The invention relates to a method for measuring
drug contents, particularly to a method for measuring danshensu, m-methyl-danshensu, protocatechualdehyde, and
protocatechuic acid in the
human blood plasma. The method comprises the following steps: 1) preparation of reference substance storing solutions: weighting reference substances of danshensu, m-methyl-danshensu, protocatechualdehyde, and
protocatechuic acid, and dissolving the reference substances with
methanol to obtain reference substance storing solutions of danshensu, m-methyl-danshensu, protocatechualdehyde, and
protocatechuic acid; 2) preparation of an
internal standard solution: dissolving an
internal standard substance of
vanillic acid with
methanol to obtain the
internal standard solution; 3) a treatment method of a
blood plasma sample: taking the
blood plasma sample, adding the internal
standard solution,
methanol,
hydrochloric acid, and
ethyl acetate into the
blood plasma sample, performing uniform mixing, performing
centrifugation of the obtained mixture to obtain a supernatant
ethyl acetate layer,
drying the
ethyl acetate layer, re-dissolving the ethyl acetate layer with methanol, and performing
centrifugation of the obtained solution to obtain supernatant liquid of the blood
plasma sample; and 4) content measurement: by adopting LC-MS / MS
chromatography, injecting the reference substance storing solutions and the supernatant liquid of the blood
plasma sample into a chromatographic instrument to obtain a chromatogram, and calculating concentrations of danshensu, m-methyl-danshensu, protocatechualdehyde, and protocatechuic acid in the
human blood plasma according to the areas of chromatographic peaks in the chromatogram.