Mixed mode chromatography via column switching for the simultaneous HPLC analysis of lonic and non-lonic nucleic acid constituents

Halfpenny, Anne P.; Brown, Phyllis R.

doi:10.1007/bf02311602

Cited by 26 publications

(6 citation statements)

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“…Mixed‐mode chromatography has been used primarily for the analysis of nucleotides, oligonucleotides and peptides (Bischoff and McLaughlin, 1984; Bischoff et al. , 1983; Halfpenny and Brown, 1986;Nogueira et al. , 2005, 2006; Progent et al.…”

Section: Introductionmentioning

confidence: 99%

“…Sato et al (2004) have reported the use of capillary electrophoresis coupled with MS for analyzing 88 structurally diverse metabolites extracted from rice leaves. Mixed-mode chromatography has been used primarily for the analysis of nucleotides, oligonucleotides and peptides (Bischoff and McLaughlin, 1984;Bischoff et al, 1983;Halfpenny and Brown, 1986;Nogueira et al, 2005Nogueira et al, , 2006Progent et al, 2006). More recently, mixed-mode materials have been applied to the analysis of pesticide metabolites in human urine (Bicker et al, 2005) and fungal mycotoxins (Apfelthaler et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

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Metabolite profiling of Calvin cycle intermediates by HPLC‐MS using mixed‐mode stationary phases

et al. 2008

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SummaryA sensitive and robust mixed-mode high performance liquid chromatography-tandem mass spectrometry method was developed for the qualitative and quantitative determination of sugar phosphates, which are notoriously difficult to separate using reversed-phase materials. Sugar phosphates were separated on a Primesep SB Ò column by gradient elution using aqueous ammonium formate and acetonitrile as mobile phases. Target analytes were identified by their precursor/product ions and retention times. Quantitative analysis was performed in negative ionization/multiple reaction monitoring mode with five different time segments. The method was validated by spiking authentic sugar phosphate standards into complex plant tissue extracts. Standard curves of neat authentic standards and spiked extracts were generated for concentrations in the low picomole to nanomole range, with correlation coefficients of R 2 > 0.991, and the degree of ion suppression in the presence of a plant matrix was calculated for each analyte. Analyte recoveries, which were determined by including known quantities of authentic standards in the sugar phosphate extraction protocol, ranged from 40.0% to 57.4%. The analytical reproducibility was assessed by determining the coefficient of variance based on repeated extractions/measurements (<20%). The utility of our method is demonstrated with two types of applications: profiling of Calvin cycle intermediates in (i) dark-adapted and light-treated tobacco leaves, and in (ii) antisense plants expressing reduced levels of the Calvin cycle enzymes glyceraldehyde-3-phosphate dehydrogenase or ribulose-1,5-bisphosphate carboxylase/oxygenase (comparison with wild-type controls). The broader applicability of our method is illustrated by profiling sugar phosphates extracted from the leaves of five taxonomically diverse plants.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Metabolite profiling of Calvin cycle intermediates by HPLC‐MS using mixed‐mode stationary phases

et al. 2008

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“…Mixed‐mode chromatography (MMC) is based on a principle that reverses the approach that chromatographic stationary phase design has traditionally followed, namely the optimization of a single retention mechanism, such as reversed‐phase or ion‐exchange interaction. A mixed‐mode approach purposefully utilizes multiple retention mechanisms on the same stationary phase 20, 21. The concept is not new but an increasing range of stationary phases are becoming available.…”

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confidence: 99%

Mixed‐mode chromatography/isotope ratio mass spectrometry

McCullagh

2010

Rapid Comm Mass Spectrometry

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Liquid chromatography coupled to molecular mass spectrometry (LC/MS) has been a standard technique since the early 1970s but liquid chromatography coupled to high-precision isotope ratio mass spectrometry (LC/IRMS) has only been available commercially since 2004. This development has, for the first time, enabled natural abundance and low enrichment delta(13)C measurements to be applied to individual analytes in aqueous mixtures creating new opportunities for IRMS applications, particularly for the isotopic study of biological molecules. A growing number of applications have been published in a range of areas including amino acid metabolism, carbohydrates studies, quantification of cellular and plasma metabolites, dietary tracer and nucleic acid studies. There is strong potential to extend these to new compounds and complex matrices but several challenges face the development of LC/IRMS methods. To achieve accurate isotopic measurements, HPLC separations must provide baseline-resolution between analyte peaks; however, the design of current liquid interfaces places severe restrictions on compatible flow rates and in particular mobile phase compositions. These create a significant challenge on which reports associated with LC/IRMS have not previously focused. Accordingly, this paper will address aspects of chromatography in the context of LC/IRMS, in particular focusing on mixed-mode separations and their benefits in light of these restrictions. It aims to provide an overview of mixed-mode stationary phases and of ways to improve high aqueous separations through manipulation of parameters such as column length, temperature and mobile phase pH. The results of several practical experiments are given using proteogenic amino acids and nucleosides both of which are of noted importance in the LC/IRMS literature. This communication aims to demonstrate that mixed-mode stationary phases provide a flexible approach given the constraints of LC/IRMS interface design and acts as a practical guide for the development of new chromatographic methods compatible with LC/IRMS applications.

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“…Some compounds present in pharmaceuticals, environment, and food cannot be separated efficiently by stationary phases possessing single separation mechanism. Instead, since the paper of Halfpenny and Brown , it has been many times demonstrated that mixed‐mode stationary phases possessing various functional groups and interaction abilities could provide better results. Thus, mixed‐mode chromatography (MMC) based on more than one retention mechanism attracts a great concern for the last several years.…”

Section: Introductionmentioning

confidence: 99%

Recent advances in mixed‐mode chromatographic stationary phases

Sýkora

Řezanka

Záruba

et al. 2018

J of Separation Science

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Mixed‐mode phases have become very popular in the last decade, and the number of new mixed/multi‐mode sorbents is growing fast. Unlike single‐mode stationary phases, perfectly suited for the separation of the analytes possessing similar physicochemical properties, for instance reversed‐phase chromatography for hydrophobic solutes, mixed‐mode sorbents providing multimodal interactions can render better separation selectivity for complex mixtures of solutes differing significantly in their physicochemical characteristics. The most frequent modern mixed‐mode stationary phases are di/tri‐mode sorbents embracing the following interactions, hydrophobic, electrostatic (coulombic), and hydrophilic. According to their structures, it is possible to distinguish silica‐based, polymer‐based, hybrid, and monolithic mixed‐mode stationary phases. Herewith, newly synthesized mixed‐mode sorbents developed within the last two and half years are categorized, discussed, and summarized. The main attention is devoted to the description of the synthetic routes and characterization methods applied for the new stationary phases.

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Mixed mode chromatography via column switching for the simultaneous HPLC analysis of lonic and non-lonic nucleic acid constituents

Cited by 26 publications

References 10 publications

Metabolite profiling of Calvin cycle intermediates by HPLC‐MS using mixed‐mode stationary phases

Metabolite profiling of Calvin cycle intermediates by HPLC‐MS using mixed‐mode stationary phases

Mixed‐mode chromatography/isotope ratio mass spectrometry

Recent advances in mixed‐mode chromatographic stationary phases

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