Total Amine Content Research Articles

Method development and cultivar-related differences of nine biogenic amines in Ontario wines

We have developed a new high-performance liquid chromatographic method using pre-column derivatization with orthophthalaldehyde, gradient elution and fluorescence detection to assay the concentrations of nine biogenic amines in wines: histamine, 1-methylhistamine, methylamine, ethylamine, tyramine, tryptamine, 2-phenylethylamine, putrescine and cadaverine. The method shows excellent analytical characteristics. It has been used to measure the concentration of these biogenic amines in 73 monovarietal wines from five red and six white cultivars. All wines were from the Niagara viticultural region of southern Ontario and were certified as to origin. Pinot noir wines had the highest content of total amines, and also of histamine, putrescine, cadaverine, ethylamine and 1-methylhistamine. Among the white wines, those from Chardonnay had the highest content of total amines and also of histamine, tyramine, methylamine and 1-methylhistamine. It is suggested that longer ageing as well as Sur-lie fermentation (Chardonnay) can account, at least in part, for these findings.

Determination of biogenic amines with an electrochemical biosensor and its application to salted anchovies

This paper describes the assembling and optimization of an electrochemical biosensor for the determination of biogenic amines (putrescine, cadaverine, histamine, tyramine, spermidine, spermine, tryptamine) commonly present in food products, and its application to salted anchovy samples. Variations of the amine content in anchovies during ripening time were measured both with the biosensor and ion chromatography with integrated pulsed amperometric detection (IC-IPAD). The probe is based on a platinum electrode which senses the hydrogen peroxide produced by the reaction catalysed by the enzyme diamine oxidase (DAO), purified from commercial seeds of cicer and immobilized on the electrode surface. Parameters such as enzyme immobilization and pH have been studied and optimised in order to obtain similar sensitivity for all the amines tested. The immobilization of the enzyme on a nylon-net membrane, using glutaraldehyde as cross-linking agent, and phosphate buffer at pH 8.0 were selected. The detection limit was 5 × 10 −7 mol litre −1. The linear range common to the amines tested was observed from 1 × 10 −6 to 5 × 10 −5 mol litre −1. The effect of potentially interfering compounds was also evaluated. Underivatized biogenic amines such as putrescine, cadaverine, histamine, tyramine and spermidine were also detected with the IC-IPAD method. Changes in the concentration of biogenic amine content in salted anchovy samples, measured with the biosensor and IC-IPAD methods, exhibited the same trend and demonstrated that the biosensor is a useful tool to monitor the variation of the total amine content in fish during storage.

INHIBITION OF THE CARDIAC SARCOPLASMIC RETICULUM Ca 2+-ATPASE BY GLUCOSE 6-PHOSPHATE IS Ca 2+ DEPENDENT

Defects in the structure or function of the cardiac sarcoplasmic reticulum (CSR) Ca 2+-ATPase presumably contribute to the Ca 2+ imbalance in the diabetic myocardium. The susceptibility to nonenzymatic protein glycation by glucose metabolites is suggested due to the relatively high percent of target lysines and arginines (approaching 15 mol%) at the ATP binding and phosphorylation domains. Brief incubations (15 min) of CSR microsomes at 24°C in the presence of 5.0 mM glucose 6-phosphate (G1c6P) inhibited Ca 2+-dependent ATPase maximal activity relative to controls. Inhibition was only observed when incubations contained 0.1 mM CaCl 2 (1.86 μmol ATP hydrolyzed · mg −1 · min −1, +Glc6P versus 2.78, control). Nonconvergent regression lines drawn from maximal velocities as a function of CSR microsome concentration indicate an irreversible mechanism of inhibition which is supported by an observed depletion in CSR amine content (2.98 μmol -NH 2 groups/mg microsomal protein, +Gl6P versus 3.34, control). Glucose 6-phosphate (5.0mM) in Ca 2+-free incubations (plus 0.1 mM EGTA) had no affect on either enzyme activity or total amine content. These data suggest that the E1 but not the E2 conformation of the CSR Ca 2+-ATPase is susceptible to Glc6P-mediated modification resulting in diminished maximal Ca 2+-dependent ATPase activity.

Formation of biogenic amines in well fermented grass silages

SUMMARYBiogenic amine formation was studied in silages made from perennial ryegrass. In 1991 two batches of grass from the same sward of the ID–DLO permanent pasture were wilted to either 250 or 450 g dry matter (DM)/kg, and ensiled in eight 1-litre laboratory silos for eachtreatment (Expt A). To induce differences in fermentation pattern, the grass was ensiled without additive (CON) or treated with formic acid (5 ml/kg; FA), cell wall degrading enzymes (2·1 ml/kg; ENZ), molasses (50 g/kg; MOL), or inoculated withLactobacillus plantarum(107colony forming units (CFU)/g; LP), a combination ofLactobacillus plantarumandStreptococcus faecium(105CFU/g; LPSF), orEnterobacter sakazakii(6×lO6CFU/g; EB). One silo for each treatment was opened after 1, 2, 4 and 7 days for pH determination and duplicate silos were opened after 10 and 90 days for pH determination and analysis of fermentation products. Two similar experiments (B and C) were performed using the CON, FA and LP treatments.Total amine content of the grass was low (0·1–0·2 g/kg DM). The well preserved silages in each experiment contained considerable amounts of amines, ranging from 0·1 g/kg DM in the wilted LP and FA silages to 7·4 g/kg DM in a low DM CONsilage. Tyramine, cadaverine, putrescine and histamine were, in descending order, the principalbiogenic amines formed, representing together 90 (S.E. 9)% of the total biogenic amine contentof the silages. Formation of amines occurred mainly during the first 10 days of fermentation, and was highest in silages with a slow acidification rate. Ensiling at high DM content, with formic acid or inoculation with large numbers of lactic acid bacteria significantly (P< 0·01) reduced the amount of amines in the silage. Total and individual amine contents of the silages were significantly correlated with concentrations of ammonia and acetic acid. It was concluded that the formation of biogenic amines in grass silage is related to protein degradation, and that amine formation can be reduced by restriction of fermentation in the silage, or by achieving rapid acidification during the first phase of ensiling.

Polyamines in Relation to Growth in Carrot Cell Cultures

Changes in polyamine metabolism were investigated in relation to growth of cell suspension cultures of carrot (Daucus carota, cv Chantenay). Changes in levels of the major amines putrescine and spermidine throughout the culture period correlated poorly with changes in fresh weight, but a closer correlation with the minor component spermine was observed. The arginine decarboxylase (ADC) inhibitor difluoromethylarginine (DFMA) strongly and specifically inhibited ADC activity in the supernatant, reduced the major amine (putrescine) by 95% and the total amine content by 80%. It had no effect on cell number and stimulated fresh weight by over 25% through increased cell expansion. Spermine content, in contrast, increased with DFMA concentration in parallel with fresh weight increases. Difluoromethylornithine strongly inhibited ornithine decarboxylase activity in the pellet, but had little effect on either polyamine levels or culture growth. It was concluded that little evidence for a correlation between free polyamines and cell number in carrot cultures could be detected, but that a possible correlation between spermine content and cell expansion was observed.

XPS analysis of NH3 plasma‐treated polystyrene films utilizing gas phase chemical modification

AbstractGas phase chemical modification (GCM) is found to be more preferable as a pretreatment for the XPS surface analysis of polymer materials than the conventional liquid phase treatment because it can circumvent problems such as solvent contamination and swelling. We have tried the quantification of the surface composition successfully by estimating the yield of the reaction from model samples. GCM was then applied to correlate the surface composition of NH3 plasma‐treated polystyrene films with their cell‐affinity. The amount of primary‐amine and that of carboxylic acid were directly determined by GCM. Although the amount of primary‐amine, 15–20% of total nitrogen, did not depend on the treatment intensity, the total amine content for the treated samples increased with the plasma treatment intensity. The quantity of carboxylic acid generated was found to be very small. All treated samples had better cell‐affinity than the control. The sample N2 (of medium treatment) showed the best cell‐affinity. The most strongly treated sample N3, with larger amine content than N2, showed worse cell‐affinity because of the interference by the sputtered SiO2 on the surface.

Biogenic amines in alcoholic beverages produced in Taiwan

AbstractThe biogenic amines tyramine, putrescine, cadaverine, 2‐phenylethylamine, histamine and tryptamine were determined in 13 kinds of alcoholic beverages produced in Taiwan. They were derivatised with dansyl chloride and analysed using a high‐performance liquid chromatographic method. Total amine content ranged from 0.23 to 11.4 μg ml−1 with Charng Chuen liqueur containing the highest level and Mei Kwei Lu the lowest. Tryptamine was found in all samples in amounts between 0.01 and 1.79 μg ml−1. The levels of the other five amines in the tested samples were 0‐7.0 μg ml−1 for 2‐phenylethylamine, 0‐3.0 μg ml−1 for putrescine, 0‐2.9 μg ml−1 for cadaverine, 0‐4.5 μg ml−1 for histamine, and 0‐1.4 μg ml−1 for tyramine. These levels seem unlikely to have adverse effects on human health.

Accumulation of agmatine in chayote (Sechium edule) leaves during development

Arginine, agmatine, putrescine and spermidine were found in the apical parts and leaves of chayote (Sechium edule Swartz) at various stages of development. The concentration of agmatine, the immediate decarboxylation product of L‐arginine, increased considerably in young leaves at the first emergence of floral buds. Young leaves always had a relatively higher content of agmatine than older ones. There was a decrease in the concentration of agmatine from the apical part to the basal leaves. Agmatine was the predominant amine in young leaves at every stage of development (50–90% of the whole amine pool). It was also predominant in mature leaves when the floral buds appeared (70% of the total amine content). An accumulation of agmatine could not be found in other Cucurbitaceae species.

Serotonin in the human infant carotid body.

By immunocytochemistry serotonin was localized in the chief cells of the carotid body in human infants. Radioenzymatic measurement of the serotonin concentration revealed that it represents a significant proportion of the total amine content of the carotid body.

Storage and turnover of histamine, 5-hydroxytryptamine and heparin in rat peritoneal mast cells in vivo.

The biogenic amines and heparin of rat peritoneal mast cells were labelled in vivo by the injection of amine precursors (3H-histidine and 3H-5-hydroxytryptophan) and 35S-sodium sulfate. Uptake of label was rapid, probably reflecting the synthesis of new granule material, but the elimination was slow. Half-lives of radiolabelled histamine (23 days) and 5-hydroxytryptamine (5-HT; 25 days) did not differ statistically from that of heparin (35 days). The slow elimination rates suggest that mast cell secretion is of little biological significance under normal conditions but are well compatible with the idea that mast cell function is related to secretion evoked by appropriate immunological stimuli. It further permitted an analysis of the amine storage by repeated injections of unlabelled 5-HT. A 15-fold increase in 5-HT content was obtained while the total amine content remained constant. The uptake of 5-HT was balanced by a reduction of histamine in a molar 1:1 ratio. A displacement of histamine by 5-HT was further indicated by increased elimination rate of radiolabelled histamine in response to 5-HT injections. The results support previous binding studies in vitro and indicate that histamine and 5-HT are bound to identical storage sites in the mast cell granules.

Effects of prenatal methylazoxymethanol treatment on the development of central monoamine neurons

The effects of prenatal treatment with the antimitotic agent methylazoxymethanol (MAM; 25 mg/kg i.v., gestation day 15) on the development of monoamine neurons and their receptors have been investigated by neurochemical techniques. The MAM treatment led to a forebrain microencephaly with an approximately 50% weight reduction of the cerebral cortex and hippocampus, 30% of the striatum while the other CNS regions were reduced by about 5–15%. Endogenous noradrenaline (NA) dopamine (DA) and 5-hydroxytryptamine (5-HT) concentrations in the cerebral cortex and hippocampus were increased by about 100% and to a similar extent for all amines, whereas the total amine content in each brain region analyzed was more or less unchanged after MAM treatment. The DA concentration in the striatum was increased by 40% without any change in the total DA content. The subcellular distribution of NA and DA in the cerebral cortex and striatum was similar in MAM-treated and control rats. The effect of the NA denervation agent DSP4 was identical in MAM-treated and controls, showing very pronounced NA reductions in the cerebral cortex, hippocampus, cerebellum and the spinal cord, while the DA levels in various brain regions were not or only to a minor degree affected. Analysis of [ 3H]NA and [ 3H]5-HT uptake in slices from the cerebral cortex in vitro, demonstrated an approximate doubling of the uptake (calculated per weight) for both amines after MAM, while total uptake was not notably changed. MAM treatment also led to a significant reduction of the in vitro binding of various radioligands for monoamine (α, β, 5-HT) receptors. The most pronounced reduction was observed for β-receptors. Administration of a tyrosine-hydroxylase ( α-methyl- p-tyrosine) or a tryptophan hydroxylase (α-propyldopacetamide) inhibitor led to marked NA/DA and 5-HT reductions after MAM in all regions analyzed, indicating that the monoamine neurons are active in synthesizing and releasing their neurotransmitter. The results suggest that

Morphological characteristics and stimulus-secretion coupling in bovine adrenal chromaffin cell cultures

Chromaffin cells isolated from adult bovine adrenal medullae were plated on collagen-coated dishes in Dulbecco's modified Eagle's medium containing 10% fetal calf serum. After 8–16 h in culture, the chromaffin cells had started to develop one, two or, more rarely, three neurites. The neurites grew at a rate of7.25 ± 0.07μm/day showing a linear correlation between process length and age of the culture. The neurites displayed one or more varicosity-like structures. Some neurites made long lasting contacts with other cell bodies and processes. Cytochemical tests indicated positive catecholamine fluorescence in boti cell bodies and processes. Fluorometric determinations indicated that by day 7 the catecholamine content was75.7 ± 1% of that found on day 1 and that noradrenaline and adrenaline represented24.6 ±1% and75.3 ± 1% of the total amine content respectively. Catecholamines were released from these cultured chromaffin cells in response to stimulation by either 56 m m KCl or acetylcholine. The stimulation-induced release of amines was Ca 2+-dependent and was blocked by increasing the extracellular c oncentration of Mg 2+. Substitution of (a) extracellular Na 2+ by either choline or sucrose or (b) Ca 2+ by Ba 2+ produced a sharp increase in catecholamine output. Acetylcholine produced a doserelated increase in catecholamine release; half maximal release was produced by3.5 × 10 −5 m acetylcholine and maximal release by 10 −4 m acetylcholine. The cholinergic receptor of cultured chromaffin cells appears to be nicotinic since catecholamine release was stimulated by nicotine and carbamylcholine but not by pilocarpine. Acetylcholine-evoked catecholamine release was blocked by d-tubocurarine (ID 50 = 5.5 × 10 −7 m), and by hexamethonium (ID 50 = 8 × 10 −6 m). However, the acetylcholine-evoked amine release was not blocked by α-bungarotoxin at a concentration (1.25 × 10 −7 m) which blocked frog rectus abdominus responses to acetylcholine (10 −6–10 −3 m). Tetrodotoxin (5 × 10 −6 m) produced47 ± 2% inhibition of the secretory response to acetylcholine (10 −4 m) but it did not modify the amine release in response to 56 m m KCl. Adult bovine chromaffin cells in culture display some of the morphological and functional characteristics of sympathetic neurones. Consequently, the bovine chromaffin cell in culture appears to be a promising model for studies on development and secretion.

AMINES IN DRY FERMENTED SAUSAGE

ABSTRACTThe concentrations of some important amines were determined in dry fermented sausages. Total amine content ranged from 23.78–195.19 mg per 100g of dry matter. Maximal amounts of 28.59 mg of histamine, 39.57 mg of putrescine and 150.63 mg of tyramine per 100g of dry matter were found. The formation of putrescine from arginine was suggested. A large variation in the amine content and composition was found, possibly related to different methods of fabrication and to the specific flora of the sausages. In view of possible harmful effects of these amines, concentrations reported in this note deserve careful reflection and further investigation.

Content of catecholamines in the adrenal medulla in hypertension and other diseases

Adrenal medullary tissue removed at autopsy from 100 patients was examined for NE, E, and DA by fluorimetry. No significant differences of total amine content were observed between specimens removed less than 12 hours and those obtained 50 hours after death. Separate analysis of each adrenal gland of 19 patients disclosed a considerable variation of NE and E content of the two glands. In 13 children up to the age of 14 years the total amine content and the proportion of E increased with age. In hypertensive patients treated with reserpine the adrenal medullary content of NE and E did not differ significantly from that of a similar group of hypertensives treated with other drugs or from that of a group of normotensive patients. However, DA was not detected in reserpine-treated subjects. Patients who died of malignant neoplasia of lung or gastrointestinal tract had a markedly higher content of E and NE than any other group.

Principal Aliphatic Secondary Amines of Burley Tobacco

AbstractQuantity and distribution of the principal aliphatic secondary amines - dimethylamine, methylethylamine, diethylamine, and methylpropylamine - varied within plant tissue and among Nicotiana spp. In Burley tobacco the stem tissue had the highest content of these amines and the leaf midrib the lowest. Leaf lamina, roots and seed were intermediate in amine content. Among Nicotiana spp. there was considerable difference in total amine content as well as among the ratios of the amine fractions measured. Amine content of tissue was positively correlated with nitrogen content of tissue and was altered by the drying process of tissue prior to analysis. The amines were not present as free amines or salts but were detected only following steam distillation from a basic medium

The simultaneous spectrophotometric determination of primary and tertiary amines in aqueous solution with copper-(ethylene dinitrilo)tetraacetic acid

The preferential coordination of primary amines to the copper-EDTA complex and the resulting fall in absorbance at 720 mμ is used as a basis for the accurate determination of both primary and tertiary amines in mixed aqueous solutions.The total amine content is first determined by spectrophotoimetric titration of the mixed amine solution with the dibasic copper-EDTA complex (or vice versa). Then the mixed solution of amines is diluted to a definite total amine concentration, and its absorbance at 720 mμ is again measured after the addition of a definite amount of the tetrasodium salt of EDTA complexed with copper, (CuY) 2-. This absorbance is located on a suitable varying concentration ratio curve, and the concentrations of both primary and tertiary amines in the solution being analyzed can be read directly from the curve.

Depletion and replacement of the adrenaline and noradrenaline contents of the rat adrenal gland, following treatment with reserpine.

The effect of reserpine on the adrenaline and noradrenaline contents of rat adrenal glands has been studied. Given subcutaneously to male albino Wistar rats in 3 daily doses of 1 mg/kg, it caused, 24 hr after the final dose, a similar percentage loss of adrenaline and noradrenaline of approximately 50%. Subsequently the adrenaline content slowly increased, until at 14 days from the begining of the experiment there was no significant difference between the reserpine-treated and the control glands. In contrast, the recovery of the noradrenaline content was rapid, exceeding the control value at 7 days by approximately 250%. This increased content of noradrenaline declined to the normal level by 21 days. The total amine content returned to normal by 7 days and remained at this level subsequently. The effect of re-depleting the glands of their amine content was investigated. It was found that re-depletion at 7 days caused a preferential release of noradrenaline, followed at 14 days by a peak of noradrenaline at least as high as that obtained following the initial depletion only. Re-depletion at 21 days caused an effect similar to that obtained initially. Denervation of the left adrenal gland did not alter the degree of depletion caused by reserpine, nor did it alter the subsequent replacement of the amines as compared with that in the innervated right gland.

The adrenaline and noradrenaline content of the adrenal gland of the cat following depletion by acetylcholine.

This paper describes the attempts that were made to obtain a replacement of adrenaline and noradrenaline in the adrenal gland of the atropinized cat, subsequent to the depletion of the gland by repeated intravenous doses of acetylcholine. In the anaesthetized animal, there was no replacement of adrenal amines within 15 hours of the depletion. The further loss of amine that occurred during this time was prevented by the de nervation of the gland. If the animal was allowed to recover from the anaesthetic there was some replacement of amines within 2 to 3 days, provided that the condition of the animal was satisfactory. By 6 to 7 days the total amine content had returned to its initial level, but there was now an alteration in the relative proportions of the two amines. Although the adrenaline was still well below the resting level, the noradrenaline was several times its initial value. By one month, the noradrenaline had decreased and the adrenaline had increased to their initial amounts and proportions. Thus this work gives evidence for the formation of adrenaline from noradrenaline.

Relationship between the amine contents of the two adrenal glands of the cat.

WORKERS1 vary in their methods of calculating the adrenaline and noradrenaline content of adrenal glands. Some express them in micrograms per unit weight of gland, some in micrograms per gland and some even in micrograms per kilogram of body-weight. Since, frequently, there is a difference between the weight of the left and of the right gland of each cat, it was thought desirable to determine whether the percentage of noradrenaline is the same in the two glands, and whether the total amine content is the same for both glands or whether the heavier gland contains more amine.