Cited by 183Diabetes Management and ResearchAdvanced Glycation End Products researchHemoglobinopathies and Related Disorders

A rapid method for the determination of glycosylated hemoglobins using high pressure liquid chromatography

Cole, R.A., Soeldner, J.S., Dunn, P.J., Bunn, H.F.·1978·Metabolism

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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Crossref · 10.1016/0026-0495(78)90109-9

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Authors
Cole, R. A., Soeldner, J. S., Dunn, P. J., & Bunn, H. F.
Affiliations
Joslin Diabetes Center (US)
Corresponding author
Cole, R.A.
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 › 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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