Akhade Meenakshi S, Agrawal Poonam A, Laddha K S
Medicinal Natural Products Research Laboratory, Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, Matunga (E), Mumbai-400 019, India.
Indian J Pharm Sci. 2013 Jul;75(4):476-82. doi: 10.4103/0250-474X.119835.
The aim of the present work was to develop and validate a reversed-phase high-performance liquid chromatography method for the simultaneous estimation of picroside I, plumbagin, and Z-guggulsterone in a polyherbal formulation containing Picrorhiza kurroa, Plumbago zeylanica, and Commiphora wightii extracts. The analysis was performed on a C18 column using the mobile phase consisting of solvent A (acetonitrile) and solvent B (0.1% orthophosphoric acid in water) with the following gradient: 0-12 min, 25% A; 12-17 min, 25-80% A; 17-32 min, 80% A; and 32-37 min, 80-25% A at a flow rate of 1 ml/min. Ultraviolet detection was at 255 nm. The method was validated for accuracy, precision, linearity, specificity, and sensitivity as per the norms of the International Conference on Harmonization. From the validation study, it was found that the method is specific, accurate, precise, reliable, and reproducible. Good linear correlation coefficients (r(2)>0.900) were obtained for calibration plots in the ranges tested. Limits of detection were 2.700, 0.090 and 0.099 μg/ml and limits of quantification were 9.003, 0.310, and 0.330 μg/ml for picroside I, plumbagin, and Z-guggulsterone, respectively. Intra and interday relative standard deviation (RSD) of retention times and peak areas was less than 3.0%. Recovery was found to be 100.21% for picroside I, 102.5% for plumbagin, and 103.84% for Z-guggulsterone. The established method was appropriate and the three markers were well resolved, enabling efficient quantitative analysis of picroside I, plumbagin and Z-guggulsterone. The method is a rapid and cost-effective quality control tool for routine quantitative analysis of picroside I, plumbagin, and Z-guggulsterone in tablet dosage form.
本研究的目的是开发并验证一种反相高效液相色谱法，用于同时测定一种含有胡黄连、白花丹和没药提取物的多草药制剂中胡黄连苷I、白花丹素和Z-古芸烯酮的含量。分析在C18柱上进行，流动相由溶剂A（乙腈）和溶剂B（0.1%的磷酸水溶液）组成，梯度如下：0-12分钟，25%A；12-17分钟，25%-80%A；17-32分钟，80%A；32-37分钟，80%-25%A，流速为1毫升/分钟。紫外检测波长为255纳米。该方法按照国际协调会议的规范进行了准确度、精密度、线性、特异性和灵敏度的验证。从验证研究中发现，该方法具有特异性、准确性、精密性、可靠性和可重复性。在所测试的范围内，校准曲线获得了良好的线性相关系数（r(2)>0.900）。胡黄连苷I、白花丹素和Z-古芸烯酮的检测限分别为2.700、0.090和0.099微克/毫升，定量限分别为9.003、0.310和0.330微克/毫升。保留时间和峰面积的日内和日间相对标准偏差（RSD）均小于3.0%。胡黄连苷I的回收率为100.21%，白花丹素为102.5%，Z-古芸烯酮为103.84%。所建立的方法适用，三种标志物分离良好，能够对胡黄连苷I、白花丹素和Z-古芸烯酮进行有效的定量分析。该方法是一种快速且经济高效的质量控制工具，可用于片剂剂型中胡黄连苷I、白花丹素和Z-古芸烯酮的常规定量分析。
