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目的:建立高效液相色谱法同时测定甜叶菊叶中新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸A和异绿原酸C6个酚酸类成分。方法:采用C18色谱柱(4.6 mm×250 mm,5μm),流动相为0.1%甲酸水溶液-乙腈,梯度洗脱,柱温30℃,检测波长327 nm。结果:6个酚酸质量浓度在10~196μg·m L-1范围内线性关系良好;6个酚酸的加样回收率分别为98.3%、95.2%、100.1%、97.3%、95.6%和96.1%,加样回收率、精密度、重复性、稳定性均符合有关规定;9批样品中6个酚酸类成分的含量分别为新绿原酸0.062%~0.725%,绿原酸0.277%~2.450%,隐绿原酸0.096%~0.694%,异绿原酸B 0.046%~0.867%,异绿原酸A0.157%~0.772%,异绿原酸C 0.135%~1.821%,6个酚酸之和在1.182%~6.668%。结论:该方法可为甜叶菊进一步开发利用提供质量控制方法。
OBJECTIVE: To establish a HPLC method for the simultaneous determination of neo-chlorogenic acid, chlorogenic acid, cryptogenic acid, iso-chlorogenic acid B, iso-chlorogenic acid A and iso-chlorogenic acid C6 phenolic acids in the leaves of Stevia rebaudiana. Methods: The mobile phase consisted of C18 column (4.6 mm × 250 mm, 5 μm) and mobile phase consisted of 0.1% formic acid in water with gradient elution at 30 ℃. The detection wavelength was 327 nm. Results: The linear range of 6 phenolic acids in the range of 10 ~ 196μg · m L-1 was good. The recoveries of 6 phenolic acids were 98.3%, 95.2%, 100.1%, 97.3%, 95.6% and 96.1 %, Sample recovery, precision, repeatability, stability are in line with the relevant provisions; 9 batches of samples of six phenolic acids content were new chlorogenic acid 0.062% ~ 0.725%, chlorogenic acid 0.277% ~ 2.450 %, Chlorogenic acid 0.096% ~ 0.694%, isooctanoic acid 0.046% ~ 0.867%, chlorogenic acid A0.157% ~ 0.772%, chlorogenic acid C 0.135% ~ 1.821%, 6 phenolic acids The sum of 1.182% ~ 6.668%. Conclusion: This method can provide quality control method for the further development and utilization of stevia.