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.
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.
| Original language | English |
|---|---|
| Article number | e24938 |
| Journal | American Journal of Biological Anthropology |
| Early online date | 16 Apr 2024 |
| DOIs | |
| Publication status | E-pub ahead of print - 16 Apr 2024 |
Bibliographical note
Acknowledgments:This research was funded by an Arts Research Board (ARB) Major Collaborative Project Seed Grant, McMaster University 2015 and was undertaken, in part, thanks to funding from the Canada Research Chairs program (grant/award number: 2231563). We thank Dr. Alex Surtees and Shelley Reid for their support with some analyses and Dr. Andy Fraser for producing Figure 1. We thank the anonymous reviewers, associate editor, and editorial board member for their useful comments on the earlier version of this paper.
Keywords
- high performance liquid chromatography‐ultraviolet
- high performance liquid chromatography‐mass spectrometry
- gout
- hydrophilic interaction chromatography
- paleopathology