Analytical and Bioanalytical Chemistry

, Volume 409, Issue 9, pp 2341–2351 | Cite as

Comparison of potential higher order reference methods for total haemoglobin quantification—an interlaboratory study

  • Claudia FrankEmail author
  • Christine Brauckmann
  • Marie Palos
  • Cristian G. Arsene
  • Joerg Neukammer
  • Maria Estela del Castillo Busto
  • Sabine Zakel
  • Claudia Swart
  • Bernd Güttler
  • Rainer Stosch
Research Paper


The total haemoglobin (Hb) concentration in blood is one of the most frequently measured analytes in clinical medicine because of its significance for evaluating the health state of a human. The spectrophotometric cyanmethaemoglobin (HiCN) method is the internationally accepted conventional reference method to determine this biomarker. It is frequently used in clinical routine diagnostics but is not traceable to the International System of Units and thus does not meet highest metrological demands. A further critical issue is the toxicity of the necessary potassium cyanide. Different methods to solve these problems are reported here. They all were validated against the HiCN method in an interlaboratory comparison by measuring the total Hb concentration present in the certified reference material JCCRM 912-2M. Methods considered were the spectrophotometric alkaline haematin detergent (AHD) method as well as several isotope dilution (ID)-based approaches. The latter include inductively coupled plasma mass spectrometry (ICP-MS), species-specific (SS) ICP-MS, organic MS and Raman spectrometry.

Graphical abstract


Haemoglobin Reference method HiCN Isotope dilution Raman spectroscopy Mass spectrometry 



We thank the Stabinger GmbH for developing and providing a prototype of a high accuracy density measurement device, applicable to small sample volumes.

The research leading to these results has received funding from the European Union on the basis of Decision No. 912/2009/EC within the European Metrology Research Programme (EMRP) HLT-05 2012. The EMRP is jointly funded by the EMRP participating countries within EURAMET and the European Union.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2016_176_MOESM1_ESM.pdf (54 kb)
ESM 1 (PDF 54 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Claudia Frank
    • 1
    Email author
  • Christine Brauckmann
    • 1
  • Marie Palos
    • 2
  • Cristian G. Arsene
    • 1
  • Joerg Neukammer
    • 3
  • Maria Estela del Castillo Busto
    • 2
  • Sabine Zakel
    • 1
  • Claudia Swart
    • 1
  • Bernd Güttler
    • 1
  • Rainer Stosch
    • 1
  1. 1.Physikalisch-Technische Bundesanstalt (PTB)BraunschweigGermany
  2. 2.Laboratoire National de Métrologie et d’essais (LNE)Paris Cedex 15France
  3. 3.Physikalisch-Technische Bundesanstalt (PTB)BerlinGermany

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