禽畜水產動物體內喹諾酮類抗生素含量檢測方法的開發研究

Abstract

為了使飼養之畜、漁產免於疾病，並刺激成長，養殖業者利用抗生素、磺胺劑、氯黴素、荷爾蒙…等藥物已甚為普遍，然而在過量使用此些藥物之際，便會使得這些藥物殘留在禽畜水產動物體內，進而影響到食用者的健康。因此，開發快速、準確之動物用藥檢驗方法，使相關衛生檢驗機關能快速做好檢測作業，便成為避免檢驗資源浪費及最佳之食品安全把關措施。喹諾酮(Quinolone)藥物依照目前行政院衛生署公告之動物用藥殘留量檢驗方法，是利用高效液相層析儀以光二極體列陣檢出器及螢光檢出器串聯使用分析檢測，共可分析7 項喹諾酮藥物，偵測極限約為20ppb。本研發方法之樣品前處理過程簡易快速，無須乙腈、正己烷等有機溶劑萃取，省略反復之減壓濃縮步驟，樣品以phosphate buffer 萃取後逕行振盪離心，並以MAX 固相萃取匣淨化，層析管選用BEH C18 2.1 × 50 mm, 1.7 μm 進行分離，以0.1 % 甲酸與90 %甲醇醋酸銨做梯度沖提，利用極致效能液相層析串聯質譜儀(UPLC/MS/MS)檢驗縮短分析時間並提昇檢測靈敏度，估計每件樣品從前處理至分析完成相較現行標準公告方法約可節省3 小時且僅需少量試劑、器材。本方法共分析11 項喹諾酮藥物：ciprofloxacin、danofloxacin、enrofloxacin、flumequine、moxifloxacin、nalidixic acid、norfloxacin、ofloxacin、oxolinic acid、piromidic acid、sparfloxacin。標準品濃度在10~50ppb 呈現良好線性關係，各項標準品添加於不同測試樣品其回收率平均在85.53 % to 98.21 %，最低檢出限量範圍可達1.50~4.66 ppb。我們利用上述建立之檢驗技術，完成國內市售禽畜水產品100件之喹諾酮藥物殘留量調查，檢出喹諾酮藥物之產品比率為28 %，不合格率為21 %。本研究建立了準確且更有效率之檢驗方法，提供全國各衛生單位、學術機構採用，並藉由公共衛生安全監測調查，提供予學術機構研究參考及養殖業者監測用藥量資訊，確保禽畜水產品品質。A multiresidue method has been developed for the confirmation and quantification of 11 quinolones(Ciprofloxacin, Danofloxacin, Enrofloxacin, Flumequine, Moxifloxacin, Nalidixic acid, Norfloxacin, Ofloxacin, Oxolinic acid, Piromidic acid, Sparfloxacin) in livestock and aquatic products by liquid chromatography triple-quadrupole mass spectrometry (LC/MS/MS). The samples were extracted with phosphate buffer, analytes separated by UPLC on a C18 column using 0.1% formic acid and 90 % methanol with ammonium acetate for a linear gradient elution program, and detected by MS/MS. The linear range was 10-50 ppb with correlation coefficients (r) more than 0.9951. The limits of detection were 1.50-4.66 ppb. Mean recoveries for each analyte in livestock and aquatic products ranged from 85.53% to 98.21% and from with relative standard deviations below 10%. The method is fast, safe, sensitive and precise, and can be used simultaneously to analyse residual quinolones.