Determination of Colistin B in Chicken Muscle and Egg Using Ultra-High-Performance Liquid Chromatography–Tandem Mass Spectrometry

Abstract

Colistin, an imperative member of the polymyxin group, is a cationic peptide antibiotic. Itis also known as polymyxin E, but this peptide antibiotic has been forbidden for human consumption due to its high toxicity. Regrettably, this antibiotic is utilized as a feed additive and veterinary drug for animals. Due to the toxicity of colistin, the presence of its residue in the animal system represents a threat to human health regarding the consumption of meat, especially chicken. A novel method was proposed for quantifying colistin B in chicken muscles and eggs using ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS). In this method, extraction of colistin B from samples was achieved by mixing the sample with acidified methanol:water (1/1, v/v), followed by centrifugation and filtration by a membrane filter excluding solid-phase extraction (SPE) clean up, as well as evaporation steps. The analysis was conducted by optimized liquid chromatography–tandem mass spectrometry (LC–MS/MS), and method performance was assessed in terms of the limit of quantitation, specificity, selectivity, precision, linearity and recovery in coherence with the guidelines of SANTE and the Commission Decision 2002/657/EC. The result obtained from the study showed the limit of quantitation (LOQ) as 10 µg Kg−1 for muscles and 5 µg Kg−1 for eggs, with acceptable recoveries along with precision. The linearity was plotted in the range of 5–25 µg L−1 (solvent) for egg and 10–50 µg Kg−1 (matrix-matched) for muscles. The result of average recoveries showed the value of 70–94% (3.3–12% relative standard deviation (RSD)) for chicken muscles and 88–107% (2.5–18.6% RSD) for egg samples, which meets the criteria for acceptability of method according to both SANTE and 2002/657/EC guidelines. This proposed protocol provides a cost-effective solution for food testing labs by reducing the cost of the sample preparation by 60% along with the time required for SPE cleanup. Further, the optimized method was also tested on real samples collected from nearby provinces in Solan city, Himachal Pradesh, India, and three out of 20 muscles were found to have colistin B in the range of 50–560 µg Kg−1.

Stability of Colistin and Carbapenems in Water and Wastewater

“…Colistin is known to exist in three different structural forms, namely, deprotonated base, protonated acid, and neutral . It is also known to be “sticky” to glassware, plastics, and other laboratory ware that can contribute to a loss of compound .…”

“…14,57−60 Colistin is known to exist in three different structural forms, namely, deprotonated base, protonated acid, and neutral. 19 It is also known to be "sticky" to glassware, plastics, and other laboratory ware that can contribute to a loss of compound. 61 Hence, studies can assess the suitability and role of laboratory materials in accurately quantitating concentrations of complex antimicrobials like colistin and carbapenems from various environmental matrices.…”

“…The stability in water and wastewater is one key chemical property of an antibiotic that can majorly dictate its presence and persistence in the environment. The chemical stability of colistin and carbapenems has been scarcely reported with respect to environmental matrices, with most reported in conjunction with pharmacokinetic research. − For instance, the stability of meropenem has been studied in intravenous solutions, biological devices, solid-state formulations, and bicarbonate solutions. , Similarly, one study reported the pharmacological stability of imipenem, meropenem, ertapenem, and doripenem in aqueous solutions, but no clear data on stability variations with temperature or pH was provided . Similarly, a previous study reported the degradation characteristics of colistin in water and buffer while studying the pH rate and temperature rate profiles .…”

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“…The limit of the colistin quantitative determination by the developed method was 10 μg/kg in poultry and 5 μg/kg in eggs. This method is more cost-effective as compared to analogs due to the reduction in the cost of sample preparation [15].…”

HPLC Determination of Colistin in Human Urine Using Alkaline Mobile Phase Combined with Post-Column Derivatization: Validation Using Accuracy Profiles

“…The advantages and disadvantages of these approaches in terms of cost, instrumentation, matrix interferences, and sensitivity are more or less well known, and they were adequately discussed in a recent dedicated review article by Gikas and coworkers [ 3 ]. Mass spectrometric detection techniques are always attractive in bioanalysis, and there have been several very recent reports on the analysis of Colistin in rats [ 9 , 10 ] or human plasma [ 11 ], intestinal matrices of poultry [ 12 , 13 , 14 ], and eggs [ 15 ]. Sample preparation prior to LC–MS analysis varies from simple protein precipitation [ 9 ] to solid-phase extraction using advanced cation exchange or hydrophilic/lipophilic balance (HLB) materials [ 10 , 14 ].…”