The invention relates to the technical field of
biological engineering, in particular to a method for detecting
cotinine,
mandelic acid and phenylglyoxalic acid in human
urine simultaneously on the basis of a
derivation method. The method is characterized in that the method comprises: step one, storing collected
urine after centrifugal process; step two, taking the
urine obtained in step one and adding the urine into internal
label benzophenone,
diphenylamine,
hydrochloric acid and
chloroform to carry out extraction through rotating
centrifugation, thereby aspirating and recording organic phase as A and taking residual solution for standby; step three, taking the residual solution obtained in step two and adding the residual solution into
sodium hydroxide solution and
chloroform to carry out secondary extraction through rotating
centrifugation, thereby aspirating and recording organic phase as B; step four, mixing the organic phase A extracted in step two and the organic phase B extracted in step three inside a
glass tube, and then blowing dry both organic phases; step five, adding
acetonitrile into the
glass tube adopted in step four for redissolution and adding derivative
reagent to carry out derivative reaction; and step six, taking the reaction solution obtained in step five to carry out
gas phase chromatography-
mass spectrometry combined measurement. The method has the characteristics of simplicity, convenience, quickness, reliable result, high degree of
automation and convenient industrialized application.