LC-MS/MS를 이용한 수산물 중 콜리스틴 분석법 개발

• Published in: Han'gug sigpum wi'saeng anjeonseong haghoeji Vol. 31; no. 5; pp. 319 - 326
• Main Authors: 신다솜, 강희승, 이수빈, 조윤제, 천소영, 정지윤, 이규식, Shin, Dasom, Kang, Hui-Seung, Lee, Soo-Bin, Cho, Yoon-Jae, Cheon, So-Young, Jeong, Jiyoon, Rhee, Gyu-Seek
• Format: Journal Article
• Language: Korean
• Published: 한국식품위생안전성학회 31.10.2016
• Subjects: 식품과학; LC-MS/MS; shrimp; analytical method; colistin; fish
• ISSN: 1229-1153; 2465-9223

Colistin is a last resort antimicrobial agent against multi-drug resistant Gram-negative bacteria. This study was conducted to develop an analytical method to determine colistin in fish and shrimp. The analytes were confirmed and quantified via liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the positive ion mode using multiple reaction monitoring (MRM). The sample was extracted with acidified 5% methanol (containing 0.5% formic acid). Then, solid phase extraction (SPE) was used for cleanup. Matrix-matched calibration curves were linear over the calibration ranges (0.05-1.2 mg/kg) for all the analytes into blank sample with $r^2$ > 0.99. All the values fulfilled the criteria requested by the Codex guidelines. Average recoveries ranged from 85.9% to 107.9%. The repeatability of measurements, expressed as the coefficient of variation (CV, %), was less than 15%. The limit of detection (LOD) was 0.02 mg/kg, and the limit of quantitation (LOQ) was 0.05 mg/kg. This improved method showed higher accuracy and acceptable sensitivity to meet the CAC guideline requirements and is applicable for the analysis of residual colistin (A+B) in fish and shrimp. LC-MS/MS를 이용하여 폴리펩타이드계 동물용의약품인 콜리스틴에 대한 시험법을 확립하여 정량성 및 정밀성을 확보하였으며, 확립된 시험법의 적용성 검증을 위해 국제식품규격위원회 기준에 따라 특이성, 정확성, 직선성, 정밀성, 검출한계, 정량한계 등을 검증하였다. 콜리스틴 표준용액을 잔류허용기준의 농도에 따라 검량선을 작성한 결과 0.99 이상의 직선성을 확인 할 수 있었으며, 본 실험에서의 평균 회수율은 85.9~107%이었다. 또한, 분석오차는 11.8% 이하로 정확성 및 재현성이 우수하였으며, 검출한계는 0.02 mg/kg, 정량한계는 0.05 mg/kg이었다. 또한, 실험실간 교차검증을 통하여 신뢰성을 확보하였다. 확립된 분석법은 양식 수산물 중 잔류할 수 있는 동물용의약품인 콜리스틴에 대한 안전관리에 활용 할 수 있을 것으로 판단된다.