利用液相層析串聯質譜儀檢測肉品中殘留胺基糖苷類抗生素之多重分析法

由於胺基醣苷類抗生素廣效的抗細菌能力，常被用來作為治療動物感染性疾病或飼料添加物，故開發一個相對快速且具備多重藥品的檢驗方法極為重要。本方法利用高效液相層析搭配串聯質譜儀（high performance liquid chromatograph/tandem mass spectrometer，HPLC/MS/MS）檢測14種胺基醣苷類抗生素，包括apramycin（安痢黴素）、dihydrostreptomycin（雙氫鏈黴素）、streptomycin（鏈黴素）、gentamicin（健牠黴素）、kanamycin（康黴素）、neomycin（新黴素）、spectinomycin（觀黴素）、tobramycin、paromomycin、hygromycin、netilmicin、amikacin、ribostamycin和sisomicin，於肉品檢體中可成功分離出胺基醣苷類抗生素。檢體經含5%三氯醋酸萃取緩衝溶液進行兩次去蛋白與萃取並利用正己烷去脂，透過3 M氫氧化鋰溶液調整溶液pH值至5.5－6.5，通過經活化後之Oasis MCX（Mix mode strong-cation exchange）固相萃取匣淨化，收集沖提液並加入10%甲酸溶液1 mL後，以氮氣吹乾至固定體積，殘留物加入25%七氟丁酸溶液，並以去離子水定容至10 mL。本方法使用高效液相層析串聯式質譜儀，移動相A為20 mM七氟丁酸，而移動相B為含20 mM七氟丁酸之乙腈可同時檢測五種不同種類的基質並可成功分離出胺基醣苷類抗生素，試驗結果為雞肉回收率80.2－100%、豬肉回收率79.9－100.5%、牛肉回收率76.7－97.0%、豬肝回收率67.0－85.9%，而豬腎回收率70.1－85.1%，檢出極限肌肉介於1－25 ppb，內臟介於5－50ppb，同日異日變異係數皆小於15%，基質對應檢量線迴歸係數（R）皆為0.993以上。此方法應可作為檢測肉品中殘留胺基醣苷類抗生素之敏感且快速的方法。

關鍵字

液相層析串聯質譜法；胺基醣苷類；肉品；內臟

並列摘要

Aminoglycosides (AMGs) are commonly used antibiotics for the treatment of infectious diseases and as an additive to animal feeds for their broad-spectrum antibacterial activity. It is important to develop a relatively fast and multiplex method to determine these antibiotics. In this study, a method using liquid chromatography tandem mass spectrometry (LC/MS/MS) was developed for determination of aminoglycosides in meat, porcine liver, and porcine kidney. Sample treatment involves extraction of the analytes with 5% trichloroacetic acid buffer, defatting with hexane, adjustment of pH to 5.5-6.5, and cleanup by a solid phase extraction cartridge (SPE) with a mix mode strong-cation exchange (MCX) cartridge. The eluent was dried with N2 gas after addition of 1 mL 10% formic acid, and the residue was adjusted to 10 mL with deionized water after adding 25% hepta-fluorobutyric acid. The chromatography was carried out in an Agilent eclipse plus C18 column, and mobile phase component A was water with 20 mM heptafluorotyric acid, while component B was acetonitrile with 20 mM heptafluorotyric acid. The overall average recoveries were 67.0-100.5% in edible tissues of chicken, pork, beef, liver and kidney. The coefficients of variation were below 15% and limit of quantification for all antibiotics are 1-50 ppb in edible tissues tested. This method is considered as a sensitive and rapid one to determine aminoglycosides in meat and riscera.

並列關鍵字

Liquid chromatography tandem mass spectrometry ； Aminoglycosides ； Meat product ； Internal organs

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利用液相層析串聯質譜儀檢測肉品中殘留胺基糖苷類抗生素之多重分析法

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