Accelerator Mass Spectrometry for Quantitative in Vivo Tracing

Vogel, John S.

doi:10.2144/05386su04

Cited by 20 publications

(29 citation statements)

References 45 publications

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“…AMS is a sensitive and precise technology for tracing 14 Clabeled compounds in biological systems that affords subpicomole to zeptomole 14 C per milligram of total carbon sensitivity with a few percent precision (15)(16)(17). Because only carboplatin bound to DNA in the form of monoadducts contains radiocarbon, as shown in Scheme 1, AMS can only detect carboplatin monoadducts from DNA samples.…”

Section: Introductionmentioning

confidence: 99%

Kinetics of Carboplatin−DNA Binding in Genomic DNA and Bladder Cancer Cells As Determined by Accelerator Mass Spectrometry

Hah

Stivers

White

et al. 2006

Chem. Res. Toxicol.

101

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Cisplatin and carboplatin are platinum-based drugs that are widely used in cancer chemotherapy. The cytotoxicity of these drugs is mediated by platinum-DNA monoadducts and intra-and interstrand diadducts, which are formed following uptake of the drug into the nucleus of cells. The pharmacodynamics of carboplatin display fewer side effects than for cisplatin, albeit with less potency, which may be due to differences in rates of DNA adduct formation. We report the use of accelerator mass spectrometry (AMS), a sensitive detection method often used for radiocarbon quantitation, to measure both the kinetics of [ 14 C]carboplatin-DNA adduct formation with genomic DNA and drug uptake and DNA binding in T24 human bladder cancer cells. Only carboplatin-DNA monoadducts contain radiocarbon in the platinated DNA, which allowed for calculation of kinetic rates and concentrations within the system. The percent of radiocarbon bound to salmon sperm DNA in the form of monoadducts was measured by AMS over 24 h. Knowledge of both the starting concentration of the parent carboplatin and the concentration of radiocarbon in the DNA at a variety of time points allowed calculation of the rates of Pt-DNA monoadduct formation and conversion to toxic cross-links. Importantly, the rate of carboplatin-DNA monoadduct formation was approximately 100-fold slower than that reported for the more potent cisplatin analogue, which may explain the lower toxicity of carboplatin. T24 human bladder cancer cells were incubated with a subpharmacological dose of [ 14 C]carboplatin, and the rate of accumulation of radiocarbon in the cells and nuclear DNA was measured by AMS. The lowest concentration of radiocarbon measured was approximately 1 amol/10 µg of DNA. This sensitivity may allow the method to be used for clinical applications.

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Section: Introductionmentioning

confidence: 99%

Kinetics of Carboplatin−DNA Binding in Genomic DNA and Bladder Cancer Cells As Determined by Accelerator Mass Spectrometry

Hah

Stivers

White

et al. 2006

Chem. Res. Toxicol.

101

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“…Thus, 8-oxodG metabolism is a useful model to test the utility of AMS for metabolic flux analysis. AMS is a well established technology for radiocarbon dating and is now used for tracing 14 C-labeled compounds in biological systems (21). AMS affords subpicomole to zeptomole sensitivity in quantifying 14 C-labeled metabolites with a few percent precision, depending on experimental conditions (21,22).…”

mentioning

confidence: 99%

“…AMS is a well established technology for radiocarbon dating and is now used for tracing 14 C-labeled compounds in biological systems (21). AMS affords subpicomole to zeptomole sensitivity in quantifying 14 C-labeled metabolites with a few percent precision, depending on experimental conditions (21,22). The method relies on measurement of 14 C in a biological sample that has been converted to solid graphite before analysis, which facilitates high measurement sensitivity because of efficient ion current production in the AMS instrument.…”

mentioning

confidence: 99%

Measurement of 7,8-dihydro-8-oxo-2′-deoxyguanosine metabolism in MCF-7 cells at low concentrations using accelerator mass spectrometry

Hah

Mundt

Kim

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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Growing evidence suggests that oxidative damage to cells generates mutagenic 7,8-dihydro-8-oxo-2 -deoxyguanosine (8-oxodG), which may initiate diseases related to aging and carcinogenesis. Kinetic measurement of 8-oxodG metabolism and repair in cells has been hampered by poor assay sensitivity and by difficulty characterizing the flux of oxidized nucleotides through the relevant metabolic pathways. We report here the development of a sensitive and quantitative approach to characterizing the kinetics and metabolic sources of 8-oxodG in MCF-7 human breast cancer cells by accelerator mass spectrometry. We observed that [ 14 C]8-oxodG at medium concentrations of up to 2 pmol/ml was taken up by MCF-7 cells, phosphorylated to mono-, di-, and triphosphate derivatives, and incorporated into DNA. Oxidative stress caused by exposure of the cells to 17␤-estradiol resulted in a reduction in the rate of [ 14 C]8-oxodG incorporation into DNA and an increase in the ratio of 8-oxodG monophosphate (8-oxodGMP) to 8-oxodG triphosphate (8-oxodGTP) in the nucleotide pool. 17␤-Estradiolinduced oxidative stress up-regulated the nucleotide pool cleansing enzyme MTH1 and possibly other Nudix-related pyrophosphohydrolases. These data support the conclusion that 8-oxodGTP is formed in the nucleotide pool by both 8-oxodG metabolism and endogenous reactive oxygen species. The metabolism of 8-oxodG to 8-oxodGTP, followed by incorporation into DNA is a mechanism by which the cellular presence of this oxidized nucleoside can lead to mutations.DNA repair ͉ nucleoside metabolism ͉ oxidative stress ͉ breast cancer

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Section: Introductionmentioning

confidence: 99%

“…5,19,20 Blood plasma drawn in early Y 2004 from humans at various US locations had average F m values of 1.086 ± 0.014. 20 13 C and 14 C levels in bloods of humans deceased in Y 2006 had an average δ 13 C of −19.13 ± 1.15 and an average F m value of 1.0607 ± 0.0069. 19 14 C levels (in units of F m ) in whole blood (WB), plasma, and blood clots of humans which were collected in Japan in Y 2007 averaged the F m of 1.0814 ± 0.0838, 1.0861 ± 0.0821, and 1.1667 ± 0.0428, respectively.…”

Section: Introductionmentioning

confidence: 99%

Carbon Isotopes Profiles of Human Whole Blood, Plasma, Red Blood Cells, Urine and Feces for Biological/Biomedical ¹⁴C-Accelerator Mass Spectrometry Applications

et al. 2011

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Radiocarbon (14C) is an ideal tracer for in vivo human ADME (absorption, distribution, metabolism, elimination) and PBPK (physiological-based pharmacokinetic) studies. Living plants preferentially incorporate atmospheric 14CO2, vs 13CO2, vs 12CO2, which result in unique signature. Furthermore, plants and the food chains they support also have unique carbon isotope signatures. Humans, at the top of the food chain, consequently acquire isotopic concentrations in the tissues and body fluids depending on their dietary habits. In preparation of ADME and PBPK studies, 12 healthy subjects were recruited. The human baseline (specific to each individual and their diet) total carbon (TC) and carbon isotope 13C (δ13C) and 14C (Fm) were quantified in whole blood (WB), plasma, washed red blood cell (RBC), urine, and feces. TC (mg of C/100μL) in WB, plasma, RBC, urine, and feces were 11.0, 4.37, 7.57, 0.53, and 1.90, respectively. TC in WB, RBC, and feces was higher in men over women, P < 0.05. Mean δ13C were ranked low to high as follows, feces < WB = plasma = RBC = urine, P < 0.0001. δ13C was not affected by gender. Our analytic method shifted δ13C by only ± 1.0 ‰ ensuring our Fm measurements were accurate and precise. Mean Fm were ranked low to high as follows, plasma = urine < WB = RBC = feces, P < 0.05. Fm in feces was higher for men over women, P < 0.05. Only in WB, 14C levels (Fm) and TC were correlated with one another (r = 0.746, P < 0.01). Considering the lag time to incorporate atmospheric 14C into plant foods (vegetarian) and or then into animal foods (non-vegetarian), the measured Fm of WB in our population (recruited April 2009) was 1.0468 ± 0.0022 (mean±SD), the Fm of WB matched the (extrapolated) atmospheric Fm of 1.0477 in 2008. This study is important in presenting a procedure to determine a baseline for a study group for human ADME and PBPK studies using 14C as a tracer.

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Accelerator Mass Spectrometry for Quantitative in Vivo Tracing

Cited by 20 publications

References 45 publications

Kinetics of Carboplatin−DNA Binding in Genomic DNA and Bladder Cancer Cells As Determined by Accelerator Mass Spectrometry

Kinetics of Carboplatin−DNA Binding in Genomic DNA and Bladder Cancer Cells As Determined by Accelerator Mass Spectrometry

Measurement of 7,8-dihydro-8-oxo-2′-deoxyguanosine metabolism in MCF-7 cells at low concentrations using accelerator mass spectrometry

Carbon Isotopes Profiles of Human Whole Blood, Plasma, Red Blood Cells, Urine and Feces for Biological/Biomedical ¹⁴C-Accelerator Mass Spectrometry Applications

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Accelerator Mass Spectrometry for Quantitative in Vivo Tracing

Cited by 20 publications

References 45 publications

Kinetics of Carboplatin−DNA Binding in Genomic DNA and Bladder Cancer Cells As Determined by Accelerator Mass Spectrometry

Kinetics of Carboplatin−DNA Binding in Genomic DNA and Bladder Cancer Cells As Determined by Accelerator Mass Spectrometry

Measurement of 7,8-dihydro-8-oxo-2′-deoxyguanosine metabolism in MCF-7 cells at low concentrations using accelerator mass spectrometry

Carbon Isotopes Profiles of Human Whole Blood, Plasma, Red Blood Cells, Urine and Feces for Biological/Biomedical 14C-Accelerator Mass Spectrometry Applications

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Product

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Carbon Isotopes Profiles of Human Whole Blood, Plasma, Red Blood Cells, Urine and Feces for Biological/Biomedical ¹⁴C-Accelerator Mass Spectrometry Applications