Simultaneous Detection of Six Urinary Pteridines and Creatinine by High-Performance Liquid Chromatography-Tandem Mass Spectrometry for Clinical Breast Cancer Detection

Abstract

Recent preliminary studies have implicated urinary pteridines as candidate biomarkers in a growing number of malignancies including breast cancer. While the developments of capillary electrophoresis-laser induced fluorescence (CE-LIF), high performance liquid chromatography (HPLC), and liquid chromatography-mass spectroscopy (LC-MS) pteridine urinalyses among others have helped to enable these findings, limitations including poor pteridine specificity, asynchronous or nonexistent renal dilution normalization, and a lack of information regarding adduct formation in mass spectrometry techniques utilizing electrospray ionization (ESI) have prevented application of these techniques to a larger clinical setting. In this study, a simple, rapid, specific, and sensitive high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method has been developed and optimized for simultaneous detection of six pteridines previously implicated in breast cancer and creatinine as a renal dilution factor in urine. In addition, this study reports cationic adduct formation of urinary pteridines under ESI-positive ionization for the first time. This newly developed technique separates and detects the following six urinary pteridines: 6-biopterin, 6- hydroxymethylpterin, d-neopterin, pterin, isoxanthopterin, and xanthopterin, as well as creatinine. The method detection limit for the pteridines is between 0.025 and 0.5 µg/L, and for creatinine, it is 0.15 µg/L. The method was also validated by spiked recoveries (81-105%), reproducibility (RSD: 1-6%), and application to 25 real urine samples from breast cancer positive and negative samples through a double-blind study. The proposed technique was finally compared directly with a previously reported CE-LIF technique, concluding that additional or alternative renal dilution factors are needed for proper investigation of urinary pteridines as breast cancer biomarkers.

Department(s)

Chemistry

Keywords and Phrases

80 And Over; Adult; Aged; Biological; Breast Cancer Detection; Breast Neoplasms; Breast Tumor; Capillary Electrophoresis; Chromatography; Clinical Settings; Comparative Study; Creatinine; Diseases; Electrospray Ionization (ESI); Electrospray Mass Spectrometry; Equipment; Female; High Performance Liquid Chromatography; High Pressure Liquid; Human; Mass; Mass Spectrometry; Method Detection Limits; Methodology; Middle Aged; Pteridine Derivative; Pteridines; Reproducibilities; Simultaneous Detection; Spectrometry; Spectrometry Technique; Tandem Mass Spectrometry; Tumor Marker; Urinalysis; Urine; Very Elderly

International Standard Serial Number (ISSN)

0003-2700

Document Type

Article - Journal

Document Version

Citation

File Type

text

Language(s)

English

Rights

© 2015 American Chemical Society (ACS), All rights reserved.

Publication Date

01 Nov 2013

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