Development and Validation of HPLC Method for Quantification of Plumbagin in Plumbago Zeylanica L. Roots

Abstract

Background An RP-HPLC (Reverse Phase-High-performance liquid chromatography) method for the quantitative estimation and validation of the plumbagin in the methanolic fraction of Plumbago zeylanica L. was developed.

Method For achieving good separation, the RP-HPLC method was carried out with reverse phase C18 column, using methanol and water as mobile phase in the ratio of 65:35 (v/v), at the flow rate of 1 mL/min. The detection wavelength was set at 265 nm.

Results The retention time of plumbagin was found at 7.5±0.2 min. The coefficient of determination of plumbagin was found to be (r²) 0.9985 and equation Y=23148x+4327. The LOD and LOQ were found to be 34.06 and 113 ng/mL, respectively.

Conclusion The developed method was accurate, specific, precise, and reproducible. This RP-HPLC may be useful for quantitative estimation of the chemical constituents present in the plant extract as well as the quality assessment of the herbal product.

Key words

Plumbagin - Plumbago zeylanica L - High-performance liquid chromatography - method validation - quality control.

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