利用凝膠過濾管柱以二維液相層析儀線上檢測肉品中喹諾酮類藥物之殘留

Simultaneous Detection of Quinolone Residues in Meats Using Gel Filtration Column under a Two-Dimensional LC System

陳啟民(Chi-Min Chen) ； 周秀冠(Hsin-Kuan Chou) ； 傅以中(Yi-Chung Fu)

《臺灣農業化學與食品科學》 50卷1期 (2012/02) Pp. 47-59

https://doi.org/10.6578/TJACFS.2012.006

摘要

喹諾酮（quinolone）類藥物因具有抗菌範圍廣、生物利用率高及組織穿透性強等特點，被廣泛的使用於畜牧及養殖業的疾病防治與治療，而家畜及家禽之飼養過程如未遵守停樂時間或是不當使用未經允許的動物用藥，都可能造成動物用樂在動物組織的殘留。動物性食品的基質極為複雜，所以藥物殘留分析之樣品前處理通常經過複雜的萃取、淨化程序。本試驗擬結合二維液相層析儀及凝膠過濾管柱，開發適合大量樣品檢測之quinolone類藥物殘留分析方法。本方法取適量虱目魚肉及豬肉樣品，加入norfloxacin為內部標準品，以0.3％偏磷酸溶液：乙腈（1:10）溶液進行萃取。二維液相層析儀之分析條件為淨化管柱：PL aquage1-OH，分析管柱：ACE CI8-AR，搭配光二極體陣列檢出器。以此分析條例，共分析26種quinolone類藥物，其中有22種呈現良好之波峰，但因檢出器之限制，僅挑選pefloxacin、fleroxacin、enrofloxacin、orbifloxacin、difloxacin、tosufloxacin、pipmidic acid、enoxacin、ciprofloxacin、danofloxacin、gatifloxacin、sarafloxacin等12種quinolone類藥物進行進一步分析。試驗結果顯示12種quinolone類藥物檢量線線性相關係數R^2均大於0.995。添加50ng/g、100 ng/g及200ng/g之同日間及異日間之再現性分析：虱目魚肉平均回收率為76.3-149.8％，RSD為1.62-9.80％；豬肉平均回收率為71.9-139.1%，RSD為4.02-9.77%，儀器偵測極限為5~10ppb。此方法之樣品前處理步驟簡單、所需時間短，且有機溶劑的使用量少，適合應用於大量樣品的quinolone類藥物殘留篩檢。

關鍵字

喹諾酮類 ； 二維液相層析儀 ； 凝膠過濾管柱

並列摘要

The quinolones, with features such as wide spectra of antibacterial activities, high bioavailability, and excellent tissue penetration, abicity, are widely used to prevent and treat livestock diseases. Animal food products have extremely complicated matrix which requires a series of pre-treatment processes of extraction and purification before analysis of the drug residues. Normally the process of purification is operated with a solid phase extraction tube, yet the device has its technical limits. We aimed to develop a new method for analysis of drug residues in animal products, which can be applied to a great amount of samples. This experiment is designed to achieve our goal by combining the two dimensional LC system and the gel filtration column. We used milkfish and pork as samples with norfloxacin as an internal standard, and began the extraction using 0.3% metaphosphoric acid: acetonitrile (1:10) solution. After that we used the two dimensional LC system in conjunction with a purification tube (PL aquagel-OH), an analysis tube (ACE CI8-AR), and a photodiode array detector to run our tests. Based on the analysis conditions, we tested 26 drugs in the quinolones family, and 22 of them showed good peaks. With limitation of our detector, we chose 12 of them (pefloxacin, fleroxacin, enrofloxacin. orbifloxacin, difloxacin, tosufloxacin, pipmidic acid, enoxacin, ciprofloxacin, danofloxacin, gatifloxacin, and sarafloxacin) for further analysis. The results showed that the standard curves of all these 12 quinolones had R^2 above 0.995. The gauge repeatability and reproducibility (supplemented at the levels of 50ng/g, 100ng/g, and 200ng/g) within a day and between days indicated that the average recovery of the tested drugs from milkfish was 76.3~149.8% and RSD was 1.62~9.80%; the average recovery rate of these drugs from pork were 71.9~139.1% and RSD was 4.02~9.77%. The method detection limit (MDL) was 5~10ng/g. The technique provided in this experiment is easy to practice, time and organic solvents-saving, and therefore can be used to test a great amount of samples in short time.

並列關鍵字

Quinolones ； Two dimensional LC system ； Gel filtration column