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A high‐performance liquid chromatography method for hemoglobin A1c
Abstract
Hemoglobin A1c (HbA1c) is a glycosylated derivative of hemoglobin and is one of a family of derivatives whose concentrations are elevated in patients with diabetes mellitus. Published methods for the measurement of HbA1c are relatively tedious and require modest amounts of blood. A high-performance liquid chromatographic (HPLC) method for the determination of HbA1c is presented. The method is rapid (20 minutes), precise (coefficient of variation of 5-10 per cent), uses small amounts of sample (3 microliter.), can be automated. A sample preparation technique using filtration was developed that shortened and simplified preparation of venous blood and allowed use of capillary samples. HbA1c was measured by this method in three age-stratified groups of controls and a group of insulin-requiring juvenile diabetics. There was clear separation of HbA1c values between all normals (5.9 +/- 1.3, 5.6 +/- 0.7, 7.1 +/- 0.9 per cent) and the diabetics (12.1 +/- 2.4 per cent). Use of this method can facilitate large-scale clinical investigations and permit biochemical investigations of the metabolism and formation of hemoglobin A1c where small sample sizes are necessary.
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“The coefficient of variation of this reference method was < 3%.”
From Twenty‐four‐hour profiles of plasma glucose, insulin, C‐peptide and free fatty acid in subjects with varying degrees of glucose tolerance following short‐term, medium‐dose prednisone (20 mg/day) treatment: evidence for differing effects on insulin secretion and action (2012) · cites this paper as [19]
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“HbA1c was measured using a highperformance liquid chromatography methodology (21,22).”
MethodKendall, D. M., Riddle, M. C., Rosenstock, J., Zhuang, D., Kim, D. D., Fineman, M. S., & Baron, A. D. (2005) · Diabetes Care · doi:10.2337/diacare.28.5.1083 · Cited by 1,177
“A1C was measured using a highperformance liquid chromatography methodology (25,26).”
MethodDrucker, D. J., Buse, J. B., Taylor, K., Kendall, D. M., Trautmann, M., Zhuang, D., & Porter, L. (2008) · The Lancet · doi:10.1016/s0140-6736(08)61206-4 · Cited by 1,070
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“As [17] we found a correlation between HbAz and age, but in contrast to [ 18 ] not for HbAIr in normals aged 20-92 years.”
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An automated "high-pressure" liquid-chromatographic assay for hemoglobin A1c.
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Glycosylated hemoglobins and diabetes mellitus
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Hemoglobin A1C separation by isoelectric focusing
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Auditory Function in Children with Diabetes Mellitus
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Endothelial dysfunction in type 2 diabetes mellitus subjects with peripheral artery disease
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Hyperinsulinemia promotes aberrant histone acetylation in triple-negative breast cancer
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Plasma levels of apoprotein B in patients with diabetes mellitus: The effect of glycemic control
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O'Neal, D. N., Dragicevic, G., Rowley, K., Ansari, M. Z., Balazs, N. D. H., Jenkins, A. J., & Best, J. D. (2003) · Diabetes Care · doi:10.2337/diacare.26.1.199 · Cited by 37
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Abnormal glucoregulation during exercise in type II (non-insulin-dependent) diabetes
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Link between glycation and lipoxidation in red blood cells in diabetes
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A field effect transistor (FET)-based immunosensor for detection of HbA1c and Hb
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Glycaemic control and familial factors determine hyperlipidaemia in early childhood diabetes
Abraha, A., Schultz, C. J., Bahu, T. K., James, T., Watts, A., STRATTON, I. M., Matthews, D. R., & Dunger, D. B. (1999) · Diabetic Medicine · doi:10.1046/j.1464-5491.1999.00109.x · Cited by 31
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“Measurement of glycated hemoglobin is useful in evaluating long-term control of blood glucose (1-3) and there are numerous methods for doing so: cation-exchange (4), liquid chromatography (5), agarose gel electrophoresis (6), colorimetric and spectrophotometric methods (7, 8), isoelectric focusing (9), immunoassays (1…”
MethodGlycohemoglobin in diabetic pregnancy: A sequential study
Widness, J. A., Schwartz, H. C., Kahn, C. B., et al. (1980) · American Journal of Obstetrics and Gynecology · doi:10.1016/0002-9378(80)90631-6 · Cited by 30
Roles of site and timing of the morning insulin injection in type 1 diabetes
WITT, M. F., White, N. H., & Santiago, J. V. (1983) · The Journal of Pediatrics · doi:10.1016/s0022-3476(83)80577-0 · Cited by 26
Retinopathy in African Americans and Whites With Insulin-Dependent Diabetes Mellitus
Arfken, C. L. (1994) · Archives of Internal Medicine · doi:10.1001/archinte.1994.00420220097011 · Cited by 28
Ali, Z., Ma, H., Wali, A., Ayim, I., Rashid, M. T., & Younas, S. (2019) · Tropical Journal of Pharmaceutical Research · doi:10.4314/tjpr.v17i12.23 · Cited by 27
“, Torrance, CA, USA), photodiode-array detector Jasco MD-915 (Tokyo, Japan) and an Agilent UV- detector model 1100, was used to analyze folate and HbA1c concentrations [19,20].”
MethodZhao, H., Qiu, X., Su, E., et al. (2021) · Analytica Chimica Acta · doi:10.1016/j.aca.2021.339379 · Cited by 27
Methods for the Analysis of Glycated Haemoglobins: What is Being Measured?
John, W. G., Bullock, D. G., & MacKenzie, F. (1992) · Diabetic Medicine · doi:10.1111/j.1464-5491.1992.tb01708.x · Cited by 27
A semi-automated method for the determination of glycosylated haemoglobin
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Recent Advances in Glycosylated Hemoglobin Measurements
Goldstein, D. E., Wiedmeyer, H.-M., England, J. D., Little, R. R., Parker, K. M., & Simon, M. (1984) · CRC Critical Reviews in Clinical Laboratory Sciences · doi:10.3109/10408368409165782 · Cited by 26
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Defective glucose counterregulation limits intensive therapy of diabetes mellitus
Santiago, J. V., White, N. H., Skor, D. A., Levandoski, L. A., Bier, D. M., & Cryer, P. E. (1984) · American Journal of Physiology-Endocrinology and Metabolism · doi:10.1152/ajpendo.1984.247.2.e215 · Cited by 24
Nonenzymatic Glycosylation of Proteins in Vitro and in Vivo
Thorpe, S. R., & Baynes, J. (1982) · Elsevier eBooks · doi:10.1016/b978-0-12-356103-9.50012-9 · Cited by 24
Nabrdalik-Leśniak, D., Nabrdalik, K., Sedlaczek, K., Główczyński, P., Kwiendacz, H., Sawczyn, T., Hajzler, W., Drożdż, K., Hendel, M., Irlik, K., et al. (2021) · Oxidative Medicine and Cellular Longevity · doi:10.1155/2021/5593589 · Cited by 22
“HbA1c was measured with the HLPC (high-performance liquid chromatography) method [25], creatinine was measured with Jaffe’s colorimetric assay [26] (Cobas Integra 800, Roche Diagnostics®), and eGFR was calculated according to CKDEPI formula [27].”
MethodEffects of dietary cholesterol on plasma lipoproteins and their subclasses in IDDM patients
Romanò, G., Tilly-Kiesi, M., Patti, L., Taskinen, M., Pacioni, D., Cassader, M., Riccardi, G., & Rivellese, A. A. (1998) · Diabetologia · doi:10.1007/s001250050889 · Cited by 22
“HbA 1c was measured by a high pressure liquid chromatography [ 27 ].”
MethodPlasma Lipids in Patients With Type I Diabetes Mellitus
Semenkovich, C. F. (1989) · Archives of Internal Medicine · doi:10.1001/archinte.1989.00390010069006 · Cited by 22
Sun, L., Zhang, X., Wang, T., et al. (2017) · Experimental and Therapeutic Medicine · doi:10.3892/etm.2017.4866 · Cited by 20
“HbA1c levels were measured using high-performance liquid chromatography as previous described (20).”
MethodKordonouri, O., Jörres, A., Muller, C. R., et al. (1992) · Scandinavian Journal of Clinical and Laboratory Investigation · doi:10.3109/00365519209088381 · Cited by 21
Comparison of hemoglobin A1c and hemoglobin A1 in diabetic patients
James, T., Davis, J. E., McDonald, J. M., Santiago, J. V., & Ladenson, J. H. (1981) · Clinical Biochemistry · doi:10.1016/0009-9120(81)90135-1 · Cited by 20
Self-citation
Metadata
- Authors
- Davis, J. E., Mcdonald, J. M., & Jarett, L.
- Affiliations
- Washington University in St. Louis (US)
- Venue
- Diabetes
- Publisher
- American Diabetes Association
- Published
- 1978-02-01
- Volume
- 27
- Issue
- 2
- Pages
- 102-107
- ISSN
- 0012-1797 (ISSN-L)
- 1939-327X
- OpenAlex
- W2013055957
- Cited by
- 203
- References
- 0
- Citations by year
- 2026: 1 · 2025: 7 · 2024: 4 · 2023: 3 · 2022: 2 · 2021: 4 · 2020: 3 · 2019: 6 · 2018: 5 · 2017: 3 · 2016: 1 · 2015: 5 · 2014: 2 · 2013: 1 · 2012: 3
- FWCI
- 10.47
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Diabetes (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Diabetes Management and Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Advanced Glycation End Products research
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical Chemistry and Chromatography
- Keywords
- HbA1c, HPLC assay, diabetes mellitus, type 1 diabetes, capillary blood samples, sample preparation, small sample volumes, automated assay
- MeSH terms
- Adolescent, Adult, Age Factors, Child, Child, Preschool, Chromatography, High Pressure Liquid, Diabetes Mellitus, Diabetes Mellitus, Type 1, Glycosides, Hemoglobins, Humans, Middle Aged
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