title: Strong anion determination in biological fluids by capillary electrophoresis for clinical diagnostics.
abstract: New methods for quantitative analysis of strong anions are required for diagnostic testing of human diseases. Current techniques suffer from poor selectivity and/or long analysis times that are not amenable for labile anions in high-saline or volume-restricted samples. We introduce a rapid assay (<5 min) based on capillary electrophoresis (CE) with indirect UV detection for simultaneous analysis of sulfate, sulfite, and chloride in human urine, plasma, and sweat specimens. Remarkable selectivity for strong anions is achieved by using an acidic background electrolyte under reversed polarity that results in electrokinetic rejection of matrix interferences at the capillary inlet. A dual co-ion probe system consisting of 5 mM naphthalene disulfonate (NDS) and 5 mM naphthalene trisulfonate (NTS) in 0.4 M formic acid, pH 2.0 is developed for detection of UV transparent anions (S/N ≈ 3, 60 μM with a 25 μm inner diameter fused-silica capillary) with good peak symmetry and baseline stability. Due to the chemical reactivity of sulfite, dilute formaldehyde is used as a reagent to form an acid-stable hydroxymethylsulfonate adduct. Method validation confirmed excellent linearity (R(2) > 0.999), good accuracy (mean bias ≈7%), and acceptable long-term reproducibility (CV < 10%) over 20 days. The assay allows for artifact-free determination of sulfate and sulfite with consistent results for chloride when compared to standard electrochemical methods (R(2) > 0.975). Preliminary data suggest that kidney-stone formers have lower urinary sulfate excretion relative to non-kidney-stone patient controls (p = 0.0261). CE offers a selective yet robust platform for routine analysis of strong anions that is needed for confirmatory testing of cystic fibrosis, sulfite oxidase deficiency, urolithiasis, and other disorders of sulfur metabolism and/or anion transport.
authors.affiliation: Department of Chemistry and Chemical Biology, ‡Department of Pathology and Molecular Medicine, §Department of Pediatrics, McMaster University , 1280 Main Street West, Hamilton, ON L8S4M1, Canada.
authors.forename: Adriana Nori
authors.initials: AN
authors.lastname: de Macedo
authors.forename: Muhammad Irfan Yasin
authors.initials: MI
authors.lastname: Jiwa
authors.forename: Joseph
authors.initials: J
authors.lastname: Macri
authors.forename: Vladimir
authors.initials: V
authors.lastname: Belostotsky
authors.forename: Stephen
authors.initials: S
authors.lastname: Hill
authors.forename: Philip
authors.initials: P
authors.lastname: Britz-McKibbin
mesh_terms: D000293:Adolescent; D001826:Body Fluids / Q000737:chemistry*; D016022:Case-Control Studies; D002648:Child; D002675:Child, Preschool; D002712:Chlorides / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005260:Female; D006801:Humans; D007674:Kidney Diseases / Q000175:diagnosis*; D008297:Male; D044967:Serum / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013431:Sulfates / Q000032:analysis*; D013447:Sulfites / Q000032:analysis*; D013542:Sweat / Q000737:chemistry; D016482:Urinalysis
chemical_list: D002712:Chlorides; D013431:Sulfates; D013447:Sulfites