UPLC-MS/MS法同时测定灵源万应茶中的黄酮类成分含量

Simultaneous Determination of Flavonoids in LingYuan WanYing Medicinal Teas by UPLC-MS/MS

目的建立一种超高效液相色谱串联质谱法(UPLC-MS/MS)同时测定灵源万应茶中5种黄酮类成分(表儿茶素、花旗松素、芦丁、木犀草素、芹菜素)的含量定。方法采用Shim-pack XR-ODSⅢ(2.0 mm×75 mm,1.6μm)色谱柱,流动相选择0.1%甲酸5 mmol·L^(-1)乙酸铵水溶液(A)-甲醇(B),梯度洗脱,流速0.25 mL·min^(-1),柱温40℃;采用ESI离子源负离子扫描。结果在上述色谱条件下,测定的5种成分在各自的线性范围内呈现良好的线性关系(r≥0.9954),其精密度、稳定性、重复性良好;平均回收率在98.77%~100.1%之间,RSD小于5%。灵源万应茶样品中5个黄酮成分的含量为:表儿茶素(71.4±1.25)μg·g^(-1);花旗松素(0.284±0.0119)μg·g^(-1);芦丁(512±39.5)μg·g^(-1);木犀草素(1.51±0.454)μg·g^(-1);芹菜素(0.371±0.104)μg·g^(-1)。结论本研究所建立的同时测定灵源万应茶中5种黄酮类成分(表儿茶素、花旗松素、芦丁、木犀草素、芹菜素)的UPLC-MS/MS定量分析方法快速准确,可为灵源万应茶的质量控制提供参考依据。

OBJECTIVE To establish a ultra high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)method for simultaneous determination of 5 flavonoids(epicatechin,taxifolin,rutin,luteolin,and apigenin)in LingYuan WanYing Medicinal Teas.METHODS The UPLC-MS/MS assay was performed on Shim-pack XR-ODS C_(18)(2.0 mm×75 mm,2.6μm)column with 5 mmol·L^(-1) ammonium acetate(containing 0.1% ammoniu hydroxide)-methanol as mobile phase by gradient elution at a flow rate of 0.25 mL·min^(-1).The column temperature was set at 40℃.MS detection was performed using negative electrospray ionization.RESULTS Under the optimized chromatographic conditions,the determined chemical composition showed good linear relationship in their linear range(r≥0.9954).Its precision,stability and repeatability also met the requirements of analysis.The average recoveries(n=6)were in the range of 98.77% to 100.1%,while RSDs were less than 3%.The contents of 5 flavonoids in LingYuan WanYing Medicinal Teas were as follows:epicatechin(71.4±1.2)μg·g^(-1),taxifolin(0.284±0.0119)μg·g^(-1),rutin(512±39.5)μg·g^(-1),luteolin(1.51±0.454)μg·g^(-1),and apigenin(0.371±0.104)μg·g^(-1).CONCLUSION The proposed UPLC-MS/MS method for simultaneous determination of 5 flavonoids(epicatechin,taxifolin,rutin,luteolin,and apigenin)in LingYuan WanYing Medicinal Teas was fast and accurate,which can be used for quality control of LingYuan WanYing Medicinal Teas.