荧光能量转移法测定病毒灭活血袋中的亚甲蓝释放量

Determination of methylene blue release in plasma virus inactivation bag by fluorescence energy transfer method

目的建立一种简单、经济、快捷的方法,用于测定病毒灭活血袋中的亚甲蓝(MB)释放量。方法利用MB与牛血清白蛋白稳定的金纳米团簇(BSA-AuNCs)建立荧光能量转移体系,通过不同浓度MB对BSA-AuNCs荧光的猝灭程度建立MB测定的标准曲线,以此实现病毒灭活血袋中MB释放量的测定。结果当荧光发射波长为620nm左右,MB浓度(c_(MB))在(0.9-36)μmoL/L时,c_(MB)与BSA-AuNCs荧光的猝灭程度呈良好的线性关系,线性关系为(I_(0)-I)/I_(0)=0.018c_(MB)+0.021(r=0.996);利用本法测定MB浓度,回收率为(98.00-101.95)%;利用本法测定高、中、低3个浓度的MB,日内变异系数分别为0.73%、0.81%与0.77%,日间变异系数分别为3.92%、3.81%与4.73%;利用本法测定病毒灭活血袋中的亚甲蓝释放量,测得的结果与使用固相萃取-分光光度法测得的结果无差异(P>0.05)。结论利用荧光能量转移法可以实现病毒灭活血袋中MB释放量的简单、快速测定,得到的结果准确、可靠。

Objective To establish a simple,economical and rapid method for the determination of methylene blue(MB)release in virus inactivation bag.Methods Based on the fluorescence energy transfer between MB and BSA-stabilized gold nanoclusters(BSA-AuNCs),the standard curve of MB determination was established by measuring the fluorescence quenching degree of MB to BSA-AuNCs in different concentrations to conduct the determination of MB release in virus inactivation bag.Results There was a good linear relationship between the MB concentration(c_(MB))and the fluorescence quenching degree of BSA-AuNCs[(I_(0)-I)/I_(0)=0.018c_(MB)+0.021(r=0.996)]when the fluorescence emission wavelength was about620 nm and thec_(MB)was in the range of(0.9-36)μmoL/L.The recovery of MB was 98.00%-101.95%when applied to determine MB at high,medium,and low concentrations,the obtained intra-day variation coefficients were 0.73%,0.81%and 0.77%respectively,and the obtained inter-day variation coefficients were 3.92%,3.81%,and 4.73%respectively.There was no significant difference between the results measured by this method and those measured by combination of solidphase extraction and spectrophotometry(P>0.05).Conclusion The fluorescence energy transfer method could achieve simple and rapid determination of MB release in virus inactivation bag with accurate and reliable results.