Determination of dopamine and its metabolites in rat cerebral microdialysates by HPLC-ECD / 中国临床药理学与治疗学

ABSTRACT

AIM: To develop a sensitive and rapid determination method of dopamine(DA) and its metabolites 3,4-dihydroxyphenylacetic acid(DOPAC) and homovanillic acid(HVA) in rat cerebral microdialysates.METHODS: Microdialysis probes were placed into the right stratums of Wistar rat brains and perfused with Ringer's solution at a rate of(1.5)(?l?min~(-1)).A reverse phase HPLC with electrochemical detection (ECD) were used to assay DA,DOPAC and HVA after the cerebral microdialysates were collected every 20 minutes from awake,freely moving rats.In order to know the reliability of this method,selectivity,linear range,precision and accuracy were tested and the contents of DA,DOPAC and HVA in rat microdialysates were determined.RESULTS:The standard curve were in good linear at the range of(12.5) to 250(?g?L~(-1)).The recovery rates of DOPAC,DA and HVA at the concentration of(0.05),(0.13),(0.25)(?g?L~(-1)) were(98.33)%,(102.67)%,(92.33)% respectively.Their with-in day RSD were(3.3)%,(3.4)%,(2.5)% and between-day RSD(4.2)%,(2.3)%,(5.6)% respectively.The contents of DOPAC,DA and HVA in rat microdialysates were(1.79)?(0.07),(0.45)?(0.02),(1.67)?(0.05)(?g?ml~(-1)) (n=6),respectively.CONCLUSION: The simple,accurate and stable method can be applied to the study of the basic researches of diseases related to DA by cerebral microdialysis in rat.