Preparation of a candidate primary reference material for the international standardisation of HbA1c determinations

被引:95
作者
Finke, A
Kobold, U
Hoelzel, W
Weykamp, C
Miedema, K
Jeppsson, JO
机构
[1] Boehringer Mannheim GmbH, D-82377 Penzberg, Germany
[2] Queen Beatrix Hosp, Dept Clin Chem, Winterswijk, Netherlands
[3] De Weezenlanden Ziekenhuis, Dept Clin Chem, Zwolle, Netherlands
[4] Malmo Univ Hosp, Dept Clin Chem, Malmo, Sweden
关键词
beta-N-terminal glycated hemoglobin A (HbA1c); non-glycated hemoglobin A (HbA0); hemoglobin; glycohemoglobin; reference material; peptides; electrospray ionisation mass spectrometry; reference method; international standardisation;
D O I
10.1515/CCLM.1998.051
中图分类号
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号
1001 ;
摘要
We prepared a candidate primary reference material for the forthcoming international standardisation of beta-N-terminal glycated hemoglobin A measurements. It consists of well-defined mixtures of purified beta-N-terminal glycated hemoglobin A and non-glycated hemoglobin A. First, beta-N-terminal glycated hemoglobin A and nonglycated hemoglobin A were isolated, purified to homogeneity, and characterised. The techniques used were cation exchange and affinity chromatography for the purification, and high performance liquid chromatography, capillary isoelectric focusing, electrospray ionisation mass spectrometry, and peptide mapping for the characterisation. Hemoglobins from blood of healthy, non-diabetic volunteers were obtained with a purity of >99.5 % for non-glycated hemoglobin A and of >98.5 % for beta-N-terminal glycated hemoglobin A. However, results from peptide mapping indicate that the beta-N-terminal glycated hemoglobin A preparations still contain some non-beta-N-terminal glycated hemoglobins, co-eluting with beta-N-terminal glycated hemoglobin A. The exact content of beta-N-terminal glycated hemoglobin A in these preparations could be determined by a procedure consisting of standard addition, enzymatic cleavage and quantification of the resulting beta-N-terminal peptides to be in the range from 95 - 97.5 %. Since the beta-N-terminal glycated hemoglobin A and non-glycated hemoglobin A content could be exactly determined in the materials prepared, mixtures of both components could be successfully used to calibrate the candidate reference methods.
引用
收藏
页码:299 / 308
页数:10
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