Analysis of Non-Volatile Chemical Constituents of Menthae Haplocalycis Herba by Ultra-High Performance Liquid Chromatography-High Resolution Mass Spectrometry

Abstract

Menthae Haplocalycis herba, one kind of Chinese edible herbs, has been widely utilized for the clinical use in China for thousands of years. Over the last decades, studies on chemical constituents of Menthae Haplocalycis herba have been widely performed. However, less attention has been paid to non-volatile components which are also responsible for its medical efficacy than the volatile constituents. Therefore, a rapid and sensitive method was developed for the comprehensive identification of the non-volatile constituents in Menthae Haplocalycis herba using ultra-high performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometry (UHPLC-LTQ-Orbitrap). Separation was performed with Acquity UPLC® BEH C18 column (2.1 mm × 100 mm, 1.7 μm) with 0.2% formic acid aqueous solution and acetonitrile as the mobile phase under gradient conditions. Based on the accurate mass measurement (<5 ppm), MS/MS fragmentation patterns and different chromatographic behaviors, a total of 64 compounds were unambiguously or tentatively characterized, including 30 flavonoids, 20 phenolic acids, 12 terpenoids and two phenylpropanoids. Finally, target isolation of three compounds named Acacetin, Rosmarinic acid and Clemastanin A (first isolated from Menthae Haplocalycis herba) were performed based on the obtained results, which further confirmed the deduction of fragmentation patterns and identified the compounds profile in Menthae Haplocalycis herba. Our research firstly systematically elucidated the non-volatile components of Menthae Haplocalycis herba, which laid the foundation for further pharmacological and metabolic studies. Meanwhile, our established method was useful and efficient to screen and identify targeted constituents from traditional Chinese medicine extracts.

Characterization and identification of the metabolites of Menthae Haplocalycis Herba water extracts in rat plasma, urine, and feces by ultra‐high performance liquid chromatography with linear ion trap‐Orbitrap mass spectrometry

“…Two fragment ions m/z 151 [M+H−H 2 O] + and 123 [M+H−H 2 O−CO] + were dominant in the ESI‐MS 2 spectra. Based on past work , they were tentatively identified as isomers of bohecineole A. M3 was detected in urine and feces as well. M4 (t R = 7.58 min) showed [M+H] + at m/z 167.10602, and it had the formula C 10 H 14 O 2 .…”

“…The ion at m/z 593 also generated predominant fragment ions at m/z 549 [M−H−CO 2 ] − and m/z 575 [M−H−H 2 O] − . Based on literature comparisons , M19 was finally deduced as didymin isomer. M20 , t R = 25.02 min, it was detected [M−H] − ion at m/z 343.08026 (C 18 H 15 O 7 ).…”

“…The ESI‐MS 2 spectra gave the fragment ion at m/z 193. M61 and M62 were deemed as rosmarinic acid isomers by comparison with authentic standards and literature . M63 ( t R = 14.36 min) and M64 (17.85 min) presented [M−H] − ions at m/z 329.06531 and 329.06564.…”

“…Phytochemical investigations of M. Haplocalycis have led to the isolation of flavonoids such as hesperetin, hesperidin, diosmin and diosmetin, monoterpenoids like bohecineole A and bohenoside A, and phenolic acid as rosmarinic acid and phloretic acid. Several pharmacological investigations demonstrate that M. Haplocalycis has a variety of biological effects , reducing vital infections, oxidation, and inflammation, which may be closely linked with its traditional efficacy.…”

Rapid Characterization of Components in Bolbostemma paniculatum by UPLC/LTQ-Orbitrap MSn Analysis and Multivariate Statistical Analysis for Herb Discrimination

“…In last several years, based on the efficient and fast separation performance of UPLC, as well as the high sensitivity of MS, UPLC tandem MS has turned into an important technology for characterization of chemical components in TCM [ 4 , 5 ]. Especially, UPLC coupled with hybrid LTQ-Orbitrap MS system is applied by more and more researchers in this field [ 6 , 7 , 8 ], because it is ideal for the identification of natural compounds by obtaining accurate molecular mass and multistage MS n fragment ions of samples to be tested.…”