Effect of HbS in the determination of HbA₂ with the Menarini HA‐8160 analyzer and comparison with other instruments

Abstract

It is known that the presence of hemoglobin S (HbS) affects the determination of hemoglobin A(2) (HbA(2)) levels in clinical samples. We quantitated this effect using the Menarini HA-8160 analyzer and compared with other instruments (HELENA beta-thal quik column, TOSOH HLC-723G7 and BIORAD Variant II) using the HELENA SAS-MX alkaline gel electrophoresis kit as the reference method. The %HbA(2) values from the HA-8160 analyzer and the alkaline gel electrophoresis show a good linear correlation in the absence of HbS. A strong positive bias in the %HbA(2) values from the HA-8160 is apparent in the presence of HbS in the samples, when compared with the alkaline electrophoresis. The analytical imprecision and bias of the three HPLC instruments are comparable both in the presence and absence of HbS. The manual column method shows a lower bias in the absence of HbS but is more affected when HbS is present in the samples.

Interlaboratory comparison of current high‐performance methods for HbA2

“…With regard to the imprecision, the data obtained in our study proven that all methods were performing better than 4.5% CV, as also recently reported (Anagnostopoulos et al. , 2009), thus confirming that the quality of the automated HPLC and CE methods is higher with respect to the imprecision obtainable years ago with the minicolumns methods (Brosius et al.…”

“…With regard to the imprecision, the data obtained in our study proven that all methods were performing better than 4.5% CV, as also recently reported (Anagnostopoulos et al, 2009), thus confirming that the quality of the automated HPLC and CE methods is higher with respect to the imprecision obtainable years ago with the minicolumns methods (Brosius et al, 1978), or by cellulose acetate electrophoresis followed by densitometry, or after elution of the HbA and HbA 2 bands and spectrophotometric quantitation (International Committee for Standardization in Haemathology 1978). However, from a recent study we have performed under rigorous conditions in agreement with the approach recommended (Fraser & Harris, 1989), HbA 2 seems to have a very narrow intra-individual biological variability, of the order of 0.4% (as CV), as for a typical genetic marker (Paleari et al, 2011).…”

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

“…A strong positive bias in the HbA 2 values from the ADAMS ™ A1C HA‐8160 T is apparent in the presence of HbS ; this spurious increment was not found in the new analyzer.…”

Hemoglobin Varyantlarının Ölçümünde Interlab-G26 Elektroforezi ve Primus Ultra2 HPLC Yöntemlerinin Karşılaştırılması

“…Hemoglobin varyantlarının tanısı için kullanılan yöntemler tarama testleri ve kesin tanı yöntemleri olarak ikiye ayrılır. Hemoglobin A2 gibi normal hemoglobin varyantlarında artma veya Hemoglobin S gibi anormal hemoglobin varyantlarının tespitinde tarama testi olarak başlıca alkali ve asit elektroforezi ile otomatize edilmiş yüksek basınçlı sıvı kromatografisi (HPLC) sistemleri kullanılır (6,8 (9). Elektroforez yöntemi ise daha fazla manuel işlem gerektirmesine, düşük konsantrasyonlardaki Hb A2 düzeyini belirlemede daha az sensitif olmasına ve uzun süren çalışma prosedürüne rağmen, nispeten düşük maliyeti ve yarı otomatize cihazların kullanıma girmesiyle tarama yöntemi olarak ülkemizde halen yaygın olarak kullanılmaktadır.…”