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目的:采用反相高效液相色谱切换波长法建立同时测定白蔹中没食子酸、原儿茶醛、儿茶素、白藜芦醇4个成分含量的方法。方法:采用Agilent Zorbax Eclipse plus色谱柱(4.6 mm×250 mm,5μm);流动相为乙腈-0.1%磷酸水溶液,梯度洗脱;流速:1.0 mL·min-1;检测波长:270 nm(0~27 min),360 nm(27.01~36 min),306 nm(36.01~40 min);柱温:30℃。结果:没食子酸、原儿茶醛、儿茶素、白藜芦醇的进样量分别在0.0522~1.044μg(r=0.9993)、0.0548~1.096μg(r=0.9992)、0.4004~8.008μg(r=0.9990)和0.0086~0.172μg(r=0.9993)范围内,与色谱峰峰面积呈良好线性响应;平均回收率分别为100.5%、98.16%、99.60%、99.80%(n=6)。结论:该方法经方法学验证可用于白蔹的质量控制。
OBJECTIVE: To establish a method for simultaneous determination of four constituents of gallic acid, protocatechuic aldehyde, catechin and resveratrol in Bletilla striata by reversed phase high performance liquid chromatography with switched wavelength method. Methods: The Agilent Zorbax Eclipse plus column (4.6 mm × 250 mm, 5 μm) was used. The mobile phase consisted of acetonitrile-0.1% phosphoric acid solution with gradient elution. The flow rate was 1.0 mL · min-1. The detection wavelength was 270 nm 27 min), 360 nm (27.01 ~ 36 min) and 306 nm (36.01 ~ 40 min). The column temperature was 30 ℃. Results: The injection volume of gallic acid, protocatechuic aldehyde, catechin and resveratrol were 0.0522-1.044μg (r = 0.9993), 0.0548-1.096μg (r = 0.9992), 0.4004-8.008μg = 0.9990) and 0.0086 ~ 0.172μg (r = 0.9993), respectively. The average recoveries were 100.5%, 98.16%, 99.60% and 99.80% (n = 6), respectively. Conclusion: This method is validated by methodology for the quality control of Bletilla striata.