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Liquid chromatography–mass spectrometry for measuring deoxythioguanosine in DNA from thiopurine-treated patients
Adverse reactions and non-response are common in patients treated with thiopurine drugs. Current monitoring of drug metabolite levels for guiding treatment are limited to analysis of thioguanine nucleotides (TGNs) in erythrocytes after chemical derivatisation. Erythrocytes are not the target tissue...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4955110/ https://www.ncbi.nlm.nih.gov/pubmed/27362994 http://dx.doi.org/10.1016/j.jchromb.2016.06.017 |
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author | Coulthard, Sally A. Berry, Phil McGarrity, Sarah Ansari, Azhar Redfern, Christopher P.F. |
author_facet | Coulthard, Sally A. Berry, Phil McGarrity, Sarah Ansari, Azhar Redfern, Christopher P.F. |
author_sort | Coulthard, Sally A. |
collection | PubMed |
description | Adverse reactions and non-response are common in patients treated with thiopurine drugs. Current monitoring of drug metabolite levels for guiding treatment are limited to analysis of thioguanine nucleotides (TGNs) in erythrocytes after chemical derivatisation. Erythrocytes are not the target tissue and TGN levels show poor correlations with clinical response. We have developed a sensitive assay to quantify deoxythioguanosine (dTG) without derivatisation in the DNA of nucleated blood cells. Using liquid chromatography and detection by tandem mass spectrometry, an intra- and inter-assay variability below 7.8% and 17.0% respectively were achieved. The assay had a detection limit of 0.0003125 ng (1.1 femtomoles) dTG and was quantified in DNA samples relative to endogenous deoxyadenosine (dA) in a small group of 20 patients with inflammatory bowel disease, all of whom had been established on azathioprine (AZA) therapy for more than 25 weeks. These patients had dTG levels of 20–1360 mol dTG/10(6) mol dA; three patients who had not started therapy had no detectable dTG. This method, comparable to previous methods in sensitivity, enables the direct detection of a cytotoxic thiopurine metabolite without derivatisation in an easily obtainable, stable sample and will facilitate a better understanding of the mechanisms of action of these inexpensive yet effective drugs. |
format | Online Article Text |
id | pubmed-4955110 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-49551102016-08-15 Liquid chromatography–mass spectrometry for measuring deoxythioguanosine in DNA from thiopurine-treated patients Coulthard, Sally A. Berry, Phil McGarrity, Sarah Ansari, Azhar Redfern, Christopher P.F. J Chromatogr B Analyt Technol Biomed Life Sci Short Communication Adverse reactions and non-response are common in patients treated with thiopurine drugs. Current monitoring of drug metabolite levels for guiding treatment are limited to analysis of thioguanine nucleotides (TGNs) in erythrocytes after chemical derivatisation. Erythrocytes are not the target tissue and TGN levels show poor correlations with clinical response. We have developed a sensitive assay to quantify deoxythioguanosine (dTG) without derivatisation in the DNA of nucleated blood cells. Using liquid chromatography and detection by tandem mass spectrometry, an intra- and inter-assay variability below 7.8% and 17.0% respectively were achieved. The assay had a detection limit of 0.0003125 ng (1.1 femtomoles) dTG and was quantified in DNA samples relative to endogenous deoxyadenosine (dA) in a small group of 20 patients with inflammatory bowel disease, all of whom had been established on azathioprine (AZA) therapy for more than 25 weeks. These patients had dTG levels of 20–1360 mol dTG/10(6) mol dA; three patients who had not started therapy had no detectable dTG. This method, comparable to previous methods in sensitivity, enables the direct detection of a cytotoxic thiopurine metabolite without derivatisation in an easily obtainable, stable sample and will facilitate a better understanding of the mechanisms of action of these inexpensive yet effective drugs. Elsevier 2016-08-15 /pmc/articles/PMC4955110/ /pubmed/27362994 http://dx.doi.org/10.1016/j.jchromb.2016.06.017 Text en © 2016 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Short Communication Coulthard, Sally A. Berry, Phil McGarrity, Sarah Ansari, Azhar Redfern, Christopher P.F. Liquid chromatography–mass spectrometry for measuring deoxythioguanosine in DNA from thiopurine-treated patients |
title | Liquid chromatography–mass spectrometry for measuring deoxythioguanosine in DNA from thiopurine-treated patients |
title_full | Liquid chromatography–mass spectrometry for measuring deoxythioguanosine in DNA from thiopurine-treated patients |
title_fullStr | Liquid chromatography–mass spectrometry for measuring deoxythioguanosine in DNA from thiopurine-treated patients |
title_full_unstemmed | Liquid chromatography–mass spectrometry for measuring deoxythioguanosine in DNA from thiopurine-treated patients |
title_short | Liquid chromatography–mass spectrometry for measuring deoxythioguanosine in DNA from thiopurine-treated patients |
title_sort | liquid chromatography–mass spectrometry for measuring deoxythioguanosine in dna from thiopurine-treated patients |
topic | Short Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4955110/ https://www.ncbi.nlm.nih.gov/pubmed/27362994 http://dx.doi.org/10.1016/j.jchromb.2016.06.017 |
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