天麻中天麻素RH-HPLC测定及提取工艺研究

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Diamonsil C18 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。

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

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 Bl.. 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 μm) 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 μg/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 Bl. were determined as follows: 50% alcohol as extraction solvent, temperature 70 ℃, ratio of solid to liquid 1:6 (g/ml), and extraction time 3 h.