Department for Life Quality Studies, University of Bologna, Corso D'Augusto 237, 47921, Rimini, Italy.
Freia Farmaceutici Srl, via Marghera 22, 20149, Milano, Italy.
J Pharm Biomed Anal. 2021 May 10;198:113985. doi: 10.1016/j.jpba.2021.113985. Epub 2021 Feb 20.
A new chromatographic method by Ultra High Performance Liquid Chromatographic (UHPLC) technology, has been developed and validated for the determination of polydatin and resveratrol, as potential metabolite, in human plasma. After the optimization of the chromatographic conditions, the method has been validated on spiked human plasma samples. The optimized extraction allowed to obtain analytes recovery up to 98.48 ± 4.03 %. Then, the isocratic elution in reversed phase mode, provides the separation of polydatin and resveratrol in less than 10.0 min. Chromatographic analysis was performed on a C18, 10 cm x 3.0 mm, 2.7 μm stationary phase, by using triethanolamine phosphate solution (0.1 M, pH = 3.7) and ACN 85:15 (v/v) as mobile phase at a flow rate of 0.5 mL/min. The UV detector was set at 306 nm for the analysis of both polydatin and resveratrol. The limit of detection (LoD) and the limit of quantification (LoQ) for polydatin in plasma samples were found to be 7.82 ± 0.38 nM and 26.06 ± 1.28 nM respectively. The method was found to be accurate and precise with a coefficient for intra- and inter-day variation below 5 %. All the reported data demonstrate how the developed method is rapid and sensitive. Moreover, results of the analysis of plasma samples, obtained from orally treated volunteers with nutritional supplements containing polydatin, have shown the method to be suitable for the pharmacokinetic characterization of polydatin and resveratrol, as metabolite, in humans.
一种新的色谱方法通过超高效液相色谱 (UHPLC) 技术，已经开发并验证了用于测定人血浆中白藜芦醇苷和白藜芦醇作为潜在代谢物。在优化了色谱条件后，该方法对人血浆样品进行了验证。优化的提取方法使分析物的回收率达到 98.48 ± 4.03%。然后，反相模式下的等度洗脱，在不到 10.0 分钟内即可分离白藜芦醇苷和白藜芦醇。色谱分析在 C18 上进行，10 cm x 3.0 mm，2.7 μm 固定相，使用三乙醇胺磷酸盐溶液（0.1 M，pH = 3.7）和 ACN 85:15（v/v）作为流动相，流速为 0.5 mL/min。UV 检测器设置为 306nm 用于分析白藜芦醇苷和白藜芦醇。在血浆样品中白藜芦醇苷的检测限（LoD）和定量限（LoQ）分别为 7.82 ± 0.38 nM 和 26.06 ± 1.28 nM。该方法被证明具有准确性和精密度，日内和日间变异系数均低于 5%。所有报告的数据均表明，所开发的方法快速灵敏。此外，从口服含有白藜芦醇苷的营养补充剂的志愿者的血浆样品中分析结果表明，该方法适用于白藜芦醇苷和白藜芦醇作为代谢物在人体中的药代动力学特征。
