牛血清白蛋白荧光猝灭法测定甲紫含量

Determination of methyl violet by fluorescence quenching method base on the interaction of bovine serum albumin with methyl violet

目的建立基于牛血清白蛋白（BSA）荧光猝灭测定甲紫（MV）含量的荧光分析法。方法选择激发波长为279 nm,发射波长为340 nm,MV与BSA相互作用使其内源性荧光产生猝灭,荧光强度的下降值△I与MV浓度呈线性相关,标准曲线法测定甲紫溶液中MV的含量。结果△I与MV的浓度在2～10μg/mL内呈线性关系,△I=11.709C＋0.95（r=0.999 2）,加样回收率为97.3%～100.1%,RSD为0.24%～1.7%。3批次医用甲紫溶液中MV的平均含量分别为1.016%、1.011%、1.029%,与标示量一致。结论该法操作简便,灵敏度高,结果准确,可用于低含量甲紫样品的测定。

Objective To establish a quantitative analysis of Methyl violet（ MV） by fluorescence spectroscopy.Methods At an excitation wavelength of 279 nm and an emission wavelength of 340 nm,the entogenous fluorescence of bovine serum albumin（ BSA） was quenched as the interaction of bovine serum albumin with methyl violet.The decrease of fluorescence △I and the MV concentration showed a linear correlation. The content of MV in Methylrosanilinium Chloride Solution was determined by calibration curve method. Results The △I showed a good liner relation with the concentration of the MV in 2 ～ 10 μg / mL,△I = 11. 709 C ＋ 0. 95（ r = 0. 999 2）. The recovery was 97. 0% ～ 100. 4%,and RSD was 0. 24% ～ 1. 7%. Three batches of medical Methylrosanilinium Chloride Solution were determined,and the average concentrations were 1. 016%,1. 011%,1. 029%,respectively,which were consistent with the labeled amounts. Conclusion The proposed method is simple,accurate and sensitive,and it can be applied to determine the trace amount of the Methyl violet.