摘要
建立了测定食品中细菌总数的ATP生物发光检测技术,考察了各种理化因素对生物发光反应的影响。反应体系最优化条件:Ln浓度为70mg/L,FL浓度为50mg/L,Mg2+浓度为0.25mmoL/L,pH为7.2,最适温度为23℃。在10-10～10-15mol/mL范围内,ATP浓度与生物发光强度之间有较好的线性关系,相关系数R2=0.978。方法检出限为10-15mol/mL,批内变异和批间变异分别小于7%和8%。将建立好的ATP生物发光反应体系应用于食品样品中细菌总数的检测,加标回收率范围为82.2%～112.4%,检测结果与平板计数结果相关性良好。因此,建立的ATP生物发光检测技术用于检测食品中细菌总数是可行的。
A bioluminescence assay method was established for the determination of bacterial count in food samples. The effects of several physical and chemical parameters on the bioluminescence assay were studied. Such conditions were optimized as the concentration of luciferin of 70 mg/L, firefly luciferase of 50 mg/L, Mg2＋ of 0.25 mmol/L, pH value of 7.2, optimal temperature of 23 %. The intensity of bioluminescent showed a linear response to ATP in a concentration range from 10^-10 mol/mL to 10^-15 moll mL, with a correlation coefficient of 0.978 and in a low minimum detectable concentration of 10^15 mol/mL. The coefficient of variation was less than 7% and 8% for intra- and inter-assay precision, respectively. This method has been successfully applied to the detection of the total count of bacterial in food samples with recoveries from 82.2% to 112.4%. It showed a good correlation with the traditional method. So it is feasible to use the bioluminescence assay method to detect bacterial count in food samples.
出处
《食品科技》
CAS
北大核心
2012年第1期275-278,282,共5页
Food Science and Technology
基金
湖北省教育厅项目(Q20081801)
武汉工业学院创新基金项目(08cx006)
