title: A revised method for determination of serum mercaptalbumin and non-mercaptalbumin by high-performance liquid chromatography coupled with postcolumn bromocresol green reaction.
abstract: BACKGROUND
  Previously, high-performance liquid chromatography (HPLC) equipped with ultraviolet or fluorescence detectors has been used for separation of human mercaptalbumin (HMA) and human non-mercaptalbumin (HNA). However, it is difficult to perform reliable chromatographic analysis due to peak interference of such serum compounds as uric acid and bilirubin. The aim of this study is to explore a selective and simple analytical method for the determination of HMA and HNA.
  
  
  METHODS
  HMA and HNA in serum sample were separated by HPLC and reacted with bromocresol green using a postcolumn reaction scheme.
  
  
  RESULTS
  A complete separation of HMA and HNA is achieved in less than 30 min by using weak anion exchange columns and isocratic elution. Within-run and between-day precisions at albumin concentration of 45 g/L were 4.2 and 1.7% for HMA and 4.5 and 4.6% for HNA, respectively. There was no interference in HMA and HNA peaks when bilirubin-, haemoglobin- or chyle-spiked pooled serum samples were analysed.
  
  
  CONCLUSIONS
  Our method is reliable and not labour-intensive and, therefore, might be applicable for clinical and epidemiological studies.
authors.affiliation: Department of Pathophysiological Laboratory Sciences, Field of Radiological and Medical Laboratory Sciences, Nagoya University Graduate School of Medicine, Nagoya, Japan ueyama@met.nagoya-u.ac.jp.
authors.forename: Jun
authors.initials: J
authors.lastname: Ueyama
authors.affiliation: Department of Medical Technique, Nagoya University Hospital, Nagoya, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Ishikawa
authors.affiliation: Department of Pathophysiological Laboratory Sciences, Field of Radiological and Medical Laboratory Sciences, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors.forename: Takaaki
authors.initials: T
authors.lastname: Kondo
authors.affiliation: Department of Pathophysiological Laboratory Sciences, Field of Radiological and Medical Laboratory Sciences, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors.forename: Megumi
authors.initials: M
authors.lastname: Motoyama
authors.affiliation: Department of Medical Technique, Nagoya University Hospital, Nagoya, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Matsumoto
authors.affiliation: Department of Transfusion Medicine, Nagoya University Hospital, Nagoya, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Matsushita
mesh_terms: D015982:Bias; D001663:Bilirubin / Q000737:chemistry; D001961:Bromcresol Green / Q000737:chemistry*; D002138:Calibration; D005591:Chemical Fractionation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D044967:Serum; D012709:Serum Albumin / Q000302:isolation & purification*; D000075462:Serum Albumin, Human; D014527:Uric Acid / Q000737:chemistry
chemical_list: D012709:Serum Albumin; C001702:mercaptoalbumin; C079732:nonmercaptalbumin; D014527:Uric Acid; D001961:Bromcresol Green; D001663:Bilirubin; D000075462:Serum Albumin, Human
keywords: Mercaptalbumin; high-performance liquid chromatography; non-mercaptalbumin