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Simultaneous quantification of urinary purines and creatinine by ultra high performance liquid chromatography with ultraviolet spectroscopy and quadrupole time‐of‐flight mass spectrometry: Method development, validation, and application to gout study
Author(s) -
Yuan Yuan,
Jiang Min,
Zhang Hongyang,
Liu Ju,
Zhang Min,
Hu Ping
Publication year - 2019
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201900170
Subject(s) - chemistry , chromatography , xanthine , mass spectrometry , purine metabolism , uric acid , hypoxanthine , creatinine , detection limit , analyte , gout , biochemistry , enzyme
We present an ultra high performance liquid chromatography with ultraviolet spectroscopy and quadrupole time‐of‐flight mass spectrometry method for the simultaneous quantification of ten purines (adenine, hypoxanthine, guanine, xanthine, deoxyadenosine, adenosine, inosine, guanosine, xanthosine, and uric acid) and creatinine in human urine. After chromatographic separation on an ACE Excel 2 AQ column, high abundant creatinine and uric acid and the other low abundant purines were sequentially detected by ultraviolet and quadrupole time‐of‐flight mass spectrometry within a single run. Method validations including specificity (improved by accurate mass measurement), linearity (correlation coefficients ≥0.9944), limit of quantification (0.002–9.756 µg/mL), intra‐ and interday precision (relative standard deviations ≤9.1 and 14.0%, respectively), accuracy (relative errors ≤13.1%), extraction recovery (between 90.3 and 109.6%), matrix effect (between 85.3 and 110.5%), and stability (relative errors ≤14.3%) were fully evaluated. This approach was applied to characterize the disordered purine metabolism in acute and chronic gout as an example. Quantitative results (normalized by creatinine) showed that an overproduction of urinary purine precursors might be involved in the gout process. The developed method represents a useful tool to investigate the purine disturbances in gout and other relevant diseases.