Capillary electrophoresis for clinical problem solving: analysis of urinary diagnostic metabolites and serum proteins.
Many clinical laboratories employ gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC) to detect abnormal compounds occurring in urine and serum due to disease. The methods, particularly GC-MS, often require laborious sample pre-treatment, and separation times may exceed an hour. We describe the use of capillary electrophoresis (CE) equipped a with a diode-array detector in an attempt to improve the efficiency of an analytical system routinely used for diagnosis of human metabolic disease. It was found that urine samples could be injected directly onto the CE instrument without any pre-treatment, and over 50 metabolites were separated in 15 min. Identification of abnormal metabolites was based on migration times and characteristic diode-array spectra. The method readily diagnosed adenolysuccinase deficiency, 5-oxoprolinuria, propionic acidemia and disorders have orotic acid as diagnostic metabolite (e.g. the HHH-syndrome). The results show that CE may become a useful additional tool for diagnosis of metabolic disease. In a different project CE was used to study sera from the Janus-bank. This large serum bank comprises samples collected at intervals from nearly 300,000 blood donors. As the sera are stored at -25 degrees C and not at a lower temperature, a major concern has been the stability of the specimens. GC-MS, 2D-protein electrophoresis, certain immunological assays and enzyme measurements have previously been used to evaluate the stability of the sera. We can now also show that the protein profile, as determined by CE, is remarkably stable even after 22 years of storage. The results moreover confirmed that the CE-method and traditional gel electrophoresis gave almost identical results, except for small amounts of fibrinogen which did not show up on the CE-pattern.
Jellum
E
E
Institute of Clinical Biochemistry, Rikshospitalet, Oslo, Norway.
Dollekamp
H
H
Blessum
C
C
D001798:Blood Proteins / Q000032:analysis*; D019411:Clinical Laboratory Techniques; D019075:Electrophoresis, Capillary / Q000379:methods*; D005260:Female; D006801:Humans; D008659:Metabolic Diseases / Q000175:diagnosis / Q000652:urine*
D001798:Blood Proteins