High-performance liquid chromatography (HPLC) as a means of assessing the presence of uric acid in archeological human remains: Challenges and future directions

Jo Buckberry; Richard Telford; Laura Castells Navarro; John Snaith; David Swinson; Andrew Healey; Megan B Brickley

doi:10.1002/ajpa.24938

High-performance liquid chromatography (HPLC) as a means of assessing the presence of uric acid in archeological human remains: Challenges and future directions

Am J Biol Anthropol. 2024 Apr 16:e24938. doi: 10.1002/ajpa.24938. Online ahead of print.

Authors

Jo Buckberry¹, Richard Telford², Laura Castells Navarro³, John Snaith⁴, David Swinson⁵, Andrew Healey², Megan B Brickley⁶

Affiliations

¹ School of Archaeological and Forensic Sciences, University of Bradford, Bradford, UK.
² School of Chemical and Biological Sciences, University of Bradford, Bradford, UK.
³ Department of Archaeology, University of York, York, UK.
⁴ School of Chemistry, University of Birmingham, Birmingham, UK.
⁵ Former Institute of Archaeology and Antiquity, University of Birmingham, Birmingham, UK.
⁶ Department of Anthropology, McMaster University, Hamilton, Ontario, Canada.

PMID: 38623788
DOI: 10.1002/ajpa.24938

Abstract

Objectives: This research aimed to replicate the Swinson, D., Snaith, J., Buckberry, J., & Brickley, M. (2010). High performance liquid chromatography (HPLC) in the investigation of gout in paleopathology. International Journal of Osteoarchaeology, 20, 135-143. https://doi.org/10.1002/oa.1009 method for detecting uric acid in archeological human remains to investigate gout in past populations and to improve the original High Performance Liquid Chromatography-ultraviolet (HPLC-UV) method by using HPLC-mass spectrometry (HPLC-MS), a more sensitive, compound-specific detection method.

Materials and methods: We used reference samples of uric acid to create a dilution series to assess the limits of quantification and detection. Samples from individuals with and without gout lesions were taken from foot bones and ribs from the English cemeteries of Tanyard, Hickleton, Gloucester, and Lincoln.

Results: We could not replicate the results of Swinson and colleagues using HPLC-UV. Tests using a dilution series of uric acid showed HPLC-MS was approximately 100× more sensitive than HPLC-UV, with the additional benefit of being compound specific. A newly developed hydrophilic interaction chromatography (HILIC) method improved retention characteristics. Fourteen samples from eight individuals, five with skeletal lesions consistent with gout, were analyzed with the final method. None showed evidence of uric acid despite the newly developed method's improved sensitivity and specificity.

Discussion: The lack of detectable uric acid extracted from these samples suggests that (1) urate crystals were not present in any of the bone samples, regardless of gout status; (2) urate crystals did not survive these specific archeological conditions; or (3) the concentration of uric acid in our bone extracts was low, and thus larger samples would be required.

Keywords: gout; high performance liquid chromatography‐mass spectrometry; high performance liquid chromatography‐ultraviolet; hydrophilic interaction chromatography; paleopathology.

Abstract

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