Analysis of hydrophilic metabolites in physiological fluids by HPLC‐MS using a silica hydride‐based stationary phase

Abstract

Aqueous normal-phase chromatography was used for the analysis of metabolites in human saliva and urine samples. The column was packed with a silica hydride type separation material. Several gradients were tested with different mobile phase additives in order to produce retention for amino acids, small organic acids, and carbohydrates. Detection was done by TOF MS. In some cases the relative concentration levels of various metabolites in human saliva were compared for normal patients and patients with pancreatic cancer or pancreatitis. The reproducibility of retention of individual metabolites in these complex matrices was tested for several compounds.

Aqueous normal phase retention of nucleotides on silica hydride‐based columns: Method development strategies for analytes revelant in clinical analysis

“…Two of the columns investigated in this study were found to be useful for the studies of retention of nucleotides: a Cogent DH HPLC column, already used in the analysis of many metabolites and polar compounds [8,9], and a UDA weak cation exchange, carboxylic acid HPLC column. A hydride-based C-18 was also tested for the sake of completeness.…”

“…In addition, the ANP analysis of polar compounds is achieved at high concentration of the organic solvent in the mobile phase (usually acetonitrile, but acetone may be used as well), which provides increased sensitivity. When MS detection is used, the analysis can be performed without derivatization of any substrates, including highly polar amino acids, sugars and presumably sugar-phosphates, which are also present in biological samples [8,9].…”

Data Processing, Metabolomic Databases and Pathway Analysis

“…In particular, LC-MS can be divided into dozens of different methods that target different compound classes, from highly hydrophilic metabolites using novel aqueous normal phase columns (Silica-Hydride columns) (Pesek et al, 2009) or zwitterionic hydrophilic interaction columns (HILIC) (Strege, 1998) to various reverse phase columns that cover wide XlogP lipophilicity ranges. Secondly, LC effluents can be used in combination with more than one detector, for example using a diode array detector for carotenoid detection in conjunction with a mass spectrometer for ionizable compounds (Lu et al, 2007).…”

The retention behaviour of amino acids in hydrophilic interaction liquid chromatography on zwitterionic stationary phases†

“…Silica hydride based stationary phases have been employed for bioanalysis, although the term aqueous NPC (ANPC) was preferred [15][16][17]. The chiral separation of drug substances was achieved using a cyclodextrin-derivatised stationary phase [18].…”