title: Development of an immunoaffinity column method using broad-specificity monoclonal antibodies for simultaneous extraction and cleanup of quinolone and sulfonamide antibiotics in animal muscle tissues.
abstract: This paper describes a novel mixed-bed immunoaffinity column (IAC) method. The IAC was produced by coupling anti-quinolone and anti-sulfonamide broad-specificity monoclonal antibodies to Sepharose 4B for simultaneously isolating 13 quinolones (QNs) and 6 sulfonamides (SAs) from swine and chicken muscle tissues, followed by antibiotic determination using liquid chromatography-tandem mass spectrometry (LC-MS/MS). A new broad-specificity Mab (B1A4E8) toward sulfonamides was produced using sulfamethoxazole as hapten that demonstrated cross-reactivities to 6 SAs in the range of 31-112%. IAC optimized conditions were found that allowed the IAC to be reused for selective binding of both SAs and QNs. Recoveries of all 19 antibiotics from animal muscle ranged from 72.6 to 107.6%, with RSDs below 11.3% and 15.4% for intra-day and inter-day experiments, respectively. The limit of quantification ranged from 0.5 to 3.0ng/g. The strategy used here for a mixed-bed IAC may be used to study other compounds and more than two classes of analytes simultaneously.
authors.affiliation: Department of Pharmacology and Toxicology, College of Veterinary Medicine, China Agricultural University, Beijing 100094, China. hhlicun@live.cn
authors.forename: Cun
authors.initials: C
authors.lastname: Li
authors.forename: Zhanhui
authors.initials: Z
authors.lastname: Wang
authors.forename: Xingyuan
authors.initials: X
authors.lastname: Cao
authors.forename: Ross C
authors.initials: RC
authors.lastname: Beier
authors.forename: Suxia
authors.initials: S
authors.lastname: Zhang
authors.forename: Shuangyang
authors.initials: S
authors.lastname: Ding
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Li
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Shen
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000276:immunology / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D002645:Chickens; D002846:Chromatography, Affinity / Q000379:methods*; D009132:Muscles / Q000737:chemistry*; D015363:Quinolones / Q000276:immunology / Q000302:isolation & purification*; D015203:Reproducibility of Results; D013449:Sulfonamides / Q000276:immunology / Q000302:isolation & purification*; D013552:Swine
chemical_list: D000900:Anti-Bacterial Agents; D000911:Antibodies, Monoclonal; D015363:Quinolones; D013449:Sulfonamides