呋喃唑酮代谢物人工抗原的制备及鉴定

Preparation and Identification of Artificial Antigen of Furazolidone Metabolite

在催化剂条件下,以碳酸二甲酯和乙醇肼为原料,四氢呋喃为溶剂,合成了高收率的呋喃唑酮代谢物3-氨基-2-恶唑烷酮(AOZ)。实验结果表明,得到的AOZ收率可达到82.7%,此时的最佳反应条件如下:催化剂用量12%,反应温度75℃,反应时间3h。随后对AOZ进行结构改造,得到AOZ半抗原衍生物,同时采用戊二醛法将AOZ及其半抗原衍生物与活化的卵血清蛋白(cOVA)和牛血清白蛋白(cBSA)交联,成功制备了免疫原和包被原;通过气相质谱联用仪(GC-MS)、元素分析、紫外分光光度计等手段对半抗原和完全抗原进行鉴定。GC-MS结果证明了AOZ的成功合成;元素分析数据表明,半抗原顺利合成;紫外扫描数据显示,半抗原与cOVA、cBSA偶联成功,免疫原I、免疫原Ⅱ的偶联比分别为16.74和24.05。

The furazolidone metabolite 3-amino-2-oxazolidinone（AOZ） was synthesized at a high yield from the raw materials dimethyl carbonate and ethanol hydrazine hydrate using a catalyst and tetrahydrofuran as a solvent. The results showed that the optimum reaction conditions for maximum AOZ yield（82.7%） were as follows： catalyst dosage, 12%; reaction temperature, 75 ℃; and reaction time, 3 h. Subsequently, a hapten derivative was obtained from the structural transformation of AOZ. Then, immunogens and coating antigens were successfully prepared using a method involving glutaraldehyde to conjugate AOZ and its hapten derivative with activated ovalbumin（cOVA） and bovine serum albumin（cBSA）. The hapten and immunogen were identified by gas chromatography-mass spectrometry（GC-MS）, elemental analysis, and UV spectrophotometry. The GC-MS data demonstrated the successful preparation of AOZ. The elemental analysis indicated successful formation of the hapten. UV spectrophotometry results showed that conjugation of the hapten with cOVA and cBSA was successful, and the conjugation ratios of immunogen I and immunogen II were 16.74 and 24.05, respectively.