High performance liquid chromatography (HPLC) is currently the mainstream technology for detecting hemoglobin. Glycated hemoglobin (HbA1c) is a gold indicator for diagnosing diabetes, however, the accuracy of HbA1c test is affected by thalassemia factor hemoglobin F (HbF)/hemoglobin A2 (HbA2) and variant hemoglobin during HPLC analysis. In this study, a new anti-interference hemoglobin analysis system of HPLC is proposed. In this system, the high-pressure three-gradient elution method was improved, and the particle size and sieve plate aperture in the high-pressure chromatography column and the structure of the double-plunger reciprocating series high-pressure pump were optimized. The system could diagnose both HbA1c and thalassemia factor HbF/HbA2 and variant hemoglobin, and the performance of the system was anti-interference and stable. It is expected to achieve industrialization. In this study, the HbA1c and thalassemia factor HbF/HbA2 detection performance was compared between this system and the world's first-line brand products such as Tosoh G8, Bio-Rad Ⅶ and D10 glycosylated hemoglobin analysis system. The results showed that the linear correlation between this system and the world-class system was good. The system is the first domestic hemoglobin analysis system by HPLC for screening of HbA1c and thalassemia factor HbF/HbA2 rapidly and accurately.
高效液相色谱法（HPLC）是目前主流的检测血红蛋白技术，糖化血红蛋白（HbA1c）是诊断糖尿病的金指标，但 HPLC 检测时 HbA1c 易受到地中海贫血因子血红蛋白 F（HbF）/血红蛋白 A2（HbA2）和血红蛋白变异体的干扰而影响检测结果的准确性。本研究基于高效液相色谱法提出一种新型抗干扰血红蛋白分析系统，本系统通过改进高压三梯度洗脱法，优化高压层析柱中微球粒径和筛板孔径，并优化双柱塞往复式串联高压泵的结构，既能检测 HbA1c 又可检测地中海贫血因子 HbF/HbA2 和血红蛋白变异体，抗干扰性好，性能稳定，有望实现产业化。本系统与日本东曹 G8、美国伯乐Ⅶ和 D10 糖化血红蛋白分析系统等世界一线品牌产品进行 HbA1c 和地中海贫血因子 HbF/HbA2 检测性能对比，结果表明，本系统与世界一流系统的线性相关性好，检测性能优异，是国内首次实现快速、精准筛查糖化血红蛋白和地中海贫血因子 HbF/HbA2 的高效液相色谱法血红蛋白分析系统。.
Keywords: glycated hemoglobin; hemoglobin variant; high performance liquid chromatography; thalassemia factor HbF/HbA2.