Determination of Catecholamines in Rat Brain Microdialysate by Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry Using Isotope-coded Derivatization Coupled with Dispersive Liquid-Liquid Microextraction
  
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KeyWord:in vivo microdialysis  Parkinson’s disease  multiple reaction monitoring(MRM)  stable isotope label
  
AuthorInstitution
QI Wei-mei,ZHAO Xian-en*,QI Yong,SUN Zhi-wei,CHEN Guang,YOU Jin-mao*,SUO You-rui 1.莱芜职业技术学院化工教研室;2.山东省生命有机分析重点实验室,山东省高校绿色天然产物与医药中间体开发重点实验室,曲阜师范大学化学与化工学院;3.中国科学院西北高原生物研究所
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Abstract:
      To achieve sensitive detection of catecholamines in trace rat brain microdialysate samples,using d0/d3-10-methyl-acridone-2-sulfonyl chloride(d0/d3-MASC) as isotope-coded derivatization, a novel ultra high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS) method was developed for the detection of dopamine(DA),norepinephrine(NE) and epinephrine(EP) by ultrasonic-assisted dispersive liquid-liquid microextraction(UA-DLLME).d0-MASC(light form) and d3-MASC(heavy form) were used as derivatization reagents for microdialysate samples and standards,respectively.The mixture of two solutions was enriched by dispersive liquid-liquid microextraction,and 3 catecholamines were detected by UHPLC-MS/MS with heavy derivatives as internal standards for corresponding light derivatives.The result indicated that the stable derivatives were obtained in pH 10.8 buffer/acetonitrile at 37 ℃ for 3.0 min,and then were separated within 3.0 min.Linearity ranges were obtained in the range of 0.02-10.0 nmol/L(r>0.995) for three catecholamines, with LODs of 0.005-0.010 nmol/L and LOQs of 0.018-0.040 nmol/L.Compared with the reported methods,the method is a convenient and validated method for the sensitive and accurate determination of catecholamines in trace rat brain microdialysates.
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