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Original Research Article | OPEN ACCESS

Effect of β-Glucuronidase on Extraction Efficiency of Silymarin from Human Plasma Samples Using Validated HPLC-UV Analysis

Muhammad Usman1 , Mahmood Ahmad1, Abdullah Dayo2, Asadullah Madni1, Liaqat Ali1, Muhammad Yousuf1, Mahtab Ahmad Khan1, Abubakar Munir1, Muhammad Sohail1, Arshad Mahmood3

1Faculty of Pharmacy and Alternative Medicine, the Islamia University of Bahawalpur-Punjab; 2Faculty of Pharmacy, University of Sindh, Jamshoro; 3Department of Pharmacy, COMSATS Institute of Information Technology, Abbotabad, Pakistan.

For correspondence:-  Muhammad Usman   Email: minhasiub@hotmail.com   Tel:+923006074181

Received: 24 May 2011        Accepted: 16 December 2011        Published: 21 February 2012

Citation: Usman M, Ahmad M, Dayo A, Madni A, Ali L, Yousuf M, et al. Effect of β-Glucuronidase on Extraction Efficiency of Silymarin from Human Plasma Samples Using Validated HPLC-UV Analysis. Trop J Pharm Res 2012; 11(1):84-90 doi: 10.4314/tjpr.v11i1.11

© 2012 The authors.
This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited..

Abstract

Purpose: To investigate the effect of β-glucuronidase on the extraction efficiency of silymarin (mainly as silybin) from spiked human plasma using a sensitive and reproducible high performance liquid chromatography (HPLC) method.
Methods: The importance of β-glucuronidase was evaluated by comparing the extraction efficiency of silymarin in β-glucuronidase-treated and untreated plasma samples. Isocratic HPLC with simple UV detection (288 nm) was applied to analyze the major silymarin components using Thermo-Electron C18 column (200 mm, 4.6 mm I.D., 5µm particle size). The mobile phase, consisting of methanol and 20 mM potassium dihydrogen phosphate buffer (50:50 v/v pH 2.8), was pumped at 1 ml/min.
Results: The mean extraction efficiency was 98.97 % (CV = 1.69 %) for treated and 40.88 % (CV = 2.77 %) for untreated plasma samples, compared with nominal concentrations.
Conclusion: The studied method showed 60 % reduced extraction efficiency of untreated samples compared to treated samples.

Keywords: Silymarin, Silybin, Extraction Efficiency, ^6;-glucuronidase, HPLC

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