Identification of chemical constituents of the extract of Cynanchum auriculatum based on UHPLC Q-Exactive Plus Orbitrap HRMS

GOU Jian; WU Zong-qin; MENG Xin; WEI Xiao-yi; LIU Chun-hua; ZHU Di; QIN Lan; SUN Jia

doi:10.3969/j.issn.1003-3734.2024.05.009

Abstract：Objective:The UHPLC Q-Exactive Plus Orbitrap HRMS technique was used to establish a comprehensive and rapid method for the analysis of the chemical composition of Cynanchum auriculatum and to provide a scientific basis for the study of the medicinal substances of C.auriculatum.Methods:A Hypersi 1 GOLD chromatographic column(100 mm x2.1 mm,1.9 pm)was employed using gradient elution with 0.1％formic acid aqueous solution(A)-0.1％formic acid acetonitrile solution(B)as mobile phase at a flow rate of 0.3 mL·min-1 and a column temperature of 40 ℃.The mass spectrometry was performed using an electrospray ionization and the data were collected in positive and negative ion modes in a scanning range m/z 100～1 500 Da.The spray voltage was set at 3.5 kV(+),2.5 kV(-),sheath gas flow rate was 35 arb,auxiliary gas flow rate was 10 arb,capillary temperature was 320 ℃,and auxiliary heater temperature was 350 ℃.According to the accurate molecular mass information of excimer peaks and fragment ions provided by high-resolution mass spectrometry,combined with reference substance comparison,related literature reports,m/z Cloud,and PUBchem database,comparison of secondary mass spectrometry was conducted to identify the chemical composition.Results:A total of 66 compounds were analyzed from the C.auriculatum extraction,including 34 C21 steroids,13 organic acids,five acetophenones,four coumarins,four alkaloids and six other compounds,among which 18 compounds were reported for the first time in C.auriculatum.Conclusion:The developed method can carry out a rapid,accurate and comprehensive qualitative analysis of the chemical composition of C.auriculatum,and provide a scientific basis for further elucidation of its pharmacodynamic material basis and quality control in the future.