Cancer disease is the second leading cause of death in the world. Epidemiology data indicate that early diagnosis of a tumour increases a patient's chance of recovery. Biomarkers are effective instruments which can potentially lead to precancer screening or precancer diagnosis and may provide useful information on the cancer type and the disease's stage of progression. The analysis of new biomarkers for cancer is currently a popular area of study in clinical and cancer research. Pteridines are a class of potential cancer biomarkers. The monitoring levels of pteridines may have prospective value for controlling the course of the malignant process. This review describes the functional employment of pteridines, as biomarkers, in cancer diagnosis. It also contains the description of application of analytical techniques such as high-performance liquid chromatography (HPLC) and capillary electrophoresis (CE) used for pteridine analysis.