高效液相色谱联用质谱法对睡莲花中4种成分的鉴定和含量测定

Identification and determination of 4 components in Nymphaea candida C.Presl by HPLC-UV-MS/MS

目的建立睡莲花中4种成分(没食子酸、没食子酸甲酯、烟花苷、槲皮素)的高效液相色谱联用质谱(HPLC-UV-MS/MS)鉴定和含量测定方法。方法采用Agilent C_(18)反相色谱柱(4.6 mm×250 mm,5μm)分析,以流动相A为0.2%的冰乙酸水溶液,流动相B为甲醇进行梯度洗脱,检测波长为270 nm,柱温为30℃,流速为1 mL·min^(-1);质谱条件:电喷雾负离子扫描模式(ESI-)。结果通过睡莲花中4种成分的一级和二级质谱与对照品对比,鉴定药材中的目标化合物,4个化合物在浓度范围内线性关系良好(R^(2)≥0.9990),平均加样回收率为98.87%~103.25%。结论本法能准确、可靠的对睡莲花中4种化学成分进行鉴定和含量测定,可用于睡莲花的质量控制,为其药效物质基础研究提供科学依据。

Objective To develop an HPLC-UV-MS/MS method for the simultaneous identification and quantification of 4 components(gallic acid,methyl gallate,nicotiflorin and quercetin)in Nymphaea candida C.Presl.Methods The chromatographic separation was carried out on an Agilent C18 reversed phase chromatography column(4.6 mm×250 mm,5μm)with the mobile phase consisting of 0.2%acetic acid aqueous solution(A)and methanol(B)in a gradient elution.The detection wavelength was 270 nm,the column temperature was 30℃,and the flow rate was 1 mL·min^(-1).The electrospray negative ion scanning mode(ESI-)was used.Results The 4 target components were identified by comparing the MS and MS/MS spectrometry information of the reference substance with that of Nymphaea candida C.Presl.The methodology validation showed good linearity of the 4 components in the concentration range(R^(2)≥0.9990),and the average recoveries ranged at 98.87%~103.25%.Conclusion The HPLC-UV-MS/MS method can be used to identify and determine the 4 chemical components in Nymphaea candida C.Presl accurately and reliably.It can control the quality of Nymphaea candida C.Presl and provide scientific basis for the study of its pharmacodynamic materials.