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Fast and sensitive HPLC-MS/MS method for direct quantification of intracellular deoxyribonucleoside triphosphates from tissue and cells

J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.. 2017-11; 
OlafssonSigurast, WhittingtonDale, MurrayJason, RegnierMichael, Moussavi-HaramiF
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Biochemicals … dNTP standards, 2′-deoxyadenosine 5′-triphosphate (dATP), 2′-deoxyguanosine 5′-triphosphate (dGTP), 2′-deoxycytidine 5′-triphosphate (dCTP), and 2′-deoxythymidine 5′-triphosphate (dTTP), were purchased as a mixture from GenScript (Piscataway, NJ, USA) … Get A Quote

摘要

Deoxyribonucleoside triphosphates (dNTPs) are used in DNA synthesis and repair. Even slight imbalances can have adverse biological effects. This study validates a fast and sensitive HPLC-MS/MS method for direct quantification of intracellular dNTPs from tissue. Equal volumes of methanol and water were used for nucleotide extraction from mouse heart and gastrocnemius muscle and isolated cardiomyocytes followed by centrifugation to remove particulates. The resulting supernatant was analyzed on a porous graphitic carbon chromatography column using an elution gradient of ammonium acetate in water and ammonium hydroxide in acetonitrile with a run time of just 10min. Calibration curves of all dNTPs ranged from 62.5 t... More

关键词

ATP,HPLC–MS/MS,dNTP p