高效液相色谱波长切换法同时测定筋骨草中5种成分含量

Simultaneous Determination of Five Constituents in Ajuga Decumbens by HPLC with Wavelength Switching

目的建立同时测定筋骨草中哈巴苷、乙酰哈巴苷、芹菜素、木犀草素和杯苋甾酮含量的高效液相色谱(HPLC)法。方法色谱柱为Kromasil C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.4%磷酸溶液(梯度洗脱),流速为1.0 m L/min,检测波长为202 nm(哈巴苷、乙酰哈巴苷)和245 nm(杯苋甾酮、木犀草素、芹菜素),柱温为30℃,进样量为10μL。结果精密度、稳定性、重复性试验的RSD均小于4.7%;哈巴苷、乙酰哈巴苷、杯苋甾酮、木犀草素和芹菜素质量浓度分别在33.44~1 672.00μg/m L、18.248 0~912.40μg/m L、5.377 6~268.88μg/m L、1.728 0~86.40μg/m L、1.507 2~75.36μg/m L范围内与峰面积线性关系良好,平均加样回收率分别为99.06%,98.89%,98.97%,99.61%,97.15%,RSD分别为0.56%,0.41%,0.77%,1.23%,1.08%(n=6)。结论该方法高效、简便、稳定性、重复性好,可用于同时测定筋骨草中的5种成分。

Objective To establish a high-performance liquid chromatography(HPLC)method for the simultaneous determination of hapagide,acetylhapagide,cyasterone,luteolin and apigenin in Ajuga decumbens.Methods The chromatographic column was Kromasil C18 column(250 mm×4.6 mm,5μm),the mobile phase was acetonitrile-0.4%phosphoric acid solution(gradient elution),the flow rate was1.0 m L/min,the detection wavelength was 202 nm for hapagide and acetylhapagide and 245 nm for cyasterone,luteolin and apigenin,the column temperature was 30℃,and the injection volume was 10μL.Results The RSDs of precision,stability and repeatability tests were less than 4.7%.The liner ranges of hapagide,acetylhapagide,cyasterone,luteolin and apigenin were 33.44-1672.00μg/m L,18.2480-912.40μg/m L,5.3776-268.88μg/m L,1.7280-86.40μg/m L and 1.5072-75.36μg/m L,respectively.The average recoveries of hapagide,acetylhapagide,cyasterone,luteolin and apigenin were 99.06%,98.89%,98.97%,99.61%,97.15%with RSDs of 0.56%,0.41%,0.77%,1.23%,1.08%(n=6),respectively.Conclusion The method is efficient,simple,stable and reproducible,which can be used for the simultaneous determination of five constituents in Ajuga decumbens.