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標題: Amoxicillin藥物殘留檢測技術之開發
A Study for Amoxicillin Residue Detection Technique
作者: 蔡岳智
關鍵字: Amoxicillin;藥物殘留;檢測技術;HPLC
出版社: 獸醫學系
本研究之目的在於嘗試以高效液相層析法 (high- performance liquid chromatography; HPLC)開發出一套簡單、快速、精確又可同時檢測雞肉、豬肉、牛肉、魚肉及牛奶中amoxicillin殘留之檢測方法,並依所建立之檢測方法來進行市售中禽畜水產品之amoxicillin殘留抽樣調查。
禽畜水產品 (雞肉、豬肉、牛肉、魚肉及牛奶)以磷酸緩衝溶液 (10 mM KH2PO4, pH4.5)進行萃取amoxicillin後,再以75% 三氯乙酸 (Trichloracetic acid, TCA)移除游離中的蛋白質,經固相萃取管柱 (solid phase extraction column)淨化及濃縮amoxicillin並以0.2 μm濾膜過濾後,以HPLC進行分析,各組織之回收率為70%-80%,變異係數 (coefficient of variation value, CV)均在6.25%以內。
實驗結果顯示,利用層析管柱 (Cosmosil 5C18-MS),配合移動相MeOH:5 mM KH2PO4, pH6.5 (10:90; v/v),流速0.8 ml/min,以波長210 nm之紫外光檢測器 (UV detector)可精確地檢測雞肉、豬肉、牛肉、魚肉及牛奶中添加的amoxicillin (5-100 ppb),且amoxicillin標準品之滯留時間為7分鐘,最低檢測極限為5 ppb (s/n≧3)。
以本方法抽驗本省市售禽畜水產品 (45件)的調查結果顯示,雞肉、豬肉、牛肉、魚肉及牛奶中之檢出率均為0% (未能檢測出amoxicillin的殘留)。

The aim of this investigation is to develop a simple, rapid, and reliable high-performance liquid chromatography (HPLC) method for the determination of the amoxicillin residues in chicken, pork, beef, fish, and milk, respectively.
The samples were extracted with 10 mM potassium phosphate buffer pH4.5, followed by trichloracetic acid (TCA) precipitation of proteins and solid-phase extraction column (SPE column, C18). The recoveries of the amoxicillin form chicken, pork, beef, fish meat, and milk were about 70%-80%. The cofficient of variation value (CV) is not over 6.25%.
The HPLC separation was carried out on a C18 column (Cosmosil 5C18-MS column; 150 mm ×4.6 mm I.D.) with 5 mM potassium phosphate buffer pH6.5-methanol (90:10) as the mobile phase at a flow rate of 0.8 mL/min. The chemical was detected by UV detection which wavelength set at 210 nm. The retentsion time and the limit of detection of the developed method was 7min and 5 ppb (s/n≧3), respectively.
The 45 samples from chicken, pork, beef, fish meat, and milk from different markets located in Taiwan area were surveyed by random for the residue of amoxicillin by the developed method. The results of the residue of amoxicillin in chicken, pork, beef, fish meat, and milk samples were below 5 ppb.
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