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Authors
Abstract(s)
Biogenic amines are neurotransmitters involved in several physiological processes.
Changes and disturbs in their function are associated to several neurological disorders or
diseases, as well as consumption of psychotherapeutic or psychotropic drugs. The detection of
these compounds in biological fluids is therefore of clinical and neurochemical interest.
The goal of this work was to develop and validate and analytical method for the
detection and quantification of the biogenic amines serotonin (5-HT), dopamine (DA) and
norepinephrine (NE), using microextraction in packed syringe (MEPS) and liquid
chromatography coupled to electrochemical detection (HPLC-ED) in both urine and plasma
samples. The internal standard used was 3,4-dihydroxybenzylamine (DHBA). The MEPS
extraction procedure was optimized using the design of experiments (DOE) tool, and the final
conditions of 8 strokes, no washing, and two elutions of 100 FL methanol were chosen.
The method was fully validated according to internationally accepted guidelines from
the Food and Drug Administration. Linearity was established between 50-1000 ng/mL for 5-HT
and between 5-1000 ng/mL for DA and NE, with determination coefficients (R2) higher than
0.99 for all compounds. The limits of detection and quantification were respectively 20 and
50 ng/mL for 5-HT, and 2 and 5 ng/mL for DA and NE. Intra- and interday precision ranged
from 1 to 10 %, while accuracy was within a ±15% interval for all compounds.
Authentic urine and plasma samples were analysed by the validated method, and the
three compounds were detectable and quantifiable in urine, while only 5-HT was detected
and quantified in plasma.
This is the first time that a commercially available MEPS column was used for the
simultaneous detection of biogenic amines in urine, and also the first time that 5-HT was
detected and quantified in plasma using MEPS. MEPS proved to be fast to perform with the
use of less solvent volumes, saving time and money and being less laborious. The validated
method is useful for the determination of biogenic amines in laboratorial routine.
Description
Keywords
Aminas biogénicas Fluidos biológicos DA (Dopamine) Biogenic amines