title: Development of hemoglobin A1c certified reference material by liquid chromatography isotope dilution mass spectrometry.
abstract: We report the development of a National Institute of Metrology (NIM) hemoglobin A(1c) (HbA(1c)) certified reference material (CRM). Each CRM unit contains about 10 μL of hemoglobin. Both hemoglobin and glycated hemoglobin were quantitatively determined by high-performance liquid chromatography (HPLC)-isotope dilution mass spectrometry (IDMS) with synthesized VHLTPE and glycated VHLTPE as standards. The mass fraction of synthesized VHLTPE or glycated VHLTPE was also quantitatively determined by HPLC-IDMS with NIM amino acid CRMs as standards. The homogeneity and stability of the CRMs were examined with a commercial HbA(1c) analyzer based on the HPLC principle. Fifteen units were randomly selected for homogeneity examination, and statistical analysis showed there was no inhomogeneity. Examination of the stability showed that the CRM was stable for at least 6 months at -80 °C. Uncertainty components of the balance, amino acid purity, hydrolysis and proteolysis efficiency, method reproducibility, homogeneity, and stability were taken into consideration for uncertainty evaluation. The certified value of NIM HbA(1c) CRM was expressed as the ratio of HbA(1c) to total hemoglobin in moles, and was (9.6 ± 1.9)%. The CRM can be used as a calibration or validation standard for clinical diagnostics. It is expected to improve the comparability for HbA(1c) measurement in China.
authors.affiliation: College of Science, Beijing University of Chemical Technology, Beijing, China.
authors.forename: Jiaming
authors.initials: J
authors.lastname: Bi
authors.forename: Liqing
authors.initials: L
authors.lastname: Wu
authors.forename: Bin
authors.initials: B
authors.lastname: Yang
authors.forename: Yi
authors.initials: Y
authors.lastname: Yang
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000592:standards*; D003920:Diabetes Mellitus / Q000097:blood; D006442:Glycated Hemoglobin / Q000032:analysis / Q000592:standards*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods / Q000592:standards*; D012015:Reference Standards
chemical_list: D006442:Glycated Hemoglobin