Development and Validation of a HPTLC Method for Rivaroxaban in Human Plasma for a Pharmacokinetic Study

The present study is concerned with the development and validation of a bioanalytical method for estimation of rivaroxaban in human plasma using high performance thin layer chromatography. The chromatographic separation was achieved on pre-coated silica gel 60F254 thin layer chromatography plate using toluene:ethylacetate:methanol (6:3:1, % v/v/v) as a mobile phase. Detection was carried out at 284 nm. A compact spot was obtained with an Rf value of 0.44±0.02. The linearity was found over the concentration range of 25-125 ng/band in human plasma. The relative standard deviation for repeatability of sample application and sample measurement was 0.60 and 1.65 %, respectively. The relative standard deviation for intraday and interday precisions was in the range of 1.37 to 1.85 % and 1.03 to 2.77 %, respectively. The limit of quantitation was 8.00 ng/band and the limit of detection was 2.64 ng/band. Percent recovery of rivaroxaban was in the range of 66.95-69.03%. This method was applied to a pharmacokinetic study of rivaroxaban. The Cmax and tmax for test and marketed formulation were found to be 63.83 and 47.3 ng/ml and 3 and 4 h respectively. The area under the curve0-t for test and marketed formulation were found to be 290 and 219.0 ng/ml.h, respectively. The t1/2 for test and marketed formulation was found to be 6.87 and 6.64 h.