Chaurasiya Narayan Das, Uniyal Girish Chandra, Lal Payare, Misra Laxminarain, Sangwan Neelam Singh, Tuli Rakesh, Sangwan Rajender Singh
Central Institute of Medicinal and Aromatic Plants, PO CIMAP, Lucknow-226015, India.
Phytochem Anal. 2008 Mar-Apr;19(2):148-54. doi: 10.1002/pca.1029.
A reversed-phase HPLC method for the simultaneous analysis of nine structurally similar withanolides, namely, 27-hydroxy withanone, 17-hydroxy withaferin A, 17-hydroxy-27-deoxy withaferin A, withaferin A, withanolide D, 27-hydroxy withanolide B, withanolide A, withanone and 27-deoxywithaferin A, has been developed using a linear binary gradient solvent system comprising methanol and water containing 0.1% acetic acid. Both photodiode array and evaporative light scattering detection were used to profile the extract compositions and to quantify the withanolides therein. Homogeneity and purity of each peak was ascertained by comparative evaluation of the on-line UV spectra of the eluted compounds with those of the reference compounds. The method has been validated with respect to various parameters of performance quality including computation regression analysis based on calibration curves, peak resolution factor, asymmetry factor, tailing factor, RSD (%) of retention time and peak area response, limit of quantivation, limit of detection, precision and recovery. The developed method has been applied to the analysis of leaf and root tissues of Withania somnifera for withanolide content.
已开发出一种反相高效液相色谱法，用于同时分析9种结构相似的睡茄内酯，即27 - 羟基睡茄酮、17 - 羟基睡茄素A、17 - 羟基 - 27 - 脱氧睡茄素A、睡茄素A、睡茄内酯D、27 - 羟基睡茄内酯B、睡茄内酯A、睡茄酮和27 - 脱氧睡茄素A，该方法使用由甲醇和含0.1%乙酸的水组成的线性二元梯度溶剂系统。利用光电二极管阵列和蒸发光散射检测对提取物成分进行分析并对其中的睡茄内酯进行定量。通过将洗脱化合物的在线紫外光谱与参考化合物的光谱进行比较评估，确定每个峰的均一性和纯度。该方法已针对性能质量的各种参数进行了验证，包括基于校准曲线的计算回归分析、峰分辨率因子、不对称因子、拖尾因子、保留时间和峰面积响应的相对标准偏差（%）、定量限、检测限、精密度和回收率。所开发的方法已应用于分析印度人参叶和根组织中的睡茄内酯含量。
