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Research Journal of Pharmacy and Technology
Year : 2016, Volume : 9, Issue : 6
First page : ( 671) Last page : ( 676)
Print ISSN : 0974-3618. Online ISSN : 0974-360X.
Article DOI : 10.5958/0974-360X.2016.00126.8

Bio-analytical method development and validation for the estimation of Clotrimazole in human plasma by RP-HPLC method

Tamilselvi N.1,*, Sinha Hempushpa1, Visakh Deepthi1, Vanathi P.2

1Department of Pharmaceutical Analysis, K.M.C.H College of Pharmacy, Kalapatti Road, Coimbatore-641048, Tamil Nadu, India

2Department of Pharmaceutical Chemistry, Karpagam College of Pharmacy, Coimbatore, Tamil Nadu, India

*Corresponding Author E-mail: tamildeiva@gmail.com

Online published on 21 July, 2016.

Abstract

A simple, reliable HPLC method was developed and validated for quantitation of clotrimazole in human plasma. Preparation was carried out using protein precipitation method. Separation was done on a Enable C18 reverse phase column with a mobile phase of 0.5% TEA in water (pH 3 adjusted with o-phosphoric acid) and acetonitrile in the ratio of 20:80%v/v and detection was done at 215nm. The retention time of ibuprofen (IS) and Clotrimazole was found to be 5.2 and 8.2mins respectively. The method was developed in terms of accuracy, precision, linearity, specificity, system suitability, and stability studies, limit of detection, limit of quantification, ruggedness and robustness. The calibration curve was linear within the range of 50-250ng/ml (r2>0.999). The lowest limit of quantification and detection was found to be 23.90ng/ml and 7.8 ng/ml respectively. System suitability test results on the retention time, area ratio and peak ratios of peak chromatogram shows the CV (%) <10%. The proposed method was validated and proved to be adequate for the determination of Clotrimazole in human plasma.

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Keywords

Clotrimazole, RP-HPLC, Validation, Bioanalytical method development in Human Plasma.

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