Development and validation of an analytical method for determination of pharmaceuticals in fish muscle based on QuEChERS extraction and SWATH acquisition using LC-QTOF-MS/MS system

Abstract

The first report on the presence of pharmaceuticals in the wastewater impacted river appeared in the mid-1970s [1-3] followed by the first systematic works in the late 1990s [4]. Studies on the accumulation of pharmaceutically active compounds (PhACs) on aquatic organisms such as fish were not initiated until the beginning of the 2121st st century [5]. Fish tissues such as muscles can be considered as a bio-indicator of water pollution due to the direct exposure to wastewater borne contaminants in rivers. The methods for PhAC extraction from fish muscle includes solid-liquid extraction [6-8], enzymatic microwave-assisted extraction [6,9], ultra-sound extraction (USE) [10], pressurized liquid extraction (PLE) [11,12], solid-phase microextraction (SPME) [13], and rotary extraction [5,14]. The clean-up step usually relies on solid phase extraction (SPE) with various types of sorbents: florisil, alumina, silica gel, hydrophilic-lipophilic balance (HLB) and mixed-mode cation-exchange (MCX) [15-17], gel permeation chromatography, washing with hexane, liquid-liquid extraction, and freezing out [15,18] An alternative and revolutionary method for the simultaneous extraction and clean-up of pesticides in foodstuff [19] which recently has been used for the determination of PhACs in biological matrices is the so-called QuEChERS (for Quick, Easy, Cheap, Effective, Rugged and Safe). Few studies are dealing with the extraction of PhACs using QUuECH hERS in different matrices such as earthworms [18], vegetables [20], and bivalves [21].QuEChERS is based on liquid-liquid partitioning of the analytes between water and acetonitrile (MeCN) using salts to produce the effect of salting out, followed by a dispersive solid phase extraction (d-SPE). This procedure offers many advantages including basic instrumentation for application, small sample amounts, rapid extraction, and low waste volume, which results in a time-efficient method.The use of QuEChERs for the determination of PhACs from complex biological matrices, such as fish muscle, generates extracts that contain material prone to adversely affect the mass spectrometric analysis (co-extracts),

Analysis of pharmaceuticals in fish using ultrasound extraction and dispersive spe clean-up on que Z-Sep/C18 followed by LC-QToF-MS detection

“…Recoveries were calculated as the ratio between the peak area in the extract from spiked fish sample and the peak area in a blank fish extract. The recovery was acceptable between 70 and 130% for majority of compounds, values that fall into the range from other reported studies of pharmaceuticals validated in fish matrix [1 , 2 , 19 , 20] . The precision of the method expressed by the intra-day repeatability was calculated as the relative standard deviation (RSD%) obtained from the relative recoveries ( n = 3) for each concentration level, while the inter-day precision was determined by analyzing of the concentration levels for three consecutive days.…”

“…Common approaches for the extraction of pharmaceutically active compounds (PhACs) from fish tissues rely on solid-liquid extraction, enzymatic microwave-assisted extraction, ultra-sound extraction (USE), QuEChERS, and pressurized liquid extraction [1 , [3] , [4] , [5] , [6] , [7] , [8] , [9] , [10] . Prior to the analysis of the extracts by LC-MS, it is critical to remove co-extracted lipids as much as possible in order to reduce matrix effects during analyte ionization in the interface.…”

“…Although this technique may be effective, the use of up to 200 mL of cleaning solvent for each sample makes it an overly expensive method. In only two previous studies, the determination of pharmaceutical residues in fish muscle was performed using high resolution mass spectrometry (HRMS) tools such as Q-Orbitrap and Q-TOF [1 , 35] . While the former had a large group of contaminants of different types and ME and recoveries varied from −327 to 54% and 40 to 160% respectively, the latter works with freeze-dried fish, less PhAC and its quantification limits range from 1.8 to 303%.…”

“… 0.03 – 5.5 40 - 160 Q-Exactive 54 - (−327) [35] 1.0 dw QuEChERS 19 EMR lipid removal 21 0.5– 91 DW 1.8 - 303 dw 10 - 139. Q-ToF 113 - (−89) [1] 0.2 dw vortex 10 filtration 42 0.01 to 2.00 0.1 - 40.2 29 - 188 QqQ 2191 -(−83) [36] 1 USE NR SPE 29 0.01–2.00 0.03 - 6.67 61 - 111 QqQ NR [32] 5 USE-McIlvain 15 SPE 27 N.R. 6–30 43 - 103 QqQ 5 - (−40) [33] 1 z.f.…”

“…Here, we present SQUEEZe ( S olid-li Q uid U ltrasound E xtraction with Qu E Z -S e p/C18 as dispersive clean-up): a fast method for analysis of the trace 47 PhACs in fish muscle. Compared to our previously reported method [1] , it offers alternatives with improvements in recoveries, number of analytes, sample volume and solvent used. The key aspects of this method are: The ultrasound extraction was performed with acetonitrile/isopropanol 0.1% V/V formic acid.…”