運用高效液相層析法對飼料中安默西林及安比西林檢測方法之建立

Abstract

本實驗目的在以高效液相層析法建立同時檢測飼料中安默西林（amoxicillin）及安比西林（ampicillin）的方法。移動相為0.01 M 磷酸二氫鉀緩衝液（pH 2.5）：乙腈（82: 18, v/v），層析管柱使用SymmetryShield® column（RP8, 5μm, I.D. 4.6 × 250 mm），螢光檢測器檢測波長設定為λex 358 nm、λem 440 nm進行檢測。兩種藥物均以三種不同濃度添加於飼料中之平均回收率介於93.28∼102.69 % 及88.35∼106.14 %；回收率之天內及天間變異係數皆在4 % 以下。安默西林及安比西林於飼料中之檢測極限分別為0.3536及0.0269 mg/kg，定量極限分別為1.1787及0.0896 mg/kg。本試驗所建立之檢測方法通過準確度、精密度、專一性、線性等各項確效，可實際應用於檢測飼料中安默西林及安比西林之含量。

The study was to develop a high performance liquid chromatography (HPLC) method for determination of amoxicillin and ampicillin in feeds. Fluorescence detector was set at excitation wavelength of 358 nm and emission wavelength of 440 nm. A SymmetryShield® column (RP8, 5 μm, 4.6 × 250 mm I.D.) with a mobile phase consisting of 0.01 M potassium dihydrogen phosphate (pH 2.5)：acetonitrile (82: 18, v/v) was used. The average recoveries of amoxicillin and ampicillin from spiked samples were 93.28 – 102.69 % and 88.35 – 106.14 % with repeatabilities below 4 % R.S.D.. The limits of detection of amoxicillin and ampicillin were 0.3536 and 0.0269 mg/kg, the limits of quantitation were 1.1787 and 0.0896 mg/kg for feed, respectively. Not only the method of validation was performed, but also specificity, reproducibility, precision and accuracy were confirmed. This assay has been applied to the measurement of amoxicillin and ampicillin in feeds.