Paleari Renata, Cannata Monica, Leto Filippo, Maggio Aurelio, Demartis Franca R, Desogus Maria F, Galanello Renzo, Mosca Andrea
Dipartimento di Scienze e Tecnologie Biomediche, Università degli Studi di Milano, Via Fratelli Cervi 93, 20090 Segrate, Milano, Italy.
Clin Biochem. 2005 Feb;38(2):159-65. doi: 10.1016/j.clinbiochem.2004.10.017.
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.
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.
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.
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.
研究一种新型自动化高效液相色谱系统（Tosoh HLV - 723 G7）用于血液中Hb A₂和Hb F定量分析的性能。
对Hb A₂和Hb F测量的不精密度、线性、残留、干扰及样品浓度效应进行研究。采用Bio - Rad Variant II高效液相色谱系统进行方法比较研究。Hb F结果还与碱性变性试验所得结果进行比较。同时研究了一些常见Hb变异体的检测。
Hb A₂批内和批间变异系数（CV）分别在0.8 - 2.2%和2.9 - 7.2%之间，而Hb F在正常情况下CV高达10.0%，在更具临床相关性的Hb F浓度（>1.5%）时CV在1.9 - 5.3%之间。与Bio - Rad Variant II进行Hb A₂测定的比较研究显示出良好的相关性，但突出了校准差异。在两个不同实验室得到以下结果：y = 1.163x - 0.52，r = 0.9918，n = 144（实验室A）；y = 1.060x - 0.40，r = 0.9920，n = 93（实验室B）。关于Hb F的测定，发现所测试方法的测量结果与碱性变性试验（y = 1.0138x - 0.36，r = 0.9842，n = 20）以及Variant II高效液相色谱系统（y = 0.812x + 0.52，r = 0.9835，n = 110）相关性良好。在0.8 - 19%范围内，Hb A₂和Hb F均呈现出出色的线性（r = 0.999）。Hb S、Hb C和Hb D可通过指定的保留时间窗口进行初步鉴定。
新型分析仪Tosoh HLV - 723 G7被发现是一种可靠的方法，可用于Hb A₂和Hb F定量分析以及一些常见Hb变异体的初步鉴定。
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