基于HPLC-Q-TOF-MS技术分析红景天颗粒的化学成分及大鼠口服后的入血成分

Chemical profiling of Rhodiola Crenulata granules and characterization of the absorbed components and metabolites in rat plasma following oral administration based on HPLC-Q-TOF-MS

目的:借助高效液相色谱-四级杆飞行时间质谱联用技术(HPLC-Q-TOF-MS)分析红景天颗粒的主要化学成分以及大鼠口服红景天颗粒后的入血成分。方法:采用HPLC-Q-TOF-MS分析技术,选用反相ODS色谱柱,以乙腈-0.1%甲酸水溶液为流动相进行梯度洗脱,采用正、负离子模式及IDA扫描模式采集质谱数据,采用PeakView软件获取色谱保留时间以及母离子和碎片离子信息,定性鉴别红景天颗粒的化学成分及大鼠口服红景天颗粒后的入血成分,分析质谱断裂规律,解析可能的代谢途径。结果:综合色谱-质谱检测信息,包括色谱保留时间、分子量、二级质谱信息,结合对照品比对,参考相关文献报道,从红景天颗粒中共鉴定出49个化学成分,包括黄酮类、苷类、酚酸类、鞣质类、原花青素类以及其他类成分;并对黄酮类、酚酸类、苷类化合物的质谱裂解规律进行解析。大鼠口服红景天颗粒后,血浆样本共鉴定出26个入血成分,包括4个原型成分(红景天苷、Rhodioloside E、Rhodiooctanoside、Kenposide A)和22个代谢产物,并对黄酮类化合物的代谢途径进行解析。结论:上述研究结果或为藏药红景天的质量控制、药效物质基础及药理学研究提供参考。

Objective: To analyze the main chemical constituents of Rhodiola Crenulata granules and the absorbed components and metabolites in rat plasma following oral administration based on HPLC-Q-TOF-MS. Methods: The chemical constituents of Rhodiola Crenulata granules and the absorbed components and metabolites in rat plasma following oral administration of Rhodiola Crenulata granules were determined for qualitative identification by HPLC-Q-TOF-MS. The chromatographic separation was performed on a reversed phase ODS column with acetonitrile-0.1% formic acid containing water as the mobile phase for gradient elution. The fragmentation patterns of target compounds were observed in both positive and negative electrospray ionization modes with information dependent acquisition(IDA) manner. The retention time, precursor ion and product ion information of the components in Rhodiola Crenulata granules and the absorbed components in rat plasma after oral administration were obtained by PeakView software, respectively. The fragmentation rule of mass spectrum and the possible metabolic pathway were analyzed. Results: By comparing chromatography and mass spectrum information of detected compounds( retention times,molecular weight,MS/MS spectra),combined with reference standards and previous literatures,a total of 49 chemical constituents were identified from Rhodiola Crenulata granules,including flavonoids,glycosides,phenolic acids,tannins,procyanidins and others,and the fragmentation pathways of flavonoids,phenolic acids and glycosides were analyzed. After oral administration of Rhodiola Crenulata granules, 26 compounds including 4 prototype compounds( Salidroside, Rhodioloside E,Rhodiooctanoside,Kenposide A) and 22 metabolites were detected in rat plasma,and the metabolic pathway of flavonoids was analyzed.Conclusion: The results may provide reference for the quality control,pharmacodynamic material basis studies and pharmacological researches of the Tibetan herb Rhodiola Crenulata.