超高效液相色谱-串联质谱法测定清音饮中5种有效成分含量研究

Simultaneous determination of five active components in Qingyin Yin by UPLC-MS/MS

关键词:  清音饮  超高效液相色谱-串联质谱法  化学成分  含量测定

Key words:  Qingyin Yin  UPLC-MS/MS  chemical component  content determination

摘要:

[目的] 建立一种超高效液相色谱-串联质谱（UPLC-MS/MS）方法，用于同时测定清音饮中橙黄决明素、绿原酸、木犀草苷、3，5-O-二咖啡酰基奎宁酸和4，5-O-二咖啡酰基奎宁酸5种有性成分的含量。[方法] 色谱分离在Waters ACQUITY UPLC BEH C₁₈（2.1 mm×100 mm，1.7 μm）色谱柱上进行。流动相由含0.1%甲酸的水溶液（A）和乙腈（B）组成，采用梯度洗脱程序，流速为0.3 mL/min。柱温箱设定为35℃，进样体积为5 μL。质谱检测在电喷雾电离源负离子模式下进行，采用多反应监测扫描方式。[结果] 方法学验证结果表明，所测5种成分在相应浓度范围内均呈现良好线性关系，相关系数r均不低于0.998 7。该方法的精密度、重复性和稳定性均符合分析要求，平均加样回收率范围为94.00%~112.55%。[结论] 研究建立的UPLC-MS/MS含量测定方法灵敏度高、稳定性好，可以为清音饮的质量控制提供参考。

Abstract:

[Objective] To establish an ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS）method for the simultaneous determination of five bioactive components：aurantio-obtusin， chlorogenic acid，luteoloside，3，5-O-dicaffeoylquinic acid，and 4，5-O-dicaffeoylquinic acid in Qingyin Yin. [Methods] Chromatographic separation was performed on a Waters ACQUITY UPLC BEH C₁₈ column（2.1 mm×100 mm，1.7 μm）with a gradient elution program using 0.1% formic acid in water（A）and acetonitrile（B）as the mobile phase at a flow rate of 0.3 mL/min. The column temperature was set at 35 ℃，and the injection volume was 5 μL. Mass spectrometric detection was carried out in negative electrospray ionization（ESI）mode with multiple reaction monitoring（MRM）. [Results] The method was validated and showed good linearity for all five components within their respective concentration ranges，with correlation coefficients（r）not less than 0.998 7. The precision， repeatability，and stability of the method were satisfactory. The average recoveries ranged from 94.00% to 112.55%. [Conclusion] The established UPLC-MS/MS method is sensitive，specific，and reproducible，providing a reliable approach for the quality control of Qingyin Yin.