Residues of Fluoroquinolone Antibiotics Induce Carbonylation and Reduce In Vitro Digestion of Sarcoplasmic and Myofibrillar Beef Proteins

Abstract

Although the impact of oxidation on human health has been of growing interest, the oxidation of proteins, major component of meat, has received little attention. This paper describes the in vitro effect of five fluoroquinolones (FQs) on carbonylation of sarcoplasmic and myofibrillar proteins of beef when found at concentrations close to the maximum residue limit (MRL). Samples were treated individually with the FQs, determining in each protein fraction the carbonyl index, protein content and oxidized proteins identification, using 2,4-dinitrophenyhydrazine (DNPH) alkaline assay, Western blot and Bradford methods, and mass spectrometry, respectively. Besides, the in vitro effect of these residues on gastric and duodenal digestion of proteins was evaluated. The carbonylation induced by FQs affected both protein fractions being significant with respect to the blank in 73.3% of cases. This damage was correlated with loss of solubility and digestibility, with sarcoplasmic proteins the most affected. Danofloxacin and enrofloxacin were the FQs with greatest oxidant effects, especially affecting glycolysis and glycogen proteins. Our results suggest that these residues induce irreversible oxidative damage on the main beef proteins and could affect their nutritional value.

Residues of tetracyclines and β-lactams antibiotics induce carbonylation of chicken breast

“…This behavior could be associated to accessibility of proteins in muscle cells: sarcoplasmic being more available than myofibrillar, which are more available than insoluble proteins; this is due to sarcoplasmic proteins being soluble in sarcolemma, while myofibrillar and insoluble proteins are stored in myofibrilla and connective tissue, respectively. 13,18 Finally, carbonylation of chicken breast proteins provides evidence for the ability of tetracyclines to promote oxidative stress, which is consistent with assays carried out with several biological models. [19][20][21][22][23][24] According to Wen et al (2012), CTC induced oxidative stress in maize root through the production of hydroxyl radicals 19 ; Pes et al (2018) reported a decrease of antioxidant enzymes in fish when these were exposed to OTC; Hang et al (2019) showed that exposure to TC induced oxidative stress in ryegrass seedlings mediated by the increase of lipid oxidation and decrease of antioxidants enzymes.…”

“…While comparing the carbonylation on three protein fractions, only DCL and OXA promoted greater oxidative damage on these (Figure 4), probably mediated by a largest number of molecules of DCL and OXA available in the muscle cell, these amounting to 6 times the concentrations of PGN and AMP. 4,13 According to the literature, the carbonylation induced by residues of tetracyclines and β-lactams at MRL concentrations are similar to those promoted by fluroquinolones MRLs on beef proteins, which was reported by Marquez et al (2020).…”

“…This procedure was described by Marquez et al (2020). 13 Chicken breast aliquots were randomly divided in control samples and samples to contaminate with tetracyclines and β-lactams. Then, samples were contaminated individually with 200 μL of each antibiotic working solutions until reaching final concentrations of 0.5, 1.0 and 1.5 MRL (Table 1), followed by vortexing for 30s and then incubation for 1 h at room temperature in the dark.…”

“…15 Determination of carbonyl content in proteins samples Protein carbonyl content was measured according to Mesquita et al (2014) with some modifications described by Marquez et al (2020). 13,16 For this, 300 μL of 2,4-dinitrophenylhydrazine (DNPH, 10 mM in 0.5M H 3 PO 4 ) was added to the same volume of protein solution (150 μg) and incubated in the dark for 10 minutes. Next, 160 μL of this solution was placed in a 96-well plate and 40 μL of 6M NaOH were added and incubation for 10 minutes was carried out.…”

In vitro oxidation promoted by chlorpyrifos residues on myosin and chicken breast proteins

“…However, chicken meat proteins are more susceptible to undergo irreversible oxidation such as carbonylation. This fact can occur directly by incorporation of a carbonyl group on lysine, threonine, arginine or proline residues; or indirectly by reaction with end products of sugar and lipid oxidation (Márquez-Lázaro, 2020;Estévez, 2015;Estévez, 2011). The impacts of carbonylation in meat have been associated to a reduction of its quality traits such as texture, flavor, tenderness, color and nutritional value (Estévez, 2011).…”