- 期刊
A Polyclonal Antibody-based Immunoassay for Determination of Growth Stimulant Ractopamine: Comparative Study with Recent Advances in Immunoassay Methods
開發萊克多巴胺（生長促進劑）免疫檢驗試劑：免疫分析方法及其研究進展
摘要
美國、加拿大、澳大利亞等國核准萊克多巴胺（Ractopamine，Rac）作為豬及牛隻之生長促進劑使用，此飼料添加物在我國、歐盟及中國大陸禁用（註：衛生署已於101年9月11日公告牛肌肉中萊克多巴胺之殘留容許量為10 μg/kg，使用萊克多巴胺且殘留符合標準的牛肉可輸入。國內則須經動物用藥品檢驗登記核准後才能使用）。於此，我們開發檢測萊克多巴胺之酵素免疫檢驗試劑，以免疫注射ractopamine-succinate-ovalbumin抗原之兔子產生anti-Rac多株抗體。此套組校正曲線線性濃度範圍0.3-24.3 μg/kg，IC50為3.2 μg/kg，針對尿液及血清樣本毋需特殊的樣品前處理，肉及飼料則使用鹽酸進行萃取。實驗結果顯示尿液、血清和肌肉等基質的檢測極限為0.3 μg/kg，飼料的檢測極限為4 μg/kg，本研究開發具有快速、靈敏及特異的檢測，可應用於肉禽場屠前自主藥物殘留監控並應用於現場例行的藥物殘留檢驗監控作業。同時，文中亦對萊克多巴胺殘留的各種免疫分析方法進行文獻整理與回顧，以期做為未來從事研究萊克多巴胺殘留檢驗的重要參考。
並列摘要
Ractopamine is a β-adrenergic agonist approved for use as a growth promoter in swine and cattle in the USA, Canada and Australia etc. but not in the European Union, China, or Taiwan. Here, we report the development of a polyclonal antibody-based enzyme-linked immunosorbent assay (ELISA) for ractopamine. Rabbits immunized with ractopamine-succinate-ovalbumin were utilized for polyclonal antibody generation. The calibration graphs of ractopamine showed linearity over the concentration ranges 0.3-24.3 μg/kg, whereas IC50 was 3.2 μg/kg, allowing the analysis of urine and serum samples without the need for sample clean-up. This paper also deals with the extraction of hydrochloric acid respect to meat and feed samples. It appears that the limit of determination of this assay for urine, serum and meat matrix was estimated to be 0.3 μg/kg, and for feed samples was 4 μg/kg. The results obtained a rapid, sensitive and specific assay to detect positive samples in routine analysis. Simultaneously, recent advances in immunoassay methods for ractopamine residues are reviewed herein, that can be considered as a basis for further research aimed at identifying the most efficient approaches for the analysis of ractopamine.