TAP CHI §) ¥ DUOC HOC QUAN SU Journal of Military Pharmaco-medicine ISSN 1859-0748 Vol 37, N°, thang 1/2012N 2 > a et @ 10. TAP CHI Y - DUGC HOC QUAN SU SO 1-2012 myc Lyc . Chiét xudt phan lap mot s6 phenolic glycoside tty Qué chi Viét Nam (Cinnamonum cassia Blume). Dinh lugng déng théi mudi hoat chat sinh hoe trong Bai hoang bang sac ky léng hiéu nang cao, Nghién c(tu bao ch& gel chita tiéu phan nano vitamin K, Nghién citu bao ché va buéc déu danh gid kha nang giai phéng in vitrro cla nha tung nano nhé mat diclofenac . Nghién citu dic diém gidi phu dong néng gitfa bap chan. . Bac diém gidi phdu nhanh lén dong mach mii dui ngoai 6 ngudi Viet Nam. . Bién déi cla déng mach gian cét sau va nh huSng t6i thiét ké vat gian cét c&ng tay sau nguge dong. . Két qua nghién ctu budc déu mot sé chi tiéu nhan tric od ban va tinh trang dinh duGng ngudi truéng thanh huyén Ba Vi. . Thyc trang kién thde, thai d6, thyc hanh cla nhan vién y té trong quan ly va xir ly chat thal y 16 tai 10 bénh vién Khu vyc pha Bac. Thye trang an ton vé sinh thyc phdm tai cAc cd 88 ché bién thuc phém & mot so tinhithanh phd. Nguyén Minh Khdi Dao Van Bon Tran Minh Ngoc Chir Van Mén Nguyén Van Long Trinh Nam Trung Va Binh Duong Ng6 Thj Thu Trang Pham Thi Phuong Dung Nguy6n Tran Linh D&ng Thi Hién Va Ngoc Mai Nguyén Trén Linh Va Hau Dang Va Nhat Binh Nguy&n Tién Binh Tran Ngoc Anh Tran Bang Khoa Nguyén Bite Nghia Trén Sinh Vuong Nguyén Van Chuyén Déng Khdc Hung Chu Bute Thanh Nguyén Van Ba Nguyén Duy Bac Tr4n Ngoc Anh Trang 241 27 33 40 46 51 87‘TAP CHI Y - DUGC HOC QUAN sy sé 1-2012 NGHIEN CUU BAO CHE GEL CHUA TIEU PHAN NANO VITAMIN K, Ng6 Thi Thu Trang*; Pham Thj Phuong Dung"; Nguyén Tran Linh* TOM TAT Khdo sat anh huting ola lipid (alcol cetylic, axit tween 80, cremophor EL) én gidi phéng in vitro ct gel. Kt qua cho thay: hn hgp 0,5% alcol cetylic, 0, cho ty 18 giai phong due cht cao nha. * Tirkhéa: Vitamin K,; Lipid ran; Tiéu phan nano. stearic, suppocire) va chat didn hoat (span 80, ja vitamin K, tir hé tiéu phn nano fipid rn va ty 8% suppocire va 0,7% span 80, 1,5% tween 80 FORMULATION OF GEL CONTAINING NANOPARTICLES OF VITAMIN K, SUMMARY A study was conducted on the influences of lipids (cetyl alcohol, stearic acid, suppocire) and sufactants (span 80, tween 80, cremophor EL) on in vitro drug release. The excipient mixture of 0.5% cetyl alcohol, 0.8% suppocire, 0.7% span 80, 1.5% tween 80 showed maximum drug release. * Key words: Vitamin K,; Solid lipid; Nanoparticles. DAT VAN DE Vai nam gan day, vitamin K, xudt hién trong my phdm va ché phém diing ngoai 48 diéu tri finh mach mang nhén va cai thién nhitng trigu ching: ban dé, ban xuat huyét, vét tham tim do nhiéu nguyén nhan khac nhau, Nhung han ché cla né khi ding ngoai da la kha nang théim va duy tri tac dung trong théi gian dai. C6 nhiéu bién phap dé cai thign tinh thdm cda duge chat, trong d6 c6 hé tigu phan nano lipid ran vai nhiéu wu diém: han ché kich (ng, kéo dai thdi gian tac dung, bao phi bé mat, tang * Trting Bai hoc Dug Hé NOI Phén bién khoa hoc: GS. Nguyén Liém PGS. TS. Nguyén Van Minh hydrat héa Iép siing va tang tinh thdm cla duge chat. Hé tigu phan nano fipid ran con 6 kha nang khéa tia tir ngoai (UV). Do vay, bao vé duge vitamin K,, m6t dude chat nhay cm véi anh sdng trong Idi lipid rn gidp tranh khdi suf phan hily cilia tia UV. Trén thé Qidi, 48 c6 nghién ctu vé bao ché tiéu phan nano chia vitamin K,, nhigu ché phdém dang gel, cream chita vitamin K, ty do va cA ché phdm chifa hé tigu phan nano vitamin K,, Nhung 6 Viet Nam, cha cé nghién oftu nao vé van dé nay. Do vay, nghién ctu bao ché gel chita tiu phan nano vitamin K, 18 cn thiét.Tae cHI ¥ DUOC HOC QUAN SY $6 1-2012 Trong bai nay, ching tdi théng bao két qua nghién cits buée dau vé bao ché hé tiu phan nano lipid ran chifa vitamin K,, gel chita hé tiéu phan nano va két qua danh gia nh hung cila cac thanh phd trong hé tigu phan nano dén kha nang gidi phéng in vitro cla vitamin K, tiv hé tiéu phan nano va gel. NGUYEN LIEU, THIET B] VA PHUONG PHAP NGHIEN CUU 4, Nguyén ligu va thiét bi. - Vitamin K,, alcol cetylic, axit stearic, suppocire, cremophor EL, span 80, tween 80, carbopo! 940, glycerin, triethanolamin, ethanol tuyét di, natriclorid, nati dihydrophosphat, natri hydroxyd theo tiéu chudn duge dién Trung Quéc hoa tigu chudn nha s&n xuat. Axit phosphoric, axit acetic, methanol tinh khiét dling cho HPLC. - May sidu am LABSONIC® M, may khudy tir IKA - WERKE (Bifc), may quang phé UV-VIS Cary 50, may do quang phé UV-VIS HITACHI U1800 (Nhat), may do pH EUTECH INSTRUMENTS pH 510, may ly tam HERMLE 2200 A, kinh hién vi dign tr truyén qua JEM 1010, may phan tich kich thu6c hat Zetasizer Nano ZS90, hé théng anh gid giai phéng thuéc qua mang Hanson Research, h@ théng HPLC THERMO- FINNIGAN va mt s6 thiét bj khac, 2, Phuong php nghién etfu, - Phutang php bao ché: + Bao ch hé tiéu phan nano vitamin K,: b&ng phuong phép sidu am két hgp véi khudy ti, Bao ché méi m&u 100 mi chita 0,25 g vitamin K,, 3 logi lipid (aicol cetylic, axit stearic, suppocire), 2 loai ch&t din hoat 22 than nudc (tween 80, cremophor EL) vay loai chat dign hoat than dau (span 89) yg 9 Ié khac nhau. + Bao ché ché phém gel ti hé tigu pha, nano vitamin A palmitat: bao ché 100 mi gy véi cong thifc: carbopol 940: 0,4 g; hé tigy phan nano vitamin K,: 50 ml (tttong duong v6i 0,125 g vitamin Ky); glycerin: 10 mi; triethanolamin: 1 ml; nuéc c&t vita dé: 100 mi - Phuong phap danh gid hé tiu phan nano: + Danh gid giai phng vitamin K, ti hg tigu phan nano bang hé théng gidi phéng qua mang Hanson Research (mang cellulose acetat 0,2 ym). Méi trudng gidi phéng la h6n hgp ethanol: dung dich dém phosphat pH 7.5 (1:4) 6 37°C. Téc 46 khudy 400 vong/phit. Thé tich mau thit: 0,5 ml, Sau 30 phit I4y mAu mét lan trong 3 gid. Xac dinh ham lugng vitamin K, gidi phong bang phuong phap quang phé hap thu UV é budc song 277 nm, + Dinh lugng ham lugng vitamin K, trong hé tiéu phan nano: sit dung hé théng HPLC Thermo Finnigan véi cdc théng sé: cdt Altech C18 (250 x 4,8 mm, 5 um), t6c 4 dong 1,2 mliphit; detector UV (buéc séng phat hién 226 nm); thé tich tiém m&u 20 pl; pha déng la hon hgp methanol: dung dich axit acetic 0,25 M (99:1), Mau thét: 2,0 ml hé tigu phan nano duge hda tan trong ethanol dun néng vita di; 50 mil. Lam lanh é lipid két tia, ly tam lay phn dich va pha loang 10 an bang pha déng. + Banh gia hinh thai va kich thuéc tiéu phan cia hé tiu phan nano bang kinh hién vi dign te truyén qua. + Danh gid kich thuée va phan bé kich thutc ti8u phan bang may Zetasizer Nano 2890,~ Phuong phap danh gia gel chifa hé tiéu phan nano vitamin K;: + Danh gié giai phéng vitamin K, tif gel: mang cellulose acetat 0,45 um, khdi lung gel thi 0,25 - 0,35 g véi diéu kién va thiét bi thi tueng tt nhu phuong phdp danh gia gi phéng vitamin K, tir hé tiéu phan nano. TAP CHI ¥ - DUOC HOC QUAN SU Sé 1-2012 + Binh luong néng do vaamin K. trong gel bang HPLC v6i thiét bi va diéu kién sdc ky tuong ty nhu phuong phap dinh fuong vitamin K, trong hé tiéu phan nano. Mau thi la 1,0 g gel phan tan trong hén hop nuéc: ethanol (25:75) & 70 - 80°C tao thanh 100 mi dung dich va pha lodng 5 ln bang pha dong. KET QUA NGHIEN CUU VA BAN LUAN 1. Khao sat anh huéng cila cac thanh phn céng thie dén Iugng vitamin K, giai phéng tir hé tiéu phan nano. Dua vao phuong phap thiét ké mat hop tit tai tm. phan mém MODDE 8.0. chang tdi the duge thiét ké thi nghiém. Bang 1: Bang thiét ké thi nghiém. cone ee THUC ‘Lugng CM] ‘Suppocire | Logi CDH | Luong CDH Span 80" Nuke od (g) (9) than nue | than nude (9) (g)__| visa Gi (rm i [95 Ooi ececmal eas 03 100 2 NS O.S ei |e 0,6 eee ees 2 i 03 | 100 3 25 | 08 cet 2 07 160 4 2508 | C&L, 7807 100 5 sgh ae oamml acer 2) 07 100 é oo st ee hLrhmhUhCLcT Chr 7 25 | 2 | ce 15 | 03 100 8 025 | AS 25 Z ca | Zi {031 100 Ea 0,25 AC 0S 08 Teo | 15 ~~! «(OF | 10 10 02 [as | os | 08 wo [2 | o7 100 1 025 | Ac | 28 08 80 2 | 63 100 12 @25 [as | 25 08 To 5 | 03 (100 3 (025m ACE | anos 2 Teo 2 03 100, [44 02 | AS | 05 2 780 1S | 03 | 100 16 025 | AC | 25 2 Te Ti 16 025 | AS | 25 2 a0 2 a7 {10 7 025 [ac] 15 14 ca 475 | 05 | 10 18 0,25, AC 45 44 4 CEL 175] 05) ) 100 | 19 ozs [ac | 45 [ 14 | ce frie (ake. sme eenvOo Ney! (AC: alcol cetylic; AS: axit stearic; CEL: Cremophor EL; T80: tween 80) Ching t6i chi tién hanh bao ché 15 céng thie trong bang thiét ké thi nghiém bing phucng phap siéu &m va bé qua cdc ong thie 4, 8, 12 va 16 (do lung axtt stearic ion anh hudng d6n thé ch&t). Sau 46, danh gia giai phéng. 23TAP CHIY - UGC HQC QUAN SY SO 1-2012 Bang 2: Két qua danh gia d6 gidi phong cua vitamin Ky ter cdc mu theo cong thic thidt qg [cone PHAN TRAM VITANIN K; GIAI PHONG (%) @ CAC GIO THUS — i 18 a" 25 | a 1 23,88 23,29 44,19 _ 36,82 seo | 4 =e) 2 43,32 59,72 51,39 53,24 53,16 ee6 | 3 oss =| 13,22 | 965 7,36 5,98 ova | 5 15,68 48,48 | 21,08 2505 | 26686 | 27.08 | 6 10,92 16,68 31,23 44,61 53,39 61.41 7 366 | re | rt 3,21 3,42 390 | “o “4.19 4 13.15 | 28,56 46,45 65,76 wo | OMo 4 472 315 | 5,29 " 010 =| 4,10 0,27 0,06 0,30 0,33 3 13,36 | 15,58 17.73 20,79 20,76 21,95 14 10,05 1244 | 44.33 12,96 13,76 14,26 | 15 1,66 361 7,08 8.94 11,47 43,01 To 2.08 4.22 737 10,85 12,82 18 Comes 530 | 716 | 900 937 19 161 aat [ae 8.47 7.76 374 Tiép tuc danh gia anh huéng ctia mot s6 thanh phén trong céng thie v6i sy tr¢ giup cba phan mém FormRules 2.0. ” 468" “ 0 ” ag mca 88 8 ety! ee Hinh 1: Mat dap clia Y, theo luong chat mang (alcol cetylic) va fugng CDH than nuéc {tween 80) (suppocire 0,8g, span 80 0.79). 24 Hinh 2: Mat dap ciia Y, theo lugng chat mang (axit stearic) va lugng CDH than nude {tween 80) (suppocire 0,8 g, span 80 0,79).; : Lugng chét mang alcol cetylic va iusng CDH than nutic (teen 80) anh huting dén ty 16 (%) vitamin K, gidi phong tir hé tiéu phan, Khi lugng chét mang alco! cetyiic tang ty 0.5 g lén 1,3 9, lugng vitamin K, gidi phong tirh@ tiéu phan nano gidm nhiéu. Trong khi 46, néu tang tis 1,3 - 2,5 g khong anh hudng t6i ty 18 vitamin K, gidi phong. Thong thudng, CDH tam tang t5c 44 gidi phong duge chit. Tuy nhién, trong trudng hgp nay. khi lugng alcol cetylic ding khoang tir 0.5 - 1.3.9 va tang khdi Iugng tween 80 thi ty 6 phan tram vitamin K, gidi phong gidm. Tuong ty nhu alcool Cetylic, lugng axit stearic cing anh hudng 16 16 vitamin K, gidi phong, nam trong gidt hen tU'0,5 - 0,9 g néu sit dung CDH than nutc la tween 80. Mé hinh anh hung tuong ty alco! cetylic, ty I@ vitamin K, giai phéng fudn nhd hon véi t&t c& cao mau (hinh 2), gz Phan tram vitamin Kigiai Phong (3) o ° 1 i ESS _—_ Tae CH Y - DUC HOC OUAN Sy $6 1.2 2 Két qua thu dugc cho thdy alco! cetyl: ph6i hop voi suppocire la chét mang tt hon cho hé tu phan nano. Chum thé két juan cremophor EL hay tween 80 cd higu qué hon, nhung miu bao ché voi tween 80 cb phan tram vitamin K, gldi phong cao nhéit ‘sau 3 gid. Do vay, chung tdi lya chon tween 80 cho nhiing nghién ctu tiép theo. 2. Dénh gid mot sé dic tinh cua he tiéu phan nano vitamin K,. Lf chon céng thie 9 l& céng thie cho mu 06 thé chat, t6c 46 gidi phong én dinh nhdt va lugng vitamin K, gidi phong lon nha sau 3 gid dé danh gid mot 86 dc tinh cla h@ tu phan nano. ~ Binh hugng ham lugng vitamin Ky, Ham Ivong vitamin K, trong hé tiéu phan nano Iipid r4n dugc xe dinh bling phuong phap HPLC det 91,2% so voi ly thuyét. + Danh gia sy gidi phéng vitamin K, tis be tiéu phan nano. 2 3 4 Hinh 3: Ty W8 (9) ca vitamin K, gidi phéng tir hé tiéu phan nano ching thie 9. 6 thi giai phéng trén cho théy: t 16 vitamin K, duge gla phéng tir hé tiéu phan trong gid du tuong 461 thp. Nguyén nhan 66 thé do thanh phén mang duigc chat trong ho tidy phan 1 céc 165i lipid rn. D8 gidi phéng khdi Ii nay, lipid phai duigc bao mon din dul the dong cia nhiet 6) va thdém Udt dung moi. Tuy nhién, sau 1 gid iugng vitamin K, gidi ph6ng ting nhanh dén, Seu 3 gid, 65,76% vitamin K, duc gidi phéng tir 1 Voi tS 6 tuong 66% dn dinh.Hinh 4: Anh chup hé tiéu phan nano cilia céng thitt 9 qua kinh hign vi dién ti truyén qua. H@ tigu phan tao dugce cé kich thudc tuong d6i nhé va déng déu. Bung kinh tiéu phan trung binh 92,64 nm. 99,8% tiéu phan 6 duéng kinh < 1.000 nm. Khoang phan bé Kich thu8c tuong déi hep, cha yéu nam trong khoang tl 10 - 300 nm. 3. Banh gié mét sé tinh chat cla gel chia hé tigu phan nano, Bao ché gel chia hé tiéu phan nano theo c6ng thifc 9 véi thanh phan da néu & tren. Ham lugng vitamin K, trong ge! duge inh lugng theo phuong phap HPLC dat 81,60% so véi ly thuyét. Béng 3: TY 18 (%) vitamin K, gidi phéng tir gel chita hé tiéu phan nano (Céng thifc 9) THacun | 1] 2 | 4] 8 | 8 Phd tim viternin K, gid phOng (%) 772 | 12,88 | 9,37 | 9,67 | 7.45 26 Luang vitamin K, giai ph6ng tang dan va dat gia tr) 16n nhat 6 thoi diém 2 gid, sau a6 duy tn va giém dan. Ty Ié vitamin K, gig phong tis gel tuong déi thap. Nguyén nhan 66 thé do ham Ivgng vitamin K; trong gel qua thap, duge chat nam su trong 16ilipia ran nén chua thé giai phéng hoan toan. KET LUAN Hé tigu phn nano lipid rn chita vitamin K, véi loai va ty 16 ta duge khac nhau da duge bao ché va danh gia mét s6 dac tinh Trén co 86 46, Ira chon céng thifc véi 0,5% alcol cetylic, 0,8% suppocire, 0,7% span 80 va 1,5% tween 80. Mau tiéu phan nano c6 thé chét, t6c dé gidi phéng én dinh nhat va ty Ié vitamin K, gidi phéng !6n nhat sau 3 gid. Hé tigu phan nano bao ché dude 6 tiéu phan hinh cu, kich thu tusng d6i nhé va déng déu. Chuing téi cing da bao ché gel tit hé tigu phan nano trén va sd b6 danh gid mét s6 dac tinh cla gel. TAI LIEU THAM KHAO. 1. Lé Thi Thém. Nghién ctu bao ché gei chifa tiéu phan nano vitamin A. Khéa tuan tot nghiép Duge sf Dai hoc khoa 2005 - 2010, Trudng Bai hoc Duoc Ha NOi. 2010. 2. Hou D, Xie C, Huang K, et al. The production and characteristics of solid fipid nanoparticles (SLNs). Biomaterials. 2003, 24 (10), pp.1781-1785. 3. Iscan Y, Wissing S. A, Hekimogiu S et al. Solid lipid nanoparticles (SLN) for topical drug delivery: incorporation of the lipophilic drugs N, N-diethyl-m-toluamide and vitamin K, Pharmazie, 2005, 60 (12), pp.905-809, __ 4. Jenning V, Gysler A, Schafer-Korting M et al. Vitamin A loaded solid lipid nanoparticles for topical use: occlusive properties and drug targeting to the upper skin. European Journal of Pharmaceutics and Biopharmaceutics. 2000, 49 (3), pp.211-218, 5. Liu C. H, Wu C. T, Fang J. Y. Characterization and formulation optimization of sold pid nanoparticles in vitamin K1 delivery. Drug Development and Industrial Pharmacy. 2010, 36 (7), pp.751-761.