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The Simultaneous Determination of Salinomycin and Lasalocid in Poultry Feed by High Performance Liquid Chromatography
|關鍵字:||high performance liquid chromatography;高效液相層析法;salinomycin;lasalocid;polyether ionophorous antibiotics;沙利黴素;拉薩羅;攜帶離子型抗生素||出版社:||獸醫學系||摘要:||
本研究的目的在於建立簡單、快速、準確的高效液相層析法，能同時檢測雞飼料中沙利黴素 (salinomycin) 與拉薩羅 (lasalocid)。實驗中將沙利黴素與拉薩羅添加於空白飼料中，配置成不同的濃度，利用acetonitrile進行萃取，再以2,4-dinitrophenylhydrazine行衍生化，過濾後注入HPLC系統檢測。分析條件為：：Symmetry® column（C18, 5 μm; 3.9x150 mm I.D.）；移動相組成是methanol和1.5% acetic acid水溶液（pH 2.7）以95：5的比例進行混合；流速0.8 mL/min；注入量20 μL；紫外光檢測器波長設為392、248 nm。實驗結果顯示，沙利黴素標準品滯留時間為7.9分鐘，在波長392 nm時最低檢測極限為20 ng (S/N≧3)，最低定量極限為60 ng (S/N≧10)；在波長248 nm時最低檢測極限為60 ng (S/N≧3)，最低定量極限為300 ng (S/N≧10)。空白飼料中含有沙利黴素(30~90 ppm)，在波長392 nm時回收率為94~110%，相對標準偏差2.7~5.5%；在波長248 nm時回收率為95~111%，相對標準偏差2.7~5.4%。而拉薩羅標準品滯留時間為4.5分鐘，在波長248 nm時最低檢測極限為100 ng (S/N≧3)，最低定量極限為500 μg (S/N≧10)。空白飼料中含有拉薩羅(30~120 ppm)，在波長248 nm時回收率為93~110%，相對標準偏差2.8~5.4%。
The aim of this study is to develop a simple, rapid and curate method for simultaneously determination the presence of salinomycin and laslocid. Salinomycin and lasalocid were extracted from poultry feed with acetonitrile. Extract derivatized with 2,4-dinitrophenylhydrazine (DNPH) in an acidic medium at 56℃. HPLC separation was carried out on a Symmetry® column(C18, 5 μm; 3.9 x 150 mm I.D.) using isocratic mobile phase : methanol : 1.5% acetic acid aqueous solution(pH 2.7)(95: 5,v/v) at a flow rate of 0.8 mL/min. The injection volume was 20 μL and Photodiode Array Detector used a quantitative wavelength of 392 and 248 nm. The retention time of salinomycin was 7.9 min. At 392 nm, the detection limit for salinomycin standard was 20 ng (S/N≧3). The quantitative limit for salinomycin standard was 60 ng (S/N≧10). At 248 nm, the detection limit for salinomycin standard was 60 ng (S/N≧3). The quantitative limit for salinomycin standard was 300 ng (S/N≧10). At 392 nm, the recoveries of spiked samples of salinomycin (30~90 ppm) were 94~110%, and the relative standard deviation were 2.7~5.5%. At 248 nm, the recoveries of spiked samples of salinomycin (30~90 ppm) were 95~111%, and the relative standard deviation were 2.7~5.4%. In the other side, the retention time of lasalocid was 4.5 min. At 248 nm, the detection limit for lasalocid standard was 100 ng (S/N≧3). The quantitative limit for lasalocid standard was 500 μg (S/N≧10). At 248 nm, the recoveries of spiked samples of lasalocid (30~120 ppm) were 93~110%, and the relative standard deviation were 2.8~5.4%.
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