ABSTRACT

Amphenicol antibiotics including chloramphenicol (CAP), florfenicol (FF) and thiamphenicol (TAP) have been widely used in aquaculture and veterinary to treat various infections. Because of their high toxicity, it is important to detect these amphenicol antibiotics simultaneously. Using the monoclonal antibody against CAP (mAb_CAP) binding CAP only and the mAb_FF recognizing both FF and TAP, the two antibodies were covalently coupled to CNBr-activated Sepharose-4B for the simultaneous extraction of CAP, FF and TAP from samples. The bound CAP, FF and TAP were eluted from the Sepharose-4B and measured by LC–MS/MS. The extraction conditions of the two-antibodies-immobilized immunoaffinity clean-up (IAC) for CAP, FF and TAP were optimized and the maximum capacity was investigated. The extraction efficiency of IAC for CAP, FF and TAP was 1.1–2.1 times higher than that of SPE. The LC–MS/MS coupled with IAC was a powerful analytical method for the detection of CAP, FF and TAP simultaneously.

摘要

包括氯霉素（CAP）、氟苯尼考（FF）和甲氧苄啶（TAP）在内的氨苄类抗生素已广泛用于水产养殖和兽医治疗各种感染。由于它们的高毒性，同时检测这些双酚类抗生素很重要。使用抗 CAP 的单克隆抗体 (mAb _CAP ) 仅结合 CAP 和 mAb _FF识别 FF 和 TAP，这两种抗体与 CNBr 激活的 Sepharose-4B 共价偶联，用于从样品中同时提取 CAP、FF 和 TAP。从 Sepharose-4B 中洗脱结合的 CAP、FF 和 TAP，并通过 LC-MS/MS 进行测量。优化了CAP、FF和TAP的双抗体固定化免疫亲和纯化(IAC)的提取条件并考察了最大容量。IAC 对 CAP、FF 和 TAP 的提取效率是 SPE 的 1.1-2.1 倍。LC-MS/MS 与 IAC 联用是同时检测 CAP、FF 和 TAP 的强大分析方法。