The alternative system of medicines like Unani and Ayurveda is preferred worldwide nowadays due to its therapeutic efficacy, lower side effects, holistic approach, psychological dimensions, and qualitative action of weather and seasonal requirement. A simple procedure is described for the simultaneous extraction and estimation of piperlongumine and piperine in a well-known Unani polyherbal formulation using reversed-phase high-performance liquid chromatography (HPLC). The chromatography was carried out on reversed-phase C18 (250 × 4.6 mm) column with a mobile phase containing acetonitrile—water (50:50 v/v). Detection was accomplished with ultraviolet (UV) detection at λ = 325 nm. The flow rate was kept as 1.0 mL−1. The proposed method was validated according to International Conference on Harmonization (ICH) guidelines for accuracy (94.4–105.0%), precision (0.37–2.17% RSD), and robustness (0.14–2.11% RSD). The limit of detection (LOD) values were found as 30 and 10 ng mL−1, while limit of quantification (LOQ) was 100 and 30 ng mL−1 for piperlongumine and piperine, respectively, which proved the sensitivity of the method satisfactory enough for accurate analysis of the both piperlongumine and piperine.
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Piperine is the bioactive constituent of black pepper (Piper nigrum) . For quality control of piperine content, expensive instruments are usually used. A simple and inexpensive TLC image-analysis method for quantification of the piperine content of the traditional medicinal preparations of Bhutan has been established in this study. An image of the TLC chromatogram, under UV light at 254 nm, was taken by use of a digital camera and was further transformed to a density profile plot, along the direction of development, by use of Scion Image software. The concentration of piperine was calculated by comparing its peak area with a calibration plot established by chromatography of piperine standards on the same TLC plate. Linear regression analysis of the calibration data revealed a good linear relationship ( R 2 = 0.9962) between response and amount of piperine in the range 0.34–1.03 μg per spot. The specificity, precision, accuracy, and recovery of the method are satisfactory. The analytical results obtained by use of the method were not significantly different from those obtained by use of densitometric TLC. A disadvantage of the method is its low sensitivity — the limits of detection (LOD) and quantification (LOQ) were 15.36 and 46.54 ng per spot, respectively.
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