一种用于测定吴茱萸中去氢吴茱萸碱、柠檬苦素、吴茱萸碱和吴茱萸次碱的超高效液相色谱-电喷雾串联质谱法的建立与验证

Development and validation of an UPLC-ESI-MS/MS method for determination of dehydroevodiamine, limonin, evodiamine, and rutaecarpine in Evodiae Fructus.

作者信息

Zhao Yang, Zhao Yunling, Zhou Xin, Gong Xiaojian

机构信息

The Research Center for Quality Control of Nature Medicine, Guizhou Normal University, Guiyang 550001, Guizhou, China ; Key Laboratory for Information System of Mountainous Area and Protection of Ecological Environment of Guizhou Province, Guizhou Normal University, Guiyang 550001, Guizhou, China.

出版信息

Pharmacogn Mag. 2014 Jul;10(39):374-83. doi: 10.4103/0973-1296.137381.

OBJECTIVE

Evodiae Fructus (EF), one of the most widely used traditional Chinese medicines, mainly consists of alkaloids, is widely used for the treatments of headache and gastrointestinal disorders. In this study, a sensitive and reliable UPLC-ESI-MS/MS method was developed for qualitative determination of dehydroevodiamine, limonin, evodiamine, and rutaecarpine.

MATERIALS AND METHODS

Chromatographic separations were accomplished on a Phenomenex Kinetex XB-C18 column (2.1 × 150 mm, 1.7 μm) by using a gradient elution profile with a mobile phase consisting of 0.5% formic acid in water (A) and acetonitrile (B). Detection was performed using multiple reactions monitoring mode under ESI in the positive ion mode.

RESULTS

The results showed good linearity over the investigated concentration ranges (R (2)>0.9900) for the analytes. The limit of quantitations (LOQs) were 6.88 ng/mL for dehydroevodiamine, 18.6 ng/mL for limonin, 6.24 ng/mL for evodiamine, and 2.56 ng/mL for rutaecarpine, respectively. Intraday and interday precisions (relative standard deviations, %) were <5% and accuracies ranged from 92% to 106%.

CONCLUSION

The validated method was successfully applied to assay the contents of the four compounds in EF samples from different regions, with which just 10 min was needed to analyze each sample.

目的

吴茱萸是应用最为广泛的传统中药之一，主要成分是生物碱，广泛用于治疗头痛和胃肠道疾病。本研究建立了一种灵敏可靠的超高效液相色谱-电喷雾串联质谱（UPLC-ESI-MS/MS）方法，用于定性测定去氢吴茱萸碱、柠檬苦素、吴茱萸碱和吴茱萸次碱。

材料与方法

采用Phenomenex Kinetex XB-C18色谱柱（2.1×150 mm，1.7μm）进行色谱分离，流动相为含0.5%甲酸的水（A）和乙腈（B），采用梯度洗脱。在电喷雾正离子模式下，采用多反应监测模式进行检测。

结果

结果表明，在所研究的浓度范围内，分析物具有良好的线性关系（R²>0.9900）。去氢吴茱萸碱、柠檬苦素、吴茱萸碱和吴茱萸次碱的定量限（LOQ）分别为6.88 ng/mL、18.6 ng/mL、6.24 ng/mL和2.56 ng/mL。日内和日间精密度（相对标准偏差，%）均<5%，准确度在92%至106%之间。