The Use Of HPLC-EC: Hydrodynamic Voltammograms (Neuropharmacology)

   High-performance liquid chromatography (HPLC) with electrochemical detection: is an analytical technique utilized to separate, quantify, and identify the contents of a mixture. For instance, chromatography is the separation of a mixture by passing it through a medium, where components flow at differing rates. In other words, a fluid solvent known as the mobile phase is continuously pumped throughout the system. For example, the mobile phase carries the differing components of the mixture/sample and passes through a column, known as the stationary phase. Moreover, sample separation is dependent on the various chemical dynamics of the mobile phase, stationary phase, and the sample itself. Additionally, adjustments and modifications can be made to both mobile and stationary phases to yield diverse responses. For example, a specific sample can potentially have a stronger interaction with the stationary phase. Whereas, a weaker interaction with the mobile phase. As a result, any modifications to the column or mobile phase can display optimal or incomplete separation of the sample. Moreover, once pumps force the mixed solvents through the column, the sample emerging from the column passes into the detector. As a result, the detector will produce a peak on the chromatogram. Additionally, the detector utilized in the laboratory process is an electrochemical detector. As a result, a redox reaction is executed at the site of the detector where the samples will lose or gain electrons as they pass between electrodes that differ in electrical potential.


    Furthermore, hydrodynamic voltammetry is a form of voltammetry that is used to discover the most efficient guidelines for the identification of analytes (a substance that is being measured for various chemical components) within the running sample. This will serve as the basis of the chromatogram. Thus, recognizing the optimal potential that should be utilized to operate the HPLC. In other words, the objective is to discover the potential which would be most effective in measuring a sample containing various neurotransmitters.

    It is believed, that the greater the potential the more optimal it will be in yielding a response. For example, in most laboratories the highest potential is 500 mV, there is significant reason to presume that 500 mV: should yield the best response in the chromatogram. Although, it is important to consider that not all of the neurotransmitters may respond to each potential in a similar manner. However, on average, the weakest responses are to be yielded between the range: 50 – 250 mV. Whereas, the greater responses were initiated and yielded through the range 300 – 350 mV.

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