Rapid separation and identification of furocoumarins in Angelica dahurica by high‐performance liquid chromatography with diode‐array detection, time‐of‐flight mass spectrometry and quadrupole ion trap mass spectrometry

Abstract

High-performance liquid chromatography with diode-array detection (HPLC/DAD), time-of-flight mass spectrometry (HPLC/TOFMS) and quadrupole ion trap mass spectrometry (HPLC/QITMS) were used for separation, identification and structural analysis of furocoumarins in Angelica dahurica. Two furocoumarins (imperatorin and isoimperatorin) in Angelica dahurica extract were identified unambiguously by comparing their relative retention times, characteristic ultraviolet information and accurate mass measurement. A formula database of known furocoumarins in Angelica dahurica was established, against which the other 21 furocoumarins were identified effectively based on the accurate extract masses and formulae acquired by HPLC/TOFMS. In order to distinguish the isomers, multi-stage mass spectrometry (MSn, ion trap mass spectrometry) was used. General fragmentation behavior of the furocoumarins in the ion trap mass spectrometer was studied by the two furocoumarin standards, and their fragmentation rules in MS(n) spectra were summarized. These deduced fragmentation rules of furocoumarins were successfully implemented in distinguishing the three groups of isomers in Angelica dahurica by HPLC/QITMS. By using the three different analytical techniques, 23 furocoumarins in Angelica dahurica were tentatively identified within 30 min. Finally, HPLC/TOFMS fingerprints of Angelica dahurica were established by which it can be concluded that a rapid and effective method based on the three analytical techniques for identification of chemical components was established. This can provide help for further quality control of Angelica dahurica and pharmacology mechanism study of furocoumarins in Angelica dahurica.