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作 者:王晶 王红丹[2] 芦云 李玲 薛晓晶 WANG Jing;WANG Hongdan;LU Yun;LI Ling;XUE Xiaojing(CAIQTEST(Beijing)Co.,Ltd.,Beijing 100123,China;Chinese Academy of Inspection and Quarantine Comprehensive Test Center,Beijing 100123,China)
机构地区:[1]中检科(北京)测试技术有限公司,北京100123 [2]中国检验检疫科学研究院综合检测中心,北京100123
出 处:《食品科技》2021年第11期280-284,共5页Food Science and Technology
基 金:中国检验检疫科学研究院基本科研业务费项目(2020JK038)。
摘 要:针对微生物法检测V_(B12)不易操作,结果平行性和重复性差等问题,对国标GB 5413.14—2010的关键步骤进行分析和优化。在乳酸杆菌培养基中加入V_(B12)标准溶液以刺激莱士曼氏乳酸杆菌生长;调整工作菌种的接菌方式,明确加菌量;验证前处理提取缓冲液中偏重亚硫酸钠对菌株的影响并确定用量;用酶标仪替代分光光度计测定结果。结果表明,菌悬液按0.05%的比例加入到测定培养基中混匀分装的方式,操作简便,线性关系好,R;值为0.9995;浓度为0.01 mg/mL偏重亚硫酸钠缓冲液适合大多数样品提取。优化后的方法加标回收率范围是98.6%~113.3%,重复性实验的RSD范围在1.17%~3.09%。微生物法影响因素多,不易掌握,方法优化后结果准确、稳定,可操作性强,可为V_(B12)检测提供优化思路和技术支持。The key steps of GB 5413.14—2010 were analyzed and optimized to solve the problems of complex operation and poor parallelism and reproducibility of V_(B12)detection by microbiological method.The growth of Lactobacillus leichmannii was stimulated by adding V_(B12)standard solution in Lactobacillus culture medium.The inoculation mode of working strains was adjusted and the amount of bacteria added was specified.The effect of sodium pyrosulfite in the pretreatment extraction buffer on the strain was verified and the dosage was determined.The results were measured with a spectrophotometer instead of a microplate reader.The results showed that the method of mixing and packing with 0.05% of bacterial suspension into the test medium was easy to operate and had good linear relationship with R;of 0.9995.The concentration of 0.01 mg/mL sodium pyrosulfite buffer was suitable for most samples.The recoveries of the optimized method were in the range of 98.6%~113.3%,and the RSDs of the reproducible experiments were in the range of 1.17%~3.09%.Microbiological method has many influencing factors and is difficult to grasp.The optimized results of the method are accurate,stable,and highly maneuverable,which can provide optimization ideas and technical support for V_(B12)detection.
分 类 号:TS252.7[轻工技术与工程—农产品加工及贮藏工程]
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