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Introduction: Plumbagin (5-hydroxy-2-methyl-1,4-naphthoquinone) is a yellowish quinonoid compound possessed various biological activities, abundantly found in roots of Plumbago indica L. or scarlet leadwort which is an herbal plant used as traditional remedies to treat a variety of illness. Objectives: The present study aimed to develop and validate an analytical method for quantification of plumbagin in the P. indica crude extract using reverse phase-high performance liquid chromatographic technique (RP-HPLC). Utilization of the validated method was applied for determination of plumbagin in the P. indica root crude extracts. Method: The RP-HPLC system consisted of a C18 column as stationary phase with a mobile phase of acetonitrile and water (50:50, v/v) at a flow rate of 1 mL/min. Chromatogram was detected at UV 254 nm. Results: No interference peak was observed at the retention time of plumbagin (tR = 6.2 min) with the good linearity (r2 = 0.99823). Limit of detection (LOD) and limit of quantification (LOQ) were 21.85 ng/mL and 72.82 ng/mL, respectively. Within-day and between-day precision expressed by % relative standard deviation were of 0.37-0.65% and 0.17-1.25%, respectively, with the accuracy as %recovery of 98.71±4.83%. Conclusion: The method was proven to be specific, sensitive, precise, and accurate for quantitative determination of plumbagin in the P. indica crude extracts. The validated method was utilized and found the content of plumbagin in the P. indica methanolic and ethanolic crude extracts of 0.15±0.00% and 0.21±0.01% dry weight, respectively.
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