Rout Kedar Kumar, Pradhan Subhalaxmi, Mishra Sagar Kumar
Utkal University, Pharmacognosy and Phytochemistry Division, University Department of Pharmaceutical Sciences, Vani Vihar, Bhubaneswar, Orissa, India.
J AOAC Int. 2008 Sep-Oct;91(5):1149-53.
A sensitive, simple, rapid, and efficient high-performance thin-layer chromatographic (HPTLC) method has been developed and validated for the analysis of berberine in marketed Ayurvedic formulations containing Berberis aristata DC for regulatory purposes. Chromatography of methanolic extracts of these formulations was performed on silica gel 60 F254 aluminum-backed TLC plates of 0.2 mm layer thickness. The plate was developed up to 66 mm with the ternary-mobile phase butanol-acetic acid-water (8 + 1 + 1, v/v/v) at 33 +/- 5 degrees C with 5 min of tank saturation. The marker, berberine, was quantified at its maximum absorbance of 350 nm. The limit of detection and limit of quantitation values were found to be 5 and 10 ng/spot. The linear regression analysis data for the calibration plot showed a good linear relationship with correlation coefficient = 0.9994 in the concentration range of 10 to 50 ng/spot for berberine with respect to peak area. The instrumental precision was found to be 0.49% coefficient of variation (CV), and repeatability of the method was 0.73% CV. Recovery values from 98.27 to 99.11% indicate excellent accuracy of the method. The developed HPTLC method is very accurate, precise, and cost-effective, and it has been successfully applied to the assay of marketed formulations containing B. aristata for determination of berberine.
为满足监管目的,已开发并验证了一种灵敏、简单、快速且高效的高性能薄层色谱(HPTLC)方法,用于分析市售含小檗(Berberis aristata DC)的阿育吠陀制剂中的黄连素。这些制剂的甲醇提取物在厚度为0.2 mm的硅胶60 F254铝背TLC板上进行色谱分析。在33±5℃下,使用三元流动相正丁醇 - 乙酸 - 水(8 + 1 + 1,v/v/v),在展开缸饱和5分钟的条件下,将板展开至66 mm。标记物黄连素在其350 nm的最大吸光度处进行定量。检测限和定量限分别为5和10 ng/斑点。校准曲线的线性回归分析数据显示,黄连素在10至50 ng/斑点的浓度范围内,相对于峰面积具有良好的线性关系,相关系数 = 0.9994。仪器精密度为变异系数(CV)0.49%,该方法的重复性为CV 0.73%。回收率在98.27%至99.11%之间,表明该方法具有出色的准确性。所开发的HPTLC方法非常准确、精密且具有成本效益,已成功应用于市售含小檗制剂中黄连素的含量测定。