Zheng Guodong, Yang Xiujuan, Liu Mengshi, Chao Yingxin, Chen Baizhong, Yang Depo, Wei Minyan
Key Laboratory of Molecular Target & Clinical Pharmacology and the State Key Laboratory of Respiratory Disease, School of Pharmaceutical Sciences & the Fifth Affiliated Hospital, Guangzhou Medical University, Guangzhou, Guangdong 511436, China.
Guangdong Xinbaotang Biological Technology Company, Ltd, Jiangmen, Guangdong 529000, China.
ACS Omega. 2020 Jan 30;5(5):2180-2186. doi: 10.1021/acsomega.9b03123. eCollection 2020 Feb 11.
A rapid and simple method based on the coupling of supercritical fluid extraction and ultra-high-performance liquid chromatography combined with Q-Exactive Orbitrap tandem mass spectrometry (SFE-UHPLC-Q-Exactive Orbitrap-MS) detection for the identification of compounds from semen (CRS) was developed for the first time in this study. Through the optimization of the SFE parameters including extractive pressure, extractive temperature, and time, most of the compounds were successfully extracted at 50 °C, 33 MPa, and 2 h without an entraining agent, among which 32 compounds were successfully identified. Moreover, the operating conditions of UHPLC-Q-Exactive Orbitrap-MS were also optimized for the analysis of the SFE extracts, and the extracts in the CRS showed good separation performance in 20 min. A total of 28 compounds from the SFE extract were identified by comparing the standard sample together with full scan and related literature data, among which esters and flavonoids were the major compounds identified in the CRS extracts. In addition, 2 phenols, 2 aldehydes, 2 triterpenes, and 5 other compounds were identified. The SFE-UHPLC-Q-Exactive Orbitrap-MS method was successfully validated and applied for the identification of compounds from the CRS.
本研究首次开发了一种基于超临界流体萃取与超高效液相色谱联用Q-Exactive Orbitrap串联质谱（SFE-UHPLC-Q-Exactive Orbitrap-MS）检测的快速简便方法，用于鉴定精液（CRS）中的化合物。通过优化超临界流体萃取参数，包括萃取压力、萃取温度和时间，在50℃、33MPa和2h且无夹带剂的条件下成功萃取了大部分化合物，其中32种化合物被成功鉴定。此外，还对超高效液相色谱-Q-Exactive Orbitrap-MS的操作条件进行了优化，用于分析超临界流体萃取提取物，CRS中的提取物在20分钟内显示出良好的分离性能。通过与标准样品以及全扫描和相关文献数据进行比较，共鉴定出超临界流体萃取提取物中的28种化合物，其中酯类和黄酮类是CRS提取物中鉴定出的主要化合物。此外，还鉴定出2种酚类、2种醛类、2种三萜类和5种其他化合物。超临界流体萃取-超高效液相色谱-Q-Exactive Orbitrap-MS方法成功得到验证，并应用于CRS中化合物的鉴定。
