Bioanalytical HPTLC Method for Estimation of Zolpidem Tartrate from Human Plasma

Author(s)

Shirode, A.R., Jadhav, B.G., Kadam, V.J.

Author's Affiliation

Abstract

A simple and selective high performance thin layer chromatographic (HPTLC) method was developed and validated for the estimation of zolpidem tartrate from human plasma using eperisone hydrochloride as an internal standard (IS). Analyte and IS were extracted from human plasma by liquid liquid extraction (LLE) technique. The Camag HPTLC system, employed with software winCATS (ver.1.4.1.8) was used for the proposed bioanalytical work. Planar chromatographic development was carried out with the help of Silica Gel 60 F₂₅₄ precoated TLC plates. The optimized mobile phase was composed of ethyl acetate: methanol: acetonitrile (7:1.5:1.5, v/v/v).The detection of spots was carried out densitometrically using a UV detector at 298nm in absorbance mode. In HPTLC densitogram well defined peak was obtained for zolpidem tartrate with peak start position at 55 hR_f, max position at 59 hR_f and end position at 63 hR_f. The optimal hR_f value for zolpidem tartrate and IS were found to be 58 and 20 hR_f respectively. Performance characteristics of HPTLC method for estimation of zolpidem tartrate from human plasma were statistically validated as per the validation protocol designed based on the recommendations given by European Medicines Agency (EMA) guidelines of bioanalytical method validation. With validated method regression analysis of the calibration data showed a good linear relationship with mean correlation coefficient (r²) 0.998. The method was found to be simple and selective. The proposed method can be effectively used for accurate and precise determination of zolpidem tartrate from human plasma and can be applied for therapeutic drug monitoring and pharmacokinetic (Pk) studies on real clinical samples.

Keywords

HPTLC, Zolpidem Tartrate, Eperisone Hydrochloride, EMA, Bioanalytical Method Validation

Cite This Article

Shirode, A.R., Jadhav, B.G., Kadam, V.J. (2016). Bioanalytical HPTLC Method for Estimation of Zolpidem Tartrate from Human Plasma, International Journal for Pharmaceutical Research Scholars (IJPRS), 5(1), 114-123.