| ObjectivesTo establish a HPLC method for determining rifampin, isoniazid, pyrazinamide concentration in human plasma and use an UV-visible spectrophotometry of urine concentration ethambutol hy drochloride, the methods was successfully to study the clinical pharmacokinetics and bioequivalence of compound rifampicin dual-release capsules in chinese healthy male volunteers.MethodsTwenty Chinese healthy male volunteers were randomLy divided into two groups, with10volunteers in each group. The observation group was given12compound rifampicin dual-release capsules and control group was given reference preparation. One week after their own cross, another kind of preparation was administrated.Single-dose group taken4mL blood sample at before the medication and after medication0.33、0.67、1、1.5、2、2.5、3、3.5、4、4.5、5、6、8、12、15、24and48h.Multi-dose group was collected samples at consecutive6days oral administration. According to the FDA analysis of biological samples methodologntly shake and immediately centrifuged to sepatated and stored in-30℃refrigerator ical guideline, the full validation was carried out to test the precision, accuracy, linearity, lower limit of quantification, biological matrix effects, stability and recovery. The concentration of rafampicin, isoniazid and pyrazinamide in plasma were determined by high performance liquid chromatography, the urine concentration of ethambutol was determined by UV-visible spectrophotometry and the pharmacokinetic parameters were calculated by3P97software.ResultsDetermination of rafampicin had good linearity in the0.24-63.29mg·L-1, lower limit of quantification was0.24mg·L-1. Determination of isoniazid had good linearity in the0.26-32.64mg·L-1, lower limit of quantification was0.26mg·L-1. Determination of pyrazinamide had good linearity in the0.32-162.48mg·L-1, lower limit of quantification was0.32mg·L-1.Determination of ethambutol had good linearity in the25.02-500.40mg·L-1, lower limit of quantification was25.02mg·L-1. Methodology confirmatory results showed that the analytical method complies with the relevant requirements of the SFDA biological sample analysis method guide applied to the human pharmacokinetics of rifampin, isoniazid, pyrazinamide and ethambutol.Single-dose group:The pharmacokinetic parameters of test formulation and reference preparation of rifampicin were as follow:tmax were (4.15±0.80) and (2.03±0.73)h, Cmax were(10.83±1.47)and(10.94±1.82)mg-L-·1, AUCo.t were (89.77±15.43)and (71.78±14.01)mg·h·L-1, AUC0-∞were (93.93±15.69) and (77.04±14.96)mg·h·L-1, respectively. F was (126.8±18.2)%and there are significant differences in tmax. The pharmacokinetic parameters of test formulation and reference preparation of isoniazid were as follow:tmax were(0.77±0.51) and (0.79±0.46)h,Cmax were(4.44±1.45)and(4.22±1.42)mg·L-1, AUCo-twere (12.40±5.32) and (11.49±5.20) mg·h·L-1,AUC0-∞were (13.33±5.47) and (13.60±9.58)mg·h·L-1, respectively. F was(109.9±14.2)%. The pharmacokinetic parameters of test formulation and reference preparation of pyrazinamide were as follow:tmax were (1.47±0.45) and (1.35±0.48)h, Cmax were (35.99±4.63) and (32.80±3.69) mg·L1, AUC0-t were (555.46±76.81) and (481.70±74.03) mg·h·L-1, AUC0-∞were(574.93±82.06) and (498.22±79.40) mg·h·L-1, respectively. F pyrazinamide was(116.8±17.5)%. The cumulative urinary excretion of test formulation and reference preraration of ethambutol hydrochloride were (68.58±21.19)%and (67.69±18.70)%, respectively. F was (107.6±47.0)%.Multi-dose group:The pharmacokinetic parameters of test formulation and reference preparation of rifampicin were as follow:tmax were (4.60±0.37) and (1.91±0.66)h, Cmax were(9.73±1.82)and(9.35±1.31)mg·L-1, AUCss were(51.20±7.84) and (40.16±5.97)mg·h·L-1, Ka were (0.39±0.11) and(1.52±1.83) h-1, t1/2were (3.91±3.21) and(2.16±0.58) h-1, respectively. The pharmacokinetic parameters of test formulation and reference preparation of isoniazid were as follow:tmax were (1.00±0.30) and (0.87±0.45)h, Cmax were(4.52±1.14)and(4.12±1.05)mg·L-1, AUCss were (12.30±5.75) and (12.28±2.05)mg·h·L-1, Ka were (5.55±7.76) and (4.54±5.54) h-1were (1.77±0.77) and(1.57±0.20) h-1respectively.The pharmacokinetic parameters of test formulation and reference preparation of pyrazinamide were as follow:tmax were (1.58±0.58) and (1.47±0.52)h, Cmax were(39.75±7.30)and(44.10±10.97)mg·L·1, Ka were (4.13±5.67) and (3.81±1.43) h-1, t1/2were (8.48±0.74) and(9.14±1.18) h-1, respectively. The cumulative urinary excretion of test formulation and reference preraration of ethambutol hydrochloride were (1.15±0.55)%and (5.74±10.24)%, respectively.ConclusionsThe results by the analysis of variance and two-sided t test indicated that the two formulations were not bioequivalent in rifampicin, there were bioequivalent in isoniazid and pyrazinamide. |
PDF Full Text Request