The systematic investigation and development of the histamine radioenzymatic assay

Thesis/Dissertation · Sun Jan 01 04:00:00 EST 1984

OSTI ID:6896119

Verburg, K M

The radioenzymatic assay for histamine is a widely used analytical procedure based on the enzymatic conversion of histamine to ({sup 3}H)tele-methylhistamine utilizing histamine N-methyltransferase (HNMT) and S-adenosyl-L-({sup 3}H-methyl) methionine (({sup 3}H)SAME). Despite numerous modifications of this method, the assay lacks the sensitivity and specificity required to quantify histamine from many important biologic samples such as human plasma. The objective of this study was to investigate systematically the radioenzymatic assay for histamine and develop a highly sensitive and specific assay for use in basic or clinical studies. HNMT was purified 260-fold from rat kidney and the use of purified HNMT in the histamine radioenzymatic assay improved specificity of this method and also improved sensitivity by eliminating the enzyme-dependent blank and permitting the inclusion of high specific activity ({sup 3}H)SAME. The adsorption of histamine to glass surfaces was characterized and strategies were developed to prevent binding. Finally, optimization of the reaction allowed the development of a simplified product isolation procedure. The histamine radioenzymatic assay developed in this study has a sensitivity of 2.0 pg and is specific for histamine as judged by direct product identification and cross-contamination studies. The assay was utilized to establish reference values for the concentration of histamine in human plasma and the 24-hour urinary excretion of histamine for normal human subjects. In summary, a sensitive and specific radioenzymatic assay for histamine was developed as a result of the systematic investigation of this methodology.

• Research Organization: Indiana Univ., Bloomington, IN (USA)
• OSTI ID: 6896119
• Country of Publication: United States
• Language: English

Related Subjects

550601* -- Medicine-- Unsealed Radionuclides in Diagnostics
62 RADIOLOGY AND NUCLEAR MEDICINE
ADSORPTION
AMINES
AZOLES
BIOLOGICAL MATERIALS
BIOLOGICAL WASTES
BLOOD
BLOOD PLASMA
BODY FLUIDS
ENZYME ACTIVITY
ENZYMES
HETEROCYCLIC COMPOUNDS
HISTAMINE
HYDROGEN COMPOUNDS
IMIDAZOLES
MATERIALS
METHYL TRANSFERASES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
RADIOENZYMATIC ASSAY
SENSITIVITY
SORPTION
TRANSFERASES
TRITIUM COMPOUNDS
URINE
WASTES