Alam Prawez, Ali Mohd, Singh Rahul, Ahmad Sayeed, Shakeel Faiyaz
Department of Pharmacognosy and Phytochemistry, Jamia Hamdard, Hamdard Nagar, New Delhi-110062, India.
J Chromatogr Sci. 2009 Nov-Dec;47(10):910-3. doi: 10.1093/chromsci/47.10.910.
A simple, economic, robust, reproducible, selective, and precise high-performance liquid chromatography (HPLC) method for estimation of cordifolioside A in both 60% methanolic extract of Tinospora cordifolia and marketed formulation is developed and validated in the present study. The mobile phase composed of acetonitrile-water (25:75%, v/v) give a sharp and well-defined peak of cordifolioside A at the retention time of 9.52 +/- 1.03 min. HPLC estimation of cordifolioside A is carried out at wavelength of 210 nm with flow rate of 1.0 mL/min. The linear regression analysis data for the calibration curve shows good linear relationship with correlation coefficient of 0.997 in the concentration range of 0.5-50 microg/mL. The linear regression equation is y = 29716x - 4417.4. The limit of detection and limit of quantification are 0.18 and 0.55 microg/mL, respectively. The developed method is validated for accuracy, precision, reproducibility, and robustness as per ICH guidelines. The proposed method with high degree of precision and accuracy is employed for the estimation of cordifolioside A in methanolic extract and in formulation. Statistical analysis proved that the method is precise, reproducible, selective, and accurate for the estimation of cordifolioside A.
本研究开发并验证了一种简单、经济、稳健、可重现、选择性好且精确的高效液相色谱（HPLC）方法，用于测定毛叶锡生藤60%甲醇提取物和市售制剂中的cordifolioside A。由乙腈-水（25:75%，v/v）组成的流动相在9.52±1.03分钟的保留时间处给出了cordifolioside A尖锐且明确的峰。cordifolioside A的HPLC测定在210 nm波长、1.0 mL/min流速下进行。校准曲线的线性回归分析数据显示，在0.5 - 50μg/mL浓度范围内，相关系数为0.997，具有良好的线性关系。线性回归方程为y = 29716x - 4417.4。检测限和定量限分别为0.18和0.55μg/mL。按照ICH指南对所开发的方法进行了准确性、精密度、重现性和稳健性验证。所提出的方法具有高度的精密度和准确性，用于甲醇提取物和制剂中cordifolioside A的测定。统计分析证明，该方法对于cordifolioside A的测定是精确、可重现、选择性好且准确的。
