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Original
Research Article
Simultaneous quantitative determination of zidovudine
and nevirapine in human plasma using isocratic, reverse
phase high performance liquid chromatography
Vibhuti Kabra1*,
Vivek Agrahari1, Chandrabose
Karthikeyan2 and Piyush Trivedi2
1Department
of Pharmacy, Shri G. S. Institute of
Technology & Science (SGSITS), Indore
452001, 2School of Pharmaceutical
Sciences, Rajiv Gandhi Technical University
(RGTU), Bhopal 462036, Madhya Pradesh,
India.
*Corresponding
author:
E-mail:
vibhuti.kabra@rediffmail.com Phone:
+91-9827500875.
Received: 30 July 2008
Revised accepted: 24
October 2008
Tropical
Journal of Pharmaceutical Research, February
2009; 8(1):
79-86
Abstract
Purpose:
To develop a sensitive and rapid reverse phase high
performance liquid chromatography (HPLC) method for the
measurement of the levels of zidovudine (ZVD) and
nevirapine (NVP) in human plasma.
Methods:
Standard stock solutions for HPLC analysis were prepared
by dissolving ZVD and NVP in methanol. In the HPLC
measurement, sample detection was carried out at 246 nm
using an ultraviolet (UV)-photo diode array (PDA)
detector. Plasma sample pretreatment consisted of
protein precipitation extraction with methanol. The
compounds were separated using a mobile phase consisting
of a pH 3.0 solution (obtained by adjusting the pH of
water with orthophosphoric acid): acetonitrile (73:27
v/v) on a Phenomenex LUNA C18, column
(250×4.6 mm i.d., 5μm) at a flow rate of 0.9 mL min-1.
The total run time for the assay was 10.2 min. The
method was validated over the range of 300-9600 ng mL-1
and 200-6400 ng mL-1 for ZVD and NVP,
respectively.
Results:
The lowest limits of quantification (LLOQ) and of
detection (LOD) were 300 and 63 ng mL-1 for
ZVD and 200 and 17 ng mL-1 for NVP,
respectively. The method was found to be accurate, with
accuracy ranging from -10.92 to +9.57 % and precise,
with intra-day, inter-day as well as analyst to analyst
precision of 0.68 to 9.38 %. Extraction recoveries of
the drugs from plasma were 91.39, 95.01, 89.51 % for ZVD
and 90.93, 93.26, 92.13 % for NVP, for LQC (low quality
control), MQC (medium quality control) and HQC (high
quality control) samples, respectively.
Stability data revealed that the drugs were stable in
plasma under various test conditions.
Conclusion:
This assay can be suitably used for the determination of
zidovudine (ZVD) and nevirapine (NVP) in human plasma
and should be useful in HIV clinical trials and clinical
therapeutic drug monitoring (TDM) programs. It would
also be potentially useful in the determination of
pharmacokinetic profiles and in bioequivalence studies
in HIV research.
Keywords:
Assay, Zidovudine, Nevirapine, Human plasma, Reverse
phase high-performance liquid chromatography. |
