Quantitative determination of chemical constituents of Piper spp. using UPLC-ESI-MS/MS

- UPLC-ESI-MS/MS method was developed for quantification of 13 compounds.
- Piperamides was detected as the major constituents in ten Piper species.
- PCA was performed to compare and discriminate the samples.
- The proposed method would serve as a useful tool in the quality control of Piper species.

Species of the genus Piper are important medicinal plants of high commercial and economic importance have been used extensively as an individual plant part in various systems of medicine. To provide chemical information for differential use, quantitative determination and comparative analysis were carried out in different plant parts (fruits & leaves) of ten Piper species. To determine the contents and distribution of thirteen compounds a new method has been established using ultra performance liquid chromatography coupled with triple quadrupole electrospray tandem mass spectrometry in multiple reaction monitoring (MRM) mode with continuous polarity switching. A good linear regression relationship for each analyte was obtained (r, 0.9986-0.9999), intraday precision (%RSD, 0.63-3.37), interday precision (%RSD, 0.91-4.09), stability (RSD, 1.59-3.14) and recovery (%RSD, 0.78-2.78). The method sensitivity expressed in terms of limit of detection (LOD), which is ≤1.34 ng/mL for all analytes. The analysis results showed that piperamides are the signature class of compounds, which were found highest in fruits of Piper longum (21,500 μg/g), while phenolics were in close proportion and highest in fruits of Piper mullesa (1129 μg/g) and leaves of P. longum (1094 μg/g). Similarly, terpenoid content was detected highest in fruits of Piper umbellatum (8600 μg/g). Principal component analysis was carried out to compare and evaluate the quality of ten Piper species based on the contents of the thirteen investigated compounds. The present comparative study is of great importance for the rational application and quality control of different Piper species.

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