Simultaneous single isotope radioenzymatic assay of plasma norepinephrine, epinephrine and dopamine

doi:10.1016/0024-3205(77)90070-4

Life Sciences

Volume 21, Issue 5, 1 September 1977, Pages 625-636

https://doi.org/10.1016/0024-3205(77)90070-4 Get rights and content

Abstract

Modification of the original single isotope radioenzymatic assay of Passon and Peuler (1) permits the direct and simultaneous analysis of norepinephrine, epinephrine and dopamine in plasma samples of 50 μl or less. Plasma or cerebrospinal fluid without prior extraction of catecholamines or deproteinization is added directly into a mixture of 100 μl. This catechol-O-methyl-transferase-catalyzed assay is sensitive to 1 pg (20 pg/ml of plasma) for norepinephrine and epinephrine and 6 pg (120 pg/ml) for dopamine. A rapid thin layer chromatographic separation of the three ³H-methylcatecholamines contributes to the excellent specificity of the differential assay of the three catecholamines. The differential analysis of 15–20 plasma samples can be completed easily within one day. A total assay which omits the chromatographic step and, thus, measures norepinephrine plus epinephrine at the same sensitivity can be completed in 20 samples in one-half a working day.

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Quantitative detection of dopamine in human serum with surface-enhanced Raman scattering (SERS) of constrained vibrational mode
2023, Talanta
Dopamine (DA) is a crucial neurotransmitter involved in the hormonal, nervous, and vascular systems being considered as an index to diagnose neurodegenerative diseases, including Parkinson's and Alzheimer's disease. Herein, we demonstrate the quantitative sensing of DA using the peak shift in surface-enhanced Raman scattering (SERS) of 4-mercaptophenylboronic acid (4-MPBA), resulting from the concentration of DA. To enable the signal enhancement of Raman scattering, Ag nanostructure was built with one-step gas-flow sputtering. 4-MPBA was then introduced using vapor-based deposition, acting as a reporter molecule for bonding with DA. The gradual peak-shift from 1075.6 cm⁻¹ to 1084.7 cm⁻¹ was observed with the increasing concentration of DA from 1 pM to 100nM. The numerical simulation revealed that DA bonding induced a constrained vibrational mode corresponding to 1084.7 cm⁻¹ instead of a C–S-coupled C-ring in-plane bending mode of 4-MPBA corresponding to 1075.6 cm⁻¹. Proposed SERS sensors depicted reliable DA detection in human serum and good selectivity against other analytes, including glucose, creatinine, and uric acid.
Multiple catechols in human plasma after drinking caffeinated or decaffeinated coffee
2021, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Citation Excerpt :
Cys-DA and Cys-DOPA were provided by the National Institute of Mental Health. Combining COMT with tracer-labelled SAMe was the basis for radioenzymatic assays that preceded LCED for measuring levels of catecholamines in biological fluids [18,19]. With this background we designed the COMT + SAMe experiments reported here.
Coffee is one of the most frequently consumed beverages worldwide. Research on effects of coffee drinking has focused on caffeine; however, coffee contains myriad biochemicals that are chemically unrelated to caffeine, including 3,4-dihydroxyphenyl compounds (catechols) such as caffeic acid and dihydrocaffeic acid (DHCA).
This prospective within-subjects study examined effects of drinking caffeinated or decaffeinated coffee on plasma free (unconjugated) catechols measured by liquid chromatography with series electrochemical detection (LCED) after batch alumina extraction. To confirm coffee-related chromatographic peaks represented catechols, plasma was incubated with catechol-O-methyltransferase and S-adenosylmethionine before the alumina extraction; reductions in peak heights would identify catechols.
Ten healthy volunteers drank 2 cups each of caffeinated and decaffeinated coffee on separate days after fasting overnight. With subjects supine, blood was drawn through an intravenous catheter up to 240 min after coffee ingestion and the plasma assayed by alumina extraction followed by LCED.
Within 15 min of drinking coffee of either type, >20 additional peaks were noted in chromatographs from the alumina eluates. Most of the coffee-related peaks corresponded to free catechols. Plasma levels of the catecholamines epinephrine and dopamine increased with both caffeinated and decaffeinated coffee. Levels of other endogenous catechols were unaffected. Plasma DHCA increased bi-phasically, in contrast with other coffee-related free catechols.
Interpretation.
Drinking coffee—whether caffeinated or decaffeinated—results in the rapid appearance of numerous free catechols in the plasma. These might affect the disposition of circulating catecholamines. The bi-phasic increase in plasma DHCA is consistent with production by gut bacteria.
Paper-microfluidics based SERS substrate for PPB level detection of catechol
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Citation Excerpt :
In another report, the levels of polyphenols are an indicator for the quality of tea [15]. Dopamine, a form of di-phenol, also appears in the neurological system, whose levels indicate the neurological status of a person [16,17]. Thus, the detection of phenolic compounds has a wide range of applications.
The application of microfluidics for largescale rapid analytics holds great promise in the pharmaceutical diagnostics and analytical chemistry. Here we report a paper based microfluidic substrate designed by the impregnation of Silver nanoparticles. This study demonstrates the achievement of a thousand-fold increase in the successful detection level up to 10 ppb (90.8 nM) by employing Surface Enhanced Raman Spectroscopy (SERS) for the detection of Catechol. The presented sensor exhibits the following main features: (i) high specificity of enzyme (Hemocyanin)-based sensing, (ii) effective SERS sensitivity, (iii) easiness and cost-effectiveness of a paper-based platform. We rationalize these findings using the ab initio DFT simulations using Gaussian 09 whose theoretical calculations reflect the observed experimental Raman peaks.
Neuromodulatory effects of ethyl acetate fraction of Zingiber officinale Roscoe extract in rats with lead-induced oxidative stress
2019, Journal of Integrative Medicine
This study investigated the ameliorative potential of Zingiber officinale Roscoe extract against lead-induced brain damage in rats.
Thirty male rats were divided into 5 groups of 6 rats each. Lead-acetate toxicity was induced by intraperitoneal injection (10 mg/kg body weight (b.w.)) in Groups B–E. Group A (control) and Group B (lead-acetate) were left untreated; vitamin C (200 mg/kg b.w.) was administered to Group C; ethyl acetate fraction from Z. officinale extract (200 and 100 mg/kg b.w.) was administered to Group D and E by oral gavage once daily for 7 days. Changes in the content of some key marker enzymes such as acetylcholinesterase (AChE), butyrylcholinesterase (BChE), monoamine oxidase (MAO), epinephrine, dopamine, Na⁺/K⁺-ATPase, catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) as well as malonaldehyde (MDA) levels were determined in serum.
Exposure to lead acetate resulted in a significant decrease (P < 0.05) in the activities of BChE, AChE, Na⁺/K⁺-ATPase, SOD, CAT and GPx with a corresponding increase in the levels of MDA, xanthine oxidase, epinephrine, dopamine and MAO relative to the control group. Levels of all disrupted parameters were alleviated by co-administration of Z. officinale fraction and by the standard drug, vitamin C.
These results suggest that ethyl acetate fraction of Z. officinale extract attenuates lead-induced brain damage and might have therapeutic potential as a supplement that can be applied in lead poisoning.
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Social stress produces behavioral alterations, and autonomic and cardiac dysfunction in animals. In addition to the well-known roles of oxytocin on birth and maternal bonding, recent evidence shows that this neuropeptide possesses cardio-protective properties. However less is known about its role in the regulation of the autonomic nervous system. The direct influence of oxytocin on the cardiac catecholamine synthesizing enzyme, transport beta-adrenoceptors and muscarinic receptors in animals exposed to chronic social isolation stress has not yet been studied. In this study, we examined the influence of peripheral chronic oxytocin treatment on anxiety-related behavior, the morphology and content of epinephrine and norepinephrine, mRNA and protein levels of tyrosine hydroxylase (TH), norepinephrine transporter (NET) and receptors <beta> 3 (β₃-AR) and muscarinic 2 (M₂ MR) in the right and left cardiac atrium and ventricle of chronically socially isolated male rats. Our results show that oxytocin treatment exhibits an anxiolytic effect, decreases the heart/body weight ratio and prevents the hypertrophy of cardiomyocytes in the wall of the left ventricle of stressed rats. Epinephrine and TH protein levels were unchanged after prolonged oxytocin treatment. Peripheral oxytocin administration led to the enhancement of gene expression of β₃-AR in both atria, NET protein in the left ventricle and gene expression of M₂ MR in the right atrium and the left ventricle of chronically socially isolated rats. The study provides evidence that oxytocin treatment in chronically socially isolated animals enhances norepinephrine uptake and expression of cardio-inhibitory receptors in cardiac tissues, which could have a beneficial effect on the cardiovascular system under the increased activity of the sympathoneural system.
Concentration gradient of noradrenaline from the periphery to the centre of the cornea - A clue to its origin
2018, Experimental Eye Research
Citation Excerpt :
Levels of DOPA, DA and DOPAC were detected in both iris and aqueous samples (Table 1 and Fig. 3b). Mean human venous plasma concentration of catecholamines measured in supine resting adults, as reported elsewhere in the scientific literature, were considered for the sake of comparison (Jason Eliot et al., 1980; Peuler and Johnson, 1977). Statistically significant were the differences between: 1) DA versus NA and AD concentrations in the central (P < .0001) and intermediate (P < .0001) corneal segments; 2) NA versus AD (P < .0001) and DA (P = .0003) concentrations in the peripheral corneal segments; 3) NA, AD and NA concentrations in the iris (P < .0001); 4) AD versus NA (P = .0039) and DA (P < .0001) concentrations in the aqueous humour; 5) NA concentrations in peripheral corneal segments versus those in either central (P < .0001) or intermediate corneal segments (P = .0002); 6) DA concentrations in peripheral corneal segments versus those in either central (P < .0001) or intermediate corneal segments (P < .0001).
We set out to demonstrate that the major source of corneal catecholamines is its neuronal release from intrinsic sympathetic nerves rather than circulating or non-neuronal local production. Three concentric segments (central, intermediate, peripheral) were obtained by double trephination (9.5–7.25 mm) performed on corneas harvested from 3 to 4 month old rabbits and human corneas rejected for transplantation, along with aqueous humour, full iris tissue and blood samples. Endogenous catecholamines were quantified by high pressure liquid chromatography with electrochemical detection (HPLC-ED), and comparison with the uptake of radio-labelled noradrenaline (3H-NA) before and after incubation with cocaine was performed. Results are means ± SEM. Ratios between enzymatic end products and their substrates were calculated. ANOVA was used for statistical analysis. Catecholamine levels were found to be about one log unit lower in the human cornea than in the rabbit cornea. In the rabbit, dopamine (DA), noradrenaline (NA) and adrenaline (AD) were identified by HPLC-ED in all corneal segments, whilst in the human cornea NA was identified only in the intermediate and peripheral corneal segments, and no AD was found. In the iris and aqueous humour only DA and NA were present. A concentration gradient for NA decreasing from the periphery to the centre of the cornea was identified in both species (NA/DA ratio higher than 1 in the periphery; low AD/NA ratio in all corneal segments), but not for DA or AD. After incubation with 3H-NA all corneal segments and iris tissue showed loading with the aforementioned gradient being reproduced, and a decrease in 3H-NA loading after cocaine was significant only in the peripheral corneal segment and in the iris of both species. Reduction in 3H-NA loading after incubation with cocaine shows that NA in the cornea is mostly of neuronal origin and demonstrates the presence of functional sympathetic nerves (also expectedly found in the iris); the existence of a gradient both for 3H-NA loading and loading reduction after cocaine points to a higher density of fibres in the peripheral cornea.

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^∗: Present address: Department of Pharmacology, Milton S. Hershey Medical Center, Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033.

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Simultaneous single isotope radioenzymatic assay of plasma norepinephrine, epinephrine and dopamine

Abstract

Analyt. Biochem.

Life Sci.

Life Sci.

J. Biol. Chem.

J. Neural. Trans.

Diabetes

J. Neurochem.

J. Neurochem.

Endocrin.

Circ. Res.

Am. J. Med. Sci.