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Abstract

RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF SILIBININ AND URSODEOXYCHOLIC ACID IN BULK AND IN THEIR COMBINED TABLET DOSAGE FORM

Sayali Salkar*, Dr. (Mrs).Vandana Jain

ABSTRACT

A novel, simple, precise, accurate RP-HPLC method was introduced for the simultaneous determination of hepatoprotective polyphenolic drug silibinin with ursodeoxycholic acid. The method was adapted to analyze drugs in their combined dosage form (tablet). The separation was achieved using a Prontosil C18 column (250 x 4.6mm, 5μm) and mobile phase comprised of acetonitrile and 0.02 M sodium dihydrogen phosphate buffer with 1.5ml Triethylamine (pH 6.5) in the ratio of 40:60 (v/v) at the flow rate 1ml/min and the detection was performed at 208nm. The retention time of silybin A, silybin B and ursodeoxycholic acid was found to be 3.847min, 5.643min and 4.453 min, respectively. The linear ranges for silibinin and ursodeoxycholic acid were 100- 200μg/ml and 100-600μg/ml, respectively. The recoveries of silibinin and ursodeoxycholic acid in pharmaceutical preparation were all greater than 97% and their relative standard deviations were not more than 2.0%. The developed method was validated in terms of accuracy, precision, linearity, limit of detection, limit of quantitation. This study aimed at developing and validating a simple, accurate and selective RP-HPLC method, and the proposed method can be used for the estimation of these drugs in combined dosage forms.

Keywords: Silibinin, Ursodeoxycholic acid, RP-HPLC, Validation.


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