基于UPLC-MS/MS快速测定中药材中6种鞣质类成分

Rapid Determination of Six Tannins in Chinese Medicinal Materials Based on UPLC-MS/MS

目的建立快速测定中药材中没食子酸、没食子酸甲酯、没食子酸乙酯、鞣花酸、3,3’-O-二甲基鞣花酸、柯里拉京6种鞣质类成分的超高效液相色谱串联质谱(UPLC-MS/MS)分析方法。方法采用Agilent RRHD Eclipse Plus C18色谱柱(2.1 mm×50 mm,1.8μm),流动相为甲醇-0.1%甲酸水溶液,梯度洗脱,流速0.3 mL·min^(–1),柱温40℃,进样量1μL;三重四级杆质谱,采用电喷雾离子源、负离子模式,通过多重反应监测模式对15种中药材中6种鞣质类成分进行定量分析。结果 6种成分在各自浓度范围内线性关系良好(r>0.999),没食子酸、没食子酸甲酯、没食子酸乙酯、鞣花酸、3,3’-O-二甲基鞣花酸、柯里拉京的加样回收率分别为99.8%,98.3%,100.1%,100.3%,100.7%,99.2%,RSD值分别为1.11%,2.23%,1.07%,1.34%,1.64%,1.24%。15种中药材中均含一定量的没食子酸和没食子酸乙酯,且以五倍子中含量最高;鞣花酸含量以覆盆子根最高;另外3种成分不均匀分布于各药材中。结论所建立的方法稳定、快速、灵敏度高,适用于含有上述6种鞣质类成分中药材的定量分析。

OBJECTIVE To develop an UPLC-MS/MS method for rapidly qualitative and quantitative determination of six tannins(gallic acid, methyl gallate, ethyl gallate, ellagic acid, 3,3’-Di-O-methylellagic acid, coriagin) in Chinese medicinal materials. METHODS An Agilent RRHD Eclipse Plus C18 column(2.1 mm×50 mm, 1.8 μm) with a gradient elution of methanol and 0.1%formic acid in water at a flow rate of 0.3 mL·min^(–1), the column temperature was 40 ℃, and the injection volume was 1 μL. Triple quadrupole mass spectrometry with electrospray ion source and negative ion mode was used to quantitatively analyze 6 tannins in 15 kinds of Chinese herbal medicines by multiple reaction monitoring mode. RESULTS Good linear relationship(r>0.999) were observed in the test ranges for 6 compounds, and the average recoveries of gallic acid, methyl gallate, ethyl gallate, ellagic acid, 3,3’-Di-O-methylellagic acid, coriagin were 99.8%(RSD=1.11%), 98.3%(RSD=2.23%), 100.1%(RSD=1.07%), 100.3%(RSD=1.34%), 100.7%(RSD=1.64%), 99.2%(RSD=1.24%), respectively. The gallic acid and ethyl gallate were detected in all 15 kinds of Chinese medicinal materials, and the content in Chinese gallnut was the maximum;the content of ellagic acid in Rubi Radix et Rhizoma was the highest;another 3 compounds were distributed in above drugs non-uniformly. CONCLUSION The established method is stable, rapid, sensitive, it can be used for the quantitative analysis of above six tannins in Chinese medicinal materials.