Automated analysis of urinary catecholamines by high-performance liquid chromatography and on-line sample pretreatment |
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| Institution: | 1. Allergy Immunology and Blood and Marrow Transplant Division, Department of Pediatrics, Benioff Children''s Hospital and University of California San Francisco, San Francisco, CA 94143, USA;2. Department of Pediatrics, Tuba City Regional Health Care Center, Tuba City, AZ 86045, USA;3. Department of Pediatrics, Chinle Comprehensive Health Care Facility, Chinle, AZ 86503, USA;4. Research and Development, PerkinElmer Genetics, Inc., Bridgeville, PA 15017, USA;1. Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA, United States;2. Clinical Laboratories, University of Pittsburgh Medical Center Presbyterian and Shadyside Hospitals, Pittsburgh, PA, United States;3. McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, United States;4. Clinical Laboratory, Children''s Hospital of Pittsburgh of UPMC, Pittsburgh, PA, United States |
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| Abstract: | A simple and automated solid-phase extraction for the selective and quantitative HPLC analysis of free catecholamines in urine is described. The urinary catecholamines react with diphenylboric acid, giving a complex at pH 8.5 which is strongly retained on a PLRP-S cartridge; elution is accomplished with the same mobile phase used for HPLC analysis. Separation is performed by ion-pair reversed-phase HPLC, with sodium heptanesulphate as counter-ion, and a totally end-capped C18 analytical column. Quantitation is achieved with an electrochemical detector. A Spark Holland Prospekt system controls the on-line solid-phase extraction, preconcentration and direct elution to the LC column. Chromatography run-time is 10 min and the total time to process one urine sample is ca. 12 min. |
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