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. 2021 Apr 2;26(7):2046.
doi: 10.3390/molecules26072046.

Development of a Multimatrix UHPLC-MS/MS Method for the Determination of Paracetamol and Its Metabolites in Animal Tissues

Konrad Pietruk  1 , Małgorzata Gbylik-Sikorska  1 , Beata Łebkowska-Wieruszewska  2 , Anna Gajda  1 , Mario Giorgi  3   4 , Irene Sartini  4 , Piotr Jedziniak  1
Affiliations

Development of a Multimatrix UHPLC-MS/MS Method for the Determination of Paracetamol and Its Metabolites in Animal Tissues

Konrad Pietruk et al. Molecules. .

Abstract

Paracetamol/acetaminophen (APAP) is one of the most popular pharmacologically active substances used as an analgesic and antipyretic agent. The metabolism of this drug occurs in the liver and leads to the formation of two main metabolites-glucuronic acid and sulfate derivate. Despite the wide use of paracetamol in veterinary medicine, a handful of analytical methods were published for the determination of paracetamol residues in animal tissues. In this paper, a multimatrix method has been developed for the determination of paracetamol and two metabolites-paracetamol sulfate (PS) and p-Acetamidophenyl β-D-glucuronide (PG). A validation procedure was conducted to verify method reliability and fit purpose as a tool for analyzing acetaminophen and metabolites in muscle, liver, lung, and kidney samples from different species of animals. Established validation parameters were in agreement with acceptable criteria laid by the European legislation. The initial significant matrix effect was successfully reduced by implementing an internal standard-4-Acetamidophenyl β-D-glucuronide-d3 (PG-d3, IS). The usefulness of the developed method was verified by analyzing samples from an experiment in which paracetamol was administrated to geese.

Keywords: UHPLC-MS/MS; heart; kidneys; liver; lungs; metabolites; muscle; paracetamol; residues.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
UHPLC-MS/MS XICs, control muscle sample from geese spiked with: (a) paracetamol (APAP), (b) p-Acetamidophenyl β-D-glucuronide sodium salt (PG), (c) Paracetamol sulfate potasium (PS), (d) and (e) 4-Acetamidophenyl β-D-glucuronide-d3 sodium salt (PG-d3) IS in negative and positive ionization, respectively. Paracetamol and metabolites were spiked at 50 µg/kg and internal standard at 25 µg/kg.
Figure 2
Figure 2
UHPLC-MS/MS XICs, blank muscle sample from geese spiked with 4-Acetamidophenyl β-D-glucuronide-d3 sodium salt (PG-d3) IS at 25 µg/kg in negative and positive ionization, respectively.
Figure 3
Figure 3
Matrix effect (mean ± SD, n = 20) in muscle, liver, lung, and kidneys samples: (a) without internal correction and (b) with internal correction.
See this image and copyright information in PMC

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