Planta Med 2013; 79 - PH11
DOI: 10.1055/s-0033-1352088

UHPLC-ESI(+)-HRMS-based metabolic profiling as a dereplication tool for the detection and identification of Acronychia-type acetophenones in crude extracts

E Kouloura 1, M Halabalaki 1, M Litaudon 2, K Awang 3, V Nguyen 4, A Skaltsounis 1
  • 1Laboratory of Pharmacognosy and Natural Products Chemistry, School of Pharmacy, University of Athens, Panepistimioupolis Zografou, 15571 Athens, Greece
  • 2Centre de Recherche de Gif, Institut de Chimie des Substances Naturelles, CNRS, 1, Avenue de la Terrasse, 91198 Gif-sur-Yvette Cedex, France
  • 3Department of Chemistry, University of Malaya, 50603 Kuala Lumpur, Malaysia
  • 4Institute of Marine Biochemistry – Vietnam Academy of Science and Technology (VAST), 18, Hoang Quoc Viet Road, Caugiay, Hanoi, Viet Nam

Acronychia-type acetophenones constitute a characteristic group of constituents of Acronychia species. Acronychia species have been traditionally used in folk medicine for their anti-inflammatory and antipyretic effects to treat asthma, ulcers and rheumatism. Previous studies have demonstrated the biological importance of Acronychia-type acetophenones as significant cytotoxic principals1. However, their analysis and identification via chromatographic and spectroscopic/spectrometric techniques have not been reported up to now. Therefore, in the present study, a sensitive and specific method using an UHPLC-ESI(±)-HRMS platform was developed for the separation, detection and structural characterization of Acronychia-type acetophenones in crude extracts. Specifically, a UHPLC apparatus hyphenated to a hybrid LTQ-Orbitrap MS equipped with an ESI ionization probe, in positive and negative mode was incorporated. Chromatographic (Rt) and spectrometric features such as suggested ECs and RDBeq. values in both full scan and MSn level as well as data dependent acquisitions were employed. Fragment ions generated in MS2 and MS3 level at m/z 341.1719, 285.1095 and 229.0469 were found characteristic for Acronychia-type acetophenones and could be utilized for the identification thereof. Moreover, diagnostic ion ratios were determined for the identification of acetophenones isomers and different substitution patterns. In order to evaluate the validity of the generated method for dereplication purposes, diverse species and organs of Achronychia genus were collected from Malaysia and Vietnam and extracted. Applying the developed method, known acetophenones were successfully detected together with unknown ones. Additionally, supervised and unsupervised data analysis methods were used to facilitate the identification procedure and support the findings.

Reference:

[1] Kouloura et al, Cytotoxic Prenylated Acetophenone Dimers from Acronychia pedunculata, J. Nat. Prod, 2012, 75 (7):1270 – 6