Dilute-and-Shoot-Liquid Chromatography-Quadrupole Time of Flight-Mass Spectrometry for Pteridine Profiling in Human Urine and Its Association with Different Pathologies

Pteridines are a group of compounds synthesised by many living organisms that are involved in the metabolism of many cofactors and vitamins. Their concentration in biological fluids may be altered by various pathologies such as cancer or inflammatory bowel disease, urine being the main route of excretion. In this study, three lumazines and ten pterins were analysed in their native oxidation state using high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry. Their high concentration in urine samples and their good ionisation behaviour allow the use of the dilute-and-shoot method by simple filtration of the urine prior to chromatographic analysis. The final method offers excellent linearity, sensitivity and precision parameters, and a total of 135 urine samples were analysed from patients with some relevant information such as faecal calprotectin (FCP) levels, common diseases such as diabetes, hypertension and dyslipidaemia and immunological diseases such as inflammatory bowel disease (IBD). The pteridine profile was related to FCP levels without showing any correlation. In addition, pteridine levels were compared between healthy subjects and IBD, diabetic, hypertensive and dyslipidaemic patients, and significant differences were found between the two groups for some of the pteridines.

Automated summary

Pteridines are compounds involved in the metabolism of cofactors and vitamins, and their concentration in urine can be altered by diseases such as cancer or inflammatory bowel disease. A method using high-performance liquid chromatography coupled to mass spectrometry was developed to analyze the concentration of lumazines and pterins in urine samples. The method showed excellent linearity, sensitivity, and precision parameters. The pteridine profile was related to faecal calprotectin levels, but no correlation was found. Significant differences in pteridine levels were observed between healthy subjects and patients with various diseases.