An improved liquid chromatographic method with electrochemical detection for direct determination of serotonin in microdialysates from Caudate-putamen and pineal gland regions of rat brain.
A liquid chromatography method coupled with electrochemical detection has been developed for the direct measurement 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) in brain microdialysates. The separation conditions have been optimized to detect only the 5-HT and 5-HIAA in dialysates and elute the other monoamines and their metabolites in the void. Linear regression analysis of chromatographic peak area as a function of concentration in the range 5-1000 pg/mL gave correlation coefficients over 0.995. Sample stability and an assay validation for precision and accuracy were also performed. The limit of detection (S/N = 3) for 5-HT was 12 femtomol/mL. The method has been applied to simultaneously measure extracellular 5-HT and 5-HIAA in brain microdialysates from the pineal gland and caudate-putamen of awake and freely-moving rats under basal conditions.
Gunaratna
P Chandrani
PC
Bioanalytical Systems Inc., West Lafayette, IN 47906, USA. prema@bioanalytical.com
Cadle
Karen K
KK
Kissinger
Candice B
CB
D000818:Animals; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D004563:Electrochemistry / Q000295:instrumentation / Q000379:methods*; D045604:Extracellular Fluid / Q000737:chemistry / Q000378:metabolism; D006897:Hydroxyindoleacetic Acid / Q000032:analysis / Q000378:metabolism; D008297:Male; D017551:Microdialysis / Q000295:instrumentation / Q000379:methods; D009043:Motor Activity / Q000502:physiology; D017072:Neostriatum / Q000737:chemistry* / Q000378:metabolism; D009448:Neurochemistry / Q000295:instrumentation / Q000379:methods*; D009474:Neurons / Q000737:chemistry / Q000378:metabolism; D010870:Pineal Gland / Q000737:chemistry* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012044:Regression Analysis; D015203:Reproducibility of Results; D012701:Serotonin / Q000032:analysis* / Q000378:metabolism; D014851:Wakefulness / Q000502:physiology
D012701:Serotonin; D006897:Hydroxyindoleacetic Acid