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Related Experiment Videos

Development of analytical methods to evaluate SFH.

K Nakai1, H Abe, N Matsuda

  • 1Hokkaido Red Cross Blood Center, Sapporo, Japan.

Biomaterials, Artificial Cells, and Immobilization Biotechnology : Official Journal of the International Society for Artificial Cells and Immobilization Biotechnology
|January 1, 1992
PubMed
Summary

Two new analytical methods, HPLC-UV and enzyme immunoassay, were developed to measure stroma content in solutions of Stroma-Free Hemoglobin (SFH). These methods confirmed that ultrafiltration and BMM-40 nm effectively purify hemoglobin, removing over 99.7% of phospholipids and nearly all blood group antigens.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Biotechnology

Background:

  • Stroma-free hemoglobin (SFH) solutions are crucial for various applications.
  • Accurate determination of residual stroma components is essential for SFH quality control.
  • Existing purification methods require robust analytical techniques for validation.

Purpose of the Study:

  • To develop and validate analytical methods for quantifying stroma content in SFH.
  • To evaluate the efficacy of different SFH purification techniques.
  • To ensure the purity and safety of hemoglobin solutions.

Main Methods:

  • High-Performance Liquid Chromatography with Ultraviolet detection (HPLC-UV) for phospholipid analysis.
  • Enzyme Immunoassay (EIA) for blood group antigen determination.

Related Experiment Videos

  • Evaluation of three SFH preparations using these analytical methods.

    • Main Results:

    • The 36,000 x g centrifugation method yielded SFH with higher stroma component levels.
    • BMM-40 nm and 100 kd ultrafiltration methods removed >99.7% phospholipids and nearly all blood group antigens.
    • The developed analytical methods proved effective in assessing SFH purity.

    Conclusions:

    • HPLC-UV and EIA are reliable tools for stroma content determination in SFH.
    • BMM-40 nm and ultrafiltration are highly effective purification methods for hemoglobin solutions.
    • These validated methods support the quality control of highly purified hemoglobin products.