以高效液相層析法建立牛肉中巨內脂環類抗生素Abamectin, Doramectin, Moxidectin, Ivermectin, Milbemectin A3及Milbemectin A4之多重殘留分析法   全文下載

A Multiresidue Method for the Determination of Abamectin, Doramectin, Moxidectin, Ivermectin, Milbemectin A3, and Milbemectin A4 Residues in Bovine Muscle Using HPLC with Fluorescence Detection

本研究目的為利用快速簡便的高效液相層析法（hlgh perforrnance liquid chromatography, HPLC）檢測牛肉中abamectin, doramectin, moxidectin, ivermectin, milbemectin A3及milbemectin A4等巨內脂環類抗生素，以建立多重殘留量分析方法。標準品以乙?溶解後，以1-methyimidazole與trifluoroacetic anhydride在60℃經30分鐘衍生化後，再利用HPLC定量。使用之層析管柱為Merck Lichrospher 100RP-18E （5μm，125 × 4 mm），移動相為乙?／水（94／6, v／v），以螢光檢出器激發波長365nm，放射波長470 nm，進行檢測。結果顯示abamectin, moxidectin, milbemectin A3及milbemectin A4在1-300 ppb範圍內，doramectin及iverrnectin則在2-300 ppb範圍內均呈現良好的線性關係，r-square均大於0.998，顯示本方法能成功地分離上述6種成分，且各定量極限均低於法定的殘留容許量。牛肉檢體以乙?萃取，經C18固相萃取匣淨化。分別添加5、10、20、30、50、100、200及300 ppb於牛肉中進行回收試驗，結果abamectin、doramectin、moxidectin、ivermectm、milbemectin A3及milbemectin A4之回收率分別介於77.7-104%、73.3-108%、72.3 -104%、74.1-106%、78.1-108.8% 與74.8-110%之間，而相對標準差則介於2.11-16.57%之間。本方法中6種成分在牛肉檢體中最低檢出限量均為5 ppb。利用已建立之分析方法檢測市售生鮮及冷凍牛肉共50件，結果均未檢出。"

Abamectin, doramectin, moxidectin, ivermectin, milbemectin A3 and milbemectin A4 are similar macrocyclic lactone chemicals used as parasiticides or acaricides. A method using high performance liquid chromatography (HPLC) with fluorescence detection is presented for the simultaneous determination of the residue amounts of these compounds in bovine muscle. Samples are extracted using acetonitrile and cleaned up with solid phase extraction using a C18 column, followed by fluorescence-derivatized with 1-methylimidazole and trifluoroacetic anhydride in acetonitrile. The analogue was measured by HPLC with fluorescence detector at 365 nm excitation and 470 nm emission wavelengths. The limits of quantification are below the stipulated Taiwan Maximum Residue Limit for each compound. The recoveries of this method in bovine muscle ranged from 73.3 to 110%, with a RSD from 2.11 to 16.57%. The detection limit of those 6 compounds in bovine muscle was 5 ppb. No any above compounds were detected in 50 samples of bovine muscle tested. Therefore, the developed method can be used for rapid screen of macrocyclic lactones in bovine muscle. 本研究目的為利用快速簡便的高效液相層析法（hlgh perforrnance liquid chromatography, HPLC）檢測牛肉中abamectin, doramectin, moxidectin, ivermectin, milbemectin A3及milbemectin A4等巨內脂環類抗生素，以建立多重殘留量分析方法。標準品以乙腈溶解後，以1-methyimidazole與trifluoroacetic anhydride在60℃經30分鐘衍生化後，再利用HPLC定量。使用之層析管柱為Merck Lichrospher 100RP-18E （5μm，125 × 4 mm），移動相為乙腈／水（94／6, v／v），以螢光檢出器激發波長365nm，放射波長470 nm，進行檢測。結果顯示abamectin, moxidectin, milbemectin A3及milbemectin A4在1-300 ppb範圍內，doramectin及iverrnectin則在2-300 ppb範圍內均呈現良好的線性關係，r-square均大於0.998，顯示本方法能成功地分離上述6種成分，且各定量極限均低於法定的殘留容許量。牛肉檢體以乙腈萃取，經C18固相萃取匣淨化。分別添加5、10、20、30、50、100、200及300 ppb於牛肉中進行回收試驗，結果abamectin、doramectin、moxidectin、ivermectm、milbemectin A3及milbemectin A4之回收率分別介於77.7-104%、73.3-108%、72.3 -104%、74.1-106%、78.1-108.8% 與74.8-110%之間，而相對標準差則介於2.11-16.57%之間。本方法中6種成分在牛肉檢體中最低檢出限量均為5 ppb。利用已建立之分析方法檢測市售生鮮及冷凍牛肉共50件，結果均未檢出。