Nghien
CLFU- Ky thuat
pho ehuin tuong duong nhau Pho IR ciia c h i t tong hgp duge cd eac dinh dae trung ciia cac nhdm chire: nhan thom Vc=c 1600-1500 cm"', 5N-H 1655-1620 cm"', Vc=o 1850-1650 em''. .
Diim chiy: 144-146 °C (tuong t u vdi felodipin nguyen lieu cung d p tir nha may Pymephareo).
Dinh luang vi mit khdi lugng do siy khd Ham lugng felodipin tinh tinh tren c h l pham s l y kho la 99,4% va do i m la 0,5%.
K§t luan
TLJ' eac nguyen lieu d i u la didorobenzaidehyd va methylaeeto acetat de tao MBI sau dd eho MBI phan irng vdi ethyl-3-aminocrotonat de tao felodipin. cae phan irng da duge nghien ciru cae d i l u kien toi uu (nhiet do, ti le moi cac c h i t phan irng, t? le moi c h i t xiic tac), d i l u kien tinh c h l . Felodipin tong hgp duge dap irng yeu c i u ciia Duge diin Anh 2007 ve djnh tinh, djnh lugng.
Qui trinh tong hgp hoan toan cd t h i triin khai san xuit thir va da dugc ap dung trong cac bai thue tap eho hoc vien ldp cao hoc tai khoa Dugc - Dai hoc Y Dugc TP.HCM trong nam 2008.
Summary
Optimization of synthetic conditions for preparing felodipine. a wellknown anti- hypertension agent.
The Hansch reaction was applied in two steps. The synthetic conditions (temperature, rate of reagents, catalyst, solvents for purification) were investigated.
A suitable synthesis procedure of felodipine was established in total yield over 45 % and the final product conformed to the specifications of felodipine by BP 2007.
Tai lieu tham khao
1. Astra Aktiebolag (1999), Manufacturing process for felodipin, US patent 5,942,624.
2. BP 2007 - CD Rom
3. Desai, Ranjit (1997), Process to prepare dihydropyridine and derivatives there of, PCr/L/S96/20383
4. Duang Chi Uy (2006), Tang huyet ap : hudng din dilu tri ciia WHO/ISH & JNC VII, Thai su tim mach hoc, 99, tr. 6-8
5. MedlMedia Asia (2003), CD-Rom
6 Pharmaceutical Press (2004), Clarke's analysis of drugs and poisons, 3"^ edition, volume II, pp. 1018- 1019.
iTng dung tac nhan doi quang 2 - hydroxypropyl beta - cyclodextrin trong phan tich dong phan amlodipin bang
phu'O'ng phap dien di mao quan
Le Thj Thu Cue", Nguyen Ngoc Vinh', Dang Van Tjnh^, Nguyen Dure Tuan^*
Vien Kiem nghiem thuoc TP Ho Chi Minh Khoa Dirge, trudng dgi hoc Y Dirge TP Ho Chi Minh Dat v^n d§
Amlodipin la d i n c h i t cua 1,4-dihydropyridin, thudc nhdm chen kenh ealei cd tae dung keo dai, duoc sir dung ehu y i u de dilu trj eao huyit ap va dau thit ngue. Phin Idn cae c h l p h i m ehira hoat chat amlodipin d dang k i t hgp hai dang TAP CHi DUQC HQC - 10/2009 (SO 402 NAM 49)
dong phan R va S ciia amlodipin hay cdn gpi la hdn hgp racemk:. Hai dang dong phan tuy cd c l u triic hda hpe tuang t u nhau nhung khi vao CO the con ngudi eho tae dung khae nhau Dong phan S-amlodipin dugc xem la ed tae dung dan maeh va ha huyit ap '''.
41
Nghien CLFU - Ky t h u a t
Biet duge ehi ehira S-amlodipin da x u i t hien tren thj trudng An Dp vdi ten Asomex®. Cy Hoa Ky S-amlodipin da duge dua vao thir nghiem lam sang nhung ehua duge luu hanh tren thj trudng. Trong mpt nghien ciru g i n day eho t h i y c h l p h i m chira 2,5 mg S-amlodipin Quae xem la tuong duong vdi c h l p h i m ehira 5 mg h6n hgp racemic eua amlodipin '''.
De phan biet cae dang dong phan la mpt d i l u khd khan nen viee phan tach cae d i n g phan quang hpe ngay cang trd thanh v i n d l quan trpng trong ITnh vuc kiem nghiem dugc p h i m . Hien nay, xu hudng eua eac nha nghien ciru tren the gidi la di tir tac nhan ddi quang (chiral selector) pho b i i n n h i t la P-CD d l tong hgp ra nhilu tac nhan dli^ quang chuyen biet (hydroxypropyl P-CD, methyl p-CD, earboxymethyl p-CD,...) ed tinh chon Igc eao, tan d l dang trong eac dung moi n h i m tao d i l u kien thuan Igi eho eac nha phan tieh trong viec lua ehpn tae nhan tach ddng phan thieh hgp '^'. Dien di mao quan la mdt ky thuat r i t thuan Igi trong viee tach eac ddng phan quang hpe '^"^'. Vdi viee sir dung cae d i n c h i t cyclodextrin, cae nha nghien ciru da tach thanh cong dong phan eiia hai amin '®', amphetamin '''', ephedrin '®',... Trong bai bao nay ehiing tdi xin gidi thieu k i t qua irng dung phuang phap dien di mao quan vdi tae nhan d i i quang 2-hydroxypropyl beta-cyclodextrin d l tach ddng phan amlodipin.
N g u y e n l i e u v a p h u ' a n g p h a p C h i t ddi c h i i u , trang thilt bj, dung mdi va hda chit
Chat ddi chieu
Amlodipin besilat - S l 16: QT145031208 - Vien K i i m nghiem thuic TP H i Chi Minh, ham lugng 99,71%; dp I m 0,11%.
Trang thiet bi
May dien di mao quan Hevi^lett Paekard^^CE, d i u dd day died quang. Cdt mao quan silica nung chay, dudng kinh trong 50 pm, ehilu dai tong epng 64 em, ehilu dai hieu qua 55,5 em.
Dung moi va hda chat
2-hydroxypropyl beta-eyelodextrin (HP P-CD) da dugc tong hgp tai Vien Kiem nghiem t h u i c TP H i Chi Minh va da duge tieu chuan hda.
Methanol dat tieu ehuan dung eho s i c ky long
(Merck). Tris (hydroxymethyl)-aminomethan (TRIS) va acid phosphoric dam dae dat tieu ehuan phan tieh (Merck). • ^ - ^
Ddi tu^ang nghien cu'u
Vien nen amlodipin 5 mg (amlodipin besilat 5 mg), s l 16 80758, han dung 03/2011, nha san x u i t : Cipla, An 0 6 .
Phu'O'ng phap nghien ciru
Trong pham vi d l tai nay, chiing tdi m u i n khao sat kha nang irng dung ciia tae nhan d i i quang HP p-CD vao cdng viee tach ddng phan amlodipin b l n g ky thuat dien di mao quan.
Trong qua trinh thuc nghiem, cae y i u t l anh hudng den qua trinh tach d i n g phan amlodipin nhu pH va n i n g dp dung djch dien ly n i n , ning dp tac nhan d i i quang da d u g e khao sat. Sau khi tim duge d i l u kien tach ddng phan amlodipin, se khao sat tinh t u a n g thich he t h i n g va tham djnh qui trinh p h i n tieh qua viec khao sat dp dung, dp lap lai, tinh tuyen tinh va tinh dae hieu.
Chuan bj mau
Dung dich miu ddi chiiu: can chinh xac khoang 25 mg amlodipin besilat, cho vao binh dinh mire 50 ml, hda tan trong 20 ml methanol, sau dd them nude d t khu" khoang vu'a du the tieh, l i e d i u . L i y chinh xac 5 ml dung dich tren, cho vao binh djnh mire 50 ml, them nude cat khir khoang vira du the tich, l i e d i u . Lpe qua mang lpe millipore 0,2 pm.
Dung dich miu thir; can 20 vien, xac djnh khii lugng trung binh ciia mdt vien, nghiin thanh bpt mjn. Can mpt lugng bpt vien tuong irng 5 mg amlodipin besilat, eho vao binh dinh mire 100 ml, them 2 ml methanol, l i e ky, sau dd them khoang 70 ml nude e l t khir khoang, sieu am khoang 15 phiit, de nguoi, th6m nude cat khir khoang vij'a dii the tieh, l i e deu. Lpe qua g i i y lpe (loai bo 20 ml djch lpe d i u ) . Djch lpe sau dd dugc Ipc t i i p qua mang lpe millipore 0,2 pm.
Dung dich miu tring: d u g e e h u i n bj tuang t u mau thir nhung khdng cd c h i t khao sat.
K i t qua va ban luan
Khao sat d i l u kien phan tich
Anh hwang pH va nong dp cua dung djch dien ly nen
42 TAP CHI DUQC HOC - 10/2009 (SO 402 NAM 49)
Nghien CLFU - Ky thuat
* — S- va R-am lodipin
2.5 3.5 4.5 5.5 pH dung djch dien ly nen
<n c Jl
a.
•o-O 2.5 1
2 1.5 1 0.5 0
- • — S- va R-am lodipin
25 50 75 N6ng do dung djch dien ly n4n (mM) Hinh 1 : Anh hudng pH dung djch dien ly nen
tren do phin giai ddng phin amlodipin.
Dieu kien dien d i : cot mao quan 64 cm (55,5 cm) x 50 Mm, 25°C, 20 kV, 50 mbar x 10 giay, 214 nm, 5 mM HP
|1-CD, cac dung dich dien ly nen TRIS 50 mM c6 gia tri pH khac nhau.
Anh hirang ndng do tic nhan ddi quang HP p-CD
Khao sat anh h u d n g nong d d HP p-CD (trong dung djch T R I S 50 m M , pH 2,5) d cae gia tri 5 m M , 10 m M , 20 m M .
Hinh 2: Anh hudng ndng do dung dich dien ly nin tren do phan giai ddng phin amlodipin.
Oieu kien dien d i : cot mao quan 64 cm (55,5 cm) x 50 nm, 25°C, 20 kV, 50 mbar x 10 giay, 214 nm, 5 mM HP H-CD, cac dung dich dien ly nen TRIS pH 2,5 c6 ndng do khac nhau.
3.5 :2 2.5 H '2 1.5
Q 0.5- -0.5
- • — S- va R-amlodipin
1 1 1
5 10 20 N6ng dp dung dich HP beta-CD (mM) Hinh 3: Anh hudng ndng do HP p-CD tren do phin giii ddng phin amlodipin. Oieu kien dien d i : ept m a o quan 64 em (55,5 em) x 50 )am, 25°C, 20 kV, 50 mbar x 10 giay, 214 n m , dung dich dien ly nen T R I S 50 m M pH 2,5.
Nhin xet:
- Khi pH dung djch dien ly nen tang thi d d phan giai g i a m . Cy pH d u d i 3,5; hai pic ddng phan tach hoan toan, t h i hien qua tn sd dp phan giai tren 1,5.
- Khi nong d6 dung djch dien ly nen tang thi dd phan giai tang. Cy nong dp dung djch dien ly nen tr§n 50 m M , hai pie dong phan tach hoan toan.
- Khi nong d d H P p - C D tang thi dp phan giai va t h d i gian di c h u y e n d i u t a n g . Cy ndng d p H P p - C D til' 10 m M , hai pie d d n g phan tach hoan toan n h u n g t h d i gian di c h u y i n tang len rd ret tij' nong dp 10 m M d i n 2 0 m M .
Qua khao sat trSn, ehpn nong d p d u n g dich di$n ly n i n 50 m M , pH 2,5 v a n o n g d p H P p- TAP CHi DUQC HQC - 10/2009 (SO 402 NAM 49)
30
" =-25-1 .5 -= 20 H
15
- • — P i c d 6 n g phan 1 - B - Pic d6ng phan 2
5 10 20 Nong dp dung djch HP beta-CD (mM) Hinh 4: Anh hudng ndng dd HP f3-CD tren thdi gian di chuyin ciia ddng phin amlodipin.
D i l u k i e n d i e n d i : giong n h u hinh 3
C D 10 m M la thieh h g p ve d p phan giai va t h d i gian phan tich ( x e m hinh 5a vi 5b trang sau).
K h a o s a t t i n h tu'O'ng t h i c h h e t h d n g
B a n g 1 : Kit qua khao sit tinh tuang thich he thdng cua phuang phip (n=6)
M i u O d n g p h a n RSD
Dp phan amlodipin Thai gian di Corr gi^j
chuyen Area
Chuan
Pic
dong phan 1 1,72% 0,89%
Pic ddng phan 2
2,28 1,73% 0,80%
- . .- •, 1.64% 0,83%
dong phan 1 „ „ „
Thu ^-^ 2,33 Pic
dong phan 2 1,63% 0,86%
43
Nghien CLFU - Ky thuat
K i t qua t h i n g ke eho thay, sau 6 l i n dien di thdng s l CorrArea va thdi gian di chuyen ed
RSD < 2%; gia trj dp phan giai Idn han 1,5. Vay phuang phap dat tinh t u a n g thich he thong.
(5a) . . , _ . ^^^^
Hinh 5a va 5b. Dien di dd dung dich miu ddi chiiu vi miu thir (hdn hgp racemic cua amlodipin).
Di4u kien dien di: dung dich dem TRIS 50 mM, pH 2,5 chira 10 mM HP |3-CD. Cot mao quan silica nung chay, chiSu dai tong cong 64 cm, chiiu dai hieu qua 55,5 cm, dudng kinh trong 50 jam. Nhiet dp cot 25°C. Di#n thi 20 kV. Kieu tiem miu 50 mbar x 10 giay. Bude song phat hien 214 nm.
Tham djnh phu'ang phap Tinh die hieu
T i l n hanh dien di m i u t r i n g , m i u d i i ehilu va m i u thir theo d i l u kien dien di nhu hinh 5.
K i t qua eho t h i y m i u t r l n g khdng ed pie trung vdi pic c h i t phan tieh. Dien di d l m i u thu g i l n g dien di dd m i u d i i ehilu. Khi them c h i t ddi ehilu amlodipin vao m i u thir thi dien tieh dinh eiia ehit them vao tang. Pho tir ngoai tai thdi gian luu ciia cae pie trong m i u thir g i l n g pho tir ngoai eiia m i u doi ehilu. Do tinh khiit cua cae pic Idn hon 99%. Hai pic eua hai dang d i n g phan cd pho UV-Vis giong nhau. Vay phuong phap ed tinh dae hieu.
Dp dung, dp lap lai va khoang tuyen tinh Bang 2: Kit qua khao sit dd dung, dd lap lai vi khoang tuyin tinh
Pic dong phan 1 Pic ddng phan 2 Phuo'ng trinh
hoi qui
y = 0,0011x- 0,0002; R =
0,9995
y = 0,0011x- 0,001; R = 0,9994 Khoang tuyen
tinh (ng/ml) 1 - 4 0 0
Do lap lai (n=6) RSD = 0,27% RSD = 0,27%
Do diing (n=9) RTB = 98,52% RTB = 100,21%
RTB: ty le hdi phuc trung binh.
K i t qua thong ke eho t h i y ty le h i i phue n l m trong gidi han eho phep ciia ty le h i i phue ly thuyit (98% - 102%) va RSD < 3% ddi vdi dp lap lai Qui trinh phan tieh co khoang t u y i n tinh rpng vdi he so tuang quan eao.
44
Ket luan
B l n g phuang phap dien di mao quan va irng dung tac nhan HP p-CD tong hgp tai eo sd, chiing tdi da xay dung duge qui trinh tach hai dang d i n g phan trong hon hgp racemic eua amlodipin. Phuang phap xay dung ed tinh dae hieu, cho k i t qua ehinh xae va ed tinh lap lai. Cac k i t qua thu dugc cung n h i m gdp p h i n irng dung phuang phap dien di mao quan vao viec kiim tra tap dong phan cua nguyen lieu amlodipin va phan biet cae dang d i n g phan trong eac thuie chira amlodipin dang luu hanh tren thj trudng.
Summary
A capillary electrophoretic method for separation of the enantiomers of amiodipine was established and validated. The two enantiomers were separated by a fused-silica capillar with TRIS running buffer (50 mmol L'\ pH 2.5) containing 10 mmol L'^ of hydroxypropyl j8- cyclodextrin (HP P-CD). The effects on the separation of the buffer pH, buffer concentration, and the concentration of HP jS-CD were investigated. The range of quantitation for both enantiomers at 214 nm was 1 - 400pgmL'\
Intra-day relative standard deviation (RSD; n = 6) was < 3%. The recovery ranged at 98% - 102%.
The method can be applicable to quantitative determination of the amiodipine enantiomers in pharmaceuticals and chiral impurities in amiodipine raw materials.
(Xem tiep (rang 8)
TAP C H i DUQC HQC - 10/2009 (SO 402 NAM 49) i
