The invention relates to a method for detecting nine
cholesterol oxides in food by
gas chromatography-
mass spectrometry, which comprises the following steps: S1, adding 19-hydroxycholesterol as an
internal standard substance to carry out sample
lipid extraction; S2, dissolving the obtained lipid extract, and carrying out
saponification extraction to remove
moisture so as to obtain a concentrated filtrate; S3, purification and concentration: adding 3mL of n-
hexane, shaking and dissolving, adding into activated NH2-SPE, washing the column, discarding waste liquid, collecting filtrate, and blow-
drying by
nitrogen; S4,
derivatization: adding 100 [mu] L of N, O-bis (
trimethylsilane) trifluoroacetamide BSTFA for
derivatization for 40 min, blow-
drying with
nitrogen, and dissolving with 1.0 mL of n-
hexane to a constant volume; and S5, taking a mixed
standard solution of nine
cholesterol oxides, detecting the mixed
standard solution under
gas chromatography conditions and
mass spectrometry conditions, and drawing a
standard curve. According to the method, a pretreatment mode suitable for complex samples is established,
saponification conditions and a
solid-phase extraction column are optimized, the accuracy of analysis and detection is improved by adopting an
internal standard substance, and the method has the advantages of accuracy, high efficiency and wide applicability.