Liu Wen-Yuan, Feng Feng, Yu Cheng-Xia, Xie Ning
Department of Pharmaceutical Analysis, China Pharmaceutical University, 24 Tongjiaxiang, 210009 Nanjing, China.
Nat Prod Commun. 2010 Jan;5(1):23-6.
A sensitive and selective high performance liquid chromatographic method coupled with DAD detection is presented for quality control of Radix Ilicis Pubescentis. By means of this analytical procedure the major individual constituents (ilexoside O, ilexgenin A, ilexsaponin A1, ilexsaponin B1, liriodendin and acanthoside B) could be quantified simultaneously. LC-ESI-MS was applied for identification of the six compounds in the plant by comparing their m/z value and retention times with those of selected standards. For quantitative analysis, the extraction procedure and the extraction solvent were optimized in order to ensure the exhaustive extraction of the plant material. The HPLC conditions were evaluated and optimized for the exact quantification of all six individual compounds. Chromatographic separation was carried out on a C18 column using gradient elution with acetonitrile and 0.1% phosphoric acid as the mobile phase. Detection was carried out using a photodiode array detector. The calibration curves for determination of the six constituents showed good linearity over the investigated ranges (r2>0.999). Measurement of intra-day and inter-day variability (expressed as RSD value) was conducted to assess precisions of the method, and RSD (%) of intra- and inter-day variation were between 1.56-3.36% and 1.61-3.58%, respectively. The recoveries of the six compounds were between 96.4-102.2%, with RSD (%) values ranging from 1.7-3.8%. These validation results demonstrated the suitability of the method for the precise and accurate determination of the main constituents in Radix Ilicis Pubescentis. The method was successfully applied for quality evaluation of 12 batches of Radix Ilicis Pubescentis obtained from different regions of southern China. The contents of the six major constituents varied significantly due to their different origins, which can be used as an aid to assessing the quality of Radix Ilicis Pubescentis.
建立了一种灵敏且具选择性的高效液相色谱法结合二极管阵列检测法用于毛冬青质量控制。通过该分析程序可同时对主要的单一成分（冬青苷O、冬青素A、冬青皂苷A1、冬青皂苷B1、鹅掌楸苷和刺五加苷B）进行定量分析。采用液相色谱-电喷雾电离质谱法，通过将植物中六种化合物的质荷比和保留时间与选定标准品的质荷比和保留时间进行比较来鉴定这些化合物。对于定量分析，对提取程序和提取溶剂进行了优化，以确保对植物材料进行完全提取。对高效液相色谱条件进行了评估和优化，以实现对所有六种单一化合物的准确定量。色谱分离在C18柱上进行，以乙腈和0.1%磷酸作为流动相进行梯度洗脱。使用光电二极管阵列检测器进行检测。测定六种成分的校准曲线在所研究范围内显示出良好的线性关系（r2>0.999）。进行日内和日间变异性测量（以相对标准偏差值表示）以评估该方法的精密度，日内和日间变化的相对标准偏差（%）分别在1.56 - 3.36%和1.61 - 3.58%之间。六种化合物的回收率在96.4 - 102.2%之间，相对标准偏差（%）值在1.7 - 3.8%之间。这些验证结果表明该方法适用于准确测定毛冬青中的主要成分。该方法成功应用于对来自中国南方不同地区的12批毛冬青进行质量评价。六种主要成分的含量因产地不同而有显著差异，这可用于辅助评估毛冬青的质量。
