For the simultaneous determination of monoamine neurotransmitters
(NTs) like dopamine, serotonin, noradrenaline, and epinephrine, and
their metabolites (metanephrine, normetanephrine, 3-methoxytyramine,
vanillylmandelic acid, 3,4-dihydroxyphenylacetic acid, homovanillic
acid, and 5-hydroxyindoleacetic acid), a robust liquid chromatography
method coupled with tandem mass spectrometry (LC–MS/MS) was
introduced as the analytical method. This analytical method proved
to be accurate for the simultaneous measurement of the amounts of
11 NTs and their metabolites in biological samples. The method proved
to be more efficient and better than the previously reported method
in terms of precision, recovery, sample requirement, and extraction
procedure. The reported method requires only 100 μL of blood
and 200 μL of urine, and the extraction procedure requires acetonitrile
precipitation, filtration, drying, and reconstitution in water. The
separation of all analytes was performed on an C18 column (4.6 mm
× 150 mm and 1.8 μm). A 10 min gradient elution program
with a mobile phase consisting of phase A (0.2% formic acid in water)
and phase B (methanol) was used. The positive ionization mode was
used for the detection of all analytes in multiple reaction monitoring
(MRM). The proposed method was validated with an internal standard
and yielded lower limits of detection and quantification ranges of
0.0182–0.0797 ng/mL and 0.0553–0.2415 ng/mL, respectively,
with a good linearity (R
2) between 0.9959
and 0.9994. The recoveries ranged from 73.37% to 116.63% in blood
and from 80.9% to 115.33% in urine. For the NTs and metabolites, the
intra- and interday % CV were 0.24–9.36 and 0.85–9.67,
respectively. The developed LC–MS/MS method was successfully
used for the determination of trace amounts of endogenous compounds
in human blood and urine samples.