A New Highly Effective Development and Validation of Trastuzumab and Hyaluronidase-Oysk in Bulk and Pharmaceutical Dosage form by Using RP-HPLC Method

Abstract

A simple, rapid, precise, sensitive, and reproducible Reverse phase High-performance liquid chromatography (RP-HPLC) method has been developed for the quantitative analysis of Trastuzumab and Hyaluronidase-OYSK in the pharmaceutical dosage form. Chromatographic separation of Trastuzumab and Hyaluronidase-OYSK was achieved on Waters Alliance-e2695by using Agilent Eclipse XDB (250x 4.6mm, 5µ) column and the mobile phase containing Acetonitrile: Ammonium formate pH-3.0/OPA in the ratio of 30:70% v/v. The flow rate was 1.0 ml/min; detection was carried out by absorption at 228nm using a photodiode array detector at ambient temperature. The number of theoretical plates and tailing factor for Trastuzumab and Hyaluronidase-OYSK were NLT 2000 and should not be more than 2 respectively. % Relative standard deviation of peak areas of all measurements always less than 2.0. The proposed method was validated according to ICH guidelines.  The method was found to be simple, economical, suitable, precise, accurate & robust method for quantitative analysis of Trastuzumab and Hyaluronidase-OYSK study of its stability.