高效液相色谱法检测鼠饲料中抗菌肽NZ2114

doi:10.16431/j.cnki.1671-7236.2023.11.015

Abstract

Abstract: 【Objective】 The aim of this study was to establish a rapid method for the determination of antimicrobial peptide NZ2114 in rat powder feed by solid phase extraction and high performance liquid chromatography (SPE-HPLC), which was used to detect the content of the test substance NZ2114 in rat powder feed samples at low, medium and high concentrations (2, 10 and 50 g/kg), and to analyze whether the antimicrobial peptide NZ2114 was evenly distributed.【Method】 The experiment mainly optimized extraction conditions, purification column and purification method, which investigated the influence of extraction agent and extraction time on extraction efficiency, the effect of SPE cleaning agent and elutriation agent on recovery rate.Feed samples were extracted by ultrasonic centrifugation, and the supernatant was enriched via WCX solid-phase extraction column, followed by purification and elution with 3 mL methanol and 5 mL of 20% ammoniated methanol.ZORBAX Eclipse Plus C18 was used as the stationary phase, and 0.1% formic acid-acetonitrile and 0.1% formic acid-water were used as mobile phases.The column temperature was 35℃, the injection volume was 20 μL, and the flow rate was 1.0 mL/min.The content of NZ2114 in feed samples was quantitatively determined by external standard method at a wavelength of 276 nm, and the linearity, limit of detection and limit of quantification, matrix effect, recovery rate, intra-day and inter-day precision of the method were investigated.【Result】 The SPE-HPLC method had a good linear relationship in the concentration range of 62.5-4 000 μg/mL, and the linear equation was y=0.9353x-32.721, whose correlation coefficient (R²) was 0.9996.The recovery rate of the method was 88.05%-111.23% at low, medium and high concentrations.The intra-day and inter-day precision were 0.58%-4.45%, 3.18%-6.37%, respectively.And the matrix effects at 125, 500 and 2 000 μg/mL concentrations were 1.02, 1.01 and 0.99, respectively.【Conclusion】 In this study, the SPE-HPLC method for the determination of antimicrobial peptide NZ2114 in rat feed was established, which met the requirements of methodological investigation, with the advantages of high sensitivity and precision, good reproducibility and rapid analysis.Thus, the method could achieve accurate quantification of NZ2114 in feed samples at three concentration levels.

Key words: high performance liquid chromatography; solid phase extraction; antimicrobial peptide NZ2114; rat feed

CLC Number:

S816

FENG Yanan, YANG Na, MAO Ruoyu, HAO Ya, MA Xuanxuan, TENG Da, WANG Jianhua. Determination of Antimicrobial Peptide NZ2114 in Rat Feed by High Performance Liquid Chromatography[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4463-4472.

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Determination of Antimicrobial Peptide NZ2114 in Rat Feed by High Performance Liquid Chromatography

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