HPLC Method Development and Validation for Quantitative Determination of Lung Cancer Biomarkers in Urine

Abstract

Lung cancer is the most common cause of cancer-related death in men and second most common in women. There are no good clinical markers that can be used to diagnose lung cancer at an early stage and predict its prognosis have been found. A recent study analyzed metabolites in plasma and serum blood samples from lung cancer patients and individuals without cancer using GC/MS and identified a set of metabolites differently expressed in cancer patients. Based on this study, we developed an high performance liquid chromatography (HPLC) diode array detection (DAD) method to detect fumaric acid, L-glutamic acid, pyruvic acid, inosine, and creatinine simultaneously in urine. Creatinine was included to account for the renal dilution. Baseline resolutions for all five biomarkers were obtained in synthetic urine matrix by using a 5% methanol and 95% of 0.6% acetic acid, using a C18 column. The developed HPLC method was validated in synthetic urine matrix using analytical method validation parameters such as linearity, accuracy, reproducibility, robustness, limits of detection and quantitation for accurate quantification. This validated method can potentially be used in a large scale clinical study. The detailed experimental conditions and results will be presented at the conference.