Contents lists available at ScienceDirect Journal of Chromatography B journal homepage: www.elsevier.com/locate/jchromb Systematic evaluation of matrix effects in supercritical fluid chromatography versus liquid chromatography coupled to mass spectrometry for biological samples Vincent Desfontaine a , Francesca Capetti a , Raul Nicoli b , Tiia Kuuranne b , Jean-Luc Veuthey a , Davy Guillarme a, ⁎ a School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, CMU - Rue Michel Servet 1, 1211 Geneva 4, Switzerland b Swiss Laboratory for Doping Analyses, University Center of Legal Medicine Lausanne-Geneva, Centre Hospitalier Universitaire Vaudois, University of Lausanne, Chemin des Croisettes 22, 1066 Epalinges, Switzerland ARTICLE INFO Keywords: Matrix effects LC-MS SFC-MS Urine Plasma ABSTRACT Matrix effects (ME) is acknowledged as being one of the major drawbacks of quantitative bioanalytical methods, involving the use of liquid chromatography coupled to mass spectrometry (LC-MS). In the present study, the incidence of ME in SFC-MS/MS and LC-MS/MS in the positive mode electrospray ionization (ESI+) was sys- tematically compared for the analysis of urine and plasma samples using two representative sets of 40 doping agents and 38 pharmaceutical compounds, respectively. Three different SFC stationary phase chemistries were employed, to highlight the importance of the column in terms of selectivity. Biological samples were prepared using two different sample treatments, including a non-selective sample clean-up procedure (dilute and shoot (DS) and protein precipitation (PP) for urine and plasma samples, respectively) and a selective sample pre- paration, namely solid phase extraction (SPE) for both matrices. The lower susceptibility to ME in SFC vs. reversed phase LC (RPLC) was verified in all the experiments performed on urine, and especially when a simple DS procedure was applied. Also, with the latter, the perfor- mance strongly varied according to the selected SFC stationary phase, whereas the results were quite similar with the three SFC columns, in the case of SPE clean-up. The same trend was observed with plasma samples. Indeed, with the PP procedure, the occurrence of ME was different on the three SFC columns, and only the 2- picolylamine stationary phase chemistry displayed lower incidence of ME compared to LC-MS/MS. On the contrary, when a SPE clean-up was carried out, the results were similar to the urine samples, with higher performance of SFC vs. LC and limited discrepancies between the three SFC columns. The type of ME observed in LC-MS/MS was generally a signal enhancement and an ion suppression for urine and plasma samples, respec- tively. In the case of SFC-MS/MS, the type of ME randomly varied according to the analyzed matrix, selected column and sample treatment. 1. Introduction Due to its very high specificity and sensitivity, chromatography coupled with mass spectrometry (MS) has become the gold standard for the quantitative analysis of pharmaceutical drugs and their metabolites in biological fluids. In particular, liquid chromatography (LC) hyphe- nated with tandem mass spectrometry (LC-MS/MS) is nowadays one of the most important analytical platform for bioanalytical [1] and doping control laboratories [2]. The success of LC-MS/MS is based on its ability to accurately and rapidly quantify very small amounts of organic compounds in complex matrices, such as plasma or urine, with a limited sample clean-up prior to injection. The use of electrospray ionization (ESI) has also extended the scope of the technique by enabling the analysis of polar molecules and peptides/proteins. Nevertheless, the great success of LC-MS/MS also comes with a few drawbacks. In the last years, many researchers have reported that the presence of endogenous compounds, extracted from the matrix and co- eluting with the target analytes, could interfere in the MS source and alter their ionization yield, leading to inaccurate quantitative results. The quantity of ions formed in the source under the influence of matrix entities could either be increased (signal enhancement) or, more fre- quently, decreased (ion suppression) compared to the case where no https://doi.org/10.1016/j.jchromb.2018.01.037 Received 12 December 2017; Received in revised form 27 January 2018; Accepted 29 January 2018 ⁎ Corresponding author at: Swiss Laboratory for Doping Analyses, University Center of Legal Medicine Lausanne-Geneva, Switzerland. E-mail address: davy.guillarme@unige.ch (D. Guillarme). Journal of Chromatography B 1079 (2018) 51–61 Available online 07 February 2018 1570-0232/ © 2018 Elsevier B.V. All rights reserved. T