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
and 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:
Email:
minhasiub@hotmail.com Tel:
+92-300-6074181; Fax: +92-62-9255565
Received: 24 May
2011 Revised
accepted: 16 December 2011
Tropical Journal of
Pharmaceutical Research, February 2012; 11(1):
84-90
http://dx.doi.org/10.4314/tjpr.v11i1.11
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, β-glucuronidase,
HPLC
