinh khilt pic cho thiy pic -103% va RSD < 2% doi vdi dp lap lai.<br /> Dinh lupng mpt sd c h i phim chii>a desloratadin cd mat tren thi tru-dng<br /> Bang 4: Kit qua dinh luong mot sd chi phim tren thj truong (n=6)<br /> Ham lupng Ham lugng<br /> Biet dugc S6 lo Han dung Nha san xuat RSD (%)<br /> (mg/vien) so vdi nhan (%)<br /> Delopedil 5 mg 060809 08/2011 OPV 5,08 101,68 ±0,09 0,12<br /> Sedno 5 mg 0021209 12/2011 BV Pharma 5,44 108,86 ± 0,17 0,21<br /> Navana Pharma Ltd.<br /> Eurodesa 5 mg 24 12/2012 4,97 99,30 ± 0,20 0,27<br /> Bangladesh<br /> Synmedic<br /> Desler 5 mg DA 902 08/2011 4,93 98,62 ±0,13 0,18<br /> Laboratories<br /> Nhan xet: Ham lupng desloratadin trong tit ca Summary<br /> cac chi phim khao sat deu nim trong khoang A reversed-phase high performance liquid<br /> 90% -110% so vdi ham lupng ghi tren nhan. chromatography method for measurement of<br /> Ket luan desloratadine in pharmaceuticals was<br /> Bing phuang phap sic ky long hieu nang developed and validated. Separation efficiency<br /> cao vdi diu dd day diod quang, chung tdi da and selectivity were optimized using mobile<br /> xay dung dupe quy trinh djnh lupng phase containing acetonitrile and either acidic<br /> desloratadin. Phuong phap cd tinh dac hieu, water or phosphate buffer of different acidic phi's<br /> dung va chinh xac, cd thi ung dung d l dinh in variable ratios. Additionally, triethylamine<br /> lupng desloratadin trong mdt so chi phim va (TEA) which was added into mobile phase, was<br /> theo ddi dp on dinh cua chit nay trong qua trinh investigated for improvement of peak tailing.<br /> xay dung cdng thuc bao chi siro va theo ddi dp Complete separation of desloratadine was<br /> on dinh cua thanh phim sau nay. achieved with an Ascentic RP C8 column (250 x<br /> <br /> TAP CHI DUOC HQC - 10/2010 (SO 414 NAM 50) 31<br /> • 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 /> <br /> <br /> <br /> 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 /> <br /> <br /> 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 /> <br /> <br /> 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 /> <br /> <br /> <br /> <br /> Trudng dai hoc Duqc Ha Noi<br /> 1<br /> <br /> <br /> <br /> <br /> Vien Kiem nghiem An toan Ve sinh Thuc pham Qudc gia<br /> 2<br /> <br /> <br /> <br /> <br /> 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 /> <br /> <br /> <br /> <br /> 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 /> <br /> <br /> <br /> <br /> 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 /> <br /> <br /> <br /> <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 /> <br /> <br /> <br /> <br /> 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 /> <br /> <br /> 32 TAP CHI DUQC HQC - 10/2010 (SO 414 NAM 50)<br />