Simultaneous determination of 8-hydroxy-2′-deoxyguanosine and 5-methyl-2′-deoxycytidine in DNA sample by high performance liquid chromatography/positive electrospray ionization tandem mass spectrometry

Hu, Junjie; Zhang, Wenbing; Ma, Huimin; Cai, Yunmei; Sheng, Guoying; Fu, Jiamo

doi:10.1016/j.jchromb.2010.08.017

Cited by 19 publications

(9 citation statements)

References 34 publications

(37 reference statements)

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“…It can be found after the fragmentation, when the main [M+H] + adduct of 5mdC appears at m/z 126.0662. This ion originates due to the cleavage of the N -glycoside bond and transfer of a hydrogen atom from the sugar molecule [24]. Because these findings are comparable to the detection of dC and dG, the precursor/product ion pairs of m/z 228.0979/112.0505 for dC, m/z 242.1135/126.0662 for 5mdC and m/z 268.1040/152.0780 for dG were used as MRM transitions.…”

Section: Resultsmentioning

confidence: 99%

Analysis of Genomic DNA Methylation Levels in Human Placenta using Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry

Putra

Neuber

Reichetzeder

et al. 2014

Cell Physiol Biochem

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Background: DNA-methylation is a common epigenetic tool which plays a crucial role in gene regulation and is essential for cell differentiation and embryonic development. The placenta is an important organ where gene activity can be regulated by epigenetic DNA modifications, including DNA methylation. This is of interest as, the placenta is the interface between the fetus and its environment, the mother. Exposure to environmental toxins and nutrition during pregnancy may alter DNA methylation of the placenta and subsequently placental function and as a result the phenotype of the offspring. The aim of this study was to develop a reliable method to quantify DNA methylation in large clinical studies. This will be a tool to analyze the degree of DNA methylation in the human placenta in relationship to clinical readouts. Methods: Liquid chromatography-electrospray ionization/multi-stage mass spectrometry (LC-ESI/MS/MS) technique was used for the quantification of the 5dmC/dG ratio in placentas from 248 healthy pregnancies. We were able to demonstrate that this method is a reliable and stable way to determine global placental DNA methylation in large clinical trials. Results/Conclusion: The degree of placental DNA methylation seen in our pilot study varies substantially from 2% to 5%. The clinical implications of this variation need to be demonstrated in adequately powered large studies.

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Section: Resultsmentioning

confidence: 99%

Analysis of Genomic DNA Methylation Levels in Human Placenta using Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry

Putra

Neuber

Reichetzeder

et al. 2014

Cell Physiol Biochem

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“…There might therefore be a connection between genetic and epigenetic alterations, possibly via oxidative stress in such patients. Several reports have investigated the relationship between oxidized DNA and DNA methylation [55–58], including the simultaneous examination of 8-OHdG and 5MedC in DNA samples [55]. 8-OHdG may induce hypomethylation of DNA by inhibiting DNA methylation at nearby cytosine bases [58].…”

Section: Discussionmentioning

confidence: 99%

Urine 5MedC, a Marker of DNA Methylation, in the Progression of Chronic Kidney Disease

et al. 2019

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Background. Alterations in DNA methylation may be involved in disease progression in patients with chronic kidney disease (CKD). Recent studies have suggested that 5-methyl-2′-deoxycytidine (5MedC) may be a marker of hypermethylation of DNA. Currently, there is no information available regarding the urine levels of 5MedC and its association with the progression of CKD. Method. We examined the urine levels of 5MedC in spot urine samples from 308 patients with CKD (median age: 56 years, male: 53.2%, and glomerulonephritis: 51.0%) using a competitive enzyme-linked immunosorbent assay and investigated the relationships among urine 5MedC, urine albumin, urine α1-microglobulin (α1MG), and the laboratory parameters associated with CKD. The patients were followed for three years to evaluate renal endpoints in a prospective manner. Results. The urine 5MedC level was significantly increased in the later stages of CKD compared to the early to middle stages of CKD. In multiple logistic regression models, urine 5MedC was significantly associated with the prediction of later CKD stages. Urine 5MedC (median value, 65.9 μmol/gCr) was significantly able to predict a 30% decline in the estimated GFR or a development of end-stage renal disease when combined with macroalbuminuria or an increased level of urine α1MG (median value, 5.7 mg/gCr). Conclusion. The present data demonstrate that the urine 5MedC level is associated with a reduced renal function and can serve as a novel and potent biomarker for predicting the renal outcome in CKD patients. Further studies will be necessary to elucidate the role of urine DNA methylation in the progression of CKD.

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“…Therefore, the 8-OHdG levels are considered to be a valid marker of oxidative DNA damage. In addition, new techniques have been introduced into this field as gas chromatography with mass spectrometry [17,25], electrospray ionization mass spectrometry (MS) [26], liquid chromatography with mass spectrometry [27][28][29][30] and capillary electrophoresis [31][32][33][34][35][36]. From the direct ones, high performance liquid chromatography (HPLC) with electrochemical detection (ED) [23,24] belongs to the most commonly used method for determination of 8-OHdG.…”

Section: Determination Of 8-ohdgmentioning

confidence: 99%

Electrochemical Study of DNA Damaged by Oxidation Stress

Zítka¹,

Křížková²,

Skaličková³

et al. 2013

Combinatorial Chemistry & High Throughput Screening

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Many compounds can interact with DNA leading to changes of DNA structure as point mutation and bases excision, which could trigger some metabolic failures, which leads to the changes in DNA structure resulting in cancer. Oxidation of nucleic acid bases belongs to the one of the mostly occurred type of DNA damaging leading to the above mentioned phenomena. The investigation of processes of DNA oxidation damage is topical and electrochemical methods include a versatile and sensitive tool for these purposes. 8-hydroxydeoxyguanosine (8-OHdG) is the most widely accepted marker of DNA damage. Oxidative damage to DNA by free radicals and exposure to ionizing radiation generate several other products within the double helix besides mentioned oxidation products of nucleic acid bases. The basic electrochemical behaviour of nucleic acids bases on various types of carbon electrodes is reviewed. Further, we address our attention on description of oxidation mechanisms and on detection of the most important products of nucleic bases oxidation. The miniaturization of detector coupled with some microfluidic devices is suggested and discussed. The main aim of this review is to report the advantages and features of the electrochemical detection of guanine oxidation product as 8-OHdG and other similarly produced molecules as markers for DNA damage.

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Simultaneous determination of 8-hydroxy-2′-deoxyguanosine and 5-methyl-2′-deoxycytidine in DNA sample by high performance liquid chromatography/positive electrospray ionization tandem mass spectrometry

Cited by 19 publications

References 34 publications

Analysis of Genomic DNA Methylation Levels in Human Placenta using Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry

Analysis of Genomic DNA Methylation Levels in Human Placenta using Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry

Urine 5MedC, a Marker of DNA Methylation, in the Progression of Chronic Kidney Disease

Electrochemical Study of DNA Damaged by Oxidation Stress

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