Eva Klapkova1, Magdalena Fortova2, Richard Prusa2, Libuse Moravcova2, Karel Kotaska2. 1. Department of Medical Chemistry and Clinical Biochemistry, 2nd Faculty of Medicine and University Hospital Motol, Charles University, Prague, Czech Republic. eva.klapkova@email.cz. 2. Department of Medical Chemistry and Clinical Biochemistry, 2nd Faculty of Medicine and University Hospital Motol, Charles University, Prague, Czech Republic.
Abstract
BACKGROUND: A simple high-performance liquid chromatography (HPLC) method was developed for the determination of albumin in patients' urine samples without coeluting proteins and was compared with the immunoturbidimetric determination of albumin. Urine albumin is important biomarker in diabetic patients, but part of it is immuno-nonreactive. METHODS: Albumin was determined by high-performance liquid chromatography (HPLC), UV detection at 280 nm, Zorbax 300SB-C3 column. Immunoturbidimetric analysis was performed using commercial kit on automatic biochemistry analyzer COBAS INTEGRA® 400, Roche Diagnostics GmbH, Manheim, Germany. RESULTS: The HLPC method was fully validated. No significant interference with other proteins (transferrin, α-1-acid glycoprotein, α-1-antichymotrypsin, antitrypsin, hemopexin) was found. The results from 301 urine samples were compared with immunochemical determination. We found a statistically significant difference between these methods (P = 0.0001, Mann-Whitney test). CONCLUSION: New simple HPLC method was developed for the determination of urine albumin without coeluting proteins. Our data indicate that the HPLC method is highly specific and more sensitive than immunoturbidimetry.
BACKGROUND: A simple high-performance liquid chromatography (HPLC) method was developed for the determination of albumin in patients' urine samples without coeluting proteins and was compared with the immunoturbidimetric determination of albumin. Urine albumin is important biomarker in diabeticpatients, but part of it is immuno-nonreactive. METHODS: Albumin was determined by high-performance liquid chromatography (HPLC), UV detection at 280 nm, Zorbax 300SB-C3 column. Immunoturbidimetric analysis was performed using commercial kit on automatic biochemistry analyzer COBAS INTEGRA® 400, Roche Diagnostics GmbH, Manheim, Germany. RESULTS: The HLPC method was fully validated. No significant interference with other proteins (transferrin, α-1-acid glycoprotein, α-1-antichymotrypsin, antitrypsin, hemopexin) was found. The results from 301 urine samples were compared with immunochemical determination. We found a statistically significant difference between these methods (P = 0.0001, Mann-Whitney test). CONCLUSION: New simple HPLC method was developed for the determination of urine albumin without coeluting proteins. Our data indicate that the HPLC method is highly specific and more sensitive than immunoturbidimetry.
Authors: E J Lewis; L G Hunsicker; W R Clarke; T Berl; M A Pohl; J B Lewis; E Ritz; R C Atkins; R Rohde; I Raz Journal: N Engl J Med Date: 2001-09-20 Impact factor: 91.245
Authors: B M Brenner; M E Cooper; D de Zeeuw; W F Keane; W E Mitch; H H Parving; G Remuzzi; S M Snapinn; Z Zhang; S Shahinfar Journal: N Engl J Med Date: 2001-09-20 Impact factor: 91.245
Authors: W Greg Miller; David E Bruns; Glen L Hortin; Sverre Sandberg; Kristin M Aakre; Matthew J McQueen; Yoshihisa Itoh; John C Lieske; David W Seccombe; Graham Jones; David M Bunk; Gary C Curhan; Andrew S Narva Journal: Clin Chem Date: 2008-11-21 Impact factor: 8.327