Orthogonal method development using hydrophilic interaction chromatography and reversed-phase high-performance liquid chromatography for the determination of pharmaceuticals and impurities

Wang, Xiande; Li, Weiyong; Rasmussen, Henrik T.

doi:10.1016/j.chroma.2005.05.082

Cited by 99 publications

(39 citation statements)

References 13 publications

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“…Because these standards are unavailable, mass spectrometry is required to identify novel metabolites. HILIC displays unique selectivity toward polar compounds (25) and employs mobile phase conditions that are amenable to mass spectrometry (26,27). Therefore, it was felt more desirable to employ HILIC in this study for the separation and purification of the intracellular sugar-nucleotides.…”

Section: Hilic-ms Purification Of Cmp-pseam (I) and Udp-diacetamidotrmentioning

confidence: 99%

Functional Characterization of the Flagellar Glycosylation Locus in Campylobacter jejuni 81–176 Using a Focused Metabolomics Approach

McNally

Hui

Aubry

et al. 2006

Journal of Biological Chemistry

110

129

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Bacterial genome sequencing has provided a wealth of genetic data. However, the definitive functional characterization of hypothetical open reading frames and novel biosynthetic genes remains challenging. This is particularly true for genes involved in protein glycosylation because the isolation of their glycan moieties is often problematic. We have developed a focused metabolomics approach to define the function of flagellin glycosylation genes in Campylobacter jejuni 81-176. A capillary electrophoresis-electrospray mass spectrometry and precursor ion scanning method was used to examine cell lysates of C. jejuni 81-176 for sugar nucleotides. Novel nucleotide-activated intermediates of the pseudaminic acid (Pse5NAc7NAc) pathway and its acetamidino derivative (PseAm) were found to accumulate within select isogenic mutants, and use of a hydrophilic interaction liquid chromatography-mass spectrometry method permitted large scale purifications of the intermediates. NMR with cryo probe (cold probe) technology was utilized to complete the structural characterization of microgram quantities of CMP-5-acetamido-7-acetamidino-3,5,7,9-tetradeoxy-L-glycero-␣-L-manno-nonulosonic acid (CMP-Pse5NAc7Am), which is the first report of Pse modified at C7 with an acetamidino group in Campylobacter, and UDP-2,4-diacetamido-2,4,6-trideoxy-␣-D-glucopyranose, which is a bacillosamine derivative found in the N-linkedproteinglycan.Usingthisfocusedmetabolomicsapproach,pseB, pseC, pseF, pseI, and for the first time pseA, pseG, and pseH were found to be directly involved in either the biosynthesis of CMP-Pse5NAc7NAc or CMP-Pse5NAc7Am. In contrast, it was shown that pseD, pseE, Cj1314c, Cj1315c, Cjb1301, Cj1334, Cj1341c, and Cj1342c have no role in the CM-P-Pse5NAc7NAc or CMP-Pse5NAc7Am pathways. These results demonstrate the usefulness of this approach for targeting compounds within the bacterial metabolome to assign function to genes, identify metabolic intermediates, and elucidate novel biosynthetic pathways.The availability of bacterial genomic sequences has provided unprecedented opportunity for comparative studies. The functional analysis of each respective genome is currently under way and often involves an integrative, multidisciplinary approach that combines bioinformatics, mutagenesis, proteomics, and microarray technologies. Most recently, metabolomics-based analyses, although limited in their number, are becoming established as an additional tool to elucidate gene function (1, 2). Metabolomics is the characterization of all low molecular weight compounds in a defined biological system and differs from classical metabolism studies by its greater breadth and speed of metabolite analysis. Although recent advances in mass spectrometry (MS), 2 such as the ultimate resolving capability of the Fourier-transform ion cyclotron resonance MS, and also in nuclear magnetic resonance spectroscopy (NMR) technologies, such as the development of higher magnetic field spectrometers and cryogenically cooled probes (cold probes), has greatly facilit...

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Section: Hilic-ms Purification Of Cmp-pseam (I) and Udp-diacetamidotrmentioning

confidence: 99%

Functional Characterization of the Flagellar Glycosylation Locus in Campylobacter jejuni 81–176 Using a Focused Metabolomics Approach

McNally

Hui

Aubry

et al. 2006

Journal of Biological Chemistry

110

129

View full text Add to dashboard Cite

show abstract

“…This technology is named hydrophilic interaction chromatography (HILIC). The quite neutral hydrophobic/hydrophilic balance of the eluent allows an easy interfacing with ESI-MS unlike the normal phase HPLC (NP-HPLC) working with totally organic solvents which are not compatible with the nebulization process [62].…”

Section: High-resolution Liquid Chromatography Coupled To Mass Spectrmentioning

confidence: 99%

Coupled Mass Spectrometric Strategies for the Determination of Carbohydrates at Very Low Concentrations: The Case of Polysaccharides Involved in the Molecular Dialogue Between Plants and Rhizobia

Poinsot¹,

Carpéné²,

Couderc³

2012

The Complex World of Polysaccharides

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“…With the development of HILIC, it has been widely used in many fields, such as metabonomic study [16], analysis of pharmaceutical and their impurities [17] and two dimensional (2D)-LC analysis [18]. It is reported that HILIC was also used to analyze cephalosporin C [19,20].…”

Section: Introductionmentioning

confidence: 99%

Analysis of cephalosporins by hydrophilic interaction chromatography

Liu

et al. 2011

Journal of Pharmaceutical and Biomedical Analysis

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Orthogonal method development using hydrophilic interaction chromatography and reversed-phase high-performance liquid chromatography for the determination of pharmaceuticals and impurities

Cited by 99 publications

References 13 publications

Functional Characterization of the Flagellar Glycosylation Locus in Campylobacter jejuni 81–176 Using a Focused Metabolomics Approach

Functional Characterization of the Flagellar Glycosylation Locus in Campylobacter jejuni 81–176 Using a Focused Metabolomics Approach

Coupled Mass Spectrometric Strategies for the Determination of Carbohydrates at Very Low Concentrations: The Case of Polysaccharides Involved in the Molecular Dialogue Between Plants and Rhizobia

Analysis of cephalosporins by hydrophilic interaction chromatography

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