A LC-MS/MS method for the analysis of intracellular nucleoside triphosphate levels

Ping Chen; Zhongfa Liu; Shujun Liu; Zhiliang Xie; Josephine Aimiuwu; Jiuxia Pang; Rebecca Klisovic; William Blum; Michael R. Grever; Guido Marcucci; Kenneth K. Chan

doi:10.1007/s11095-009-9863-9

A LC-MS/MS method for the analysis of intracellular nucleoside triphosphate levels

Ping Chen, Zhongfa Liu, Shujun Liu, Zhiliang Xie, Josephine Aimiuwu, Jiuxia Pang, Rebecca Klisovic, William Blum, Michael R. Grever, Guido Marcucci, Kenneth K. Chan

Research output: Contribution to journal › Article › peer-review

53 Scopus citations

Abstract

Purpose: To simultaneously quantify intracellular nucleoside triphosphate (NTP) and deoxynucleoside triphosphate (dNTP) pools and to assess their changes produced by interfering with ribonucleotide reductase (RNR) expression in leukemia cells. Methods: A HPLC-MS/MS system was used to quantify intracellular NTP and dNTP pools. Results: The assay was linear between 50 nM, the lower limit of quantification (LLOQ), and 10 μM in cell lysate. The within-day coefficients of variation (CVs, n∈=∈5) were found to be 12.0-18.0% at the LLOQ and 3.0-9.0% between 500 and 5,000 nM for dNTPs and 8.0-15.0% and 2.0-6.0% for NTPs. The between-day CVs (n∈=∈5) were 9.0-13.0% and 3.0-11.0% for dNTPs and 9.0-13.0% and 3.0-6.0% for NTPs. The within-day accuracy values were 93.0-119.0% for both NTPs and dNTPs. ATP overlapped with dGTP and they were analyzed as a composite. This method was applied to measure basal intracellular dNTPs/NTPs in five leukemia cell lines exposed to the RNR antisense GTI-2040. Following drug treatment, dCTP and dATP levels were found to decrease significantly in MV4-11 and K562 cells. Additionally, perturbation of dNTP/NTP levels in bone marrow sample of a patient treated with GTI-2040 was detected. Conclusions: This method provides a practical tool to measure intracellular dNTP/NTP levels in cells and clinical samples.

Original language	English (US)
Pages (from-to)	1504-1515
Number of pages	12
Journal	Pharmaceutical research
Volume	26
Issue number	6
DOIs	https://doi.org/10.1007/s11095-009-9863-9
State	Published - Jun 2009
Externally published	Yes

Bibliographical note

Funding Information:
This work was supported in part by the National Cancer Research Institute, Bethesda, MD, RO1 CA102031 (P.I. GM) and by the Biomedical Mass Spectrometry Laboratory, The Ohio State University.

Keywords

DNTP/NTP levels
GTI-2040
LC-MS/MS

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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title = "A LC-MS/MS method for the analysis of intracellular nucleoside triphosphate levels",

abstract = "Purpose: To simultaneously quantify intracellular nucleoside triphosphate (NTP) and deoxynucleoside triphosphate (dNTP) pools and to assess their changes produced by interfering with ribonucleotide reductase (RNR) expression in leukemia cells. Methods: A HPLC-MS/MS system was used to quantify intracellular NTP and dNTP pools. Results: The assay was linear between 50 nM, the lower limit of quantification (LLOQ), and 10 μM in cell lysate. The within-day coefficients of variation (CVs, n∈=∈5) were found to be 12.0-18.0% at the LLOQ and 3.0-9.0% between 500 and 5,000 nM for dNTPs and 8.0-15.0% and 2.0-6.0% for NTPs. The between-day CVs (n∈=∈5) were 9.0-13.0% and 3.0-11.0% for dNTPs and 9.0-13.0% and 3.0-6.0% for NTPs. The within-day accuracy values were 93.0-119.0% for both NTPs and dNTPs. ATP overlapped with dGTP and they were analyzed as a composite. This method was applied to measure basal intracellular dNTPs/NTPs in five leukemia cell lines exposed to the RNR antisense GTI-2040. Following drug treatment, dCTP and dATP levels were found to decrease significantly in MV4-11 and K562 cells. Additionally, perturbation of dNTP/NTP levels in bone marrow sample of a patient treated with GTI-2040 was detected. Conclusions: This method provides a practical tool to measure intracellular dNTP/NTP levels in cells and clinical samples.",

keywords = "DNTP/NTP levels, GTI-2040, LC-MS/MS",

author = "Ping Chen and Zhongfa Liu and Shujun Liu and Zhiliang Xie and Josephine Aimiuwu and Jiuxia Pang and Rebecca Klisovic and William Blum and Grever, {Michael R.} and Guido Marcucci and Chan, {Kenneth K.}",

note = "Funding Information: This work was supported in part by the National Cancer Research Institute, Bethesda, MD, RO1 CA102031 (P.I. GM) and by the Biomedical Mass Spectrometry Laboratory, The Ohio State University.",

year = "2009",

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doi = "10.1007/s11095-009-9863-9",

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AU - Chen, Ping

AU - Liu, Zhongfa

AU - Liu, Shujun

AU - Xie, Zhiliang

AU - Aimiuwu, Josephine

AU - Pang, Jiuxia

AU - Klisovic, Rebecca

AU - Blum, William

AU - Grever, Michael R.

AU - Marcucci, Guido

AU - Chan, Kenneth K.

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N2 - Purpose: To simultaneously quantify intracellular nucleoside triphosphate (NTP) and deoxynucleoside triphosphate (dNTP) pools and to assess their changes produced by interfering with ribonucleotide reductase (RNR) expression in leukemia cells. Methods: A HPLC-MS/MS system was used to quantify intracellular NTP and dNTP pools. Results: The assay was linear between 50 nM, the lower limit of quantification (LLOQ), and 10 μM in cell lysate. The within-day coefficients of variation (CVs, n∈=∈5) were found to be 12.0-18.0% at the LLOQ and 3.0-9.0% between 500 and 5,000 nM for dNTPs and 8.0-15.0% and 2.0-6.0% for NTPs. The between-day CVs (n∈=∈5) were 9.0-13.0% and 3.0-11.0% for dNTPs and 9.0-13.0% and 3.0-6.0% for NTPs. The within-day accuracy values were 93.0-119.0% for both NTPs and dNTPs. ATP overlapped with dGTP and they were analyzed as a composite. This method was applied to measure basal intracellular dNTPs/NTPs in five leukemia cell lines exposed to the RNR antisense GTI-2040. Following drug treatment, dCTP and dATP levels were found to decrease significantly in MV4-11 and K562 cells. Additionally, perturbation of dNTP/NTP levels in bone marrow sample of a patient treated with GTI-2040 was detected. Conclusions: This method provides a practical tool to measure intracellular dNTP/NTP levels in cells and clinical samples.

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A LC-MS/MS method for the analysis of intracellular nucleoside triphosphate levels

Abstract

Bibliographical note

Keywords

UN SDGs

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