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A precise catecholamine assay for small plasma samples.

J Diamant, S O Byers

The Journal of Laboratory and Clinical Medicine

|April 1, 1975

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This study developed a precise method for measuring norepinephrine (NE) and epinephrine (E) in plasma, enabling accurate catecholamine analysis in humans and rats. The optimized assay requires minimal blood samples, improving diagnostic capabilities.

Area of Science:

Biochemistry
Analytical Chemistry
Endocrinology

Background:

Accurate measurement of plasma catecholamines like norepinephrine (NE) and epinephrine (E) is crucial for understanding physiological and pathological states.
Existing methods often suffer from interference between NE and E, requiring large sample volumes and complex procedures.

Purpose of the Study:

To develop and validate a highly precise and stable fluorescence assay for simultaneous determination of NE and E in plasma.
To establish reliable basal plasma catecholamine levels in humans and rats using minimal sample volumes.

Main Methods:

Optimized the trihydroxyindole reaction conditions (time, temperature, pH) to minimize NE-E interference.
Streamlined alumina elution and instrumental factors for efficient analysis of small plasma volumes (5 ml for humans, 8-11 ml for rats).

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Incorporated duplicate recovery measurements with radiolabeled catecholamines for precise quantification and reliability estimation.

Main Results:

Achieved minimal cross-fluorescence (<1% for NE in E, 0.5-10% for E in NE) with superior assay stability.
Determined mean basal plasma NE levels of 0.38 ng/ml (range 0.15-0.54) and E levels of 0.09 ng/ml (range 0.01-0.27) in adult men.
Established basal rat plasma NE (0.26 ng/ml) and E (0.36 ng/ml) levels, significantly lower than those under less controlled anesthesia.

Conclusions:

The developed fluorescence assay offers high precision (±10% for NE) and stability for plasma catecholamine measurement.
The method's efficiency and low sample volume requirement facilitate reliable basal catecholamine level determination in both human and animal models.
This advancement supports further research into catecholamine function and dysregulation in various physiological conditions.