| Abstrak/Abstract |
Objective: This study aimed to develop and validate an high-performance liquid chromatography method coupled with an ultraviolet detection for determination of ciprofloxacin in spiked human plasma and subsequently used for bioequivalence (BE) study.
Methods: The chromatographic separation was achieved on an RP-C18 column (Lachrom Hitachi, 250×4.6 mm, 5 ?m), utilizing a mobile phase of phosphate buffer/acetonitrile (87:13, v/v, pH 3.0±0.1) at a flow rate of 1.5 ml/minutes. Detection is carried out at 276.6 nm using a spectrophotometer. The developed method is statistically validated for the linearity, accuracy, limit of detection (LOD), limit of quantitation, precision, and specificity. The specificity of the method is ascertained by comparing chromatograms of blank plasma and plasma containing ciprofloxacin.
Results: Plasma matrix and ciprofloxacin can be separated very well using the validated method. The LOD and limit of quantification (LOQ) were 0.07 ?g/ml and 0.26 ?g/ml, respectively. The regression curve of the standard was linear (R>0.999) over a range concentration of 0.01-3.00 ?g/ml. The mean recovery of the method ranged between 95.27% and 104.45%. Both intra- and inter-day precision data showed reproducibility (relative standard deviation ?8.0, n=9). The validated method is successfully applied in pharmacokinetic and BE study of ciprofloxacin tablet in a small number of subjects. The mean values of Cmax, Tmax, area under curve (AUC)(0-12), and AUC(0-?) for the test (generic) formulation of ciprofloxacin were 2.25±0.29 ?g/ml, 1.0 hr, 8.60±0.69 ?g/ml/h, and 9.52±0.70 ?g/ml/h, respectively. Meanwhile, for the reference formulation (ciproxin tablet), the values were 2.30±0.27 ?g/ml, 1.0 hr, 8.54±0.85 ?g/ml/h, and 9.38±0.89 ?g/ml/h, respectively. From BE study, the 90% confidence intervals for the
test formulation/reference formulation ratio for the logarithmic transformations of Cmax, AUC(0-12), and AUC(0-?) were within the BE limit of 80-125% (96.08%, 101.00%, and 102.02%, respectively).
Conclusion: According to the validation result, the method was rapid, simple, and reliable. It can be used for routine analysis of ciprofloxacin in human plasma and BE study. The test formulation (tablet “X”) met the regulatory criteria for assuming BE in healthy subjects. |
