Rare endemic ethnomedicinal plants that are indigenous to the Piperaceae family have an ancient history as traditional medicine in the treatment of diverse ailments like gastrointestinal disorders. The present research work highlights a digitally optimized, rapid, accurate and highly sensitive high-performance thin-layer chromatography (HPTLC) method for the simultaneous screening and quantification of piperine from ex-situ cultured callus, a kind of endangered Piper trichostachyon species. Analysis of piperine was performed on thin-layer chromatography (TLC) aluminum plates pre-coated with silica gel F₂₅₄ as the stationary phase. Linear ascending development was carried out in a twin-trough glass chamber with a saturation pad. Developing phase comprising of n-hexane‒ethyl acetate (5:15, V/V) was put to task. Spectrodensitometric analysis of the plate was performed in absorbance mode at 342 nm using a CAMAG TLC Scanner IV. Compact bands for piperine with R_F value of 0.50 ± 0.010 were recorded exhibiting a good linear relationship with R = 99.98% in the concentration range of 100‒700 ng/spot with respect to the peak area. The proposed HPTLC method was validated in accordance with the International Council for Harmonization (ICH) guidelines for linearity, accuracy, precision, repeatability, reproducibility, limit of detection, limit of quantification, sensitivity, and specificity. The total piperine content was found to be 62.39 μg/mL in the wild plant and 41.57 μg/mL in the callus of P. trichostachyon. The chromatographic analysis concluded that the method developed and validated as a novel effort for the concurrent evaluation and comparative quantification of piperine in species of P. trichostachyon is reproducible and selective for the analysis of piperine.

胡椒科本土的稀有地方民族药用植物作为治疗胃肠道疾病等多种疾病的传统药物有着悠久的历史。目前的研究工作重点介绍了一种数字优化、快速、准确和高灵敏度的高效薄层色谱 (HPTLC) 方法，用于从异地培养的愈伤组织中同时筛选和定量胡椒碱，这是一种濒临灭绝的毛尖角胡椒物种。胡椒碱的分析在薄层色谱 (TLC) 铝板上进行，该铝板上预涂有硅胶 F ₂₅₄作为固定相。在带有饱和垫的双槽玻璃室中进行线性上升显影。开发阶段包括Ñ己烷-乙酸乙酯（5:15，V / V）中的溶液投入到任务。使用CAMAG TLC Scanner IV以吸光度模式在342nm处进行板的分光光度计分析。记录到胡椒碱的致密谱带，其R _F值为 0.50 ± 0.010， 在 100-700 ng/点的浓度范围内与峰面积显示出良好的线性关系，R = 99.98%。提议的 HPTLC 方法根据国际协调委员会 (ICH) 的线性、准确度、精密度、重复性、再现性、检测限、定量限、灵敏度和特异性指南进行了验证。野生植物中胡椒碱总含量为62.39 μg/mL，愈伤组织中胡椒碱含量为41.57 μg/mL。旋毛虫。色谱分析得出的结论是，作为同时评估和比较定量P. trihostachyon物种中胡椒碱的一种新方法而开发和验证的方法对于胡椒碱的分析具有重现性和选择性。