LC–MS/MS determination of d-mannose in human serum as a potential cancer biomarker

Abstract

Several metabolites in human serum have been identified as potential cancer biomarkers for early detection. This study focuses on the LC–MS/MS method development and validation of D-mannose in human serum. Surrogate blank serum, coupled with stable isotope D-mannose-13C6, as internal standard, was used for generating standard curves ranging from 1 to 50 μg/mL. Separation was achieved by an Agilent 1200 series HPLC equipped with a SUPELCOGELTM Pb, 6% Crosslinked column with HPLC water as a mobile phase at flow rate of 0.5 mL/min at 80 °C. Mass detection was performed under negative ionization electrospray. Inter- and intra-day accuracy and precision were <2%. The extraction recovery and matrix effect were 104.1%–105.5% and 97.0%–100.0%, respectively. This method was successfully applied for the quantification of D-mannose in the serum samples of 320 esophageal cancer patients and 323 healthy volunteers. We report a simple, specific and reproducible LC–MS/MS method for the quantification of D-mannose in human serum as a potential cancer biomarker.

Quantification of d-mannose in plasma: Development and validation of a reliable and accurate HPLC-MS-MS method

“…Chromatography of D-mannose in biological samples is complicated by the challenging separation from its endogenous epimers D-glucose and D-galactose, which often requires ad hoc strategies and specific materials [10]. In a recently published HPLC-MS-MS method, White et al [4] proposed the use of a Supelcogel Pb (Sigma-Aldrich, Saint Louis, MO, USA) column capable of separating sugars epimers by means of an ion exchange resin, which interacts with the analytes by both size exclusion and ligand exchange, the latter mechanism probably prevailing in the case of D-mannose.…”

“…In particular, biochemical and genetic data have shown that genes involved in the O-mannose glycosylation pathway may cause forms of congenital muscular dystrophy in humans [2]. Moreover, mannose has been implicated in cancer metabolism and is considered to be a reliable biomarker for esophageal adenocarcinoma (EAC) [3,4]. Moreover, serum concentrations of D-mannose have been reported to be higher in metastatic breast cancer patients than in early-stage breast cancer [5].…”

“…Moreover, serum concentrations of D-mannose have been reported to be higher in metastatic breast cancer patients than in early-stage breast cancer [5]. Use of this metabolite as a marker for the early detection of both breast and esophageal cancer may improve the clinical outcome of these diseases [4,5]. Recent studies have also shown that elevated plasma mannose levels are related with future risk of several chronic diseases, including type 2 diabetes (T2D), cardiovascular disease (CVD), and renal impairment (albuminuria) [6].…”

“…We developed and validated an accurate D-mannose assay by tandem mass spectrometry coupled to liquid chromatography (HPLC-MS-MS), capable of discriminating mannose from Dglucose, a C2 epimer of D-mannose, which in humans circulates at a concentration at least 100-fold higher [4]. Compared to traditional techniques, tandem mass spectrometry has enhanced selectivity, limited cost of analysis, and allows the determination of compound of interest in biological fluids.…”

Serum Biomarker Identification by Mass Spectrometry in Acute Aortic Dissection

“…Although such studies offer effective information for the early diagnosis and prognosis of AD and also provide constructive suggestions for AD pathogenesis elucidation, due to the intrinsic particularity of protein macromolecules, their dissemination in clinical detection for AD faces many challenges, including determining the preservation and testing conditions, diagnosis interval, and cost. Peripheral blood metabolic biomarkers have gained more attention with the development of HPLC-MS technology, our increasing understanding of the metabolic pathways of small molecules, and the gradual perfection of small molecule databases [20-22]. In the present study, metabolites in the peripheral sera of patients with AD were examined by HPLC-MS and compared with corresponding metabolite levels in healthy subjects and hypertensive patients.…”