An evaluation of the Diamat HPLC analyserfor simultaneous determination ofhaemoglobins A 2 and F

• Published in: Journal of analytical methods in chemistry 1989-01, Vol.11 (6), p.273-279
• Main Authors: Mosca, Andrea, Carpinelli, Assunta, Majavacca, Rita, Cantu′-Rajnoldi, Angelo, Garatti, Massimo, Paleari, Renata, Ferrari, Maurizio, Agape, Vittorio, Maccioni, Liliana, Pisano, Sandra, Galanello, Renzo
• Format: Article
• Language: English
• ISSN: 2090-8865; 2090-8873
• DOI: 10.1155/S1463924689000520

The authors describe a modification of the instrumental parameters of the Diamat fully automated HPLC system for Hb A 2 assay (Bio‐Rad Laboratories, Milan, Italy) in order to obtain simultaneous determination of Hb A 2 and Hb F. Hb A 2 and Hb F measurements are reproducible (within‐run CV 2.6%, with Hb A 2 2.7%; 5.1%, with Hb F 1.3%) and accurate (from a comparison with two microchromatographic techniques for Hb A 2 : r = 0.9639 and 0.9755; with two alkali denaturation procedures for Hb F: r = 0.9990 and 0.9952; with radial immunodiffusion, r = 0.9877). Assay linearity has been confirmed for Hb A 2 concentrations between 0 and 6.0%, and for Hb F between 0 and 60%. The data obtained from the analysis of some pathological samples for Hb Bart′s, Hb H, Hb J Sardegna, Hb Lepore and Hb S are in agreement with cellulose acetate electrophoresis analysis. The Hb A 2 reference intervals for normals (N = 597) and Beta‐thalassemia carriers (N = 200) are respectively (95% limits) 2.02‐3.27 and 3.92‐5.90 in % units. Hb F values measured in normals (N = 968), in β‐thal carriers (N = 302) and in δβ‐thal carriers (N =3) have been found to be consistent with the usual diagnostic parameters. Some minor limitations emerged: the most relevant concerns Hb A 1c , which is overestimated with respect to a reference method (y = 1.217x + 0.16; N = 79; r = 0.9235). A probable interference from labile fractions is responsible for this Hb A 1c inaccuracy.