Abstract

ISOLATION, DEVELOP AND VALIDATION OF AN RP-HPLC METHOD FOR ESTIMATION OF LEVODOPA FROM MUCUNA PRURIENS AND ITS HEAVY METAL PROFILING BY ICP-OES

Harmeet Kaur*, Gurpreet Singh Sandhu, Bhupinder Kaur and Supandeep Singh Hallan

ABSTRACT

Purpose: In the present research two different experiments was performed on Mucuna pruriens seeds (black and white seed). First one, is to isolate, develop and validate RP-HPLC method for estimation of Levodopa. The second one, its heavy metal profiling by ICP-OES. Methods: The RP-HPLC method composed extract with Methanol, water, Ascorbic acid for the detection of levodopa and some physiochemical parameters. The chromatographic separation was achieved on C18 (250 X 4.6 mm, 5μ) analytical column with the mobile phase consisting ratio mixture of Sodium lauryl sulfate, Tartaric acid at flow rate-1 ml/min and detector wavelength 280 nm. Results: The retention time of levodopa was found to be 5.573 min. Linearity was observed in the range of 62.5-375 ppm with the correlation coefficient of 0.997. The detection limit was 0.86 ppm and quantification limit was 4.09 ppm. Recovery values from 100.27% to 102.75%, indicates the best accuracy of the method. In ICP/OES, heavy metal contents from Mucuna pruriens (white) was found to be Arsenic-0.68ppm, Mercury-0.14ppm, Cadmium-not dect., Lead-0.58ppm and Mucuna pruriens (black) Arsenic-0.54ppm, Mercury-0.23 ppm, Cadmium-not dect., Lead-1.08ppm. Conclusions: The above experiment offers rapid multi-element analysis in water samples (by using ICP/OES method) as well as less time consuming, cost-effective, accurate, specific, precise, and can be used for the identification and quantitative determination of L-dopa in herbal extracts. HPLC-High Performance Liquid Chromatography; ICP-OES-Inductively Coupled Plasma Optical Emission Spectrometry; RP- Reverse Phase; LOD- Limit of Detection, LOQ-Limit of Quantification; S/N- Signal to Noise ratio; RSD- Relative Standard Deviation, DADDiode Array Detection.

Keywords: RP-HPLC (Reversed phase High-performance liquid chromatography), ICPOES (Inductively coupled plasma optical emission spectrometer), levodopa, method development, Mucuna pruriens, Validation and heavy metal.