md. Phuong phap da xay dung sii dung cgt Apollo C18 (250 X 4,6 nun, 5 pm) dugc duy tri d 50°C trong qua trinh phan tich. Pha dOng la hdn hgp Acetonitril - Methanol - Dung dich acid formic 0,1 % (10:75:15), tdc do ddng 1 ml/phut, the tich tiem mlu 20 (il, detector PDA d buoc s6ng 210 nm. Phuong phap da dugc thim dinh vd do die hieu, khoang tuyen tinh, dg dung, do lap lai, dugc xac dinh gidi han phit hien, gidi ban dinh lugng. Ket qua tham dinh cho thay phuong pfa^
dugc xay dung hoin toan phii hgp cho viec djnh lugng tacrolimus trong ch6 pham thudc md Protopic va co thd ap dung dinh lugng ho^it chat nay tiong cac thuoc md cd thanh phan boat chat tuong tu.
3.2.5. Giai han dinh luong, giai han phat hien Gidi han dinh lugng, gidi han phat hien ciia phuong phap dugc xac dinh qua thuc nghiem bang each tiem cac dung dich chuan tacrolimus cd ndng do giam dan vao he thdng sac ky cho den khi khdng con dap ling pic tren sac ky do. Ket qua thuc nghiem cho thay, d ndng do 15 |ig/ml, pic tacrolimus tren sac ky dd cao hon khoang 10 Ian duong nen vi d ndng do 5 pg/ml, pic tacrolimus tren sac ky dd cao hon khoang 3 lin so vdi duong nen. Nhu vay phuong phap da xay dung cd gidi han dinh lugng vi gidi han phat hien vdi tacrolimus Ian lugt la 15 ^ig/ml vi 5 jig/ml.
4. Ket luan
Phuong phap sac ky long pha dao da dugc thiet lap de dinh lugng tacrolimus trong che pham dang thudc
Tai lieu tham khao
1, A.Dasgupta, T. G. Taamervaan, Journal of Liquid Chromatograhy & Related Technologies, tr.95-100, 1995.
2. A. Fujiwara, N. Kihara, S. Koda, K Hane and T. Yasuda, Journal of Pharmaceutical and Biomedical Analysis.
tr 278-281,2006.
SUMMARY
A reversed-phase liquid chromatography method was developed for assay of Tacrolimus in ointment. The method was carried out by using a Apollo CIS column maintained at 50°C during analysis procedure as stationary phase and a mixture of Acetonitrile, Methanol and 0.1% solution of Formic acid (10:75:15) as mobile phase. Flow rate of mobile phase is maintained at 1 0 ml per minute, injection volume: 20 pi, analyte was detected by a PDA detector set at 210 nm The method was validated in specificity, linear range, accuracy, LOQ and LOD. Validation results proved that the developed method was suitable for assay of Tacrolimus in ointment and was used to determine this active principle in Protopic ointment
X A Y Dl/NG PHl/OfNG P H A P XAC DjNH HOAT Ll/C CLPA STREPTODORNASE TRONG CHE PHAM HON HOfP C H I J A STREPTOKINASE VA STREPTODORNASE
HA THI MINH CHAU, THAN THI THANH HUE NGUYEN THI KIM HUONG. DOANCAOSON
Vien Kiem nghiem thuoc Trung uong
1. Dat vin de
Streptodomase li enzym thudc nhdm deoxyribonuclease vdi co chat dac hieu la acid deoxynbonuclease (ADN). Enzym Streptodomase la each viet tat ciia enzym deoxyribonuclease tu vi khuan tan huyet Streptococcus sp. (Streptococcus deoxyribonuclease) [4], [5]. Trudc day cac che phim thudng chiia don thanh phan Streptokinase nhung khi phat hien ra vai ti-d tuong hd nhau trong tang tac dung chdng viem va tieu huydt khoi cua Streptokinase v i
Streptodomase thi hai enzym niy thudng dugc phoi hgp vdi ty le 1:4 trong bio chd [1], [2], [3]. Tren th8 gidi cd nhieu phuong phap xac dinh hoat lye Streptokinase da dugc ip dung nhung chua thuc su c6 phuong phap xac dinh hoat luc cua Streptokinase va Streptodomase trong che pham hdn hgp dugc dua vao cac dugc dien.
(3 Viet Nam, khi d^ing thudc niy dugc nhap khiu thi cac phuong phap co so da dugc yeu ciu thim dinh. Tuy vay, vdi dieu kien thi nghiem chua diy du thi cac tieu
T9p chi KIEM NGHI$M THUOC - S6 4.2011; TSp 9.(34)
chuin CO sd cua cic che phim niy chi dugc thim djnh tren nguyen tic mi chua the tien hinh thyc nghiem.
Trudc nhu ciu thuc t% chiing tdi tien hinh de tai "Xdy dung phuang phdp xdc dinh hoat luc Streptodomase trong vien chua hon hap 2 enzym" nham ning cao nang luc kiem tra, giim sit chit lugng ddi vdi cac san phim thuoc cd ngudn gdc sinh hgc vdi dieu kien trang thi6t bi hi?n cd cua Vien KiSm nghigm thudc Trung uong va cac Trung tim Kilm nghiem thudc, my pham tren toin qudc.
2. NguySn v§t li^u v^ phuwng phap nghiSn cihi 2.1. Thietbi, d^ngctf, hoa chdt, chdtchudn 2.1.1. Thietbi, difngcyi
Da dugc hieu chuin theo ISO/EC 17025.
- Can phan tich SarKirius CP224S, do chinh xac 0,1 mg;
- Be di6u nhiet Grant ± 0 , r C ; - Nhdt ke Ostwald, No.22972, k = 0,0122;
- Ddng hd bim giiy;
- May do pH Hanna;
- Micro pipet;
- Cac dng nghiem, pipet thuy tinh chinh xic cac loai va mgt so dyng cy thiiy tinh can thiet.
2.1.2. Hoa chat, thuoc thic, chat chudn - Chat chuan Streptokinase: Chuan lam viec, Id:
118K1115, ham lugng 3578 lU/mg x 14mg/lg (Sigma Aldrich).
Chat chuan hdn hgp Streptokinase vi Streptodomase: gdm Streptokinase 1122 lU/mg, Streptodomase 268 lU/mg cua Reyon Pharma.
- Dung dich Pepton - TS: Cho 1 gam hSn hgp pepton vio 100ml dung dich thudc thu Barbital 0,025M, lie v i trdn deu. Chinh pH bing dung dich natri hydroxyd 0,2M vd 7,4, lgc. Bio quan dung dich niy d dieu kign 2 - 8°C, CO the sii dyng dung dich niy trong thdi gian 2 man.
- Dung djch natri Deoxyribonucleat acid - TS (dung dich co chit ADN): Hoa tan khoang 0,15g natri deoxyribonucleat acid trong 75ml dung dich thudc thii pepton - TS trong khoing thdi gian 75 phut, d didu ki?n 55 - 60°C. Lam lanh, them dung dich magnesi sulfat thuoc thir 12,0% (kl/tt). Bio quin dung dich niy d dieu ki$n - 12°C den - S^C vi sii dung trong khoang thdi gian 6 tuan.
2.2. D6i tufng vdphuomgphdp nghien cuu 2.2.1. Doi tu0}g nghien cuu
Vien nen HC - Tonase, sd Id M06, sin xuat boi Korean Dmg Co.Ltd (Streptokinase 10.OOOIU, Streptodomase 2.500 lU).
2.2.2. Phifongphdp nghien cuu
Streptodomase thugc nhdm deoxyribonuclease theo CO che la xuc tic thuy phan lien ket phosphodiester trong mach ADN dya tren dac tinh phan cat cua Streptodomase. Hien nay, phuong phap phd bien nhat de xac dinh boat tmh ciia Streptodomase li tren co che thuy phan co chit ADN.
Trong cic tai Ueu tham khio thi phuang phap xac dinh hoiit lye Streptodomase chua dugc dua vio dugc dien mi chi cd trong cic tieu chuan co sd. Co sd ciia phuong phip xac dinh boat luc nhu sau:
Streptodoi
ADN {c6 dp nhcft cao) *• Dung djch co do nhdt giam
Vdi CO sd tren, hoat tmh ciia Streptodomase dugc xac dinh tren khi nang lam giam do nhdt theo thdi gian khi cho Sh-eptodomase phin ling vdi co chit ADN.
Vdi cic trang thiet hi va hda chat hi?n cd tai Vien, chung tdi da cii tien phuong phap tham khio trong cic tieu chuan co sd de cd the ap dyng dugc trong dieu ki?n thi nghiem trong nude va tham dinh phuong phap dua tren chi tieu do d^ic hieu, dg tuyen tinh, dd dung vadglaplai.
2.2.2.1. Chuin hi dung dich thu
Can xac dinh khoi lugng trung binh ciia 20 vien (bdc vd bao phim neu cin) vi nghien thinh hot min.
Can chinh xac mgt lugng hot vien tuong ling voi 2.500 lU Streptodomase. Hda tan hoin toin trong dung dich nude mudi sinh 1^ vua du 50ml. Hiit 5,0 ml dung dich nay pha loang thinh 25,0 ml vdi dung dich thudc thit pepton - TS dS thu dugc dung dich thii cd ndng dg tuong ling khoang 10 lU/ml.
2.2.2.2. Quy hinh tiiu
- De dn dinh nhdt ke trong bd dn nhi^t d di§u ki?n 30 ± 0,3°C vi dim bao toin bO dng mao quin dugc ngap hoan toan trong mdi truong nude.
- Kiem tra dO dn dinh cua he thdng: Cho mgt lugng the tich can thidt ciia dung dich co chit ADN, de dn dinh d nhiet do quy dinh. Cho dung dich co chit niy chay hi diem xuat phat den didm cuoi h-ong dng mao ijuin ciia nhdt kd, dung dich niy phii cd thdi gian chiy dn dinh. Neu thdi gian chiy cua dung dich nay trong
Tap chi KIEM NGHI$M THU6c - So 4.2011; T^p 9.(34)
mao quan cua nhdt ke khdng dn dinh thi can nia lai dng mao quin v i pha l^i dung dich co chit ADN vi dung djch thudc thii nay da bj nhilm bin.
- Xac dinh khi nang lam giam dg nhdt cua dung dich chuan v i dung dich thir ddi vdi dung dich co chat ADN: Cho chinh xac 0,4 ml dimg djch chuan hoac dung dich thii vio dung dich co chat, xac dinh chinh xic thdi gian chiy trong mao quan ciia nhdt ke d cic thdi diem sau 5,10, 15, 20,25 va 30 phut sau khi cho dung dich chuin vi dung dich thir thu dugc cac gia tri tuong ling Bl, B2, B3, B4, B5 v i B6 (tinh tiieo don vi giay).
- Xac dinh thdi gian chiy ciia dung djch ddi chung (A giay): Thay dung dich chuan bang 0,4 ml hdn hgp dung dich thudc thii peptone, tien hanh xac dinh thdi gian chay trong mao quin tuong tu nhu cich xic dinh thdi gian chay dn dinh cua nhdt ke va dung dich thudc thit ADN.
2.2.2.3. Tinh ket qua
- Xac dinh he sd lam giam do nhdt ciia dung dich chuan so vdi dung dich ddi chiing: he sd Xgj, Xgj, Xg,, Xg^, Xgj va Xgg li gia tri thdi gian ciia dung dich ddi chiing (A giay) chia cho cac gii tri do dugc d cac thdi diem 5,10, 15, 20, 25 va 30 phiit tuong ling cua dung dich chuin (Bg,, B^^, Bgj, B^^, B^, vi Bg^ giay).
- Xic dinh h? sd lim giim do nhdt ciia dung dich thii so vdi dung dich ddi chung: he sd Xj.,, Xj^, Xpj, Xj.^, Xp5 va Xpg la gia tri thdi gian ciia dung dich doi chiing (A giay) chia cho cac gia tri do dugc d cac thdi diem 5,10,15, 20, 25 v i 30 phut tuong ling ciia dung dich thii (B^i, B-^^' BT3' ^^-4, B^.^ va B^.^ giay).
- Xay dyng dudng thing tuydn tinh giira he sd giam dg nhdt cua dung dich chuan va dung dich thii vdi thdi diem xac dinh dg nhdt tuong ling 0 5, 10, 15, 20, 25 v i 30 phiit.
- Tii dudng thing xiy dung dugc, xic dinh gii tri Cg, Cj. la gii tri dugc tuong ling d thdi didm 0 phut tir dd thi cua dung dich chuin va dung dich thir va xic dinh gia tri Dg, D^ li gia tii tuong ling 0 tiidi didm 25 phiit tu dd thi cua dung dich chuan va dung dich thir.
- Tinh gii hi K^sr = D,. - C^ va Kjjg = Dg - Cg.
- Hoat tinh Sh-eptodomase ti^n vien dugc tinh theo cdng thiic sau:
Wg X Pg Kj- DT-
X (lU) = X X X W Dg Kg W^.
Trong dd:
- Wg: khdi lugng chuan
- Pg: Ham lugng chuin Streptodomase - Dg: He s6 pha loang ciia dung dich chuan - W^: khdi lugng mlu thii
- Dp He so pha loang ciia dung dich thii - W: khdi lugng trung binh vien
3. K^t qua va ban luSn
3.1.03 dac hieu « Dung dich chuan 1: Pha dung djch chuifl Streptokinase trong dung djch nude mudi sinh ly de dm dugc dung dich chuin co nong dd khoang 400 lU/ml.
Pha loang dung djch chuan Streptokinase trong hdn hgp dung dich pepton dd thu dugc dung dich chuin Streptokinase cd ndng dg khoang 40 lU/ml
Dung dich chuan 2: Can chinhxac mgt lugng khoing 80 mg chuan hdn hgp Streptokinase v i Strqjtodomase (4:1) pha trong nude mudi sinh ly vira du 200 ml Pha loang dung dich chuan gdc 10 lin trong hdn hop dung dich pepton de thu dugc dung dich fliii cd nong dd Streptokinase khoang 40 lU/ml v i Streptodomase khoing 10 lU/ml.
Tien hanh: Xic dinh boat tinh Streptodomase trong dung djch chuin 1 v i dung djch chuin 2 theo myc 2.2.2.2. So sanh hoat tinh Streptodomase trong 2 dung dich tren.
Kit gud:
3.1.1. Xdc dinh khd ndngphdn ung Idm gidm dg nhat cua dung dich 1
Thdi gian chay cua dung dich ADN ddi chung va pepton la: 245 giay.
Thdi gian chiy trong nhdt ke cua dung dich chuin Streptokinase dugc trinh bay d Bang 1.
Bang 1. Thai gian chay trong nh&t ki cua dung dich chudn 1
Stt 1 2 3 4 5 6
Thoi ^im 5 phut 10 phut 15 phut 20 phfit 25 phut 30 phut
Thifi gian chay (gi^) 244 giay 244 giay 243 giSy 244 giay 243 giay 242 giay
H§s6(A/B^
1,0041 1,0041 1,0080 1,0041 1,0080 1,0120 ' ]
12 T?p chi KlfcM NGHIEM THU6c - S6 4.2011; T|p 9.{3«
Kit qua cho thay Streptokinase khdng lim giim do nhdt dung dich co chit ADN, dieu nay cd ngMa la ADN khdng phii li co chit thich hgp cua Streptokinase.
3.1.2. Xdc djnh khd nang phan img lam gidm do nhat cua dung djch 2
Thdi gian chiy cua dung dich ADN ddi chiing va pepton la: 245 giay.
Thdi gian chiy trong nhdt kl ciia dung dich chuin 2 dugc trinh biy trong Bing 2 v i Hmh 1.
Bang 2. Thdi gian chay trong nhat ke cua dung dich chuan 2
Bang 3. Kit qua khao sdt do tuyen tinh ciia phuangphdp
Stt 1 2 3 4 5 6
ThUi mim 5 phut 10 phut 15 phiit 20 phut 25 phiit 30 phut
Thdigian chdy (giay) 231 216 208 196 188 183
H$ so (A/B^
1,0606 1,1343 1,1779 1,2500 1,3032 1,3389
Hinh 1. Ehf&ng tuyen tinh bieu dien gidm do nhat theo thai gian cua dung djch chudn 2
Nhu vay dung dich chuin 2 lim giam do nhdt cua dung djch CO chat ADN, trong khi dd dung dich chuan 1 khdng cd tic dyng thuy phan ADN nen phuong phap dinh lugng niy cd thih d^ic hifu vdi Streptodomase do dung djch chuan 1 chi cd mat ciia enzym Streptokinase khdng cd bo^t tinh ddi vdi co chat la ADN tai lidn ket phosphodiester trong m^ich ADN.
3.2. BQ tuyen tinh
Sau qui hinh khio sat, chung tdi da lua chgn dugc khoang nong do phu hgp vdi dung dich co chit va phuong phap do dg nhdt bing mao quin cd he sd k = 0,0122. Tir dd, chiing tdi tien hanh xay dyng dudng chuin d 5 ndng dg dugc pha tir dung dich chuan gdc cd ndng dg dd thu dugc 5 nong dg khoang tii 5,0 den 16,0 lU/ml va dinh lugng theo phuong phap d myc 2.2.2.2. Ket qui dugc hinh biy d Bing 3 va Hinh 2.
Ndng dd dung dich chu^ (IV/ml)
Gid tri K2^ Thong ke
Phuong trinh hSi quy tuyen tinh.
y = 0,027 X + 0,0001 He s6 tuyen tinh.
R^ = 0,9959
Ndng 0 4 dd c h u i n (lUhnI)
Hinh 2. Do thi bieu dien mdi tuong quan giiia nong dp vd he so gidm dp nhdt ^^jj
Kdt qua thu dugc cho thiy, trong khoang ndng do dung dich chuan khao sat tii khoang 5 - 1 6 lU/ml cd su tuong quan tuyen tinh giua ndng dg va he so giam do nhdt K25S. Vay ndng dd dung dich Strqjtodomase ndn chgn trong khoing 10 lU/ml li phii hgp.
3.3. Dp diing
Tien hinh phuong phip them chuin:
- Chuan hi dung dich chuan: Can chinh xic 50,2 mg chuin hdn hgp streptokinase (1122 lU) - streptodomase (268 lU). Hda tan trong dung dich NaCl 0,9% vira dii 50 ml. Pha loang dung dich chuan niy 20 lin bang dung dich thudc thii pepton de thu dugc dung dich chuin cd ndng dg streptodomase khoang 10 lU/ml.
- CSiuin bi dimg dich thii: Can chinh xic mgt lugng miu thu tuong duong khdi lugng trung binh cua 1 vien (293,8 mg), hoa tan bing dung dich NaCl 0,9% vira dii 50 ml. Pha loang dung dich mau thu nay vdi dung djch thudc thu pepton de tiiu dugc dung dich thii cd nong dg streptodomase khoang 10 lU/ml.
- Chuan hi dung dich thii cd them dung dich chuin streptodomase:
Day tini 1: Lay chinh xac 20ml dung dich thii ndng do khoang 50 lU/ml va cho them 3 ml dung dich chuin hdn hgp Sti-eptokinase - Sti-eptodomase de thu dugc dung dich thii cd them chuin.
Tfp chf KIEM NGHI$M THU6C - S6 4^011; T^p 9.(34)
Day thii 2: Lay chinh xac 25 ml dung djch thu ndng do 50 lU/ml va cho them 5 ml dung dich chuan hdn hgp Streptokinase - Streptodomase de thu dugc dung dich thir cd them chuan.
Pha loang cac dung dich mau thir nay vdi dung dich thudc thir pepton de thu dugc dung djch thii them chuan cd ndng dd streptodomase khoing 10 lU/ml.
Thdi gian chay ciia dung djch ADN ddi chiing v i pepton (A) xac dinh dugc li 228 giay. Tien hanh xac dinh kha nang lam giim do nhdt ciia dung dich chuan sft-eptodomase theo muc 2.2.2.2. Kdt qua thu dugc trinh biy trong Bing 4.
Bang 4. Thai gian chdy trong nh&t ke cua dung dich chudn
(272 lU). Hoa tan h-ong dung dich NatTl 0,9 % v i | du 100 ml. Pha loang dung dich nay 10 lin bing dui^
djch hdn hgp thuoc thii pepton. Tiln hanh dinh lugng theo muc 2.2.2.2.
Thdi gian chiy cua dung dich ADN ddi chiing va pepton li: 228 giay.
Thdi gian chay trong nhdt ke cua dung dich chua|
thu dugc ttmh biy trong Bing 6
Bang 6. Thai gian chdy trong nhat ke cua dung dich chudn
Stt 1 2 3 4 5 6
Th&i diem 5 phiit 10 phiit 15 phiit 20 phiit 25 phut 30 phut Phu'crag trinh hoi
quy tuyen tinh
Thdi gian chdy (giay) 217 203 191 180 174 166
Hf s6 (A/B^
1,0507 1,1232 1,1937 1,2667 1,3103 1,3735
y = 0,0128x + 0,9949 (R^= 0,9944) Tir dudng hdi quy tuyen tinh tinh dugc K^jg = 0,3200.
Tien hanh xac dinh hoat luc cua Streptodomase trong dung dich thir va dung dich thii them chuan theo muc 2.2.2.2. Ket qui dugc ghi trong Bang 5.
Bang 5. Kit qua khao sdt dp diing STT
1 2 3 4 5 6
Khoi luong mdu thu-
287,3 295,8 286,7 294,1 292,8 288,5
Lu^g cliuan them vdo (IV)
1345,4 1345,4 1345,4 807,2 807,2 807,2
Lu0ig tim thay (W)
1302 1322 1353 795 821 788
Ty /p dm Iqi
(%)
96,8 98,3 100,6 98,5 101,7 97,7 Trung binh 98,9%; SD = I,S3; RSD = 7,95%
Ket qua cho thay phuong phap cd do dung cao vdi khi nang tun lai la 98,9 % vi RSD = 1,95%.
3.4. Do lap lai
Dung dich chuan: Can chinh xac 40,8 mg chuin hdn hgp Streptokinase (1.100 lU) - Streptodomase
stt 1 2 3 4 5 6
Thdi diem 5 phut 10 phut 15 phut 20 phut 25 phiit 30 phut
Thefigian chay (giay) 210 201 188 181 171 165
H^sim^\
1,0857 ;; 1 1,1343 3 ! 1,2128 J ; 1,2597 - ' 1,3333 - 1,3818 • Phirong trinh hoi quy tuyen tinh ^ y-0,l)121s +1,0222 (R=-0,9958) - Tir dudng tuyen tinh bieu dien giam do nhdt theo thdi gian ciia dung dich chu&i tinh dugc K^jg = 0,3025.
- Dung dich thir: Cin chinh xac mgt lugng mau thii bang khdi lugng trung binh vien tuong duong 1 vien, hoi tan trong dung djch NaCl 0,9% vira du 50 ml. Pha loang dung djch mlu thir nay bang hon hgp dung dich thudc thu pepton dd thu dugc cic dung dich thu cd ndng do khoing 10 lU/ml. Tien hinh dinh lugng Streptodomase frong miu thu theo myc 2.2.2.2, ap dyng tren 6 lugng miu thu rieng biet. KLTB vien=
293,8 mg. Ket qua thu dugc trinh bay trong Bing 7.
Bang 7. Ket qua khao sdt do lap lai ciia phuangphdp STT
1 2 3 4 5 6
Khoi lucmg mauthif(mg)
279,3 283,9 290,5 275,6 285,4 296,1
'^IST
0.3250 0,3200 0,3400 0,3194 0,3237 0,3457
Hdm luffng
(%)
100,3 97,2 100,9 99,9 97,8 100,7
Thdng lu
TB = 99,4V.
n - 6 SD •= 1,58 RSD-I,59«
Ket qua thu dugc cho thay phuong phip cho (1$
l^p lai tdt. Kdt qua djnh lugng Streptodomase hong 6 lin thii nghiem cbdnh lech nhau khdng dang ke vdi RSD = 1,59 % la phu hgp vdi phep tiiu djnh lipig trong sinh hgc.
T?p chi KIEM NGHI$M THUOC - So 4.2011; T$p9.{3<)
4. K^t lu^n
Da xay dyng dugc cac quy trinh xac djnh boat tinh ciia Sh^todomase tixing vien chua hdn hgp Streptokinase va Stt^ptodomase cd tuih d^c hieu trdn co chit ADN vi cho do tuyen tinh, do lap lai tdt, do dung phii hgp vdi phep thii dinh lugng. Qay trinh phu hgp vdi didu kien hi?n cd ttong phong thi nghiem vdi cic ttang thidt bi, hda chit thdng dung, ky tiiuit khdng phiic tap cd the thuc thi dugc d cic co sd kiem nghiem cap tmh, tiiinh pho va cac doanh nghi^. Tuy nhidn dd quy trinh chiing minh dugc tinh dac hieu mdt cic tdt nhit can phai thir vdi chuan don thanh phin chiia Streptodomase nhung vdi dieu kien hien nay chung tdi chua cd chuan don thinh phan Streptodomase nen chua tidn hinh thuc hien dugc mgt each day dii vi can hoin thien trong thdi gian tdi.
Tii lieu tham Idiao 1. Tieu chuin co sd HCTONASE Korean Dmg Co.Ltd.
2. Tieu chuin co sd Hawon - Strepase, Hawon pharmaceutical corporation.
3. TiSu chuin co sd Strase, Green Cross Corporati.
4. Deoxyribonuclease I,website: http://en.wikipedia.org/wiki/Deoxyribonuclease_I.
5. Ian C. Locke; Brian G. Carpenter. Functional characteristics of the streptococcal deoxynbonuclease "Sueptodomase", a protein with Dnase activity present in the medicament Varidase. Enzyme ADN Microbial Technology 35 (2004) 67-73.
SUMMARY
A simple and rapid method for analyzing of Streptodomase in combined pharmaceutical products containing 2 compounds of Streptokinase and Streptodomase have been developed and validated. The method is based on the digestion ofDNA nature to the lower viscid solution. And this change is determined by Ostwald viscometer. The activity of sample IS calculated with Streptodomase reference standard. This method showed linearity within the range of Streptodomase concentration of 5.0-16.0 lU/mi The experimental results proved that it is precise, accurate and suitable for the quality control of enzyme products containing Streptodomase combined of Streptokinase and Streptodomase.
XAY Dl/NG PHl/dNG PHAP C H I ^ T V A PHANTI'CH ATROPIN, S C O P O L A M I N
TRONG 2 LOAI THU6c HdTRd CAI NGHIEN
BANG PHl/ONG PHAP SAC KY LONG HlfU N A N G CAO (HPLC)
Dd THI BiCH THU4N, NGUYEN TUAN ANH, PH^M THJ GIANG Vien Kiem nghiem thuoc Trung uong
1. Dat van de
Nghien hiit li mgt t$ nan giy suy giim siic khde vi nhu Cedemex, Balok. Ngoai viec kidm tra chit lugng dao diic ciia con ngudi va la mgt van de biic xiic cua theo tieu chuan co sd, chiing tdi cdn kiem tra phat hi$n toin xa hgi. Vi vay vi?c phdng chong te nan ma hiy cic chit khdng dugc phep trong cac thudc fren nhu da vi dang la nhiem vy chung cua tit ci cic qu6c gia rotundm, naltrexol, clorpromazin, levomepromazin va tren the gidi. Ben canh viec quin 1;^ buon bin, sii dyng mgt sd chat an than gayngii thudng dung (phenobarbital, chat ma hiy, giio dye cho cgng ddng vd tic hai ciia ma diazepam, bromazepam, Zolpidem),
hiy, mdt h-ong nhung bien phap lam giam ganh nang De tiip tyc dinh gii chit lugng cac loai tiiudc cho gia dinh va xa hgi la giiip ngudi nghien ma ttiy cai hd ti-g cai nghien, nam 2009, ngoai vi$c kidm tra cic nghien bing tiiuoc cai va hd tra cai nghien, Ud ve vdi chit tren, chung tdi da tien hinh nghien ciiu xay dyng cugc song bmh tiiudng phuong phip chidt va phan tich afropin, scopolamin vi Hi?n nay, da cd mgt sd lo^ thudc hd trg cai nghien cic alcaloid cua li ca dgc dugc trong cac thudc hd frg cd ngudn gdc tii dugc li?u da dugc Bg Y td cip sd dieu tii cai nghien Cedemex va Balok [2].
dang ky sin xuit luu hinh. Tii nam 2006 d ^ nay, Yietx Trong bai bao nay, chiing tdi xm gidi thieu ngi Kidm nghi?m thudc Trung uong hen hac liy miu kidm dung xay dyng phuong phap chiet xuat vi phan tich tra, danh gia chat lugng cac loai thuoc hd trg cai nghien atropin va scopolamin bing phuong phip sic ky long
T9P chi Kl^M NGHIEM THUOC - So 4.2011; T^p 9.(34) 15
