Nghien CCFU - Ky thuat
Ket luan
Khi bao ehl he phan tan rin eua curcumin vdi cac chit mang, dp hda tan eiia eurcumiri duge eai thien dang k l so vdi nguyen ligu ban diu, cae phugng phap bao ehl he phan tan rin anh hudng din mCre dp hda tan cua duge chit. So sanh dp hda tan cua eae cdng thCre he phan tan rin thu dugc kit qua edng thCre N_F3 bao ehl bing phuang phap nghiin udt eho dp hoa tan cao nhit vdi ty le curcumin:PVP K30:Tween 80 la 1:4:0,22 va HPTR nay on djnh trong 2 nam d diiu kien nhiet do mdi trudng ty nhien.
Tai lieu tham khao
1. Aggawal (2007), "Biovailability of curcumin:
Problems and promises", Molecular Pharmaceutics,
4(6), 807-818.
2. Ambike. A. Ambike et al. (2004), "Characterization of curcumin-PVP solid dispersion obtained by spray drying", Intemational Joumal of Phannaceutics, 271, 281-286.
3. Damian F. et al. (2002), "Physical stability of solid dispersions of the antiviral agent UC-781 with PEG 6000, Gelucire 44/14 and PVP K30", International Joumal of Phannaceutics, 244, 87-98.
4. Loftsson T. et al. (2002), "Studies of curcumin and curcuminoids XXVII cyclodextrin complexation: solubility, chemical and photochemical stability", intemational Joumal of Phannaceutics, 244, 127-135.
5. Vedamurthy Joshi, Wlohammed Gular Ahmed et al. (2010), "A comparative study: solution stability and dissolution behavior of solid dispersions curcumin", Indian Joumal of Novel Dmg delivery, 2(3), 88-95.
{Ngay nhan bar 13/09/2016 - Ngay phan bien: 28/09/2016 - Ngay duyet dang: 02/12/2016)
Xay drnig qui trinh phan tich dong phan quang hoc
A r
(iS)-ibuprofen bang phu'ong phap sac ky long hieu nang cao
V
Le Thi Thu Ciic', Cao Thi Thu HaS Nguyin Dire Tuka^'
'Vien Kiem nghiem thuoc TP Ho Chi Minh -Khoa Duac, Dgi hoc Y Duac TP Ho Chi Minh 'E-mail: [email protected]
Summary
A high perfonmance liquid chromatographic (HPLC) method for separation of the ibuprofen enantiomers was established and validated. The two enantiomers were separated by a Lux"^ Ceilulose-1 column with mixture of n-hexane-isopropanol-trifluoroacetic acid (98:2:0.1 as mobile phase with UV detection at 220 nm.
The resolution of enantiomers was more than 3.0. The linearity range of quantification for both enantiomers was 50-1000 pgmLi Intra-day relative standard deviation (RSD) was below 2%. The recovery range was 98 • 102%. The validated method showed applicable to quantitative determination of (S)-ibuprofen enantiomers in several phannaceutical preparations and chiral impurity in ibuprofen raw materials.
Keywords: Ibuprofen, chiral separation, Lux™ Ceilulose-1 column.
Dat van d l
Ibuprofen la thule khang vigm khdng steroid (NSAID), cd tac dung giam dau, ha s i t va khang viem. Do eiu triie ed 1 carbon bit dii nen ibuprofen ed hai ding phan dii quang la (S)-ibuprofen va (R)-ibuprofen, nhung chi cd dang (S)-ibuprofen la dang ed hoat tinh, edn (f?)-ibuprofen thi khdng. Mat khac (R)-ibuprofen lai gay tich tru trong md md do trao dii vdi
cac acyl eua triaeylglyeerol ndi sinh trong qua trlnh chuyen d i i nguge thanh (S)-ibuprofen. Cho d i n nay, tac dgng lau dai cua sy tich luy nay va nguy co gay hai cua nd v i n ehua dugc bilt •''.
Tilp theo eae edng trlnh nghign eiru v l ITnh vu-c phan tach ding phan quang hpe, trong bai bao nay, chung tdi gidi thieu k i t qua phan tich dong phSn quang hgc ('Sj-ibuprofen bing phuang phap s i c ky long hieu nang cao (HPLC).
22 TAP CHI DUOC F '
Nghien
CLFU- Ky thuat
Nguyen lieu va phuxyng phap
Chit ddi chilu, trang thilt bj, dung moi va hoa chit
Chat doi chieu
Ibuprofen: S l Id QT026 100715 do Vien Kiem nghiem thulc TP H i Chi Minh cung cip, ham lugng 99,67% tinh trgn ehl phim nguygn trang.
(S)-ibuprofen: S l Id SIB1308073 do Cdng ty TNHH Lien doanh Stada Viet Nam cung cip, ham lugng 99,80% tinh tren ehl phim nguygn trang.
Trang thiet bj
May sie ky Idng hieu nang cao Shimadzu UFLC 20A, diu dd day died quang SPD-M20A;
Cdt Lux™ Cellulose-1 (cellulose tris(3,5- dimethylphenyl earbamat) (250 x 4,6 mm; 5 |jm) va ept Lux™ Cellulose-2 (cellulose tris(3-eloro-4- methylphenyl earbamat) (250 x 4,6 mm; 5 pm);
Can phan tieh dien tu- Mettler Toledo AT200, b l sigu am Hwashin va eae dung cu thuy tinh loai A thudng diing trong phdng thi nghiem. Cac thilt bj phan tieh va dung eu phan tieh da duge hieu ehuin dat quy djnh theo GLP va ISO/lEC 17025.
Dung moi va hoa chat
n-Hexan, 2-propanol (IPA), ethanol (EtOH) va acid trifluoroacetie (TFA) dat tieu ehuan diing cho s i c ky Idng (J. T. Baker).
Ddi tu-o-ng nghien cu'u Chephim
Vien nen Stada fS)-ibuprofen 200 mg Vien nen (Sj-ibuprofen 200 mg Vien nang nngm (Sj-ibuprofen 300 mg
Nha san xuat
Cong ty TNHH Lien doanh Stada Viet Nam CongtyX
CongtyY
Solo 03/001 ZM411 15114008
Ngay san xuit 15/02/2016 21/09/2015 29/07/2014
Han ddng 15/02/2019 20/09/2018 28/07/2016 Phu-o-ng phap nghien cu'u
Dya vao eiu triic hda hgc cua ibuprpfen la mpt acid hO-u ca va qua tai lieu tham khao irng dung hgp chit polysaceharid lam pha tTnh bit dii d l tach ding phan quang hgd^^i. cdt Lux™ Cellulose-1 (cellulose tris(3,5-dimethylphenyl earbamat) (250 x 4,6 mm; 5 pm) va edt Lux™ Cellulose-2 (cellulose tris(3-cloro-4-methylphenyl earbamat) (250 x 4,6 mm; 5 pm) da dugc lya chgn. Trong qua trinh thye nghiem, cac y l u t l anh hudng din qua trinh phan tieh ding phan nhu cdt sic ky, thanh phin va ty le dung mdi pha ddng da dugc khao sat nhim chgn dugc diiu kien phan tich thieh hgp dap Crng cac tigu ehi sau: hai pic ding phan phai cd he s l bit dii nim trong khoang 0,8 - 1,5 va tach hoan toan vdi dp phan giai Rg > 1,5, dap irng diu dd eao, thdi gian phan tich ngin. Sau khi tim duge diiu kien sic ky thieh hgp, tiln hanh thim dinh qui trinh phan tieh thee hudng din cua ICH^i va So tay hudng din dang ky thuld^', bae gom khao sat tinh phil hgp cua hg thing, tinh dac hieu, khoang tuyin tinh va gidi han phat hign, do ehinh xac va do diing. Qui trinh sau khi dugc thim djnh se duge irng dung d l djnh lugng va xae djnh tap dong phan trong mdt s l c h i phim chira ("Sj-ibuprofen tren thj trudng.
Chuan bi mau
Dung dich mau chuin ibuprofen (1000 pg/ml):
Can ehinh xac khoang 25 mg ibuprofen chuin, cho vao binh djnh mire 25 ml, them khoang 10 ml n-hexan, sjgu am 1 phut, lie diu, d l ngudi, thgm n-hexan din vach, lie diu, Igc qua mang Ipe PTFE 0,45 pm.
Dung dich chuin (S)-ibuprofen (500 pg/ml):
CSn ehinh xac khoang 25 mg ('Sj-ibuprofen chuin, cho vao binh djnh mirc 50 ml, them khoang 10 ml n-hexan, sigu am 1 phut, lie diu, d l ngugi, them n-hexan din vach, lie diu, Igc qua mang Ipe PTFE 0,45 pm.
Dung dich miu thd (S)-ibuprofen: Can 20 vign ('Sj-ibuprofen, xac djnh khii lugng trung binh vien.
Nghiin 20 vign thanh bdt min, can lugng bpt vien tugng irng vdi khoang 50 mg ("Sj-ibuprofen, eho vao binh djnh mire 100 mi, thgm khoang 50 ml n-hexan, sigu am 10 phiit, lie diu, d l ngudi, them n-hexan vCra du din vach, lie diu, Igc qua mang Ipc PTFE 0,45 pm.
Dung dich gia duuc: Chuin bj nhu dung djeh miu thu- nhung thay lugng bdt thule bing lugng ta dugc tuong Crng trong miu thir.
Dung dich miu trang: Dung mdi hda tan miu n-hexan.
K i t qua nghien CLFU Khao sat diiu kien sic ky
Bang 1 tdm t i t cac dllu kien s i c ky khao sat.
Hinh 1 minh hpa anh hudng ciia cac dllu kien khao sat lgn do phan giai va thdi gian luu cua ding phan ibuprofen.
Bang 1: Cac diiu kien sac ky khao sat
Bieu kien 1 Bieu kien 2 Bieu kien 3 Bieu kien 4
Pha dgng n-tiexan-EtOH (99:1) n-hexan-EtOH (98:2) n-tiexan-EtOH (98:2) n-tiexan-lPA (98:2)
Cof sac ky Opt Lux™ Cellulose-2 Cpt Lux™ Cellulose-2 Cot Lux™ Cellulose-1 Cpt Lux™ Cellulose-1
TAPPHf m r g g H o r - i7/7ni<; rgn ASH N A M sfi)
23
Nghien
CIFU- Ky thuat
Bieu kien 5 Bieu kien 6 Bieu kien 7 Bilu kien 8 Bieu kien 9
n-hexan-IPA(95:5) n-hexan-EtOH-TFA (98:2:0,1)
n-hexan-IPA-TFA (98:2:0,1) n-hexan-IPA-TFA (98:2:0,2) n-fiexan-IPA-TFA (98:2:0,05)
Cpt Lux™ Cellulose-1 Cot Lux™ Cellulose-1 Cot Lux™ Cellulose-1 Cot Lux™ Cellulose-1 Cpt Lux™ Cellulose-1
D K l DK2 B K 3 DK 4 DK 5 DK 6 BK 7 DK
Hinh 1: Dd thi biiu dien anh huung cua cac diiu kien khao sat len do phan giai va thdi gian luu ciia hai ddng phan (S)_-ibuprofen va (R)-ibuprofen trong
miu chuin ibuprofen
Nhan xet: Diiu kien 1: xuit hien 2 pic ding phan vdi dp phan giai R^ = 1,0 va thdi gian luu cua pic ding phan thir 1 khoang 18,4 phiit; Biiu kien 2:
xuit hign 2 pie ding phan vdi do phan giai R^ = 1,0 va thdi gian luu cua pic ding phan thCr 1 khoang 8,7 phut; Diiu kien 3: xuit hien 2 pie ding phan vdi dp phan giai R^ = 2,4 va thdi gian luu cua pie ding phan thu-1 khoang 7,3 phut; Diiu kien 4: xuit hien
Bang 2: Kit qua khao sat tinh phu hap
2 pie ding phan vdi do phan giai R^ = 2,4 va thdi gian luu eua pie ding phan thCr 1 khoang 7,7 phut;
Biiu kien 3 va 4 diu eho dp phan giai kha eao, thdi gian luu ngin nhung khdng I n djnh, chi cho kit qua taeh hoan toan trong mot s l lin tiem diu;
Biiu kien 5: xuit hien 2 pie dong phan vdi do phan giaiR^= 1,1 va thdi gian luu eua pic ding phan thir 1 khoang 5,1 phCit; Biiu l<ien 6, 7, 8 va 9: khi cho thgm TFA thi qui trlnh on djnh, lam tang dp phan giai va cai thien he s l bit dii. Vdi pha dgng cd n-hexan- IPA (98:2) {diiu kien 7, 8 va 9), he s l bit dii cua 2 pie ding phan n i m trong khoang 1,0 - 1,1. Khi them TFA 0,1% vao hai he pha ddng n-hexan-lPA (98:2) {diiu kien 7) va n-hexan-EtOH (98:2) (d/lu kiien 6), thi dp phan giai cua 2 pic ding phan cao han han d dllu kien 7, ding thdi he s l bit dii cua 2 pie ding phan gin bing 1,0. Kit qua khao sat ning dp TFA thgm vao pha dgng {diiu kien 7, 8 va 9) eho thiy ning dp TFA 0,1% {diiu kien 7) la thich hgp nhit vi cho dp phan giai 2 pie ddng phan cao nhit vdi thdi gian luu khoang 8,2 phCiL
Nhu vay, diiu kien thieh hgp d l phan tich ding phan quang hgc fSj-ibuprofen la: edt sic ky Lux™ Cellulose-1 (250 x 4,6 mm; 5 pm), pha dpng n-hexan-IPA-TFA (98:2:0,1), tie do ddng 1 ml/phut, nhiet do opt 30°C, t h i tich tiem miu 10 pi, budc sdng phat hien 220 nm.
Thim dinh phu'o-ng phap
Khao sat tinh phu hgp cua he thong he thing cita phuang phap (n = 6)
Mauchuan
(S)-ibuprofen Piel Pic 2 Piel Pic 2
Dien tich pic (pV Taing binh
11744021 11819430 51306 12274086
xgiay) RSD (%)
0,10 013 1,26 017
Thai gian luv (phut) Tmng binh
9,229 10,560
9,223 10,570
RSD (%) 0,07 014 0,04 0,05
1,0
1,0 "'^
1,0
1,0 -"'^
Nhan xet: Kit qua thing kg eho thiy, sau 6 lin sic ky, dien tieh pic va thdi gian luu cd RSD < 2%;
gia trj do phan giai (RJ Idn han 1,5 va he s l bit ddi cua cac pic nim trong khoang 0,8 - 1,5. Vay phuang phap dat tinh phii hgp cua he thing.
Tinh dac hieu
Tiln hanh sic ky miu chuin (hinh 2 va 3), miu thir (hinh 4), miu gia duge (hinh 5) va miu tring (hinh 6). Kit qua cho thiy miu gia duge va miu tring khong cd pic triing vdi pic chit phan tieh.
Thdi gian luu eua pic ehinh trong miu thCr tuang irng vdi thdi gian luu cua pic chinh trong miu ehuin fSj-ibuprofen. Khi them chuin (Sj-ibuprpfen vap miu racemic ibuprofen thi pie cd thdi gian luu dai hgn (khoang 10,56 phiit) cd dien tieh tang eao
la pic tuong irng vdi pie ding phan ('Sj-ibuprpfen.
Pho tu ngoai tai thdi gian luu cua eae pic trong miu thu gilng pho tu- ngoai cua miu chuin. Dp tinh khiit cua eae pie Idn hon 99%. Hai pic cua hai dang ding phan cd phi UV gilng nhau. Vay phuang phap ed tinh dae hieu.
Hinh 2: Sic ky dd miu chuin ibuprofen
24 TAP CHI DX/QC r t'^ l^f\t £ fCT
Nghien
CIPU- Ky thuat
Hinh 3: Sac ky dd miu chuin (S)-ibuprofen
I ' I
Hinh 4: Sic ky dd mau thu (S)-ibuprofen
Hinh 5: Sac ky dd miu gia duac
Hinh 7: Sic ky dd mau chuin ibuprofen 0,2 )s.g/mi ((S)-ibuprofen 0,1 [xg/ml va (R)-ibuprofen 0,1 [xg/ml) Khoang tuyen tinh va gioi han phat hien, do chinh xac va do dung
Bang 3: Kit qua khao sat khoang tuyin tinh, gidi han phat hien, do chinh xac va do dung
Phu-ong trinh hoi qui
Miu chuan ibuprofen (Rj-ibuprolen ISi-ibuprofen
y = 22000x y = 22539x R'=0,9984 R'=0,9991
Mau tfii>
IS)-ibuprofen y = 23028x R'=0,9987 Khoang tuyen tinh
Gioi han phat hien (Hinh 81
50-1000 |jg/ml
0,1 Mg/ml 0,1 pg/ml
66 lap lai (n = 6 RSD = 0,83%
Bo chinh xac trung gian (IE1S)_
RSD = 0,21%
Dp diing (n = tylethuhSi, RSD
10010%-101,51%:
0,46%
Hinh 6: Sic ky dd miu trang
Bang 4: Kit qua d'inh luang (S)-ibuprofen va
K i t qua t h i n g kg eho thay qui trlnh phan tich cd khoang t u y i n tinh rdng vdi he s l tuang quan cao, ty le h i i phuc n i m trong khoang 98% - 102% va RSD < 2 % d i i vdi do chinh xae.
N h u vay qui trlnh phan tieh d i n g phan quang hgc (S)-ibuprofen b i n g phuang phap s i c ky Icng hieu nang cao dat tinh phii hgp hg t h i n g , cd tinh dac hieu, ed khoang t u y i n tinh rdng vdi he s l t u g n g quan cao, gidi han phat hien t h i p , dp chinh xac va dp dung dat yeu c i u .
LPng dung qui trinh d l phan tich mot s6 c h i p h i m chu-a (S)-ibuprofen tren thj tru'O'ng
Bang 4 trinh bay k i t qua djnh lugng (Sj- ibuprofen va xae djnh djnh tap d i n g phan (R)- ibuprofen trong mgt s l e h l p h i m (S)-ibuprofen.
Hinh 9 minh hga s i c ky d l m i u t h u tir eae e h l p h i m khao sat.
(R)-ibuprofen trong mdtsd chi phim (n = 3) Chepham
Vien nen Stada (Sj-ibuprofen 200 mg
Vien nen (Sj-ibuprofen 200 mg
Vien nang m6m (Sj-ibuprofen 300 mg
G/a tri thong ke TB RSD
TB RSD
TB RSD
Ham luvng trong 1 207,77 0,26%
216,64 0,15%
289,25 0,64%
(S)-ibuprofen
vien (mg) Ham luung phan tram so vdi nhan 103,81
0,26%
108,32 0,15%
96,42 0,64%
%tap dong phan (R)- ibuprofen
0,42 0,77%
0,26 0,79%
3,46 0,49%
TAP CHI n i r ^ ^ H O f ] - ]7/->mA r s n .<«« NAM ^fi) 25
• Nghien
CIPU- Ky thuat
(a) (b) (c) Hinh 8: SSc ky dd dung dich miu thu tif chi phim vien nen STADA (S)-ibuprofen 200 mg (a), vien nen (S)-
ibuprofen 200 mg (b), vien nang mim (S)-ibuprofen 300 mg (c) Nhan xet: K i t qua djnh lugng ba m i u ehe
p h i m (S)-ibuprofen eho t h i y ham lugng hoat c h i t d i u n i m trong khoang tir 90-110% so vdi ham lugng ghi trgn nhan. Trong khi dd c h i pham vien nang m i m (S)-ibuprofen 300 mg ed lugng tap (Rj- ibuprpfen eao n h i t 3,46%.
Kit luan
Qui trlnh phan tieh d i n g phan quang hgc (Sj-ibuprofen b i n g phuang phap HPLC, s u dung ept pha tTnh b i t d i i da d u g e xay dyng va tham dinh. Qui trinh cd tinh dac hieu, gidi han phat hign t h i p , eho k i t qua chinh xae va lap lai. Qui trlnh da duge ap dung d l dinh lugng ('Sj-ibuprofen, xac djnh tap d i n g phan (Rj-ibuprofen trong mdt s l e h l p h i m chira ('Sj-ibuprofen. K i t qua thu d u g c gdp p h i n vao viec phan biet eao dang dong phan va cd t h i d u g c irng dung d l xae djnh tap d i n g phan trong eae e h l pham (S)-ibuprofen dang luu hanh trgn thj trudng.
Tai lieu tham khao
1. P.O. Carvaltio, Q.B.Cass, S.A. Calafatti, F.J.
Contesini, R. Bizaco (2006), "Review - alternatives for ttie separation of drug enantiomers: ibuprofen as a model compound", Brazilian J. of Chemical Engineering, 23, 291-300.
2. Fumio Toda (2004), Enantiomer Separation Fundamentals and Practical Methods, Kluwer Academic Publishers, The Netherlands, pp. 308, 312, 316.
3. Satinder Ahuja (2007), "A strategy for developing HPLC methods for Chiral Drugs", LC GC North America, 25, 1118-1128.
4. Xiaoming Chen, Chiyo Yamamoto, Yoshio Okamoto (2007), "Polysaccharide derivatives as useful chiral stationary phases in high-performance liquid chromatograptiy", PureAppl. Chem., 79(9), 1564-1566,
5. ICH Harmonised tripartite guideline (2005), Validation of analytical procedures: text and methodology, pp.1-13.
6. Quyit djnh cua Cue truo-ng Cue Quan ly Du-gc, Bg Y t l s6 07/QD-QLD ngay 11 thang 01 nam 2013 \ii viec ban tianh So tay himng din dang ky thuoc, Ptiu luc 8, tr 30-35.
{Ngay nhan bai: 10/10/2016 - Ngay phan bien: 14/11/2016 - Ngay duyet dang: 02/12/2016) r r r
I Cai tien phirong phap chiet xuat rotundin 1/ tir cu binh voi
Tran Trong Bien'*, Nguyen Quoc Huy^, Nguyen Van Han' 'Truang Dgi hoc Duo'c Hd Noi -Cue Khoa hoc Cong nghe vd Ddo tgo, Bo Y te 'E-mail: trantronghien@gmail. com Summary
Crude L-tetrahydropalmatine (rotundin) was extracted from the fresh tubers of Binh voi (Stephanie brachyandra Diels) by cold maceration and further refined conventionally with 96% ethanol and novelly with a mixture of aceton - water or ethyl acetate. Practically these novel extractions showed superior to the conventional one in simplicity, efficiency and rather ecological safety. The product obtained by ail these solvents was of rather high purity (rotundin content > 95%,). But the novel extractions remarkably raised the yield from 71 % (with 96% ethanol) to 96.6 %> (with mixture of aceton - water) and the highest 98.5 % (with ethyl acetate).
In practical tests, 5 kg of fresh material (rotundin content of 1.02 %,), by such extraction and purification yielded 48.13 g purified rotundin (rotundin content of 99.42 %,); the whole extraction efficiency of rotundin was 93.3 %.
Keywords: Extraction, L-tetrahydropalmatlne, rotundin.
26 T A P C H i D l / O C " " ' '
