The Study On Immunoaffinity Chromatography For Multiple Fluoroquinolone Antibiotics And Brombuterol In Food Samples

The saying "Bread is the staff of life" simply illustrate the importance of food. With the development of society and economy, after solving the problem of insufficient foods for comsuming, people pay more attention to food safty. To ensure the health of the consumers, it is required to effectively monitor the possible contaminants in foods. Therefore, the developemnt of the simple, convenient, fast, accurate and reliable analytical method for the detection of various contaminants in foods are necessary. In this study, the immunoaffinity chromatographies respectively for simultaneously determining four fluoroquinolone antibiotics and for one β-agonist(e.g. brombuterol) detection in food samples were described.In this study, the immunoaffinity colomn was used for extraction of target analyte from samples, while high performance liquid chromatography(HPLC) was used as detection method. In the proposed immunoaffinity chromatography, the procedures of sample extraction are simple, while the detection is accurate and reliable with short analysis time. An immunoaffinity chromatography(IAC) for simultaneously selective extraction of four fluoroquinolone antibiotics, e.g. norfloxacin(NOR), pefloxacin(PEF), lomefloxacin(LOM) and enrofloxacin(ENRO) from swine and chicken meat samples was developed. The IAC column was constructed by covalently coupling polyclonal antibody(p Ab) against NOR to CNBr-activated Sepharose 4B and packed into a common solid phase extraction(SPE) cartridge. It was found that IAC column was able to separately capture NOR, PEF, LOM and ENRO with the maximum capacity of 200, 336, 352 and 421 ng, respectively. The prepared IAC column can be used in the analysis of real samples. The extraction conditions including loading, washing and eluting solutions were carefully optimized. Under optimal conditions, the standard solution extraction recoveries of the IAC column for NOR, PEF, LOM and ENRO at two different contents were within 85.8-105.9%. The IAC column was also applied to real sample extraction. Swine and chicken meat samples were collected and spiked with NOR, PEF, LOM and ENRO at the content of 20 ng g-1 and 35 ng g-1, respectively. The spiked and unspiked samples were extracted by IAC column and measured by high performance liquid chromatography(HPLC). It was found that there was no detectable NOR, PEF, LOM and ENRO in the blank samples, and the extraction recoveries of the IAC for NOR, PEF, LOM and ENRO from the spiked samples were within 75.3-99.8% with the relative standard deviation(RSD) of 4.3-12.1%. The stability of the column was also tested. It was showed that after 30 times repeated usage, 65% of the maximum capacity was remained. The results demonstrated that the proposed immunoaffinity chromatography was able to extract multiple fluoroquinolone antibiotics from food samples with the advantages of high column capacity, selectivity, extraction efficiency and stability.Brombuterol is one kind of β-agonists which may be illeagally used in animal as food additive. People consuming brombuterol-contaminated food may diaplay a series of physiological reactions such as palpitation, vomiting, trembling and rapid heartbeat. More serous physiological reactions will be appeared for the people with high blood pressure, heart disease, diabetes and hyperthyroidism. Therefore, the establishment of fast detection method for the detection of brombuterol is also required. In this study, the preparation and characterization of the immunoaffinity column for one component(e.g. brombuterol) extraction was finalized. However, due to the limited time, the experiment of the extraction and detection of brombuterol in food samples by immunoaffinity chromatography is still going on.