Study On Quality Standard On Indinavir Sulfate Capsules And Their Bioequivalence

Indinavir sulfate is a new inhibitor immunodeficiency virus (HIV) protease. HIV protease is a enzyme required for the proteolytic cleavage of viral polyprotein into individual functional proteins found in infections HIV, so HIV protease can help the replication of HIV. Indinavir binds to the protease active site and inhabits the activity of enzyme. This inhibition prevents cleavage of viral polyproteins resulting in the formation of immature non-infections viral particles. It is very important to establish a standard to control the quality for safety and effect of patient.In this study, indinavir sulfate was identified specifically using the retention time of principal peak and ultraviolet absorption. The related substance was detected by using HPLC method. Chromatographic condition: Alltima C₈ column (250×4.6mm, 5μm) was employed as analytical column, the mobile phase consisted of 0.02mol.L^-1 citric acid buffer(pH4.0)-acetonitrile(45:55,v/v);the related substance was eluted linear gradiently using acetonitrile(A) and 0.02mol.L^-1 citric acid buffer (pH4.0)(B):A-B (20:80) keep for 2.5 min in 1 min A-B (80:20) keep for twice the retention time of principal peak in 1 min A-B (20:80) keep for 5 min;the detection wavelength was set at 260 nm and flow rate was 1.0ml/min. Standard curve was linear at the concentration range of 0.1534⁰.4602mg.mL^-1 and the correlation coefficient was 1.0000.The average recovery was 99.6%(RSD=0.39%, n=9).The apparatus of method I was used for dissolution test with water as the dissolution medium. The rotational speed of the basket was set at 50 rpm. The solution was withdrawed after 30 minutes andfiltered. The absorption of the resulting solution was measured at 260 nm. The dissolution was calculated, tolerances was not less than 75% of the labelled amount is dissolved. Standard curve was linear at the concentration range of 16.16 129.31u.g-mL"1 and the correlation coefficient was 0.9995.The average recovery was I00.1%(RSD=1.0% n=12).The established standard could be used for the quality control of indinavir sulfate capsules.For study indinavir bioequivalence, we establish a LC/MS/MS method for the determination of indinavir in the human plasma. Ritonavir was used as the internal standard. Indinavir was chromatographed by using a ZORBAX Eclipse XDB-C8 column.The mobile phase consisted of lOmmol-L"1 ammonium acetate buffer(pH = 4.10)- acetonitrile contained 0.1% formic acid (20:80).Electrospray ionization(ESI) source was applied and operated in the positiv ion mode.Selected reaction monitoring(SRM) mode with the transitions of m/z614.4-?421.2 and m/z721.3-?296.0 were used to quantify indinavir and the internal standard , respectively. The linear calibration curve was obtained in the concentration range of 0.009³0.0ng-mLI. The limit of quantitation was 0.009±0.001jug-mL"1(n=5). The inter- and intra-day precision(RSD) was less than 4% .The average recoveries were above 90%.The main pharmacokinetics data of referance prepartion and test prepartion A(indinavir sulfate capsules), B(indinavir sulfate tablets): /max were (36.67±13.5) min,(36.11±11.19)min and (38.06±13.95) min;cmaxwere (15.662±4.242) ug-mL"1, (15.988±4.289) ug-mL1 and (38.06±13.95) fig-mL'1;AUC0-t were (2055.5±537.0) ug-mL"'Tnin,(2004.1±453.8) ug-mL'1?min,(2144.3±600.8)ug-mL"1?min;AUC^ were (2071.9±543.4)ug-mL'l?min> (2018.2±463.1) ug-mL"1 ?min,(2156.8±606.4) ng-mL^min . The method was proved to be robust, covenient, sensitivity and rapidity. Two test prepartions were found to be bioequivalent to referance preparation.