ICAR-Directorate of Medicinal and Aromatic Plants Research, Anand, 387310, Gujarat, India.
Department of Botany, Faculty of Science, The M.S. University of Baroda, Vadodara, 390002, Gujarat, India.
J AOAC Int. 2021 May 21;104(2):485-497. doi: 10.1093/jaoacint/qsaa133.
Phyllanthus species exhibit a wide range of in vitro and in vivo pharmacological activities; however, little is known about the compounds present in the extracts that are responsible for such actions.
Development and validation of a simple reversed phase HPLC-PDA method for profiling of phyllanthin, hypophyllanthin, nirtetralin, and niranthin in extracts of Phyllanthus species was carried out.
Separation was achieved using an XBridge column® (150 × 4.6 mm, 5.0 µm id) in an isocratic elution mode with mobile phase comprising of a mixture of acetonitrile and water with TFA (0.05%, v/v, pH = 2.15) at ambient temperature with a flow rate of 1 mL/min.
Phyllanthin, hypophyllanthin, nirtetralin, and niranthin were eluted at mean retention times of 10.47, 11.10, 13.67, and 14.53 min, respectively. LOD and LOQ for all four analytes were 0.75 and 3.00 μg/mL, respectively. RSDr values for intraday and interday precision for phyllanthin, hypophyllanthin, nirtetralin, and niranthin were 0.38-1.32 and 0.45-1.77%; 0.22-3.69 and 0.24-3.04%, 0.73-2.37 and 0.09-0.31%, and 1.56-2.77 and 0.12-0.68%, respectively.
The developed and validated HPLC-PDA method was applied for identification and quantification of phyllanthin, hypophyllanthin, nirtetralin, and niranthin in extracts of different plant parts of selected Phyllanthus species. The outcome of the present investigation could be useful for selection of best species to promote its commercial cultivation and suitable extraction solvent for preparation of lignan-enriched fractions. This HPLC-PDA method could be useful for quality control of herbal formulations containing plants from Phyllanthus species.
叶下珠属植物表现出广泛的体外和体内药理活性；然而，对于负责这些作用的提取物中的化合物知之甚少。
开发并验证了一种简单的反相高效液相色谱-光电二极管阵列法，用于分析叶下珠属植物提取物中叶下珠素、隐叶下珠素、尼替拉宁和尼拉宁的分布。
采用 XBridge 柱（150×4.6mm，5.0μm id）在等度洗脱模式下，以乙腈和水的混合物为流动相，含 TFA（0.05%，v/v，pH=2.15），在环境温度下以 1mL/min 的流速进行分离。
叶下珠素、隐叶下珠素、尼替拉宁和尼拉宁分别在 10.47、11.10、13.67 和 14.53min 的平均保留时间洗脱。所有四种分析物的检出限和定量限分别为 0.75 和 3.00μg/mL。叶下珠素、隐叶下珠素、尼替拉宁和尼拉宁的日内和日间精密度的 RSDr 值分别为 0.38-1.32%和 0.45-1.77%；0.22-3.69%和 0.24-3.04%、0.73-2.37%和 0.09-0.31%、1.56-2.77%和 0.12-0.68%。
所建立的 HPLC-PDA 方法已应用于不同叶下珠属植物部分提取物中鉴定和定量分析叶下珠素、隐叶下珠素、尼替拉宁和尼拉宁。本研究的结果可用于选择最佳物种以促进其商业种植和合适的提取溶剂制备富含木脂素的馏分。该 HPLC-PDA 方法可用于含有叶下珠属植物的草药制剂的质量控制。
