Simultaneous Quantitation of Piperine and Piperlongumine in the Fruit of Piper longum Linn. by Validated High-Performance Thin-Layer Chromatography—Densitometric Method

Summary

The fruit of Piper longum Linn. (family: Piperaceae), known as pippali in India, is a reputed drug of Ayurveda (the Indian system of medicine). It contains many amides of piperic acid among which piperine and piperlongumine are important from biological point of view. In the present work, we quantified these two marker compounds from the fruits of P. longum by high-performance thin-layer chromatography (HPTLC) with densitometry. The method was found to be precise. Relative standard deviation (RSD) values for intra-day analyses were in the range of 1.05 to1.07% (piperine) and 1.12 to 1.16% (piperlongumine) and, for inter-day analyses, the values were in the range of 1.11 to 1.21% (piperine) and 1.02 to1.18% (piperlongumine). Instrumental RSD values were 0.64 and 0.78% for piperine and piperlongumine, respectively. Accuracy of the method was evaluated by carrying a recovery study at three different levels for the two compounds. Average recoveries were found to be 99.23% for piperine and 99.26% for piperlongumine. P. longum fruits obtained from Bombay market showed 0.213 ± 0.009% w/w piperine and 0.364 ± 0.014% w/w piperlongumine when analyzed by the proposed method. Similarly, sample fruits obtained from Ahmedabad market had 0.85 ± 0.026% piperine and 2.70 ± 0.065% of piperlongumine. This TLC—densitometric method was found to be simple, precise, specific, sensitive, and accurate. So it can be used in routine quality control for the simultaneous analysis of piperine and piperlongumine from P. longum fruits and piperine alone from Piper nigrum fruits.