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A rapid method for the determination of glycosylated hemoglobins using high pressure liquid chromatography
Abstract
Hemoglobin (Hb) Alc is a minor component of Hb found in normal individuals but elevated two or threefold in patients with diabetes mellitus. Limited studies have suggested that the level of Hb Alc is proportional to the integrated concentration of glucose over time. Thus it could serve as an index of hyperglycemia. Its measurement may enable a more objective approach to assessing whether or not the control of hyperglycemia can be correlated with the severity of complications of diabetes. Large scale clinicab studies of Hb Alc have not been undertaken for lack of a rapid assay system. This article describes a method of high pressure liquid chromatography (HPLC) which enables the isolation of Hb Alc in 27 min using only 12 microgram of Hb (100 microliter of blood) and a second method for the isolation of total fast Hb components (also elevated in diabetes) in 11 min. Using the first method, a total of 36 assays were performed on the blood of a single normal volunteer over a one month period. the mean level of Hb Alc was 4.95 +/- 0.12% (SD) +/- 0.02% (SEM), while the coefficient of variation (C.V.) was 2.4%. The mean Hb Alc & b level was 1.65 +/- 0.06% +/- 0.01% (C.V. = 3.6%). Values for Hb Alc in 10 normal individuals were 5.06 (mean) +/- 0.32% (SD) +/- 0.01% (SEM). Hb Alc values in 15 patients with diabetes mellitus ranged from 6.8 to 20.0%. The second method was designed to assay Hb Ala, Hb Alb, and Hb Alc as a single peak and yielded results identical to the sum of these components as determined by the first method ( r = 0.98; p less than 0.001).
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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 [18]
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Boukhors, Y., RABASA-LHORET, R. P. R., Langelier, H., Soultan, M., Lacroix, A., & Chiasson, J. L. (2003) · Diabetes & Metabolism · doi:10.1016/s1262-3636(07)70078-2 · Cited by 25
Blackshear, P. J., Shulman, G. I., Roussell, A. M., Nathan, D. M., Minaker, K. L., Rowe, J. W., Robbins, D. C., & Cohen, A. M. (1985) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jcem-61-4-753 · Cited by 25
Increased prevalence of fetal haemoglobin in Type 1 (insulin-dependent) diabetes mellitus
Mullis, P. E., Schuler, J., & ZUPPINGER, K. A. (1989) · Diabetologia · doi:10.1007/bf00285288 · Cited by 25
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Nonenzymatic Glycosylation of Proteins in Vitro and in Vivo
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Metadata
- Authors
- Cole, R. A., Soeldner, J. S., Dunn, P. J., & Bunn, H. F.
- Affiliations
- Joslin Diabetes Center (US)
- Corresponding author
- Cole, R.A.
- ORCID
- Bunn, H.F.
- Venue
- Metabolism
- Publisher
- Elsevier BV
- Published
- 1978-03
- Volume
- 27
- Issue
- 3
- Pages
- 289-301
- ISSN
- 0026-0495 (ISSN-L)
- OpenAlex
- W2053464136
- Cited by
- 183
- References
- 24
- Citations by year
- 2026: 2 · 2025: 1 · 2024: 1 · 2023: 1 · 2022: 1 · 2020: 1 · 2019: 2 · 2015: 1 · 2014: 1
- FWCI
- 16.51
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Metabolism (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
- Health Sciences › Medicine › Genetics (clinical) › Hemoglobinopathies and Related Disorders
- Keywords
- HbA1c, diabetes mellitus, HPLC assay, hemoglobin variants
- MeSH terms
- Chromatography, High Pressure Liquid, Diabetes Mellitus, Hemoglobin A, Hemoglobins, Humans, Microchemistry
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