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Abstract

BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR ESTIMATION OF TOLPERISONE HYDROCHLORIDE IN HUMAN PLASMA BY HPLC

Sonali Mahaparale*, Indrajeet Gonjari

ABSTRACT

A simple, rapid, selective, sensitive, accurate and precise High Performance Liquid Chromatography (HPLC) with UV detection method has been developed and validated for determination of Tolperisone hydrochloride in human plasma. Tolperisone hydrochloride is a piperidine derivative, centrally-acting muscle relaxant used in the treatment of acute muscle spasms in back pain. Eperisone hydrochloride is used as internal standard. The chromatographic separation of Tolperisone hydrochloride and internal standard were achieved with Chromasil C18 column using a mobile phase Acetonitrile: water pH adjusted to 3.0 with O- Phosphoric acid (60:40 v/v) at flow rate 1.0 ml/min. The measurements were made with injection volume 20 μl and UV detection at 260.0 nm. Plasma samples were processed using acetonitrile as precipitating agent to extract drug. The linearity for Tolperisone hydrochloride was found to be 50-2000 ng/ml with regression coefficient (r2) 0.9990. The limit of quantification in plasma for Tolperisone hydrochloride was found to be 50 ng/ml. The mean recovery was obtained at 74.22 %. The method has been used to perform pharmacokinetic and bioequivalence studies in human plasma.

Keywords: Tolperisone hydrochloride, Eperisone hydrochloride, Human plasma, Method validation, HPLC-UV detection.


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