Catecholamine Derivatives Research Articles

Application of mass fragmentography to the quantitation of endogenous catecholamines

Publisher Summary This chapter discusses the application of mass fragmentography to the quantitation of endogenous catecholamines. The gas chromatography (GC) serves to resolve the various neurotransmitters from each other, while the MS is used for quantitation by measuring the ion density of specific characteristic fragments of each compound. Prior to analysis it is necessary to prepare catecholamine derivatives with the appropriate vapor pressure for GC. This is done by combining the catecholamines with pentafluoropropionic anhydride resulting in the acylation of the phenoxy, and β-hydroxy, and primary, and secondary amino groups. Using an LKB 9000 GC-MS, the structures of the derivatized catecholamines were confirmed by mass spectral analysis. For quantitation, the ion density of the most abundant fragment is recorded at the compounds retention time. The most direct approach for determination of the neurotransmitter content of the SIF cells would be to dissect out the SIF cells, and measure the content directly.

Noradrenaline binding and the search for catecholamine receptors.

The binding of 3H-noradrenaline to intact cells and microsomes seems not to measure catecholamine receptor interactions, but rather a membrane catechol-binding protein which may be related to the enzyme, catechol-O-methyl transferase. True receptors are probably scarce and detection will require catecholamine derivatives of much higher specific activity than are at present available.

Formation in vitro of N-acetyl-3,4-dimethoxyphenethylamine by pineal hydroxy-indole- O-methyl transferase

The pineal hydroxyindole- O-methyl transferase (HIOMT) catalysed formation of N-acetyl-3,4-dimethoxyphenethylamine (NADMPEA) from both N-acetyl-4-hydroxy-3-methoxyphenethylamine and N-acetyl-3-hydroxy-4-methoxyphenethylamine have been demonstrated in vitro. Dimethylation was not achieved. The formation of NADMPEA is shown also to be inhibited by haloperidol. It is suggested that, in schizophrenia, pineal HIOMT might get out of phase with its normal substrate and so act on other substrates, thus producing abnormally methylated catecholamine derivatives.

Loss of catecholamine derivatives during gas-liquid Chromatography

Loss of catecholamine derivatives during gas-liquid Chromatography

Procion yellow and catecholamine derivatives. Chemical relationships.

The use of Procion Yellow (M-4RS, ICI America, Inc.) as a fluorochrome in conjunction with catecholamine-derived isoquinoline fluorescence is examined by chemical techniques. From a series of chemical studies it has been found that: (a) Procion Yellow does not interfere with the formation of isoquinoline derivatives of catecholamine; (b) commercially available Procion Yellow is a composite of four species, all of whose emission-excitation maxima are distinguishable from that of catecholamine-derived isoquinohines; and (c) the emission-excitation maxima of Procion Yellow and of the catecholamine-derived isoquinolines are distinguishable one from another in solution. It is suggested that Procion Yellow can be a valuable fluorochrome to use in conjunction with the histofluorometric method. The data support the possibility that single adrenergic neuronal cells after being tagged with Procion Yellow can subsequently be shown to contain biogenic amines.

A new concept and further support for the importance and occurrence of 3,4-dimethoxyphenylethylamine (DMPEA) in urine of schizophrenic patients.

The biogenic formation of certain methylated derivatives of catecholamines and serotonin have been discussed in causal connection with schizophrenia. Positive and negative results by various research groups in detecting 3,4-dimethoxyphenylethylamine (DMPEA) in urine of schizophrenics have led to contradictory interpretations of the importance of DMPEA in urine as an indicator of schizophrenia. Based on the fact that most hallucinogens rapidly develop tolerance in man, we postulate that in schizophrenics endogenously formed psychotoxins must be produced at greatly altering rates in order to affect the mental state over a given period of time. Accordingly, there must be significant alterations in the excretion of endogenously formed psychotoxins or their metabolites in urine. Our first and preliminary investigations of the daily excretion of DMPEA in urine of schizophrenics over a 10-day period (horizontal study) clearly indicate the altering excretion patterns of DMPEA. These results support our hypothesis and demonstrate the necessity for further and more extensive studies of DMPEA excretion in urine of schizophrenics over a continuous period of 10–30 days.

Aldehyde derivatives of indoleamines and the enhancement of their binding onto brain macromolecules by pentobarbital and acetaldehyde.

ALDEHYDE derivatives of catecholamines and of indoleamines, for example, of serotonin, are formed in the brain by the action of monoamine oxidase (MAO)1,2. Further metabolic transformation proceeds via an NAD-dependent aldehyde dehydrogenase (ADH)3,4 and an NADPH-dependent aldehyde reductase (ADR)5, which has been shown to be inhibited by low concentrations of barbiturates, both in vitro6,7 and in vivo8. Aldehyde dehydrogenase, on the other hand, has a wide substrate specificity and a given substrate may compete for oxidation with others (such as competition between acetaldehyde and 5-hydroxyindoleacetaldehyde9). These metabolic relationships would be expected, in proper circumstances (that is, in the presence of barbiturates or acetaldehyde), to increase the steady state concentration of aldehyde intermediates of biogenic amines.

Guaiacol-0-methyltransferase: a mammalian enzyme capable of forming di-O-methyl catecholamine derivatives.

On a extrait et characterisé une enzyme capable de transférer aux catécholamines 4-O-méthyles le group méthyle provenant de l'S-adénosylméthionine et produire des substances diméthoxyques. Cette enzyme est active dans différents tissus de Mammifères.

A convenient method for the chromatographic analysis of norepinephrine, dopamine, and serotonin

Norepinephrine was completely separated from dopamine by their elution as complexes with 0.7% boric acid from a column of a weak cation exchanger. The neutral eluates were suitable for both radioactive counting and quantitation of amine content by either native fluorescence or specific fluorimetric assays. The investigation of overall catecholamine metabolism with radioactive tracers is readily carried out with this method because the amines and their metabolites are collected sequentially in these fractions: monoamine oxidase products, norepinephrine, dopamine, O-methylated derivatives of catecholamines plus serotonin.

Heptafluorobutylation of catecholamines and its application to gas-liquid chromatographic analysis by electron capture detector.

A method for formation of heptafluorobutyl derivatives of catecholamines and its use to gas-liquid chromatographic (GLC) analysis by electron capture detector (ECD) have been studied.When 0.01ml of heptafluorobutyric anhydride is used for the reaction to catecholamines in 0.1ml of ethyl acetate at 60°C for 30min, formations of heptafluorobutyl derivatives occur for all hydroxyl and amino groups of them. Heptafluorobutyl derivatives of catcholamines have good GLC properties and are more sensitive to ECD than other derivatives of them, i.e. trifluoroacetyl or trimethylsilyl ether-heptafluorobutyl derivatives. Separations of epinephrine, dopamine and norepinephrine are almost satisfactory, when 2% XF-1105 are used as stationary phase at 170°C.It has been recognized that heptafluorobutyl derivatives are applied to GLC analysis of catecholamines in biological fluids.

Influence of substituted tetrahydroisoquinolines and catecholamines on lipolysis, in vitro—II : Stereoselectivity

Utilizing glycerol release as an index of lipolysis in rat epididymal fat tissue, intrinsic activity and affinity (pD 2 values) constants were calculated for various tetrahydroisoquinoline (THI) and catecholamine derivatives. Differences were noted in the pD 2 values and to a lesser degree in the intrinsic activity constants of the d- l-isomers and deoxy derivatives of the agonists studied. In all cases, the l-isomers proved to be more active than the d-isomers or deoxy derivatives in the release of glycerol from adipose tissue, in vitro. Significant differences in the pD 2 values for the more potent l-isomers were calculated and the rank order was observed to be 1-(3′,4′,5′-trimethoxybenzyl)-6,7-dihydroxy-1,2,3,4-THI > isoproterenol > norepinephrine ≥ 1-benzyl-6,7-dihydroxy-1,2,3,4-THI. The deoxy derivatives of isoproterenol and norepinephrine possessed nearly identical pD 2 values to their respective d-isomers. N-isopropyldopamine and dopamine, however, were unable to elicit a maximum lipolytic response and did not possess equivalent intrinsic activity constants to d-isoproterenol and d-norepinephrine. Propranolol inhibited the lipolysis induced by 1-(3′,4′,5′-trimethoxybenzyl)-6,7-dihydroxy-1,2,3,4-THI and norepinephrine in a competitive manner, whereas a noncompetitive inhibition was observed in the presence of the alpha-antagonist, phentolamine. The activity-differences for the isomers of 1-(3′,4′,5′-trimethoxybenzyl)-6,7-dihydroxy-1,2,3,4-THI (795-fold) and isoproterenol (316-fold) were clearly greater than the isomeric-activity differences observed for norepinephrine (100-fold) and 1-benzyl-6, 7-dihydroxy-1,2,3,4-THI (25-fold). These results indicate: (a) that the THI derivatives which do not possess an alcoholic (μ-hydroxyl) group on the ethylamino-side chain are potent lipolytic agonists, and (b) that the tetrahydroisoquinoline derivatives act at a receptor system in adipose tissue similar, if not identical, to the interaction observed with norepinephrine. It may be suggested, therefore, that the μ-hydroxyl group is not a necessary requirement for potent lipolytic activity as previously believed.

Urinary excretion of catecholamine derivatives in essential hypertension.

Urinary excretion of norepinephrine (NE), epinephrine (E), normetanephrine (NMN), metanephrine (MN) and vanillyl mandelic acid (VMA) were measured in 10 hypertensive and 10 normotensive subjects at walking and in sleep. No difference in urinary excretion of each of NE, E, NMN, MN and VMA was observed in sleep between the hypertensive and normotensive subjects. Urinary excretions of NE, E, NMN, MN and VMA at walking were 3 to 6 times higher than those in sleep in both groups. The rate of increase in catecholamine excretion at walking was higher in the hypertensive subjects. At walking, urinary excretion of VMA in the hypertensive group was significantly higher. The ratio of VMA to total catecholamine was lower at walking. Elevated urinary catecholamine excretion in the hypertensive suggests that the substance plays a role as a causative factor of hypertension.

Action of certain catechol and oxedrine sympathomimetics on lipid mobilization in vitro

In experiments in vitro on rat epididymal adipose tissue the dose-response curves of the lipid mobilizing effects of catecholamine and oxedrine derivatives N-substituted by H-, Me-, Et-, nPr-, nBu- iPtr-, tBu- and F tBu- (phenyl- t-butyl) were investigated. In the catecholamine derivatives, the affinities were almost equal showing only a slightly rising trend towards derivatives substituted with richly branched radicals; with oxedrines, the pD 2-values rose with progressive N-substitution approximately twice as rapidly as with catecholamines. The sequence of affinities iPr > H > Me, unusual in adrenergic reactions, was observed in both homologous series.

The effect of convulsant drugs on coaxially stimulated guinea-pig ileum.

British Journal of Pharmacology and ChemotherapyVolume 31, Issue 1 p. 132-137 Free Access THE EFFECT OF CONVULSANT DRUGS ON COAXIALLY STIMULATED GUINEA-PIG ILEUM M. J. NEAL, M. J. NEAL University Department of Pharmacology, Downing Street, CambridgeSearch for more papers by this author M. J. NEAL, M. J. NEAL University Department of Pharmacology, Downing Street, CambridgeSearch for more papers by this author First published: September 1967 https://doi.org/10.1111/j.1476-5381.1967.tb01983.xCitations: 1AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL References Alving, B. O. (1961). The action of strychnine at cholinergic junctions. Archs int. Pharmacodyn. Ther., 131, 123– 150. Beleslin, D. & Polak, R. L. (1965). Depression by morphine and chloralose of acetylcholine release from the cat's brain. J. Physiol., Lond., 177, 411– 419. Beleslin, D., Polak, R. L. & Sproull, D. H. (1965). The effect of leptazol and strychnine on the acetylcholine release from the cat brain. J. Physiol., Lond., 181, 308– 316. Brown, D. A. & Quilliam, J. P. (1964). The effects of some centrally-acting drugs on ganglionic transmission in the cat. Br. J. Pharmac. Chemother., 23, 241– 256. Curtis, D. R. (1959). Pharmacological investigations upon inhibition of spinal motoneurones. J. Physiol., Lond., 145, 175– 192. Curtis, D. R. (1966). Synaptic transmission in the central nervous system and its pharmacology. In Nerve as a Tissue, ed. K. Rodahl & B. Issekutz, pp. 321– 328. New York: Harper & Row. Eccles, J. C. (1957). The Physiology of Nerve Cells, p. 195. Baltimore: Johns Hopkins Press. Eccles, J. C., Fatt, P. & Koketsu, K. (1954). Cholinergic and inhibitory synapses in a pathway from motor-axon collaterals to motoneurones. J. Physiol., Lond., 126, 524– 562. Harry, J. (1962). Effect of cooling, local anaesthetic compounds and botulinum toxin on the responses of and the acetylcholine output from the electrically transmurally stimulated isolated guinea-pig ileum. Br. J. Pharmac. Chemother., 19, 42– 55. Krnjević, K. & Philli, J. W. (1963). Pharmacological properties of acetylcholine-sensitive cells in the cerebral cortex. J. Physiol., Lond., 166, 328– 350. Lanari, A. & Luco, J. V. (1939). The depressant action of strychnine on the superior cervical sympathetic ganglion and on skeletal muscle. Am. J. Physiol., 126, 277– 288. McKINSTRY, D. N. & Koelle, G. B. (1967). Inhibition of release of acetylcholine by strychnine and its implications regarding transmission by the olivo-cochlear bundle. Nature, Lond., 213, 505– 506. Mitchell, J. F. (1963). The spontaneous and evoked release of acetylcholine from the cerebral cortex. J. Physiol., Lond., 165, 98– 116. Neal, M. J. (1967). In preparation. Paton, W. D. M. (1957). The action of morphine and related substances on contraction and on acetylcholine output of coaxially stimulated guinea-pig ileum. Br. J. Pharmac. Chemother., 12, 119– 127. Setnikar, I., Murmann, W., Magistretti, M. J. & Da Re, P. (1960). Amino-methylchromones, brain stem stimulants and pentobarbital antagonists. J. Pharmac. exp. Ther., 128, 176– 181. Takagi, K. & Takayanagi, I. (1966). Effects of strychnine, derivatives of phenyl acetate and catecholamines on contraction and acetylcholine output from the cholinergic nerve ending of guinea-pig ileum. Jap. J. Pharmac., 16, 211– 216. Citing Literature Volume31, Issue1September 1967Pages 132-137 ReferencesRelatedInformation

Tryptophan metabolism during the mid-period of embryonic development (chick)

While studying the catecholamine derivatives in the allantoic fluid of the chick embryo by gas-chromatographic techniques, numerous large peaks were seen; two of the more prominent were identified as kynurenic and xanthurenic acids. When the allantoic fluid was examined at daily intervals, it was seen that the concentration of kynuneric and xanthurenic acids reached extraordinarily high values between 7 and 9 days and then underwent a 40-fold reduction at 10 days. An increase in the concentration of anthranilic acid occurred between 9 and 10 days. There was an increase in the concentration of 5-hydroxytryptamine in the allantoic fluid after 10 days, and the assayable levels of liver kynureninase increased significantly between 8 and 9 and 9 and 10 days. Additional studies included the measurement of pyridoxine derivatives in the allantoic fluid, as well as the concentration of nicotinamide-adenine dinucleotide in the liver. The qualitative changes in tryptophan metabolites between 9 and 10 days coincided temporally with an increased concentration of pyridoxal in the allantoic fluid; and an association seemed to exist between the high concentration of oxidative derivatives in the allantoic fluid at 9 days and a relatively large concentration of nicotinamide-adenine dinucleotide present in the liver at that time.

EFFECTS OF STRYCHNINE, DERIVATIVES OF PHENYL ACETATE AND CATECHOLAMINES ON CONTRACTION AND ACETYLCHOLINE OUTPUT FROM THE CHOLINERGIC NERVE ENDING OF GUINEA PIG ILEUM

We have already reported that picric acid (1, 2) and phenyl acetate (3) provoke a contraction of the guinea pig ileum through acetylcholine liberation from the cholinergic nerve ; the effect of picric acid is inhibited by morphine but the response to phenyl acetate is not affected by morphine. So we speculated that two mechanisms could be responsible for the acetylcholine liberation: one is concerned with the actions of 5-hydroxytryptamine and picric acid and is affected by morphine, the other is brought in the action by phenyl acetate and is resistant to the inhibitory action of morphine (3). In this report the mechanisms concerned with acetylcholine liberation from the cholinergic nerve element of guinea pig ileum are studied, using strychnine, derivatives of phenyl acetate, catecholamines and reserpine.

A New Fluorescence Method for Detection and Possible Quantitative Assay of some Catecholamine and Tryptamine Derivatives on Paper

A New Fluorescence Method for Detection and Possible Quantitative Assay of some Catecholamine and Tryptamine Derivatives on Paper

The storage of norepinephrine and some of its derivatives in brain synaptosomes

The subcellular localization of norepinephrine and some of its derivatives was determined in the rat brain after their intraventricular administration. The catecholamine derivatives, norepinephrine, epinephrine and dopamine, were highly localized in the synoptosomal fraction. The methoxyamine metabolites were also localized in synaptosomes but to a lesser extent. Octopamine, the β-hydroxylated metabolite of tyramine was more highly concentrated by the synaptosomal fraction than tyramine itself.

Enzymatic Behaviour of the Product of Coenzyme A-Norepinephrine Reaction

REACTION of the sulphydryl group of coenzyme A in vitro with oxidation products of epinephrine and nor-epinephrine has been demonstrated by chemical measurement of the residual —SH groups1. The coenzyme A–catecholamine compounds may well represent an important stage in intermediary metabolism, since their breakdown may lead to regeneration of coenzyme A and formation of physiologically active catecholamine derivatives. Furthermore, if coenzyme A is inactivated by these reactions, an explanation may be at hand for some of the physiological effects of the catechol amines. The work reported here evaluates the reactivity and stability of the coenzyme A–catecholamine derivatives in the light of their enzymatic activity in vitro.