ICSH recommendations for assessing automated high‐performance liquid chromatography and capillary electrophoresis equipment for the quantitation of HbA₂

Abstract

Automated high performance liquid chromatography and Capillary electrophoresis are used to quantitate the proportion of Hemoglobin A 2 (HbA 2 ) in blood samples order to enable screening and diagnosis of carriers of b-thalassemia. Since there is only a very small difference in HbA 2 levels between people who are carriers and people who are not carriers such analyses need to be both precise and accurate. This paper examines the different parameters of such equipment and discusses how they should be assessed.

Evaluation of the suitability of the World Health Organization International Reference Reagent for Hb A2 quantitation (89/666) for continued use

“…An acceptable performance limit of ±0.25% of Hb A 2 of the assigned value was applied to the results, based on suggested published performance assessment limits . With the exception of the Bio‐Rad Variant II analysers running the β‐thalassaemia Program or the Dual Program and the Menarini HA8180T analyser, the median results for the WHO IRR from all analysers included in the exercise fell within these acceptable limits.…”

“…An acceptable performance limit of ±0.25% of Hb A 2 of the assigned value was applied to the results, based on suggested published performance assessment limits. 8,9 With the exception of the Bio-Rad…”

Analytical evaluation of the ADAMS™ A1c HA 8180 thalassemia mode high‐pressure liquid chromatography analyser for the measurement of HbA2 and HbF

“…The different items were evaluated according to the International Council for Standarization in Hematology (ICSH) recently published document for HPLC systems measuring HbA 2 , Clinical and Laboratory Standards Institute (CLSI) , or manufacture's guidelines.…”

“…Carry over and within run precision were evaluated according to ICLH guideline . Carryover was assessed analyzing a sample with high HbA 2 values three times (H1, H2, and H3) followed immediately by a sample with low HbA 2 levels (L1, L2, and L3); the percentage of carryover was calculated using the following formula (L1‐L3)/(H3‐L2)*100.…”

Comparison between capillary zone electrophoresis and capillary isoelectric focusing for thalassemia screening in southern China

“…According to the data from summary of HG which is a proficiency test program of CAP (College of American Pathologists), in 2016, users equipped with CE analyzers (from Sebia Company) firstly outnumbered users equipped with high‐performance liquid chromatography (HPLC) analyzers (from Bio‐Rad Company). And in recent years, researchers have proved in several studies that capillary zone electrophoresis (CZE), the most widespread, well‐known, and automated CE method, has equal or better analytical performance than HPLC in thalassemia screening and diagnosis, even though the latter 1 is still been considered as the gold standard for hemoglobin analysis …”