Quantitative High-performance Liquid Chromatography Analysis of Modified Nucleosides in Physiological Fluids, tRNA, and DNA

Gehrke, Charles W.; Zumwalt, Robert W.; McCune, Roy A.; Kuo, Kenneth C.

doi:10.1007/978-3-642-81947-6_26

Cited by 6 publications

(4 citation statements)

References 4 publications

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“…The most widely used method for the analysis of urinary nucleosides is based on HPLC methods that were developed by Gherke et al (Gehrke et al, 1983;Kuo et al, 1987). Prior to the development of those HPLC methods, various techniques, including paper-, thin-layer-, and open tubular column chromatography, were used for the separation and isolation of nucleosides from the other urinary components.…”

Section: Sample Preparation Separation and Analysis Of Urinarymentioning

confidence: 99%

Urinary nucleosides

Schram

1998

Mass Spectrom. Rev.

100

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Section: Sample Preparation Separation and Analysis Of Urinarymentioning

confidence: 99%

Urinary nucleosides

Schram

1998

Mass Spectrom. Rev.

100

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“…The introduction of high performance liquid chromatography (HPLC) , and, later, mass spectrometry (MS), and reviewed in ref , to nucleotide analysis has greatly increased the resolution of nucleoside separations and purifications. Guided by some basic principles, empirical methods can be employed to determine buffer systems giving optimal separation during HPLC.…”

Section: Introductionmentioning

confidence: 99%

Biosynthesis and Function of Modified Bases in Bacteria and Their Viruses

2016

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Naturally occurring modification of the canonical A, G, C, and T bases can be found in the DNA of cellular organisms and viruses from all domains of life. Bacterial viruses (bacteriophages) are a particularly rich but still underexploited source of such modified variant nucleotides. The modifications conserve the coding and base-pairing functions of DNA, but add regulatory and protective functions. In prokaryotes, modified bases appear primarily to be part of an arms race between bacteriophages (and other genomic parasites) and their hosts, although, as in eukaryotes, some modifications have been adapted to convey epigenetic information. The first half of this review catalogs the identification and diversity of DNA modifications found in bacteria and bacteriophages. What is known about the biogenesis, context, and function of these modifications are also described. The second part of the review places these DNA modifications in the context of the arms race between bacteria and bacteriophages. It focuses particularly on the defense and counter-defense strategies that turn on direct recognition of the presence of a modified base. Where modification has been shown to affect other DNA transactions, such as expression and chromosome segregation, that is summarized, with reference to recent reviews.

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“…Gas chromatography (GC) and liquid chromatography (LC) have both been used to separate and identify nucleosides from urine , and hydrolyzed DNA. ,,− These two methods can be combined with on-line MS analysis to identify and quantify isotopically enriched deoxyribonucleosides (dRNs), which has proven to be essential for the study of T-cell proliferation in vivo. − Macallan et al developed a GC/MS method for the quantification of isotopically labeled dRNs . The first step described in this approach is the administration of glucose labeled with deuterium to the subject of interest (e.g., cell lines, rats, humans, etc.).…”

mentioning

confidence: 99%

“…2 Gas chromatography (GC) and liquid chromatography (LC) have both been used to separate and identify nucleosides from urine 3,4 and hydrolyzed DNA. 5,6,[10][11][12] combined with on-line MS analysis to identify and quantify isotopically enriched deoxyribonucleosides (dRNs), which has proven to be essential for the study of T-cell proliferation in vivo. [7][8][9] Macallan et al developed a GC/MS method for the quantification of isotopically labeled dRNs.…”

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confidence: 99%

A Comparison of μLC/Electrospray Ionization-MS and GC/MS for the Measurement of Stable Isotope Enrichment from a [²H₂]-Glucose Metabolic Probe in T-Cell Genomic DNA

et al. 2003

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Measurement of the proliferation of lymphocytes and other high-turnover cell populations in vivo can be accomplished through the incorporation of an isotopically labeled DNA precursor into actively dividing cells and the subsequent determination of the isotope enrichment in the isolated genomic DNA from selected cell populations. Two published gas chromatography/mass spectrometry (GC/MS) methods were successfully modified by our laboratory whereby a postinjection methylation reaction, rather than silylation or acetylation, was used to form a volatile derivative of deoxyadenosine (dA). We also developed a second robust microcapillary liquid chromatography-electrospray ionization (microLC-ESI)/MS method that is faster and more sensitive than the GC/MS method and does not require sample derivatization. Following administration of [6,6-(2)H(2)]-glucose to human immunodeficiency virus-infected patients, peripheral blood was drawn; cells were obtained by lymphapheresis and fractionated. DNA was isolated from the desired cell subtypes and enzymatically hydrolyzed to the free deoxyribonucleosides. The digest was analyzed using both capillary GC/MS and microLC/ESI-MS to measure the levels of the dA and [(2)H(2)]-dA or their reaction products. Sample enrichments were calculated by comparison to standard curves prepared from dA and [(2)H(2)]-dA. The microLC/ESI-MS method required fewer cells, less sample preparation, shorter analysis times, and a single calibration curve. Overall, the microLC/ESI-MS method is superior to the GC/MS method in terms of precision and accuracy, while providing a 4-fold increase in sensitivity (from 20 pmol at 0.2% [(2)H(2)]-dA enrichment to 5 pmol at 0.1% [(2)H(2)]-dA enrichment).

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Quantitative High-performance Liquid Chromatography Analysis of Modified Nucleosides in Physiological Fluids, tRNA, and DNA

Cited by 6 publications

References 4 publications

Urinary nucleosides

Urinary nucleosides

Biosynthesis and Function of Modified Bases in Bacteria and Their Viruses

A Comparison of μLC/Electrospray Ionization-MS and GC/MS for the Measurement of Stable Isotope Enrichment from a [²H₂]-Glucose Metabolic Probe in T-Cell Genomic DNA

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Quantitative High-performance Liquid Chromatography Analysis of Modified Nucleosides in Physiological Fluids, tRNA, and DNA

Cited by 6 publications

References 4 publications

Urinary nucleosides

Urinary nucleosides

Biosynthesis and Function of Modified Bases in Bacteria and Their Viruses

A Comparison of μLC/Electrospray Ionization-MS and GC/MS for the Measurement of Stable Isotope Enrichment from a [2H2]-Glucose Metabolic Probe in T-Cell Genomic DNA

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Product

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A Comparison of μLC/Electrospray Ionization-MS and GC/MS for the Measurement of Stable Isotope Enrichment from a [²H₂]-Glucose Metabolic Probe in T-Cell Genomic DNA