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Development of an improved standard reference material for folate vitamers in human serum

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Abstract

The US National Institute of Standards and Technology (NIST) developed a Standard Reference Material® (SRM®) 3949 Folate Vitamers in Frozen Human Serum to replace SRM 1955 Homocysteine and Folate in Human Serum. The presence of increased endogenous levels of folic acid and 5-methyltetrahydrofolate (5mTHF) in SRM 3949, enhanced folate stability via addition of ascorbic acid, and inclusion of values for additional minor folates are improvements over SRM 1955 that should better serve the clinical folate measurement community. The new SRM contains folates at three levels. To produce SRM 3949, pilot sera were collected from 15 individual donors, 5 of whom were given a 400-µg folic acid supplement 1 h prior to blood draw to increase serum levels of 5mTHF and folic acid for the high-level material. To stabilize the folates, 0.5% (mass concentration) ascorbic acid was added as soon as possible after preparation of serum. These pilot sera were screened for five folates plus the pyrazino-s-triazine derivative of 4-α-hydroxy-5-methyltetrahydrofolate (MeFox) at the US Centers for Disease Control and Prevention (CDC) by isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC–MS/MS). Based on these results, a blending protocol was specified to obtain the three desired folate concentrations for SRM 3949. ID-LC–MS/MS analysis at the CDC and NIST was utilized to assign values for folic acid and 5mTHF, as well as several minor folates.

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Funding

This study was supported in part by funding provided by the Office of Dietary Supplements at the National Institutes of Health (NIH-ODS) to the National Institute of Standards and Technology (NIST).

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Authors and Affiliations

  1. National Institute of Standards and Technology, Gaithersburg, MD, USA

    Johanna E. Camara, Jeanita S. Pritchett, Yasmine C. Daniels, Mary Bedner, Michael A. Nelson, Mark S. Lowenthal, Karen W. Phinney, Katherine E. Sharpless, Lane C. Sander, Katrice A. Lippa & James H. Yen

  2. Centers for Disease Control and Prevention, National Center for Environmental Health, Atlanta, GA, USA

    Zia Fazili & Christine M. Pfeiffer

  3. Office of Dietary Supplements, National Institutes of Health, Bethesda, MD, USA

    Adam J. Kuszak & Stephen A. Wise

Authors
  1. Johanna E. Camara
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  2. Jeanita S. Pritchett
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  3. Yasmine C. Daniels
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  4. Mary Bedner
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  15. Stephen A. Wise
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Corresponding author

Correspondence to Johanna E. Camara.

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Human subjects ethics

The National Institute of Standards and Technology Research Protections Office reviewed the protocol for this project and determined it is “not human subjects research” as defined in 15 CFR 27, the Common Rule for the Protection of Human Subjects.

Conflict of interest

S.A. Wise is an Editor of the journal of Analytical and Bioanalytical Chemistry and was not involved in peer reviewing this manuscript. There are no financial or nonfinancial competing interests for any of the remaining coauthors.

Disclaimers

The findings and conclusions in this report are those of the author(s) and do not necessarily represent the official position of the Centers for Disease Control and Prevention. Certain commercial equipment or materials are identified in this paper to specify adequately the experimental procedure. Such identification does not imply recommendation or endorsement by the National Institute of Standards and Technology, nor does it imply that the materials or equipment identified are necessarily the best available for the purpose.

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Camara, J.E., Pritchett, J.S., Daniels, Y.C. et al. Development of an improved standard reference material for folate vitamers in human serum. Anal Bioanal Chem 415, 809–821 (2023). https://doi.org/10.1007/s00216-022-04474-w

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  • DOI: https://doi.org/10.1007/s00216-022-04474-w

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