Hossein Danafar
1, Mehrdad Hamidi
2*1 Department of Medicinal Chemistry, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran.
2 Department of Pharmaceutics, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran.
Abstract
Purpose: A simple
and available reversed-phase high performance liquid chromatography (HPLC)
method with UV detection has been developed and validated for mycophenolic acid
(MPA) assay in human plasma.
Methods: MPA was extracted from plasma with protein precipitation
method by acetonitrile: percholeric acid: methanol (75:5:20 v/v/v). The drug
separation was achieved using a C8 analytical column and a mobile
phase of 0.1M triethylammonium phosphate (pH=5.4)-acetonitril (65:35, v/v),
with a flow rate of 1.5 ml/min. The detection wavelength was 304 nm. Limit of
detection (LOD) of the method was determined as the lowest MPA concentration
producing a signal-to-noise (S/N) ratio of about 3. Limit of quantitation (LOQ)
was determined as the lowest MPA concentration capable of being quantitated
with enough accuracy and precision.
Results: The
method showed significant linear response-concentration relationship throughout
the MPA concentration range of 0.2-10 µg/ml. A typical linear regression
equation of the method was: y = 8.5523 x + 0.094, with x and y representing MPA
concentration (in µg/ml) and peak height respectively, and the regression
coefficient (r) of 0.9816. The average within-run and between-run variations of
7.81 and 4.78 percent. The average drug recovery from plasma was 95.24 percent
throughout the linear concentration range. The limits of detection (LOD) and
quantitation (LOQ) of the method were 0.05 and 0.2 µg/ml, respectively. The
practical applicability of the method was proven throughout a bioequivalence study.
Conclusion: The results showed the acceptable degree of linearity,
sensitivity, precision, accuracy and recovery for the method. The method was
used successfully for quantitation of MPA in plasma samples of healthy
volunteers throughout a bioequivalence study.