Application of High Performance Liquid Chromatography to the Determination and Validation of Levodopa in Mucuna Pruriens L.

Application of High Performance Liquid Chromatography to the Determination and Validation of Levodopa in Mucuna Pruriens L.


Upadhyay V, Tiwari AK, Sharma N, Joshi HM, Singh BP and Kalakoti BS


Abstract

The seeds of Mucuna pruriens L. were extracted with 40% ethyl-alcohol, 60% water with 1.5% glacial acetic acid for the detection of levodopa and some physio-chemical parameters. A sensitive, precise and accurate high-performance liquid chromatographic (HPLC) method has been developed for the quantitative estimation of levodopa in Mucuna pruriens extract. The method utilises sample preparation step followed by separation on a Phenomenex C18, 250 x 4.6 mm, 5 µm particle size column, using 0.1 M KH2PO4 (pH 3.0 by ortho-phosphoric acid) as the mobile phase. Analysis of levodopa was carried out in the absorbance mode at 283 nm. The method was validated in terms of linearity, precision (inter and intraday), accuracy, limit of detection (LOD) and limit of quantification (LOQ). Linearity was observed in the range of 20-100 ppm with correlation coefficient of 0.9991. Detection limit was 5.6 ppm and quantification limit was 8.5 ppm. The repeatability of the method was found to be 0.49% and recovery values from 101.17 to 100.18% indicates best accuracy of the method. The results indicated that the extract obtained was 50% Levodopa content and 38% protein content. The proposed HPLC method was also found to be precise, specific, accurate and can be used for the identification and quantitative determination of levodopa in herbal extracts.


Keywords

High-performance liquid chromatography, levodopa, method development, Mucuna pruriens L., validation.


Cite This Article

Upadhyay V, Tiwari AK, Sharma N, Joshi HM, Singh BP and Kalakoti BS, Application of High Performance Liquid Chromatography to the Determination and Validation of Levodopa in Mucuna Pruriens L., International Journal for Pharmaceutical Research Scholars, 2012, 1(4), 135-140.

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