天麻中天麻素RP-HPLC测定及提取工艺研究

Determination of Gastrodin in Gastrodia elata Bl. by Reverse Phase (RP)-HPLC and Study on Its Extraction Technology

通过考察不同检测波长、流动相和溶剂对天麻素出峰效果的影响,改进了2005年版《中国药典》收载的天麻药材中天麻素含量测定的RP-HPLC方法,并探讨了提取溶剂、提取温度、料液比、提取时间对天麻素提取工艺的影响。结果表明:采用DiamonsilC18柱(150mm×4.6mm,5μm),乙腈-水(3:97,V/V))溶液为流动相,流速1ml/min,柱温为室温,检测波长220nm,并用乙腈-0.05%磷酸溶液(3:97,V/V))作为溶剂,天麻素在10～80μg/ml浓度范围内呈现良好的线性关系,其回归方程和相关系数分别为:y=4.0264x+0.6311,R2=0.9992,最低检测限为2ng。精密度RSD为0.41%(n=6),重现性实验RSD为0.86%(n=6),回收率为98.4%,RSD为0.84%(n=6);天麻素最佳提取工艺条件为:提取溶剂50%乙醇,提取温度70℃,料液比1:10(g/ml),提取时间6h。

By investigating the effects of different detection wavelength, mobile phase and solvent on the peak profile of gastrodin, the present study improved the traditional RP-HPLC method for determining gastrodin in Gastrodia elata B1.. Also, the effects of extraction solvent, extraction temperature, solid-liquid ratio and extraction time on the extraction rate of gastrodin were discussed. The results showed that the optimal working conditions of RP-HPLC were as follows： using Diamonsil C18 column （150 mm × 4.6 mm, 5 μ m） at ambient temperature, acetonitrile-water （3：97, V/V） as the mobile phase, flow rate 1.0 ml/min, detection wavelength 220 nm, and acetonitrile-0.05% phosphoric acid （3：97, V/V） as the solvent of gastrodin. The calibration curve was well linear in the range of 10 to 80 μ g/ml, the regression equation was y = 4.0264x ＋ 0.6311 with a determination coefficient of 0.9992. The limit of detection of this method was 2 ng. The relative standard deviations （RSDs） of precision and repeatability were 0.41% and 0.86%, respectively （n = 6）, and the average spike recovery rate was 98.4% with a RSD of 0.84% （n = 6）. The optimum extraction conditions of gastrodin from Gastrodia elata B1. were determined as follows： 50% alcohol as extraction solvent, temperature 70℃, ratio of solid to liquid 1：10 （g/ml）, and extraction time 6 h.