Simultaneous Detection of Monoamine and Purine Molecules using HPLC with a Boron-Doped Diamond Electrode
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چکیده
Most biochemical analyses typically examine molecules from the same neurochemical family, like dopamine and serotonin (monoamines), and GABA and glutamate (amino acids). In this present study, two different families, monoamines and purines, are being investigated using high-performance liquid chromatography (HPLC) with a boron-doped diamond (BDD) electrode. There is considerable data demonstrating the neurotransmitters dopamine and adenosine are tightly interconnected. However, little attention has been given to simultaneously measuring both of these neurotransmitters using HPLC methods, with the main reason the large gap between their respective oxidation potentials. Dopamine oxidizes at ~ +600 mV and adenosine at ~ +1500 mV versus a Ag/AgCl reference electrode [1]. Furthermore, the largest hurdle regarding the routine detection of extracellular adenosine levels is the lack of robust analytical methods that accurately measures basal levels. Most studies using microdialysis sampling to detect adenosine with other purines use HPLC coupled with UV or fluorescence detection [2-4]. Within these methods, the basal range for detection of adenosine is between 50 200 nM, which is too wide and reflects the poor sensitivity of the available detection methods. Consequently, it is imperative to develop a method that is specific and selective to adenosine to more accurately determine its extracellular levels. Recently, a commercially available BDD working electrode has been developed to use with HPLC (figure 1). Advantages of using BDD versus a carbon electrode are that BDD electrodes have a higher working potential, less electrode fouling, and decreased noise making them preferred for compounds with higher redox potentials. The objective of this work is to evaluate the commercially available BDD working electrode by analyzing the neurotransmitters adenosine, dopamine and their respective family members.
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تاریخ انتشار 2012