LC-MS/MS法测定脑微透析液中乙酰胆碱的浓度___

Objective:To develop a sensitive and reliable method for determination of acetylcholine in microdialysates from rats brainMethods:The detection was performed by MRM mode via electrospray ionization(ESI) source operating in the positive ionization mode;The precursor-to-product ion transitions of the analyte acetylcholine is m/z 146→87 with internal standard(acetylcholine-d9) at m/z 155→87The optimal ionization and fragmentation conditions,as well as liquid chromatographic ones, to detect acetylcholine were developed and validated.The new developed analytical method was further applied in determination of acetylcholine in microdialysates from rats brain after intracerebral perfusion of 6-hydroxydopamine(6-OHDA).Results:The method was linear over the concentration range of 0.0292-1.46 ng·mL^-1(r=0.9979);The limit of quantification is 0.00292 ng·mL^-1;The accuracy was 96.1%-111.7%;The intra- and inter-day precision values were 1.8%-7.8%(n=6) and 2.0%-19.2%(n=3), respectively.Conclusion:The fully validated LC-MS/MS method has been successfully applied to determination of acetylcholine in microdialysates from rats brain.