Simple, rapid and sensitive determination of epinephrine and norepinephrine in urine and plasma by non-competitive enzyme immunoassay, compared with HPLC method.

BACKGROUND Measurement of the concentration of the free catecholamines epinephrine (adrenalin) and norepinephrine (noradrenalin) in humans is used for the diagnosis of pheochromocytoma and related diseases. METHODS A non-competitive enzyme-linked immunosorbent assay (CatCombi ELISA) kit for the measurement of epinephrine and norepinephrine concentrations in plasma and urine was developed and validated. The assay procedure consists of sample extraction, chemical and enzymatic derivatization and immunological reaction steps. A sample volume of 10 microL urine or 300 microL EDTA plasma is required for duplicate determinations of both catecholamines. For method comparison we used a reversed phase HPLC by Chromsystems after extraction by aluminium oxide with electrochemical detection for the determination of epinephrine and norepinephrine. RESULTS The CatCombi ELISA is accurate, sensitive, specific, and precise. Linear regression analysis of epinephrine and norepinephrine concentrations measured with the ELISA and with HPLC yielded highly significant correlations. CONCLUSIONS The CatCombi ELISA kit as an alternative to HPLC methods is very useful for clinical applications as well as for basic research where a simple, rapid, accurate and reproducible assay for epinephrine and norepinephrine determinations is required.

[1]  B. Mendonca,et al.  Pheochromocytoma: study of 50 cases. , 1997, The Journal of urology.

[2]  M A Pollock,et al.  Method Comparison—A Different Approach , 1992, Annals of clinical biochemistry.

[3]  Thomas G Rosano,et al.  Advances in catecholamine and metabolite measurements for diagnosis of pheochromocytoma. , 1991, Clinical chemistry.

[4]  E. Kaplan,et al.  Sensitivity of diagnostic and localization tests for pheochromocytoma in clinical practice. , 2000, Archives of internal medicine.

[5]  S. Sheps,et al.  Prevalence of clinically unsuspected pheochromocytoma. Review of a 50-year autopsy series. , 1981, Mayo Clinic proceedings.

[6]  D. Chauveau,et al.  Malignant hypertension secondary to pheochromocytoma in a hemodialyzed patient. , 1993, American journal of kidney diseases : the official journal of the National Kidney Foundation.

[7]  K. Lam,et al.  Adrenal pheochromocytoma remains a frequently overlooked diagnosis. , 2000, American journal of surgery.

[8]  R. M. Smith.  Practical aspects of modern HPLC : edited by Imre Molnar, Walter de Gruyter, 1983. DM (cloth) 170 (viii + 449 pages) ISBN 3 11 008892 4 , 1984 .

[9]  M. Mannelli,et al.  Pheochromocytoma in Italy: a multicentric retrospective study. , 1999, European journal of endocrinology.

[10]  M. Fakeeh,et al.  Investigation of phaeochromocytoma. , 1995, Clinical endocrinology.

[11]  Nisse Kalkkinen,et al.  Purification and partial characterization of rat liver soluble catechol‐ O‐methyltransferase , 1990, FEBS letters.

[12]  J. Dutton,et al.  Evaluation of a new method for the analysis of free catecholamines in plasma using automated sample trace enrichment with dialysis and HPLC. , 1999, Clinical chemistry.

[13]  R. Bender,et al.  A General Regression Procedure for Method Transformation. Application of Linear Regression Procedures for Method Comparison Studies in Clinical Chemistry, Part III , 1988, Journal of clinical chemistry and clinical biochemistry. Zeitschrift fur klinische Chemie und klinische Biochemie.

[14]  N. Musso,et al.  Reversed-phase HPLC separation of plasma norepinephrine, epinephrine, and dopamine, with three-electrode coulometric detection. , 1989, Clinical chemistry.

[15]  P. Parrilla,et al.  Differences between sporadic and multiple endocrine neoplasia type 2A phaeochromocytoma , 1998, Clinical endocrinology.

[16]  J. Kane,et al.  Freedom from drug interference in new immunoassays for urinary catecholamines and metanephrines. , 1999, Clinical chemistry.

[17]  E. Gerlo,et al.  Urinary and plasma catecholamines and urinary catecholamine metabolites in pheochromocytoma: diagnostic value in 19 cases. , 1994, Clinical chemistry.

[18]  G. Smythe,et al.  Laboratory Diagnosis of Phaeochromocytoma: Which Analytes Should We Measure? , 1993, Annals of clinical biochemistry.

[19]  D. Wild.  The Immunoassay Handbook , 2001 .

[20]  H. Geiger,et al.  Clonidine suppression test revisited. , 1998, Blood pressure.

[21]  S. Sheps,et al.  Analysis for urinary catecholamines by liquid chromatography with amperometric detection: methodology and clinical interpretation of results. , 1979, Clinical chemistry.

[22]  J. Harvey,et al.  Death from Phaeochromocytoma: Lessons from a Post-Mortem Survey , 1995, Journal of the Royal College of Physicians of London.

[23]  T. J. Panholzer,et al.  Coupled-column liquid chromatographic analysis of catecholamines, serotonin, and metabolites in human urine. , 1999, Clinical chemistry.

[24]  S. S. Deshpande.  Reagent Formulations and Shelf Life Evaluation , 1996 .