The fluorimetric estimation of adrenaline and noradrenaline in plasma

Weil‐Malherbe, H.; Bone, A. D.

doi:10.1042/bj0670065

Cited by 48 publications

(13 citation statements)

References 14 publications

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“…The sensitivity of the THI method is such that there may be considerable error introduced if small plasma samples are employed. Our findings in plasma of normal subjects are in agreement with those of WeilMalherbe, who reports identical results with both methods (14).…”

Section: Discussionsupporting

confidence: 83%

Evaluation of Catechol Amine Levels in Renal Insufficiency

Zileli¹,

Hamlin²,

Reutter³

et al. 1958

J. Clin. Invest.

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The estimation of epinephrine (E) and norepinephrine (NE) in plasma and urine has become important as a procedure in physiological research and as an aid in the diagnosis of pheochromocytoma. With the development of fluorometric procedures the chemical estimation of E and NE in biological fluids and tissues was made possible.The fluorometric procedures used most commonly are modifications of the trihydroxyindole method of Lund (1) and the ethylenediamine (EDA) condensation method of Weil-Malherbe and Bone (2, 3). The trihydroxyindole method is not sensitive enough to permit measurements of E and NE in small amounts of normal peripheral plasma (1), and its practical usefulness is limited to the assay of urine. The sensitivity of the EDA condensation method is such that extremely small amounts of E and NE can be measured; however, it is not as specific as the trihydroxyindole method and will detect phenolic compounds other than E and NE (4).While employing the EDA condensation method as an aid in the diagnosis of pheochromocytoma, we have observed, in some individuals with renal insufficiency who later were found not to have a pheochromocytoma, levels of E and NE much higher than those which occur in normal patients. The purpose of the present study was to determine whether the high levels of E and NE frequently observed in patients with renal insufficiency are true values or are due to the presence of substances which interfere with the EDA method and produce false high values of E and NE.The presence of large amounts of these substances may be due to an abnormal or increased production of E, NE or interfering compounds, or an inability on the part of the kidneys to excrete the normal amounts of these substances present in the blood. Among possible interfering substances which occur in abnormal amounts in the blood of patients with renal insufficiency are urea, creatinine, uric acid and phenolic compounds. Of these, the phenolic compounds are most likely to be involved.The phenols are excreted in both free and conjugated forms, with a normal daily output in the urine of 200 to 500 mg. (5). They are derived mainly from bacterial putrefaction of aromatic amino acids in the intestine. A minor portion comes from the presence of sex hormones, epinephrine, norepinephrine and tyrosine in the blood. The normal blood levels calculated as phenol range from 1 to 2 mg. per 100 ml., whereas in uremic patients they may reach 6 to 10 mg. per 100 ml. (6). The importance of these compounds in the pathogenesis of uremic symptoms has been reviewed by Harrison and Mason (7).In order to determine whether the presence of large amounts of phenolic compounds in the plasma of patients with renal insufficiency can account for the high levels of E and NE as measured by the EDA condensation method, the effect of various phenolic compounds on this method and the trihydroxyindole method was investigated, and the specificity of these two fluorometric procedures for estimating catechol amines was compared. Plasma E and NE levels were assayed by...

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Section: Discussionsupporting

confidence: 83%

Evaluation of Catechol Amine Levels in Renal Insufficiency

Zileli¹,

Hamlin²,

Reutter³

et al. 1958

J. Clin. Invest.

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“…Dopamine, like norepinephrine and epinephrine, is found in a conjugated form in both plasma and urine (24)(25)(26). Whereas free norepinephrine is readily detectable in human plasma (27,28) ,2 free dopamine frequently cannot be detected (25,12),2 but when it is measurable, the values are usually less than those for free norepinephrine (24). As the excretion of free dopamine is severalfold greater than norepinephrine, however, glomerular filtration could contribute importantly to urinary dopamine only if hydrolysis of the filtered conjugate occurred during storage and assay of the catecholamines.…”

Section: Discussionmentioning

confidence: 99%

Effects of Dietary Sodium and of Acute Saline Infusion on the Interrelationship between Dopamine Excretion and Adrenergic Activity in Man

Alexander¹,

Gill²,

Yamabe³

et al. 1974

J. Clin. Invest.

265

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A B S T R A C T The effects of dietary sodium and of saline infusion on urinary dopamine and norepinephrine and on the relationship of these catecholamines to adrenergic activity were determined. In seven normal subjects on a 9-meq sodium intake, urinary dopamine and norepinephrine were 136±18 (SE) and 37.4±5.3 Ag/day, respectively. When sodium intake was increased to 209 or 259 meq/day, urinary dopamine increased to 195±20 Ag/day (P < 0.01) whereas urinary norepinephrine decreased to 21.1±3.0 Ag/day (P < 0.01). Infusion of saline in seven subjects increased sodium excretion and urinary dopamine (from 2.18±0.22 to 2.72 ±0.19 jg/20 min, P < 0.01), but decreased plasma dopamine-fl-hydroxylase by 33% and urinary norepinephrine insignificantly. The clearance of inulin and paminohippurate did not change significantly and filtration fraction was the same. The data indicate that an increase in dietary sodium or infusion of saline results in an apparent decrease in adrenergic activity and an increase in urinary dopamine. Dopamine excretion would thus appear to relate inversely to adrenergic activity and to parallel sodium excretion. These findings suggest a possible role for dopamine and norepinephrine in the regulation of renal sodium excretion.

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“…The platelet CA content has been reported for Ad (Born et al, 1958), for Ad plus NA (Weil-Malherbe & Bone, 1958) and for total CA (Markwardt, 1967). A comparison of our data with those of the two pioneer studies dealing with CA content in platelets of man and cattle (Weil-Malherbe & Bone, 1957; as well as that reporting concentrations in pig platelets (Born & Homykiewicz, 1957;Bom et al 1958) (Da Prada, unpublished data). The inadequacy of the fluorimetric methods for measuring CA in blood is further supported by the fact that the same authors (Weil-Malherbe & Bone, 1957) reported values for Ad and NA in human and bovine plasma which are at least 20 times higher than those obtained by improved radioenzymatic assays (Callingham, 1975;Da Prada & Ziircher, 1976).…”

Section: Catecholamines In Plateletsmentioning

confidence: 99%

“…A comparison of our data with those of the two pioneer studies dealing with CA content in platelets of man and cattle (Weil-Malherbe & Bone, 1957; as well as that reporting concentrations in pig platelets (Born & Homykiewicz, 1957;Bom et al 1958) (Da Prada, unpublished data). The inadequacy of the fluorimetric methods for measuring CA in blood is further supported by the fact that the same authors (Weil-Malherbe & Bone, 1957) reported values for Ad and NA in human and bovine plasma which are at least 20 times higher than those obtained by improved radioenzymatic assays (Callingham, 1975;Da Prada & Ziircher, 1976). More recent experiments with improved fluorimetric techniques (Markwardt, 1967) have given values for human and rabbit platelet CA (about 8 and 50 pmol/mg protein, respectively) which were only 5 to 7 times higher than those found in the present study (1.86 and 7 pmol/mg protein respectively).…”

Section: Catecholamines In Plateletsmentioning

confidence: 99%

Content and Subcellular Localization of Catecholamines and 5‐hydroxytryptamine in Human and Animal Blood Platelets: Monoamine Distribution Between Platelets and Plasma

Prada

Picotti

1979

British J Pharmacology

151

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The content of adrenaline (Ad), noradrenaline (NA) and dopamine was measured in human, guinea‐pig, cat, rabbit and rat blood platelets by a highly sensitive and specific radioenzymatic method. In all platelet specimens analyzed, the content of the three catecholamines (CA) was several thousand times lower than that of 5‐hydroxytryptamine (5‐HT). In basal conditions, the NA concentration in platelets and plasma always exceeded that of Ad and dopamine. In rat and rabbit platelets, Ad, NA and dopamine were present only in the free (unconjugated) form. Platelets of rats with storage pool deficiency (Fawn‐hooded) contained much less 5‐HT and CA than normal rat platelets. Following restraint stress, platelets of Fawn‐hooded rats, in contrast to normal rat platelets, did not accumulate CA in spite of a dramatic rise in plasma CA. Reserpine, a monoamine depletor, released CA as well as 5‐HT from rabbit platelets in vivo. Subcellular fractionation experiments with rabbit platelets indicate that both CA and 5‐HT are most concentrated in the fraction consisting of pure 5‐HT organelles. Both in humans and rabbits the concentration gradient between platelets and plasma was much lower for CA than for 5‐HT, indicating that a high affinity transport mechanism operates in vivo for 5‐HT but not for CA. In conclusion, the present data show that both human and animal platelets contain Ad, NA and dopamine. The bulk of the CA seems to be stored as unconjugated amines together with 5‐HT, histamine and p‐octopamine in a multitransmitter storage site, namely the 5‐HT organelle.

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The fluorimetric estimation of adrenaline and noradrenaline in plasma

Cited by 48 publications

References 14 publications

Evaluation of Catechol Amine Levels in Renal Insufficiency

Evaluation of Catechol Amine Levels in Renal Insufficiency

Effects of Dietary Sodium and of Acute Saline Infusion on the Interrelationship between Dopamine Excretion and Adrenergic Activity in Man

Content and Subcellular Localization of Catecholamines and 5‐hydroxytryptamine in Human and Animal Blood Platelets: Monoamine Distribution Between Platelets and Plasma

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