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 - Paleontology
- Archeology
- Genetics
- Anthropology
- Anatomy
- Epidemiology
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.
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.