Aqueous normal-phase retention of nucleotides on silica hydride columns

Pesek, Joseph J.; Matyska, Maria T.; Hearn, Milton Thomas William; Boysen, Reinhard I

doi:10.1016/j.chroma.2008.12.046

Cited by 50 publications

(31 citation statements)

References 20 publications

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“…Bajad et al (2006). Similarly, silica hydride was used in the normal phase mode (Pesek et al, 2009). Currently, only one HILIC-MS method for the determination of a nucleotide analog has been described (Pucci et al, 2009).…”

Section: Quantitative Nucleotide Analysis With Msmentioning

confidence: 99%

Mass spectrometry in the quantitative analysis of therapeutic intracellular nucleotide analogs

Jansen

Rosing

Schellens

et al. 2011

Mass Spectrometry Reviews

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Nucleoside analogs are widely used in anti-cancer, anti-(retro)viral, and immunosuppressive therapy. Nucleosides are prodrugs that require intracellular activation to mono-, di-, and finally triphosphates. Monitoring of these intracellular nucleotides is important to understand their pharmacology. The relatively involatile salts and ion-pairing agents traditionally used for the separation of these ionic analytes limit the applicability of mass spectrometry (MS) for detection. Both indirect and direct methods have been developed to circumvent this apparent incompatibility. Indirect methods consist of de-phosphorylation of the nucleotides into nucleosides before the actual analysis. Various direct approaches have been developed, ranging from the use of relatively volatile or very low levels of regular ion-pairing agents, hydrophilic interaction chromatography (HILIC), weak anion-exchange, or porous graphitic carbon columns to capillary electrophoresis and matrix-assisted light desorption--time of flight (MALDI-TOF) MS. In this review we present an overview of the publications describing the quantitative analysis of therapeutic intracellular nucleotide analogs using MS. The focus is on the different approaches for their direct analysis. We conclude that despite the technical hurdles, several useful MS-compatible chromatographic approaches have been developed, enabling the use of the excellent selectivity and sensitivity of MS for the quantitative analysis of intracellular nucleotides.

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Section: Quantitative Nucleotide Analysis With Msmentioning

confidence: 99%

Mass spectrometry in the quantitative analysis of therapeutic intracellular nucleotide analogs

Jansen

Rosing

Schellens

et al. 2011

Mass Spectrometry Reviews

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“…The wide range of chromatography (LC) options allow physical separation of most molecules to reduce signal suppression [16]. While reversephase methods tend to utilize C18 columns, a wide range of normal phase materials are utilized [16,17] e.g., popular new materials, including hydrophilic interaction chromatography (HILIC) and aqueous normal phase (ANP) have recently been developed for improved separation of polar metabolites [18].…”

Section: Lc/ms Based Metabolomicsmentioning

confidence: 99%

Dealing with the unknown: Metabolomics and Metabolite Atlases

Bowen

Northen

2010

J. Am. Soc. Mass Spectrom.

182

158

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Metabolomics is the comprehensive profiling of the small molecule composition of a biological sample. Since metabolites are often the indirect products of gene expression, this approach is being used to provide new insights into a variety of biological systems (clinical, bioenergy, etc.). A grand challenge for metabolomics is the complexity of the data, which often include many experimental artifacts. This is compounded by the tremendous chemical diversity of metabolites. Identification of each uncharacterized metabolite is in many ways its own puzzle (compared with proteomics, which is based on predictable fragmentation patterns of polypeptides). Therefore, effective data reduction/prioritization strategies are critical for this rapidly developing field. Here we review liquid chromatography electrospray ionization mass spectrometry (LC/MS)-based metabolomics, methods for feature finding/prioritization, approaches for identifying unknown metabolites, and construction of method specific 'Metabolite Atlases'.(J Am Soc Mass Spectrom 2010, 21, 1471-1476) © 2010 Published by Elsevier Inc. on behalf of American Society for Mass Spectrometry N ature utilizes a tremendous diversity of metabolites, and it is estimated that there are Ͼ200,000 plant metabolites alone [1]. Metabolomics is a rapidly growing field for studying the small molecule composition of a biological system. Liquid chromatography coupled to electrospray ionization mass spectrometry (LC/MS) is becoming a method of choice for profiling metabolites in complex biological samples. This is due to its ability to effectively ionize a breath of metabolites with minimal fragmentation (versus gas chromatography-mass spectroscopy (GC/MS), robustness, and ability to scale-up to support tandem mass spectrometry based structural studies (versus capillary electrophoresis-mass spectrometry (CE-MS)) [2]. However, known metabolites are typically a small portion of the data obtained in a LC/MS metabolomics experiment (ϳ10%) [3] where the bulk are MS-artifacts and uncharacterized metabolites.Identification of unknown metabolites is cost and effort intensive, often requiring preparative scale isolation for nuclear magnetic resonance (NMR) studies or extensive chemical synthesis to enable structural comparisons using tandem mass spectrometry (MS/MS) [4]. Therefore, it is not surprising that the majority of reports focus on changes in metabolites that have authentic standards or at a minimum are found in metabolite databases. Yet, there are only a few thousand commercially available analytical standards due to lack of demand and the inherent instability of many metabolites. These standards and the endogenous metabolites contained in databases represent only a portion of endogenous metabolites. Therefore, effective methods for prioritizing, studying, and ultimately identifying uncharacterized metabolites is a critical development for LC/MS based metabolomics [5][6][7][8][9][10]. Here we discuss current LC/MS metabolomic approaches (detailed elsewhere [2,4,11,12]) and their inte...

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“…The versatility of hydride-based stationary phases for the retention of polar metabolites, particularly the low carbon-loaded material, has been demonstrated in previous studies [20,21]. One fundamental property of these materials that has not been studied to date is their efficiency when retaining hydrophilic compounds with respect to mobile phase velocity.…”

Section: Properties Of Hydride-based Stationary Phasesmentioning

confidence: 85%

“…This investigation has utilized a particular type of silica hydride separation material with a low-carbon content in order to provide the best ANP retention capabilities for metabolites. Previous investigations have shown that several different types of metabolites [20] and nucleotides [21] can be efficiently analyzed on this stationary phase. The designation of ANP for the retention of polar compounds is used in this report to distinguish silica hydride materials from HILIC stationary phases.…”

Section: Introductionmentioning

confidence: 99%

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

Pesek¹,

Matyska

Loo

et al. 2009

J of Separation Science

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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.

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Aqueous normal-phase retention of nucleotides on silica hydride columns

Cited by 50 publications

References 20 publications

Mass spectrometry in the quantitative analysis of therapeutic intracellular nucleotide analogs

Mass spectrometry in the quantitative analysis of therapeutic intracellular nucleotide analogs

Dealing with the unknown: Metabolomics and Metabolite Atlases

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

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