HPLC Method for Analysis of Neurotransmitters on Amaze HD Column

Application description

Four neurotransmitters - pseudoephedrine, norephedrine, phenylephrine and norphenylephrine - were separated in hydrogen-bonding and cation-exchange mode. Pseudoephedrine is sympathomimetic drug used either as a single ingredient or part of the composition of OTC or Rx medications. Phenylephrine is a hydrophilic basic drug which is used as decongestant in OTC medication composition, such as Sudafed, Benadryl, Advil, Nyquil, etc. Norphenylephrine is an adrenergic agent used as sympathomimetic drug. Norephedrine is naturally occurring endogenous trace amine that plays a role as neurotransmitter in the brain, and is used as decongestant in many OTC compositions as well as an appetite suppressant. All four drugs are hydrophilic and basic in nature, and have no retention on traditional reversed-phase column without an ion-pairing reagent. Method provides an efficient and robust separation for all drugs. The method can be used in analysis of neurotransmitters in biofluids and other sample matrices. The method is compatible with all detection techniques: MS, RI, UV, ELSD and CAD.

Conditions of Experiment
Column: Amaze HD
Separation Modes: hydrogen-bonding and cation-exchange
Column Dimenstions: 3.2 x 50 mm, 3 um
Mobile Phase: MeCN/MeOH (95/5) with 0.5% HCOOH and 0.05% AmFm
Detection: UV 275 nm
Sample: 0.3 mg/ml
Injection: 1 uL
Flow rate: 1.0 mL/min
Class of compounds: Amino acid, Aromatic base, Catecholamine, Drug, Neurotransmitter, Stimulant, Supplement
Nature of compounds: Basic, Hydrophilic, Polar, Zwitterionic
Compounds: Pseudoephedrine, Norepinephrine, Phenylalanine, Norphenylephrine