Abstract
A rapid, sensitive method for the separation of catecholamine biomarkers (CAs), of importance in traumatic brain injury (TBI) and in Parkinson’s disease (PD), has been successfully developed using hydrophilic interaction liquid chromatography (HILIC). Dopamine (DA), epinephrine (EPI), and norepinephrine (NE) are known to be three to fivefold elevated above normal in traumatic brain injury (TBI) patients. HILIC facilitates the rapid and efficient separation of these polar biomarkers, which can be poorly retained by reversed-phase liquid chromatography (RPLC), while electrochemical detection (ECD) at the boron-doped diamond (BDD) electrode provides enhanced nanomolar detection. Three HILIC columns were compared, namely the superficially porous (core-shell) Z-HILIC column and the Z-cHILIC and Z-HILIC fully porous columns. The core-shell Z-HILIC showed the highest efficiency with a rapid separation within 60 s. The HILIC method utilizing the core-shell Z-HILIC column was initially optimized for the simultaneous analysis of DA, EPI, and NE using UV detection. The advantages of using the BDD electrode over UV detection were explored, and the improved limits of detection (LODs, S/N = 3) measured were 40, 50, and 50 nM for DA, EPI, and NE, respectively. Method validation is reported in terms of the linearity, repeatability, reproducibility, and LODs. Furthermore, the proposed method was successfully applied to the real sample analysis of urinary CAs following phenylboronic acid (PBA) solid phase extraction (SPE) pretreatment.
Highlights
Catecholamines (CAs) are biogenic amines consisting of a catechol group with an amine side chain, which are synthesized from the amino acid tyrosine [1]
The direct electrochemical responses of DA, EPI, and NE at the bare boron-doped diamond (BDD) electrode were recorded in 85% ACN with 10 mM ammonium formate buffer at pH 3
Rapid, and sensitive separation method for dopamine (DA), epinephrine (EPI), and norepinephrine (NE) has been optimized, combining the superior performance of a core-shell Z-hydrophilic interaction liquid chromatography (HILIC) column coupled to nanomolar electrochemical detection (ECD) at a BDD electrode
Summary
Catecholamines (CAs) are biogenic amines consisting of a catechol group with an amine side chain, which are synthesized from the amino acid tyrosine [1]. Among the most critical CAs in biological systems are dopamine (DA), epinephrine (EPI), and norepinephrine (NE) [2] They are released from the adrenal glands and function as neurotransmitters that facilitate intercellular communication within the nervous system [3]. They have an essential biological role for many physiological functions, and their levels can be regarded as key biomarkers for several diseases and neurological disorders such as some forms of cancer, Parkinson’s disease (PD), and Alzheimer’s disease (AD) [2,4,5]. The overproduction of specific CAs in urine and plasma is utilized for biomarking for types of tumors, such as phaeochromocytomas, neuroblastomas, and ganglioneuromas [9,10]
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