Catecholamine Quantification in Body Fluids Using Isocratic, Reverse Phase HPLC‐CoulArray Multi‐electrode Chemical Detector System: Investigation of Sensitivity, Stability, and Reproducibility

Kumar, Adarsh; Fernandez, Benny; Antoni, Mike; Eisdorfer, Seth; Kumar, Mahendra

doi:10.1081/jlc-120025600

Cited by 5 publications

(5 citation statements)

References 15 publications

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“…A linear relationship between a range of concentration of dopamine (50 -500 pg/mL) and their respective response factors (RF) demonstrate a high degree of precision in the overall performance of the system and its capability for quantification of as low as 2 pg of DA, a 10 fold increase from 20 pg achieved in the earlier studies. [23,28,29] Although, DA could be detected at as low as 0.2 -1.0 pg/ mL (0.008 -0.04 pg in 40 mL injection volume) at the level of sensitivity and optimum potential of the detector system (0 -2.0 nA, 220 mV) used in this study, this limit of 0.2 pg was, however, found to be an outlier in the linear relationship between various concentrations of dopamine standards and the corresponding RF values. In earlier studies using a single electrode HPLC-ECD system, the detection limit for DA was 100 pg and 80 pg, respectively, when the system was set at the highest sensitivity of detection, [18,22] whereas with the CoulArray HPLC-ECD system, we reported the detection of 8.0 pg of DA, [23] which was a 10 -12 fold increase from the studies cited above.…”

Section: Discussionmentioning

confidence: 97%

“…[23] Briefly, each aliquot of aqueous solution containing DA standard was mixed with alumina and shaken for 15 minutes on a mechanical rocker and centrifuged. The supernatant was discarded and alumina was washed twice with 1:100 dilution of 2 M Tris-HCl, PH 8.7.…”

Section: Recovery Of Dopamine Standards From Spiked Aqueous Solution:mentioning

confidence: 99%

“…[17 -19] In order to achieve higher recovery and sensitivity, we have frequently reported modifications in the A. M. Kumar et al 778 methods of extraction, as well as detection, using the single channel and single electrode HPLC-ECD system for measurement of biogenic amines including catecholamines in plasma, [20] urine, [21] whole blood, [22] and CSF. [11] In a recent study, we used the multi-channel and multi-electrode CoulArray HPLC-ECD system and achieved higher sensitivity of detection, greater stability, and reproducibility for catecholamine quantification in human body fluids [23] than was achieved with the earlier methods. [18,19] However, since dopamine and the other neurotransmitter concentrations are variable in different brain regions of animals and humans, and studies have shown deficits of DA associated with aging and neurodegenerative diseases, [7,11,14] there is a persistent ongoing quest for achieving higher sensitivity and recovery in order to quantify picogram, or even lower concentrations of dopamine in low tissue weights from different regions of the human brain.…”

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confidence: 96%

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Ultra Micro Quantification of Dopamine and Homovanillic Acid in Human Brain Tissue: Quest for Higher Recovery and Sensitivity with CoulArray HPLC‐ECD System

Kumar

Fernandez

Gonzalez

et al. 2006

Journal of Liquid Chromatography & Related Technologies

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Dopamine (DA), a biogenic amine neurotransmitter is involved in regulating a number of functions of the central and peripheral nervous systems, including voluntary and involuntary motor functions and cognitive behaviors. Since the concentration of dopamine in different parts of the brain is extremely small and decreases further in neurodegenerative diseases, such as HIV-1 infection, a highly sensitive and specific methodology is required to quantify DA and its metabolite, homovanillic acid (HVA) in small weights of tissues from different regions of the brain. The method presented in this report used a highly sensitive multielectrode CoulArray HPLC-ECD system for quantification of ,4 pg of DA and ,10 pg of HVA in homogenates of human brain tissues.A linear relationship in the calibration curves was observed between a wide range of concentration of standards of DA (50-500 pg/mL) and HVA (0.1-20.0 ng/mL) and the corresponding response factors. Separate aliquots from the same pool of brain tissue homogenate were used to extract DA and HVA by their specific extraction procedure and were quantified separately. Recovery of both DA and HVA from homogenate preparation and aqueous extraction were in the range of 88-98%. Stability of the system is evident from the consistency of retention times of analytes in different experiments. The reliability of the method is shown from the reproducibility of the values of DA and HVA obtained in different aliquots of the brain tissue homogenates.

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Section: Discussionmentioning

confidence: 97%

Section: Recovery Of Dopamine Standards From Spiked Aqueous Solution:mentioning

confidence: 99%

mentioning

confidence: 96%

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Ultra Micro Quantification of Dopamine and Homovanillic Acid in Human Brain Tissue: Quest for Higher Recovery and Sensitivity with CoulArray HPLC‐ECD System

Kumar

Fernandez

Gonzalez

et al. 2006

Journal of Liquid Chromatography & Related Technologies

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“…7 ), and a maximum concentration value of 5906 nmol/L (1165 ) was registered. For DOPAC determination the average value of concentration was around 2992 nmol/L (503 ), with a maximum value of approximately 28471 nmol/L (4787 ).…”

Section: Analysis Of Urine Samples From Patients With Ds By Of Biomentioning

confidence: 99%

Optical Fiber Bioanalyzer Based on Enzymatic Coating Matrix for Catecholamines and Their Metabolites Assessment in Patients With Down Syndrome

et al. 2012

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The urinary levels of catecholamines [adrenaline (AD), noradrenaline (NA), and dopamine (DA)] and their metabolites [L-3,4-dihydroxyphenylalanine (L-DOPA), and 3,4-dihydroxyphenylacetic acid (DOPAC)], as indicators of physiological stress, were assessed in 40 patients with Down syndrome (DS).The analysis was performed by an optical fiber (OF) bioanalyzer; which was constituted by two main components: a miniaturized chromatographic system and a detection system based on an OF coated with an enzymatic matrix. In this study some working conditions such as, number of fibers in the miniaturized chromatographic column, number of dips for sensitive coating deposition, temperature and time of cure of the sensitive matrix, were optimized in order to achieve higher analytical performance. After tested for calibration the bioanalyzer was applied to urine samples analysis of catecholamines and their metabolites, comparing the results with those obtained by both, a classical analytical method, namely high performance liquid chromatography coupled to an electrochemical detector (HPLC-ED), and an OF biosensor based on a sensitive cladding of laccase (LacOF biosensor). The results of catecholamines in patients with DS revealed that 15% and 22.5% of the analyzed subjects showed DA and AD concentrations, respectively, above the pathological levels. In the determinations of NA, the 40 samples showed concentration values below the normal levels, while in the determination of catecholamines metabolites Manuscript 5% of the urine samples showed values above 118 and 1681 g L for L-DOPA and DOPAC, respectively.

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“…Electrochemical detection represents a cost-effective solution in the analysis of electroactive compounds such as various biomarkers, neurotransmitters, , nucleic acids, antioxidants, saccharides, drugs, etc. As ECD probes only species that are electroactive at the applied potential, it often offers higher selectivity than spectrophotometric detection − and much lower running costs compared with MS. , Nevertheless, the determinations of low concentrations of analytes that require the use of high detection potentials are particularly challenging.…”

Section: Introductionmentioning

confidence: 99%

Electrochemical HPLC Determination of Piperazine Antihistamine Drugs Employing a Spark-Generated Nickel Oxide Nanoparticle-Modified Carbon Fiber Microelectrode

Belbasi,

Petr,

Sevcik

et al. 2024

ACS Omega

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In this work, we demonstrate a sensitive highperformance liquid chromatography (HPLC) method for the determination of piperazine antihistamine drugs employing innovative electrochemical detection based on a spark-generated nickel oxide nanoparticle-modified carbon fiber microelectrode built into a miniaturized electrochemical detector. The direct carbon fiber-tonickel plate electrode spark discharge, carried at 0.8 kV DC, with the nickel electrode connected to the negative pole of the high-voltage power supply, provides extremely fast (1 s) in situ tailoring of the carbon fiber microelectrode surface by nickel oxide nanoparticles. It has been found that nickel oxide nanoparticles exhibit an electrocatalytic effect toward the piperazine moiety electrooxidation process, as confirmed by voltammetric experiments, revealing the shift in the peak potential from 1.25 to 1.09 V versus Ag/AgCl. Cetirizine, cyclizine, chlorcyclizine, flunarizine, meclizine, and buclizine were selected as sample piperazine antihistamine drugs, while diclofenac served as an internal standard. The isocratic reversed-phase separation of the above set was achieved within 15 min using an ARION-CN 3 μm column with a binary mobile phase consisting of 50 mM phosphate buffer (pH 3) and methanol (45/55, v/v). The limits of detection (LOD) were within the range of 3.8−120 nM (for cyclizine and buclizine) at E = +1500 mV (vs Ag/AgCl), while the response was linear within the concentration range measured up to 5 μmol L −1 . The method was successfully applied to the determination of piperazine antihistamine drugs in spiked plasma samples.

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Catecholamine Quantification in Body Fluids Using Isocratic, Reverse Phase HPLC‐CoulArray Multi‐electrode Chemical Detector System: Investigation of Sensitivity, Stability, and Reproducibility

Cited by 5 publications

References 15 publications

Ultra Micro Quantification of Dopamine and Homovanillic Acid in Human Brain Tissue: Quest for Higher Recovery and Sensitivity with CoulArray HPLC‐ECD System

Ultra Micro Quantification of Dopamine and Homovanillic Acid in Human Brain Tissue: Quest for Higher Recovery and Sensitivity with CoulArray HPLC‐ECD System

Optical Fiber Bioanalyzer Based on Enzymatic Coating Matrix for Catecholamines and Their Metabolites Assessment in Patients With Down Syndrome

Electrochemical HPLC Determination of Piperazine Antihistamine Drugs Employing a Spark-Generated Nickel Oxide Nanoparticle-Modified Carbon Fiber Microelectrode

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