The application belongs to the field of analysis and detection, and particularly relates to a method for detecting the content of directly absorbed exogenous
glycine of microorganisms in soil, which comprises the following steps: firstly, mixing a double-labeled
glycine solution with soil, adding
sodium laureth
sulfate and
chloroform for
fumigation, then extracting, deriving by hydrocarbonation reaction of ethyl bromoformate, and completing the extraction of
glycine in microorganisms after stable layering by using
sodium bicarbonate solution; and after the extraction is cracked by a
stable isotope ratio mass spectrometer, exogenous glycine and original glycine in microorganisms are distinguished by characteristic ions to be determined. The method can effectively detect the content of directly absorbed exogenous glycine of microorganisms, is simple in operation, strong in specificity, has a good
linear relationship, high sensitivity and accuracy, and effectively solves the problem that the content of directly absorbed exogenous glycine of microorganisms cannot be directly quantified. Thus, the direct utilization pathway of soil microorganisms to exogenous
amino acid substances is quantitatively judged.