A High-Resolution and Rapid HPLC Method for Etamsylate Estimation from Human Plasma Escorted by The Application to a Pharmacokinetic Study

Abstract The high polarity of Etamsylate hardens the estimation of Etamsylate-related substances by the liquid-phase detection method So, an accurate and sensitive High-performance liquid chromatographic (HPLC) method was created to provide a rapid analysis and high recovery percent of the Etamsylate from volunteers’ plasma. The method relied on protein precipitation by using 100% (wt/v) TCA (Tri-chloroacetic acid). Separation of Etamsylate was achieved by using a Fortis C18 column (150 x 4.6 mm, 5μm) and the mobile phase consisted of orthophosphoric acid 0.8% and acetonitrile with a ratio of 98:2 v/v and delivered to the system at a flow rate 1ml/min with injection volume 100μl. The wavelength was fixed at 302 nm. Paracetamol was used as the internal standard. A linear calibration curve of Etamsylate in plasma was obtained with a correlation coefficient r² < 0.999 while using a wide range of concentrations (0.6 - 40μg/ml). Retention times for both Etamsylate and Paracetamol were identified at 2.80 and 4.55 minutes respectively. The method is conducted on healthy Egyptian volunteers and proved to be precise. Many pharmacokinetic parameters were determined such as t1/2, tmax, Cmax, AUC0-t, AUC0-∞, and 90% CI.