尿中高香草酸的酶荧光光度测定法

Enzymatic fluorometric determination of homovanillic acid in urine

用三氯醋酸处理尿样后,离心分离沉淀物。上清液的 pH 用 Tris 缓冲液控制在8.5,加一定量的 H(?)O_2及过氧化物酶,在30±0.1℃反应30min 后于930型荧光光度计上,以λ(?)=330 nm,λ(?)=450 nm 波长测定产物的荧光强度就可测出尿样中 ppm 级高香草酸的含量。方法的选择性好,操作简便。

The urine sample is treated with several gramsof trichloroacetic acid,and neutralized withNaOH solution to precipitate the proteins.Aftercentrifugation,an aliquot of the clear solutionis taken,the pH is controlled to 8.5 with trisbuffer solution,and heated to 30±0.1℃ ina water bath,30 min.after H_2O_2 and peroxidasesolutions are added,the fluorescence of the pro-duct is measured with a model 930 fluorometerλ_(e(?))=330m;λ_(em)=450nm.This method can be usedto determine homovanillic acid in urine in ppmrange.It is very selective and easy to operate.