The Adrenergic Amines of Human Blood

Malherbe, H.Weil; Bone, A. D.

doi:10.1016/s0140-6736(53)92117-5

Cited by 98 publications

(15 citation statements)

References 6 publications

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“…The levels of "E" and "N" obtained by Aronow and Howard (10, 11) and Manger and his coworkers (12) using the EDA methods are in substantial agreement with our own, but differ greatly from those estimated by Weil-Malherbe and Bone (8). Further, we have been unable to confirm the finding of Weil-Malherbe and Bone that there is an association of "E" and "N" with platelets (16) and we have observed an increase in plasma "E" during insulin hypoglycemia in man (17) rather than a decrease which they (18) have reported.…”

Section: Specificitysupporting

confidence: 87%

“…Simultaneously, Natelson, Lugovoy, and Pincus (6) described the condensation of E with ethylenediamine (EDA) to form a highly fluorescent product. In 1952 and1953, Weil-Malherbe and Bone (7, 8) extended the use of this procedure to the simultaneous estimation of E and N in blood by measuring the intensity of fluorescence at two different wave lengths. Modifications (9, 10) have been reported, and the method appears to be in fre- quent use.…”

mentioning

confidence: 99%

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The Chemical Estimation of Epinephrine and Norepinephrine in Human and Canine Plasma. I. A Critique of the Ethylenediamine Condensation Method 1

Valk¹,

Price²

1956

J. Clin. Invest.

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The recovery of epinephrine (E) and norepinephrine (N) from plasma on activated alumina is reproducible and this adsorption technique can be made reasonably specific for catechols. The development of chemical methods for the assay of E and N in peripheral plasma has been hampered, however, by: 1) the low concentrations of E and N normally present (1 ,jg. per L.-or less); 2) the lack of specificity of the methods for E and N in the presence of other catechols which are excreted in urine (1, 2) and thus presumably are present in blood; and 3) difficulty in obtaining "blanks" from plasma extracts or eluates, a condition usually met in methods designed to measure endogenous substances.In 1949 and 1950, Lund (3-5) described a method based on the conversion of E and N to their fluorescent trihydroxyindole derivatives. Only dihydroxyphenyl compounds possessing a side chain which contains both an hydroxyl and an amino group appear to be detected by this method, and "blanks" can be obtained by allowing oxidation of the trihydroxyindole derivatives of E and N to proceed to a non-fluorescent state. However, the sensitivity of the method appears insufficient to permit measurements of E and N in small quantities of normal peripheral plasma (5). Simultaneously, Natelson, Lugovoy, and Pincus (6) described the condensation of E with ethylenediamine (EDA) to form a highly fluorescent product. In 1952 and1953, Weil-Malherbe and Bone (7, 8) extended the use of this procedure to the simultaneous estimation of E and N in blood by measuring the intensity of fluorescence at two different wave lengths. Modifications (9, 10) have been reported, and the method appears to be in fre- quent use. It is less specific than the Lund (trihydroxyindole) method, and will detect the presence of dihydroxyphenyl compounds other than E and N (7).The levels of E and N found in normal human plasma by Weil-Malherbe and Bone (1.3 and 5.2 micrograms per liter, respectively) differ from those observed by others (10-12) using essentially the same method, and from the levels obtained by bioassay (13). In view of the fact that a reliable method for estimation of E and N can assume importance in the interpretation of physiological changes in man, it was considered necessary to re-evaluate the EDA method and to compare the results obtained with this technique with those obtained by the trihydroxyindole method. Our results with both dog and human plasma indicate that the EDA method is not specific for N, and may not be so for E. METHODSBlood was collected in heparinized syringes, the cells separated by centrifugation, and to a measured amount of the plasma one-half its volume of sodium fluoride (2 per cent)-sodium thiosulfate (3 per cent) solution was added.Removal of E and N from plasma was accomplished by adsorption of the catechol amines on activated alumina at pH 8.0 to 8.5, followed by elution from the alumina column with acetic acid as described by Weil-Malherbe and Bone (7). Woelm alumina, non-alkaline, Grade I (Alupharm Chemical Co., Elmont, L. I.) was ...

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Section: Specificitysupporting

confidence: 87%

mentioning

confidence: 99%

The Chemical Estimation of Epinephrine and Norepinephrine in Human and Canine Plasma. I. A Critique of the Ethylenediamine Condensation Method 1

Valk¹,

Price²

1956

J. Clin. Invest.

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“…of the arterial blood from the femoral artery of the other side was collected for determination of the concentration of catecholamine by the method of Weil-Malherbe and Bone. 5) In several dogs, ligation of the adrenal veins or section of the greater splanchnic nerves was performed. They were approached from the back with paravertebral incision caudal to the twelfth rib.…”

Section: Methodsmentioning

confidence: 99%

Role of Catecholamine in Pressor Responses to Stimulation of the Central Nervous System

et al. 1963

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To clarify the contribution of humoral factors to blood pressure changes produced by stimulation of the central nervous system, successive determination of arterial catecholamine (CA) concentration by fluorimetric method was performed. In the case of prolonged maintenance of blood pressure elevation after the stimulation, CA increased remarkably. By the experiments of ligation of the adrenal veins, section of the greater splanchnic nerves, stimulation of the greater splanchnic nerve, and intravenous injection of epinephrine, it was concluded that during stimulation of the central nervous system the neural mechanism affected the vascular system to elevate blood pressure and also affected the adrenals through the greater splanchnic nerves to secrete humoral factors, CA especially epinephrine, which maintained the rise in blood pressure after the stimulation. Despite the prolonged elevation after stimulation, the increase in CA was transient. The secretion of CA was remarkable in the cases of stimulation of the central gray stratum of the midbrain.

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“…The trihydroxyindole (THI) method of Lund (1)(2)(3)(4) and the ethylenediamine (EDA) method of Weil-Malherbe and Bone (5)(6)(7) are the two methods currently considered suitable for analysis of plasma NE and E. Neither method is specific. In this respect, the EDA method is inferior to the THI method (6,8,9), a fact which is emphasized by the false high readings of the former in uremia (10).…”

mentioning

confidence: 99%

Arteriovenous Differences of the Norepineprine-Like Material From Normal Plasma and Infused Norepinephrine*

Wallace¹,

García²,

Stead³

1961

J. Clin. Invest.

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All physiological stimuli result in changes in the level of activity of the autonomic nervous system. A method of monitoring the over-all activity of the adrenergic nerves and adrenal medullae would greatly simplify the interpretation of many physiological events. The present work began with the assumption that blood levels of norepinephrine (NE) and epinephrine (E) might reflect adrenergic activity. Therefore, our initial objective was to modify one of the existing chemical methods for the measurement of NE and E so that it would be suitable for our purposes. However, we became impressed with the controversy in the literature over the chemical identification of plasma NE and E and, as a result, adopted the dual criteria that plasma material must have not only the chemical properties of NE and E but must be removed by the tissues like NE and E to justify identification as these substances.The trihydroxyindole (THI) method of Lund (1-4) and the ethylenediamine (EDA) method of Weil-Malherbe and Bone (5-7) are the two methods currently considered suitable for analysis of plasma NE and E. Neither method is specific. In this respect, the EDA method is inferior to the THI method (6,8,9), a fact which is emphasized by the false high readings of the former in uremia (10). However, the THI method may measure dopamine (3, 4), dihydroxyphenylalanine (2-4), isopropylnorepinephrine (2-4, 9), serotonin (2) or aureomycin (11) modification used. To a greater (EDA method) or lesser degree (THI method), the proponents of both methods have depended upon the fact that none of the possibly interfering analogs of NE and E has as yet been demonstrated in normal blood by fluorometry, paper chromatography or bioassay. Cohen and Goldenberg (3) cited the results of infusion experiments as important evidence that their method (THI) did not measure the metabolites of NE or E. Steady infusions produced steady blood levels, which would not be expected if accumulating metabolites were also determined. We have had a similar experience with the EDA method.The difficulty in preparing reagent blanks which fluoresce with the same intensity each day indicates that contaminants are capable of producing fluorescence which may be interpreted as NE and E. Both methods are subject to this difficulty. In our opinion, this type of nonspecificity has interfered more with the quantitative aspects of the methods than has nonspecificity due to possible interference by the metabolites of NE and E or other closely related compounds.Whether or not either method as reported to date can accurately measure known NE and E in the amounts said to be present in normal plasma is also open to question. Price and Price (2), using the THI method, reported normal venous levels of 0.01 ,ug E per L and 0.34 ug NE per L. They used 15-ml samples of plasma but gave no data on their ability to distinguish unmodified plasma from plasma with added E and NE in amounts of 0.00015 ptg E per 15 ml or of 0.0051 ,Mg NE per 15 ml. They stated that differences in replicates of a single samp...

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The Adrenergic Amines of Human Blood

Cited by 98 publications

References 6 publications

The Chemical Estimation of Epinephrine and Norepinephrine in Human and Canine Plasma. I. A Critique of the Ethylenediamine Condensation Method 1

The Chemical Estimation of Epinephrine and Norepinephrine in Human and Canine Plasma. I. A Critique of the Ethylenediamine Condensation Method 1

Role of Catecholamine in Pressor Responses to Stimulation of the Central Nervous System

Arteriovenous Differences of the Norepineprine-Like Material From Normal Plasma and Infused Norepinephrine*

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