Fast liquid chromatography separation and multiple-reaction monitoring mass spectrometric detection of neurotransmitters.
We describe here the fast LC-MS/MS separation of a mixture of neurotransmitters consisting of dopamine, epinephrine, norepinephrine, 3,4-dihydroxybenzylamine (DHBA), salsolinol, serotonin, and gamma-aminobutyric acid (GABA). The new UltiMate 3000 Rapid Separation system (RSLC) was successfully coupled to the 4000 QTRAP mass spectrometer operating in multiple-reaction monitoring (MRM) mode. The separation was attained using a 100 mm length, 2.2 microm particle size Acclaim column at a flow rate of 0.5 mL/min. The column back pressure was 350 bar, while the total run time including column re-equilibration was 5.2 min. The peak resolution was minimally affected by the fast separation. The RSLC-MRM separation was found to have a precision range based on peak area for 50 replicate runs of 2-5% CV for all analytes, and the reproducibility of the retention time for all analytes was found to range from 0-2% CV. The described method represents an almost seven times shorter analysis time of neurotransmitters using LC/MRM which is very useful in screening large quantities of biological samples for various neurotransmitters.
Hammad
Loubna A
LA
METACyt Biochemical Analysis Center, Department of Chemistry, Indiana University, Bloomington 47405, USA.
Neely
Matthew
M
Bridge
Bob
B
Mechref
Yehia
Y
D002853:Chromatography, Liquid / Q000379:methods*; D004298:Dopamine / Q000031:analogs & derivatives / Q000032:analysis; D004837:Epinephrine / Q000032:analysis; D007546:Isoquinolines / Q000032:analysis; D015394:Molecular Structure; D018377:Neurotransmitter Agents / Q000032:analysis*; D009638:Norepinephrine / Q000032:analysis; D010316:Particle Size; D015203:Reproducibility of Results; D012701:Serotonin / Q000032:analysis; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013997:Time Factors; D005680:gamma-Aminobutyric Acid / Q000032:analysis
D007546:Isoquinolines; D018377:Neurotransmitter Agents; D012701:Serotonin; C022133:3,4-dihydroxybenzylamine; D005680:gamma-Aminobutyric Acid; C036617:salsolinol; D004298:Dopamine; D009638:Norepinephrine; D004837:Epinephrine