title: Analytical evaluation of the Tosoh HLC-723 G7 automated HPLC analyzer for hemoglobin A2 and F determination.
abstract: OBJECTIVE
  The analytical performance of a new automated HPLC system (Tosoh HLV-723 G7) for Hb A(2) and Hb F quantification in blood was studied.
  
  
  METHODS
  Hb A(2) and Hb F measurements were studied for imprecision, linearity, carry-over, interferences and sample concentration effect. Method comparison study was performed with the Bio-Rad Variant II HPLC system. Hb F results were also compared with those obtained by the alkaline denaturation test. The detection of some common Hb variants was also studied.
  
  
  RESULTS
  Hb A(2) within-run and between-run CVs were found between 0.8-2.2% and 2.9-7.2%, respectively, while CVs for Hb F were up to 10.0% in normal and between 1.9-5.3% at more clinically relevant Hb F concentrations (>1.5%). Comparison study with Bio-Rad Variant II for Hb A(2) determination showed good correlation but highlighted calibration differences. The following results were obtained in two different laboratories: y = 1.163x - 0.52, r = 0.9918, n = 144 (Lab A); y = 1.060x - 0.40, r = 0.9920, n = 93 (Lab B). With regard to the determination of Hb F, the measurements performed by the tested method was found to correlate well with the alkaline denaturation test (y = 1.0138x - 0.36, r = 0.9842, n = 20) and with the Variant II HPLC system (y = 0.812x + 0.52, r = 0.9835, n = 110). An excellent linearity (r = 0.999) was found for both Hb A(2) and Hb F in the range 0.8-19%. Hb S, Hb C and Hb D can be presumptively identified by the assigned retention time windows.
  
  
  CONCLUSIONS
  The new analyzer Tosoh HLV-723 G7 was found to be a reliable method for Hb A(2) and Hb F quantification and for the presumptive identification of some common Hb variants.
authors.affiliation: Dipartimento di Scienze e Tecnologie Biomediche, Università degli Studi di Milano, Via Fratelli Cervi 93, 20090 Segrate, Milano, Italy.
authors.forename: Renata
authors.initials: R
authors.lastname: Paleari
authors.forename: Monica
authors.initials: M
authors.lastname: Cannata
authors.forename: Filippo
authors.initials: F
authors.lastname: Leto
authors.forename: Aurelio
authors.initials: A
authors.lastname: Maggio
authors.forename: Franca R
authors.initials: FR
authors.lastname: Demartis
authors.forename: Maria F
authors.initials: MF
authors.lastname: Desogus
authors.forename: Renzo
authors.initials: R
authors.lastname: Galanello
authors.forename: Andrea
authors.initials: A
authors.lastname: Mosca
mesh_terms: D001331:Automation; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation* / Q000379:methods; D005319:Fetal Hemoglobin / Q000032:analysis*; D014644:Genetic Variation; D006443:Hemoglobin A2 / Q000032:analysis*; D006455:Hemoglobins, Abnormal / Q000032:analysis / Q000235:genetics; D006801:Humans; D015203:Reproducibility of Results
chemical_list: D006455:Hemoglobins, Abnormal; D006443:Hemoglobin A2; D005319:Fetal Hemoglobin