epinephrine-sulfate and noradrenaline-sulfate

This study investigated the effect of cocaine abuse on peripheral catecholamines. Specifically, we measured the concentration of free dopamine, dopamine sulfate, free norepinephrine, norepinephrine sulfate, free epinephrine and epinephrine sulfate in plasma samples obtained from the blood of a group of patients with cocaine addiction (N = 15). The concentrations of free and sulfoconjugated catecholamines in plasma were measured by a radioenzymatic technique. The results of this study revealed significant (P < 0.0001) elevation in plasma dopamine sulfate (8926 +/- 1204 pg/mL) of cocaine addicts upon admission to an in-patient treatment facility when compared with the level of this dopamine metabolite in plasma of control subjects (2356 +/- 121 pg/mL). Furthermore, there was a significant (P < 0.0001) relationship between elevation in plasma dopamine sulfate levels and severity of cocaine use among these patients, and in the majority of cases the plasma levels of dopamine sulfate declined appreciably in time with abstinence from cocaine. In contrast, no appreciable difference was observed in the concentrations of either free or sulfate-conjugated norepinephrine and epinephrine in plasma of cocaine addicts as compared with controls. Differences in plasma dopamine sulfate among these patients versus controls may be interpreted as a reflection of activation of extracellular dopamine metabolism associated with chronic cocaine exposure in humans.

Topics: Adult; Cocaine; Dopamine; Epinephrine; Female; Humans; Male; Norepinephrine; Substance-Related Disorders

A method is described to measure catecholamine sulfates from human plasma and urine by isocratic reversed-phase high-performance liquid chromatography with electrochemical detection. For this measurement we use catecholamine 3-sulfate isomers as internal standards and determine the sulfoconjugates only after eliminating the catecholamines. Catecholamines that have previously been used as internal standards are shown to cause a significant overestimation (P less than 0.05) of the catecholamine sulfates--by 10% to 25% and 20% to 42% in human plasma and urine, respectively. The detection limits (signal-to-noise ratio greater than 3) in plasma and urine samples were about 80 pmol/L for each analyte. The intra-assay and interassay CVs were less than 4.0% and 10.6% in human plasma and less than 6.6% and 12.8% in human urine, respectively. The calibration curves for all catecholamine sulfates in human plasma and urine were linear (r greater than 0.96; P less than 0.001) over the respective concentration ranges of 0.1-100 nmol/L and 5-1000 nmol/L.

Topics: Adrenal Gland Neoplasms; Chromatography, High Pressure Liquid; Dopamine; Epinephrine; Humans; Norepinephrine; Pheochromocytoma; Reference Standards

Plasma, platelet and erythrocyte contents of free and conjugated norepinephrine, epinephrine and dopamine were determined by radioenzymatic assay in 12 resting healthy volunteers. Mean platelet/plasma concentration ratios were 533 for free norepinephrine, 502 for free epinephrine and 149 for free dopamine. Corresponding erythrocyte/plasma ratios were 1.04, 1.13 and 4.5, respectively. The presence of conjugated catecholamines in platelets and erythrocytes could be confirmed; however, their relative proportion within these cells, particularly in platelets, was lower than that in plasma. Upon intravenous infusion of dopamine for 3 hr at 5 micrograms kg-1 min-1, concentrations of free dopamine in plasma increased rapidly (280-970-fold), whereas conjugated dopamine only reached maximal values (14-19-fold increase) at 30 to 60 min after cessation of the infusion. The relative distribution of unconjugated dopamine in whole blood between plasma, platelets and erythrocytes changed from mean values of 1:0.33:3.7 at rest to 1:1.1:0.5 at the end of the infusion. As a result of the subsequent rapid decrease of dopamine in plasma and erythrocytes, this distribution was 1:17:1 shortly thereafter and remained constant up to the end of the investigation period. The relative distribution for conjugated dopamine of 1:0.001:0.5 at rest changed to about 1:0.2:0.1 at the termination of the infusion. Oral administration of norepinephrine and dopamine led to increases in the plasma concentrations of these amines in their conjugated forms only, whereas epinephrine concentrations remained constant. These elevations were not accompanied by corresponding increases in platelet and erythrocyte norepinephrine, epinephrine and dopamine contents.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Administration, Oral; Adult; Blood Platelets; Catecholamines; Dopamine; Epinephrine; Erythrocytes; Female; Humans; Injections, Intravenous; Male; Norepinephrine

The origins of conjugated catecholamines remain poorly known. The aim of the present study was to see whether a major contribution comes from the sympathetic nervous system. We have assumed some kind of parallelism between the activity of the sympathetic nervous system, the amount of catecholamines released and taken up, and the amount of conjugated catecholamines circulating in plasma. Accordingly, an increase in sympathetic activity should be followed by an increase in the plasma level of conjugated catecholamines. The plasma levels of sulfoconjugated and glucuroconjugated catecholamines were measured in 10 patients with mental disease resistant to drug treatment, before and after electroconvulsive therapy. As expected, blood pressure, norepinephrine concentration, and epinephrine concentration in plasma were transiently increased. Neither sulfoconjugated nor glucuroconjugated catecholamines were significantly changed. Conjugated catecholamines were measured in 10 volunteers before and at the nadir of insulin-induced hypoglycemia. As expected, plasma levels of norepinephrine and epinephrine were drastically increased. Plasma levels of sulfoconjugates were decreased and glucuroconjugates increased; these were narrow but statistically significant variations. Data reported in the present article do not support a major role for the activity of the sympathetic system in fixing the level of conjugated catecholamines in human plasma. This is a negative, but nonetheless important, observation. In human subjects, currently available information suggests an important role for the intestinal wall and renal function in determining the level of circulating sulfoconjugates.

Topics: Blood Glucose; Blood Pressure; Catecholamines; Electroconvulsive Therapy; Epinephrine; Glucuronates; Humans; Hypoglycemia; Insulin; Mental Disorders; Norepinephrine; Sulfates; Sympathetic Nervous System

Both isomers of epinephrine sulfate were synthesized, unequivocally identified by 1H-NMR and highly purified from catecholamines (less than 90 ppm). Bacterial as well as pig liver arylsulfatase A and B demonstrated a higher substrate turnover of epinephrine-4-sulfate, norepinephrine-4-sulfate and dopamine-4-sulfate as compared to the 3-sulfate isomers. The arylsulfatase B however, is less important for the deconjugation of these sulfoconjugates than arylsulfatase A. Since arylsulfatase A occurs in most human tissues, it might be of physiological significance in the deconjugation of the catecholamine sulfate isomers. Furthermore the kinetic data at pH 7.4 and 6.9 suggest the increased cleavage of the sulfate group, e.g. during exercise-induced acidosis. In contrast to results reported in the literature, dopamine sulfates were no substrates of dopamine beta-hydroxylase.

Topics: Animals; Arylsulfatases; Dopamine; Dopamine beta-Hydroxylase; Epinephrine; Hydrogen-Ion Concentration; Isomerism; Kinetics; Liver; Norepinephrine; Sulfatases; Swine

Previous research established a relationship between circulating sulfoconjugated norepinephrine (NE-SO4) and oxygen consumption at various exercise intensities. In this study, the stability of the NE-SO4 response was examined during sustained exercise at a constant relative intensity. Seven trained men bicycled at 78 +/- 3% of their maximal O2 consumption for 28 min and then rested on the ergometer for a comparable duration. After a 30-min rest, plasma samples were collected through an indwelling catheter at 7-min intervals during the exercise and recovery periods. Free NE and epinephrine increased sixfold during exercise. These changes were accompanied by increases in sulfoconjugated catecholamines, but only NE-SO4 achieved statistical significance (rest, 712 +/- 602; exercise, 1,329 +/- 1,163 pg/ml). This occurred at three collection periods (14, 21, and 28 min). Approximately 35, 52, and 95% of NE, epinephrine, and dopamine, respectively, existed as sulfoconjugated during exercise. Subject variation was present in the sulfoconjugated catecholamine response that could not be attributed to corresponding differences in circulating free catecholamine release. These findings implicate blood flow as a factor in the sulfoconjugation of NE, but not epinephrine or dopamine.

Topics: Adult; Catecholamines; Dopamine; Epinephrine; Exercise; Humans; Male; Norepinephrine; Oxygen Consumption

Plasma concentrations of free and sulfoconjugated catecholamines were measured in healthy infants and children under resting conditions. Free norepinephrine and epinephrine levels were up to three times higher in healthy children under 2 years than in adults, even under true resting conditions. In contrast, free dopamine concentrations of all age groups fell within the normal range for adults. The levels of sulfoconjugation were in the adult range.

Topics: Adolescent; Child; Child Development; Child, Preschool; Dopamine; Epinephrine; Humans; Infant; Infant, Newborn; Norepinephrine; Reference Values

The physiologic significance of the racemic 3-O-sulfate esters of epinephrine (EPI-3-O-S) and norepinephrine (NE-3-O-S) as well as 4-O-sulfoconjugated dopamine (DA-4-O-S) was evaluated. For this purpose these conjugated catecholamines (CA) were synthesized and investigated with respect to their alpha 2- and beta 2-adrenoceptor affinities and their biological activity in three different human cell systems: in mononuclear leukocytes (MNL), platelets, and fat cells. The unequivocal identification and the minimal degree of contamination of the synthesized sulfoconjugates with free CA was proved by 1H-NMR and by high-performance liquid chromatography with amperometric detection (HPLCA) respectively. In isolated human MNL, beta-adrenoceptor affinities of these conjugated CA were determined in competition experiments with the lipophilic nonspecific radioligand (-) 125I-cyanopindolol (ICYP) and, in addition, with the hydrophilic ligand 3H-CGP12177. With both ligands the affinity constants (KD) of the sulfoconjugated CA under investigation were about 100- to 1000-fold higher when compared with the respective free amines. Moreover, these sulfoconjugated CA per se induced no intracellular production of cyclic adenosine monophosphate (cAMP) in MNL. In comparison with the free amines, metanephrine (MN) and normetanephrine (NMN) showed a highly reduced competitive potency on the MNL beta-adrenoceptors labelled with 3H-CGP or ICYP. The KD values for MN and NMN in competition studies with ICYP were 10- and 5-fold higher than in those with 3H-CGP respectively, indicating a restricted access of MN and NMN to intracellular receptors. The adenylate cyclase system was not stimulated at all by MN or by NMN. In human platelets EPI-3-O-S and NE-3-O-S neither competed with the specific alpha 2-adrenoceptor antagonist 3H-yohimbine nor elicited any aggregation response at all. MN and NMN exhibited an about 40-fold reduced affinity for alpha 2-adrenoceptors in platelets when compared with the respective free amines and elicited no aggregation response at all. However, in the presence of MN and NMN the EPI- and NE-induced platelet aggregation was dose-dependently attenuated. These findings reveal an alpha 2-adrenoceptor antagonistic potency of MN and NMN. In human adipocytes EPI-3-O-S and NE-3-O-S were 100- to 1000-fold less potent to inhibit lipid mobilization via alpha 2-adrenoceptors as well as to stimulate the beta-adrenoceptor mediated lipolysis when compared with free CA.

Topics: Adipose Tissue; Adult; Binding, Competitive; Blood Platelets; Catecholamines; Cyclic AMP; Dopamine; Epinephrine; Humans; Leukocytes, Mononuclear; Lipolysis; Norepinephrine; Platelet Aggregation; Receptors, Adrenergic, alpha; Receptors, Adrenergic, beta

The determination of free and sulfoconjugated catecholamines (CA) in serum and urine with amperometric detection after HPLC separation is described. The reliability of this method has been extensively investigated. Since in men 69-90% of the total CA are sulfoconjugated, an enzymatic hydrolysis of these conjugates with arylsulfatase VI has been elaborated and optimized for a routine assay. The reproducibility with coefficients of variation between 1% and 3% for free and 5% and 10% for conjugated CA and recovery rates of 65%-75% for both were found. The calibration plots were linear between 10 and 5000 ng/l and the smallest detectable amount of CA was 0.1 nmol/l. The sample amount of 0.75 to 1.0 ml for free and 0.2 ml for conjugated CA was lower than with the extraction method (18) which needs 3 ml of serum. Using an automatic sampler, the sampling rate was 50-60 p.d. With the Al2O3 adsorption and the same buffer eluent system, L-dopa can also be detected at the same voltage of 700 mV. The values obtained for the HPLCA described method correlated well with those of the radio enzyme assays according to Da Prada et al. The HPLCA detection of free and sulfoconjugated CA in urine was carried out after an ion exchange column procedure on Biorex 70. The validity of the urine assay was at least as reliable as the determination of free and conjugated CA in plasma.

Topics: Catecholamines; Chromatography, High Pressure Liquid; Dopamine; Epinephrine; Humans; Male; Norepinephrine; Reference Values; Reproducibility of Results

With respect to the growing interest in sulfoconjugated catecholamines (CAS), reliable syntheses of those substances including high purification and unequivocal identification are required. For the syntheses of the 3-O-sulfates of norepinephrine (NE) and epinephrine (EPI), modifications of the methods of Stolz (12) and Arakawa et al. (1) were performed. Noradrenalone and adrenalone were prepared according to the method of Stolz (12) and sulfated by reaction with pyridine-sulfurtrioxide complex in dry pyridine at 60 degrees C. After reduction of these ketosulfates by sodium borohydride in dry pyridine, NE-3-O-S and EPI-3-O-S were obtained respectively. We synthesized dopamine-4-O-sulfate (DA-4-O-S) by reaction of DA hydrochloride with pyridine-sulfurtrioxide complex in dry dimethylformamide at 20 degrees C (Harbeson et al., 1983). The highly purified products (DA-4-O-S, NE-3-O-S, EPI-3-O-S) were characterized by their melting points (mp), infrared spectra (IR), thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), elemental analysis, and 1H-nuclear magnetic resonance spectroscopy (1H-NMR).

Topics: Dopamine; Epinephrine; Magnetic Resonance Spectroscopy; Norepinephrine; Structure-Activity Relationship

This study was designed to re-examine the circadian profiles of dopamine, noradrenaline and adrenaline in the plasma and to investigate the influence of daytime noise stress on these profiles. Twelve subjects participated in two experimental series of 60 h; during one they were exposed to 85 dB(A) industrial noise from 9:00 to 21:00 h. Blood samples were taken for a period of 24 h at 20 min intervals and the plasma levels of the free and the sulfoconjugated catecholaminergic compounds were simultaneously measured. A significant (p less than 0.001) circadian rhythm was found for free noradrenaline, free adrenaline and conjugated dopamine. Noise stress significantly increased (p less than 0.001) free adrenaline levels during the first 6 h of exposure and lowered (p less than 0.001) free noradrenaline values during the whole time of exposure; dopamine did not respond. It was concluded that strong noise load has effects on the sympatho-adrenomedullary system but the actual release pattern of each catecholaminergic metabolite is controlled by separate determinants.

Topics: Adult; Dopamine; Epinephrine; Humans; Male; Noise; Norepinephrine; Sleep; Stress, Physiological

Plasma free catecholamines rise during exercise, but sulfoconjugated catecholamines reportedly fall. This study examined the relationship between exercise intensity and circulating levels of sulfoconjugated norepinephrine, epinephrine, and dopamine. Seven exercise-trained men biked at approximately 30, 60, and 90% of their individual maximal oxygen consumption (VO2max) for 8 min. The 90% VO2max period resulted in significantly increased plasma free norepinephrine (rest, 219 +/- 85; exercise, 2,738 +/- 1,149 pg/ml; P less than or equal to 0.01) and epinephrine (rest, 49 +/- 49; exercise, 555 +/- 516 pg/ml; P less than or equal to 0.05). These changes were accompanied by consistent increases in sulfoconjugated norepinephrine at both the 60% (rest, 852 +/- 292; exercise, 1,431 +/- 639; P less than or equal to 0.05) and 90% (rest, 859 +/- 311; exercise, 2,223 +/- 1,015; P less than or equal to 0.05) VO2max periods. Plasma sulfoconjugated epinephrine and dopamine displayed erratic changes at the three exercise intensities. These findings suggest that sulfoconjugated norepinephrine rises during high-intensity exercise.

Topics: Adult; Catecholamines; Dopamine; Epinephrine; Humans; Male; Norepinephrine; Oxygen Consumption; Physical Exertion

Simultaneous plasma and cerebrospinal free and sulfoconjugated norepinephrine, epinephrine, dopamine and homovanillic acid determinations in 38 patients with various neurological disorders have shown consistently lower concentrations of dopamine sulfate, norepinephrine sulfate, and epinephrine sulfate in the cerebrospinal fluid than in plasma. An approximately three fold increase of plasma dopamine sulfate concentrations following banana ingestion did not result in increased dopamine sulfate concentrations in cerebrospinal fluid. There were positive correlations between plasma and cerebrospinal fluid concentrations of free norepinephrine and epinephrine as well as norepinephrine sulfate, dopamine sulfate and free homovanillic acid. Cerebrospinal fluid and plasma are apparently separated by the blood-brain barrier impermeable to catecholamine sulfates from the blood. Some other data and the observed positive correlation between cerebrospinal fluid and plasma dopamine and norepinephrine sulfates suggests however that catecholamine sulfates may pass from cerebrospinal fluid to the blood and reflect events in the brain.

Topics: Adolescent; Adult; Aged; Catecholamines; Dopamine; Epinephrine; Female; Homovanillic Acid; Humans; Male; Middle Aged; Norepinephrine

Normotensive recumbent subjects exhibit an early (11 p.m.) nocturnal increase in plasma dopamine, norepinephrine and epinephrine sulfates. In individual patients, this peak value is followed by a smaller nocturnal peak of catecholamine sulfates, while free catecholamine levels change in a direction opposite to catecholamine sulfates. This reciprocity of changes cannot however be demonstrated in the whole group. The origin of the nocturnal peaks of catecholamine sulfates is unknown. It may be due to a nocturnal decrease of the renal clearance of catecholamine sulfates, an increased generation of sulfates for reasons other than an increase in the free catecholamine substrate, or possibly a release of catecholamine sulfates from the brain.

Topics: Adult; Catecholamines; Circadian Rhythm; Dopamine; Epinephrine; Female; Humans; Male; Middle Aged; Norepinephrine; Sleep

Using a radioenzymatic technique, the highest concentrations of free catecholamines were found in the duodenum, and the lowest in the liver of untreated rats. When compared to the antrum, the concentration of free dopamine was higher, and that of norepinephrine lower in the fundus. As far as conjugated catecholamines are concerned, the tissue concentrations of both sulfo- and glucurono-conjugates were usually low, and often non detectable, with an exception: the concentration of glucurono-conjugated dopamine was very high in the duodenum, ileum, and liver of untreated rats.

Topics: Animals; Dopamine; Duodenum; Epinephrine; Glucuronidase; Ileum; Intestine, Small; Liver; Norepinephrine; Rats; Stomach; Sulfatases; Tissue Distribution

The study examined the plasma concentration of free and sulfate-conjugated norepinephrine (NE) and epinephrine (E) at rest and after vigorous bicycle exercise. Free and total catecholamines were measured by a modified radioenzymatic assay using external standards. This assay was less costly and, in subjects with normal renal function, plasma levels were highly comparable to those obtained using internal standards. Exercise was associated with a predictable rise in plasma free NE and E concentration, which correlated with the hemodynamic changes, and a significant decrease in the levels of catecholamine sulfate conjugates. There was an inverse relationship between the degree of conjugation and the free levels of NE and E both at rest and after exercise. This suggests that the degree of conjugation is another factor determining the concentration of free amines in plasma.

Topics: Adult; Catecholamines; Epinephrine; Exercise Test; Hemodynamics; Humans; Male; Middle Aged; Norepinephrine; Physical Exertion

We investigated the extent of catecholamine (CA) conjugation in plasma and accumulation inside red blood cells (RBCs) after forced immobilization of the rat. A control blood sample was obtained from undisturbed rats resting in home cages via an indwelling aortic catheter. Then rats were immobilized for 2 h, and blood samples were taken during stress at 15 min and 2 h and at 30 min poststress. Both sulfate and glucuronide conjugates were deconjugated by enzymatic hydrolysis and measured by radioenzymatic thin-layer chromatographic methods. In plasma of resting rats, the major conjugate of norepinephrine (NE) is sulfate and that of epinephrine (E) and dopamine (DA) is glucuronide; total conjugates in plasma are 53, 57, and 97% of total NE, E, and DA, respectively. During stress, sulfate conjugates of NE, E, and DA increased significantly along with free NE, E, and DA. DA glucuronide unexpectedly declined at 2 h of stress and 30 min poststress. Concentrations of free CAs rose in RBCs during stress, as did NE sulfate, the only conjugate present in RBC lysate. We conclude that both CA conjugation and RBC accumulation are enhanced when large amounts of free CA enter the circulation as a result of immobilization stress and that the relative importance of these inactivation pathways is different for the three CAs.

Topics: Animals; Catecholamines; Chromatography, Thin Layer; Dopamine; Epinephrine; Erythrocytes; Glucuronates; Immobilization; Male; Norepinephrine; Rats; Rats, Gunn; Rats, Inbred Strains; Stress, Physiological

Topics: Adult; Clinical Enzyme Tests; Dopamine; Epinephrine; Humans; Male; Movement; Norepinephrine; Posture; Sodium Chloride