Development of a novel UHPLC-MS/MS-based platform to quantify amines, amino acids and methylarginines for applications in human disease phenotyping

Abstract

Amine quantification is an important strategy in patient stratification and personalised medicine. This is because amines, including amino acids and methylarginines impact on many homeostatic processes. One important pathway regulated by amine levels is nitric oxide synthase (NOS). NOS is regulated by levels of (i) the substrate, arginine, (ii) amino acids which cycle with arginine and (iii) methylarginine inhibitors of NOS. However, biomarker research in this area is hindered by the lack of a unified analytical platform. Thus, the development of a common metabolomics platform, where a wide range of amino acids and methylarginines can be measured constitutes an important unmet need. Here we report a novel high-throughput ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) platform where ≈40 amine analytes, including arginine and methylarginines can be detected and quantified on a molar basis, in a single sample of human plasma. To validate the platform and to generate biomarkers, human plasma from a well-defined cohort of patients before and after coronary artery bypass surgery, who developed systemic inflammatory response syndrome (SIRS), were analysed. Bypass surgery with SIRS significantly altered 26 amine analytes, including arginine and ADMA. Consequently, pathway analysis revealed significant changes in a range of pathways including those associated with NOS.

Renal Function Underpins the Cyclooxygenase-2: Asymmetric Dimethylarginine Axis in Mouse and Man

“…ADMA, arginine, and citrulline were measured, within a panel of amines, in human and mouse plasma by ultra-high performance liquid-chromatography tandem mass spectrometry following derivatization with AccQTag as described previously. 36 ADMA was below the limit of detection in 2 of 55 samples and inputted at the assay limit of quantification. ADMA was also measured in the same samples using enzyme-linked immunosorbent assay (ELISA).…”

A Mini-review on Ultra Performance Liquid Chromatography

“…For quantify amines and amino acids in human disease phenotyping 65 . d) UHPLC-UV was applied for the analysis of total amino acid in infant formulas and adult nutritionals 66 .…”

Advanced Microsamples: Current Applications and Considerations for Mass Spectrometry-Based Metabolic Phenotyping Pipelines

“…The invasive nature of venepuncture and its resultant frontend sample yields an excess of 400-2000-times the volumes required for metabolic phenotyping analyses (for typical phlebotomy collections of 2-10 mL) [2]. However, metabolic phenotyping assays that employ LC-MS are amenable to low sample volumes; for example, LC-MS assays routinely require <25 µL to perform metabolic phenotyping for both discovery profiling analysis and targeted pathway analysis [2,[12][13][14][15][16][17][18][19][20]. Such methods have been applied to metabolic phenotyping experiments that have assessed outcomes of health and disease including inflammation [17], cardiometabolic diseases [21], dementia [17], and nutrition [22]; they are commonly targeted for many clinical and human physiology studies.…”