Rapid validated HPTLC method for estimation of piperine and piperlongumine in root of Piper longum extract and its commercial formulation

Abstract

Piperine and piperlongumine, alkaloids having diverse biological activities, commonly occur in roots of Piper longum L., Piperaceae, which have high commercial, economical and medicinal value. In present study, rapid, validated HPTLC method has been established for the determination of piperine and piperlongumine in methanolic root extract and its commercial formulation 'Mahasudarshan churna ® ' using ICH guidelines. The use of Accelerated Solvent Extraction (ASE) as an alternative to conventional techniques has been explored. The methanol extracts of root, its formulation and both standard solutions were applied on silica gel F 254 HPTLC plates. The plates were developed in Twin chamber using mobile phase toluene: ethyl acetate (6:4, v/v) and scanned at 342 and 325 nm (λ max of piperine and piperlongumine, respectively) using Camag TLC scanner 3 with CATS 4 software. A linear relationship was obtained between response (peak area) and amount of piperine and piperlongumine in the range of 20-100 and 30-150 ng/spot, respectively; the correlation coeffi cient was 0.9957 and 0.9941 respectively. Sharp, symmetrical and well resolved peaks of piperine and piperlongumine spots resolved at R f 0.51 and 0.74, respectively from other components of the sample extracts. The HPTLC method showed good linearity, recovery and high precision of both markers. Extraction of plant using ASE and rapid HPTLC method provides a new and powerful approach to estimate piperine and piperlongumine as phytomarkers in the extract as well as its commercial formulations for routine quality control.

APPLICATION OF ACCELERATED SOLVENT EXTRACTION (ASE) AND THIN LAYER CHROMATOGRAPHY (TLC) TO DETERMINATION OF PIPERINE IN COMMERCIAL SAMPLES OF PEPPER (PIPER NIGRUML.)

“…Up to our best knowledge, this type of extraction combined with quantification of piperine by means of TLC has never before been applied to the Piper nigrum L. fruit (but to the roots of Piper longum only [6] ). To this effect, the calibration curve was first developed, by plotting the chromatographic peak heights for the piperine standard against the sample aliquots spotted on to the chromatographic plate.…”

“…[4] As thin layer chromatography (TLC) can be performed easily and inexpensively, many applications are already reported. Piperine was determined in the plant material, for example, in fruits of Piper nigrum [5,6] and roots of Piper longum, [7] as well as in the other matrices (such, as biological tissues), for example, in liver and serum. [8] Generally, in these methods, glass plates precoated with silica gel were used, whereas in the course of the separation of piperine isomers, application of the silver nitrate impregnated plates was recommended.…”

Piperine—The Bioactive Compound of Black Pepper: From Isolation to Medicinal Formulations

“…The achieved results approved such premise when a piperine extraction yield of 3.9% was attained in 12 h with a cycle time of 8 min using DBSA, although the extraction yield of 3.8% was obtained within 22 h and cycle time of 16 min using the conventional Soxhlet apparatus. Rajopadhye and others () extracted piperine from the root of Piper nigrum by Soxhlet extraction with methanol in 8 h and reached a piperine content of 9.56 ± 0.83 mg/g of root. Yamaguchi and others () extracted piperine from black pepper corns by 3 methods, namely, solvent extraction, Soxhlet extraction, and supercritical carbon dioxide extraction.…”

Simultaneous quantification of piperlongumine and piperine in traditional polyherbal formulation using validated HPLC method

“…Literature review reveals that there are few high-performance liquid chromatography (HPLC) [17][18][19], high-performance thin-layer chromatography (HPTLC) [20], and liquid chromatography-mass spectrometry (LC-MS) [21,22] methods available for the quantification of piperine and piperlongumine individually; however, only few studies have been reported on the simultaneous analysis of these components. Pundarikakshudu et al (2014) [23] and Rajopadhye et al (2012) [24] in herbal formulations using HPTLC methods.…”

“…Since the analysis was not performed at the λ max of the compounds, the sensitivity of the components can become an issue. The second HPTLC method reported by Rajopadhye et al (2012) used a mobile of toluene-ethyl acetate (6:4, v/v) and scanned at 342 and 325 nm. The samples were extracted using accelerated solvent extraction (ASE) method; however, the developed method was found to have achieved poor recoveries ranging from 93.72 to 96.67% only.…”