Anti-interference hemoglobin analysis system by high performance liquid chromatography_Journal of Biomedical Engineering

Authors：

XU Yan ¹ , YAO Tiantian ² , HU Wenyong ² , ZHANG Bo ² ,  GUO Xingming ¹

1. College of Bioengineering, Chongqing University, Chongqing 400044, P.R.China;
2. Shenzhen Lifotronic of Technology Co., Ltd., Shenzhen, Guangdong 518055, P.R.China;

Corresponding author：

GUO Xingming, Email: guoxm@cqu.edu.cn

Keywords：

high performance liquid chromatography; glycated hemoglobin; thalassemia factor HbF/HbA2; hemoglobin variant

DOI：

10.7507/1001-5515.202101069

Video：

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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.

Citation： XU Yan, YAO Tiantian, HU Wenyong, ZHANG Bo, GUO Xingming. Anti-interference hemoglobin analysis system by high performance liquid chromatography. Journal of Biomedical Engineering, 2021, 38(5): 940-950. doi: 10.7507/1001-5515.202101069 Copy

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2.	Williams R, Karuranga S, Malanda B, et al. Global and regional estimates and projections of diabetes-related health expenditure: results from the International Diabetes Federation Diabetes Atlas, 9th edition. Diabetes Res Clin Pract, 162: 108272-108285.
3.	Little R R, Roberts W L. A review of variant hemoglobins interfering with hemoglobin A1c measurement. J Diabetes Sci Technol, 2009, 3(3): 446-451.
4.	Taher A T, Weatherall D J, Cappellini M D. Thalassaemia. Lancet, 2018, 391(10116): 155-167.
5.	Rahim F. Microcytic hypochromic anemia patients with thalassemia: genotyping approach. Indian J Med Sci, 2009, 63(3): 101-108.
6.	Guzelgul F, Seydel G S, Aksoy K. β-Globin gene mutations in pediatric patients with β-Thalassemia in the region of Çukurova, Turkey. Hemoglobin, 2020(6): 1-5.
7.	World Health Organization. Use of glycated haemoglobin (HbA1c) in diagnosis of diabetes mellitus: abbreviated report of a WHO consultation. J Autom Reasoning, 2011, 14(2): 123-125.
8.	American Diabetes Association. Diagnosis and classification of diabetes mellitus. Diabetes Care, 2013, 36(Suppl1): S67-S74.
9.	Malkani S, Mordes J P. Implications of using hemoglobin A1C for diagnosing diabetes mellitus. Am J Med, 2011, 124(5): 395-401.
10.	Randie R. Glycated hemoglobin standardization-National Glycohemoglobin Standardization Program (NGSP) perspective. Clin Chem Lab Med, 2003, 41(9): 1191-1198.
11.	Lenters-Westra E, English E. Evaluating new HbA1c methods for adoption by the IFCC and NGSP reference networks using international quality targets. Clin Chem Lab Med, 2017, 55(9): 1426-1434.
12.	Koga M, Inada S, Yoshino K, et al. IFCC-HbA1c may be more useful than NGSP-HbA1c in the comparison between HbA1c values in variant hemoglobin with a mutation on α chain and β chain. Ann Clin Lab Sci, 2019, 49(5): 650-655.
13.	Gupta S, Chauhan N, Jain U. Laboratory diagnosis of HbA1c: A review. J Nano Res, 2017, 5(4): 120-135.
14.	赵锐, 黄新成, 吴丽琴, 等. 糖化血红蛋白检测方法的研究进展. 实用医技杂志, 2014, 21(2): 167-168.
15.	聂庆东, 刘君廷, 祁瑞环. 高效液相色谱法检测全血糖化血红蛋白的稳定性分析及其应用价值. 国际检验医学杂志, 2012, 33(8): 967-968.
16.	Koga M, Okuda M, Inada S, et al. HbA1c levels measured by enzymatic assay during off-site health checkups are lower than those measured by on-site HPLC assay. Diabetol Int, 2020, 11(1): 67-71.
17.	Jalali M T, Bavarsad S S, Hesam S, et al. Assessing agreement between the three common clinical measurement methods of HbA1c. J Diabetes Met Dis, 2020, 19(1): 273-279.
18.	Fellows S E, Cipriano G C. Variation between point-of-care and laboratory HbA1c testing in clinical practice. P R Health Sci J, 2019, 38(3): 189-191.
19.	Altawallbeh G, Makky V F, Saenger A K, et al. Evaluation of an ion-exchange HPLC device for HbA1c measurement. J Appl Lab Med, 2020, 5(4): 695-703.
20.	Okamoto T, Shima H, Noma Y, et al. Hereditary spherocytosis diagnosed with extremely low glycated hemoglobin compared to plasma glucose levels. Diabetol Int, 2020, 12(2): 213-219.
21.	Fabella T D. High performance liquid chromatography (HPLC) screening among Filipinos with suspected thalassemia. Acta Med Phil, 2020, 12(4): 53-60.
22.	唐娟, 李卿, 邓爽, 等. 对 α 地中海贫血 Hb H 病糖化血红蛋白不同检测方法结果的分析. 中国糖尿病杂志, 2018, 26(3): 193-196.
23.	Reeve J, Blake L, Griffin D, et al. Incidental detection of haemoglobin (Hb) variants during high performance liquid chromatography (HPLC) analysis of HbA_1c: is it time for a standardised approach to reporting?. Ir J Med Sci, 2015, 184(2): 353-355.
24.	邹蓓. 地中海贫血对糖化血红蛋白测定的影响. 当代医学, 2012, 18(10): 102-103.
25.	温冬梅, 张秀明, 索明环, 等. 血红蛋白变异体对糖化血红蛋白测定结果的干扰及处理. 中华检验医学杂志, 2014, 37(2): 123-126.
26.	Li R, Tang H, Kan L, et al. Evaluation on the separated effect of 13 hemoglobin variants by a new automatic Hba1c analyzer. J Clin Lab Anal, 2020, 34(10): e23446.
27.	曹新玲, 苏琳, 曹光明, 等. 高效液相色谱法检测色素时梯度洗脱条件的改进. 中国计量, 2019, 288(11): 95-96.
28.	Liu L F, Xu Z. Study of chromatographic fractions from carbon dots isolated by column chromatography and a binary gradient elution via RP-HPLC. Anal methods, 2019, 11(6): 12121-12131.
29.	邹明宇. 聚合物基质阴离子交换固相萃取填料的制备及应用. 齐齐哈尔: 齐齐哈尔大学, 2018.
30.	Zhang C Q, Peng Q H, Wang S, et al. Preparation of poly (KH570-co-DVB) microspheres and application in HPLC for separation of benzene and benzene homologs. Ferroelectrics, 2020, 562(1): 145-151.
31.	Peng Q, Wu Y, Cong H, et al. Preparation of monodisperse porous polymeric ionic liquid microspheres and their application as stationary phases for HPLC. Talanta, 2019, 10(208): 120462-120474.
32.	Clinical and Laboratory Standards Institute. Evaluation of precision performance of quantitative measurement methods: EP5-A2. Zambia: CLSI, 2004: 1.
33.	Clinical and Laboratory Standards Institute. Evaluation of the linearity of quantitative measurement procedures: EP6-A. Zambia: CLSI, 2003: 3.
34.	Clinical and Laboratory Standards Institute. Method comparison and bias estimation using patient samples: EP9- A3. Zambia: CLSI, 2013: 6.

1. Bi Y, Yu X, Li M, et al. Prevalence and Control of Diabetes in Chinese adults: The China Metabolic Risk Factor Study// Scientific Sessions of the American-Heart-Association on Epidemiology. Shanghai: IEEE, 2013: 1-4.
2. Williams R, Karuranga S, Malanda B, et al. Global and regional estimates and projections of diabetes-related health expenditure: results from the International Diabetes Federation Diabetes Atlas, 9th edition. Diabetes Res Clin Pract, 162: 108272-108285.
3. Little R R, Roberts W L. A review of variant hemoglobins interfering with hemoglobin A1c measurement. J Diabetes Sci Technol, 2009, 3(3): 446-451.
4. Taher A T, Weatherall D J, Cappellini M D. Thalassaemia. Lancet, 2018, 391(10116): 155-167.
5. Rahim F. Microcytic hypochromic anemia patients with thalassemia: genotyping approach. Indian J Med Sci, 2009, 63(3): 101-108.
6. Guzelgul F, Seydel G S, Aksoy K. β-Globin gene mutations in pediatric patients with β-Thalassemia in the region of Çukurova, Turkey. Hemoglobin, 2020(6): 1-5.
7. World Health Organization. Use of glycated haemoglobin (HbA1c) in diagnosis of diabetes mellitus: abbreviated report of a WHO consultation. J Autom Reasoning, 2011, 14(2): 123-125.
8. American Diabetes Association. Diagnosis and classification of diabetes mellitus. Diabetes Care, 2013, 36(Suppl1): S67-S74.
9. Malkani S, Mordes J P. Implications of using hemoglobin A1C for diagnosing diabetes mellitus. Am J Med, 2011, 124(5): 395-401.
10. Randie R. Glycated hemoglobin standardization-National Glycohemoglobin Standardization Program (NGSP) perspective. Clin Chem Lab Med, 2003, 41(9): 1191-1198.
11. Lenters-Westra E, English E. Evaluating new HbA1c methods for adoption by the IFCC and NGSP reference networks using international quality targets. Clin Chem Lab Med, 2017, 55(9): 1426-1434.
12. Koga M, Inada S, Yoshino K, et al. IFCC-HbA1c may be more useful than NGSP-HbA1c in the comparison between HbA1c values in variant hemoglobin with a mutation on α chain and β chain. Ann Clin Lab Sci, 2019, 49(5): 650-655.
13. Gupta S, Chauhan N, Jain U. Laboratory diagnosis of HbA1c: A review. J Nano Res, 2017, 5(4): 120-135.
14. 赵锐, 黄新成, 吴丽琴, 等. 糖化血红蛋白检测方法的研究进展. 实用医技杂志, 2014, 21(2): 167-168.
15. 聂庆东, 刘君廷, 祁瑞环. 高效液相色谱法检测全血糖化血红蛋白的稳定性分析及其应用价值. 国际检验医学杂志, 2012, 33(8): 967-968.
16. Koga M, Okuda M, Inada S, et al. HbA1c levels measured by enzymatic assay during off-site health checkups are lower than those measured by on-site HPLC assay. Diabetol Int, 2020, 11(1): 67-71.
17. Jalali M T, Bavarsad S S, Hesam S, et al. Assessing agreement between the three common clinical measurement methods of HbA1c. J Diabetes Met Dis, 2020, 19(1): 273-279.
18. Fellows S E, Cipriano G C. Variation between point-of-care and laboratory HbA1c testing in clinical practice. P R Health Sci J, 2019, 38(3): 189-191.
19. Altawallbeh G, Makky V F, Saenger A K, et al. Evaluation of an ion-exchange HPLC device for HbA1c measurement. J Appl Lab Med, 2020, 5(4): 695-703.
20. Okamoto T, Shima H, Noma Y, et al. Hereditary spherocytosis diagnosed with extremely low glycated hemoglobin compared to plasma glucose levels. Diabetol Int, 2020, 12(2): 213-219.
21. Fabella T D. High performance liquid chromatography (HPLC) screening among Filipinos with suspected thalassemia. Acta Med Phil, 2020, 12(4): 53-60.
22. 唐娟, 李卿, 邓爽, 等. 对 α 地中海贫血 Hb H 病糖化血红蛋白不同检测方法结果的分析. 中国糖尿病杂志, 2018, 26(3): 193-196.
23. Reeve J, Blake L, Griffin D, et al. Incidental detection of haemoglobin (Hb) variants during high performance liquid chromatography (HPLC) analysis of HbA_1c: is it time for a standardised approach to reporting?. Ir J Med Sci, 2015, 184(2): 353-355.
24. 邹蓓. 地中海贫血对糖化血红蛋白测定的影响. 当代医学, 2012, 18(10): 102-103.
25. 温冬梅, 张秀明, 索明环, 等. 血红蛋白变异体对糖化血红蛋白测定结果的干扰及处理. 中华检验医学杂志, 2014, 37(2): 123-126.
26. Li R, Tang H, Kan L, et al. Evaluation on the separated effect of 13 hemoglobin variants by a new automatic Hba1c analyzer. J Clin Lab Anal, 2020, 34(10): e23446.
27. 曹新玲, 苏琳, 曹光明, 等. 高效液相色谱法检测色素时梯度洗脱条件的改进. 中国计量, 2019, 288(11): 95-96.
28. Liu L F, Xu Z. Study of chromatographic fractions from carbon dots isolated by column chromatography and a binary gradient elution via RP-HPLC. Anal methods, 2019, 11(6): 12121-12131.
29. 邹明宇. 聚合物基质阴离子交换固相萃取填料的制备及应用. 齐齐哈尔: 齐齐哈尔大学, 2018.
30. Zhang C Q, Peng Q H, Wang S, et al. Preparation of poly (KH570-co-DVB) microspheres and application in HPLC for separation of benzene and benzene homologs. Ferroelectrics, 2020, 562(1): 145-151.
31. Peng Q, Wu Y, Cong H, et al. Preparation of monodisperse porous polymeric ionic liquid microspheres and their application as stationary phases for HPLC. Talanta, 2019, 10(208): 120462-120474.
32. Clinical and Laboratory Standards Institute. Evaluation of precision performance of quantitative measurement methods: EP5-A2. Zambia: CLSI, 2004: 1.
33. Clinical and Laboratory Standards Institute. Evaluation of the linearity of quantitative measurement procedures: EP6-A. Zambia: CLSI, 2003: 3.
34. Clinical and Laboratory Standards Institute. Method comparison and bias estimation using patient samples: EP9- A3. Zambia: CLSI, 2013: 6.

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Anti-interference hemoglobin analysis system by high performance liquid chromatography

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