Capillary liquid chromatographic–high-resolution mass spectrometric analysis of ribonucleotides

Cai, Zongwei; Song, Fengrui; Yang, M. S.

doi:10.1016/s0021-9673(02)01155-x

Cited by 26 publications

(52 citation statements)

References 17 publications

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“…Cai et al on the other hand could reduce the N,N-DMHA concentration to 10 and 5 mM by changing the column diameter to 1.0 and 0.5 mm, respectively. Concomitantly, the MS signal increased 5-to 10-fold (Cai, Song, & Yang, 2002). In contrast to previous reports (Lai, Wang, & Cai, 2008), a clear relation between column diameter and applied N,N-DMHA …”

Section: Separation and Detectioncontrasting

confidence: 96%

“…N,N-dimethylhexylamine (N,N-DMHA) has also caused the formation of adducts (Tuytten et al, 2002). These adducts have been used as quantifying ions by some (Cai, Song, & Yang, 2002;Qian, Cai, & Yang, 2004), whereas others, using 1,5-DMHA, found that the product ions of these adducts were not sufficiently specific because only the adduct was lost upon fragmentation . Unless the intensity of the non-adduct is unacceptable, the use of adducts should be avoided in quantitative bioanalysis.…”

Section: B Ionization Suppressionmentioning

confidence: 99%

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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: Separation and Detectioncontrasting

confidence: 96%

Section: B Ionization Suppressionmentioning

confidence: 99%

Mass spectrometry in the quantitative analysis of therapeutic intracellular nucleotide analogs

Jansen

Rosing

Schellens

et al. 2011

Mass Spectrometry Reviews

View full text Add to dashboard Cite

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“…The latter has been used for many years with a broad application range in nucleotide analysis [2,5,11,[16][17][18][19]. The addition of cationic reagents such as alkylamines leads to formation of adducts between negatively charged nucleotides and positively charged ion pair reagent, increasing the retention factors for these compounds and thus improving retention.…”

Section: Introductionmentioning

confidence: 99%

Quantitation of intracellular purine intermediates in different Corynebacteria using electrospray LC-MS/MS

et al. 2012

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Intermediates of the purine biosynthesis pathway play key roles in cellular metabolism including nucleic acid synthesis and signal mediation. In addition, they are also of major interest to the biotechnological industry as several intermediates either possess flavor-enhancing characteristics or are applied in medical therapy. In this study, we have developed an analytical method for quantitation of 12 intermediates from the purine biosynthesis pathway including important nucleotides and their corresponding nucleosides and nucleobases. The approach comprised a single-step acidic extraction/quenching procedure, followed by quantitative electrospray LC-MS/MS analysis. The assay was validated in terms of accuracy, precision, reproducibility, and applicability for complex biological matrices. The method was subsequently applied for determination of free intracellular pool sizes of purine biosynthetic pathway intermediates in the two Gram-positive bacteria Corynebacterium glutamicum and Corynebacterium ammoniagenes. Importantly, no ion pair reagents were applied in this approach as usually required for liquid chromatography analysis of large classes of diverse metabolites.

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“…Nucleotide analyses are performed in biochemical [11][12][13][14][15][16][17][18][19][20][21], medical [11, 13-15, 20, 21] and pharmacological [11,13,16] studies as well as in the food industry [11,22]. Nucleotides are not only the precursors and break-down products of DNA and RNA.…”

Section: Nucleotidesmentioning

confidence: 99%

“…Also enzyme assay, bioluminescence, 31 P nuclear magnetic resonance spectroscopy, and GC can be applied to the determination of nucleotides [20,26]. The high sensitivity and selectivity of MS and tandem MS (MS/MS) techniques provide great separation and detection power, making LC-MS and LC-MS/MS [12,27,28] as well as CE-MS and CE-MS/MS [11,[17][18][19][29][30][31][32][33][34] attractive alternatives for the analysis of nucleotides.…”

Section: Nucleotidesmentioning

confidence: 99%

Analysis of nucleic acid constituents by on‐line capillary electrophoresis‐mass spectrometry

et al. 2005

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This review is focused on the capillary electrophoresis-mass spectrometric (CE-MS) analysis of nucleic acid constituents in the broadest sense, going from nucleotides and adducted nucleotides over nucleoside analogues to oligonucleotides. These nucleic acid constituents play an important role in a variety of biochemical processes. Hence, their isolation, identification, and quantification will undoubtedly help reveal the process of life and disease mechanisms, such as carcinogenesis, and can also be useful for antitumor and antiviral drug research to provide valuable information about mechanism of action, pharmacokinetics, pharmacodynamics, toxicity, therapeutic drug level monitoring, and quality control related to this substance class. Fundamental investigations into their structure, the search for modifications, the occurrence and biochemical impact of structural variation amongst others, are therefore of great value. In view of the related bioanalytical procedures, the coupling of CE to MS has emerged as a powerful tool for the analysis of the complex mixtures of nucleic acid constituents: CE confers rapid analysis and efficient resolution, while MS provides high selectivity and sensitivity with structural characterization of minute amounts of compound. After an introduction about the biochemical and analytical perspectives on the nucleic acid constituents, the different modes of CE used in this field of research as well as the relevant CE-MS interfaces and the difficulties associated with quantitative CE-MS are briefly discussed. A large section is finally devoted to field-oriented applications.

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Capillary liquid chromatographic–high-resolution mass spectrometric analysis of ribonucleotides

Cited by 26 publications

References 17 publications

Mass spectrometry in the quantitative analysis of therapeutic intracellular nucleotide analogs

Mass spectrometry in the quantitative analysis of therapeutic intracellular nucleotide analogs

Quantitation of intracellular purine intermediates in different Corynebacteria using electrospray LC-MS/MS

Analysis of nucleic acid constituents by on‐line capillary electrophoresis‐mass spectrometry

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