UPLC法测定羊红膻药材中9个活性成分的含量

目的： 采用超高相液相色谱法同时测定羊红膻药材中3个黄酮苷成分木犀草素-7-O-β-D-葡萄糖醛酸苷、槲皮素-3-O-β-D-葡萄糖醛酸苷、芹菜素-7-O-β-D-葡萄糖醛酸苷和6个有机酸类成分新绿原酸、绿原酸、隐绿原酸、异绿原酸A、异绿原酸B和异绿原酸C的含量。方法： 采用Waters Acquity UPLC BEH C₁₈色谱柱（2.1 mm×100 mm，1.7 μm），以甲醇-0.1%甲酸水为流动相梯度洗脱，流速0.3 mL·min^-1，检测波长345 nm，柱温30℃，进样量2 μL。结果： 3个黄酮类成分木犀草素-7-O-β-D-葡萄糖醛酸苷、槲皮素-3-O-β-D-葡萄糖醛酸苷、芹菜素-7-O-β-D-葡萄糖醛酸苷质量浓度分别在109.6~2 192、38.42~768.4和126.2~2 524 μg·mL^-1范围内具有良好线性关系（r ≥ 0.999 6，n=5），3批羊红膻药材中上述3个成分含量分别在3.73~4.69、1.02~1.29和4.42~5.64 mg·g^-1之间；6个有机酸类成分新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸A、异绿原酸C质量浓度分别在17.88~357.6、110.0~2 300、16.98~339.6、8.420~168.4、127.6~2 552、32.20~644.0 μg·mL^-1范围内线性关系良好（r ≥ 0.999 7，n=5），3批羊红膻药材中上述6个成分含量范围分别为0.51~0.88、3.28~6.02、0.44~1.02、0.21~0.37、3.67~5.38和1.35~1.72 mg·g^-1。结论： 本方法简便、快速，可同时测定羊红膻药材中9个活性成分的含量，测定结果可用于羊红膻药材的质量评价和质量控制。

Objective: To establish an UPLC method for the simultaneous determination of three flavonoid glycoside(luteolin-7-O-β-D-glucuronopyranoside,quercetin-3-O-β-D-glucuronpyrano-side and apigenin-7-O-β-D-glucuronopyranoside)and six organic acids(neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,isochlorogenic acid A,isochlorogenic acid B and isochlorogenic acid C)in Yanghongshan,the whole plant of Pimpinella thellungiana Wolff. Methods: Chromatographic separation was performed on a Waters Acquity UPLC BEH C₁₈ column(2.1 mm×100 mm,1.7 μm)at column temperature of 30℃.Methanol and 0.1% formic acid were adopted as the mobile phase with gradient elution at a flow rate of 0.3 mL·min^-1.The detection wavelength was 345 nm and the injection volume was 2 μL.Results: Luteolin-7-O-β-D-glucuronopyranoside,quercetin-3-O-β-D-glucuronopyranoside and apigenin-7-O-β-D-glucuronopyranoside had good linear relationship in the ranges of 109.6-2 192 μg·mL^-1,38.42-768.4 μg·mL^-1 and 126.2-2 524 μg·mL^-1,respectively(r ≥ 0.999 6,n=5),the contents of these three components in 3 batches of Yanghongshan were 3.73-4.69,1.02-1.29 and 4.42-5.64 mg·g^-1 respectively.Good linearity of the six organic acids,including neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,isochlorogenic acid B,isochlorogenic acid A and isochlorogenic acid C,were in the ranges of 17.88-357.6,110.0-2 300,16.98-339.6,8.420-168.4,127.6-2 552 and 32.20-644.0 μg·mL^-1,respectively(r ≥ 0.999 7,n=5).The contents of the six components in 3 batches of Yanghongshan were 0.51-0.88,3.28-6.02,0.44-1.02,0.21-0.37,3.67-5.38 and 1.35-1.72 mg·g^-1,respectively.Conclusion: This method is simple and fast,which can be used for the simultaneous determination of 9 active components in Yanghongshan.The determination results can be used to evaluate and control the quality of this herb.

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