Quantitative determination of cyclic guanosine monoposphate (c-GMP) in rat tissues using liquid chromatography and tandem mass spectrometry

Relevance. Cyclic 3′,5′-guanosine monophosphate is a secondary intracellular messenger that plays a key role in many physiological processes.Quantitative determination of the level of c-GMP in the tissues of laboratory animals is an urgent task of experimental pharmacology and physiology.Purpose of...

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Main Authors: N. S. Popov, V. Yu. Balabanyan, N. Yu. Kolgina, G. A. Petrov, S. A. Donskov, I. B. Atadzhanov
Format: Article
Language:Russian
Published: LLC “Publisher OKI” 2023-11-01
Series:Фармакокинетика и Фармакодинамика
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Online Access:https://www.pharmacokinetica.ru/jour/article/view/379
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author N. S. Popov
V. Yu. Balabanyan
N. Yu. Kolgina
G. A. Petrov
S. A. Donskov
I. B. Atadzhanov
author_facet N. S. Popov
V. Yu. Balabanyan
N. Yu. Kolgina
G. A. Petrov
S. A. Donskov
I. B. Atadzhanov
author_sort N. S. Popov
collection DOAJ
description Relevance. Cyclic 3′,5′-guanosine monophosphate is a secondary intracellular messenger that plays a key role in many physiological processes.Quantitative determination of the level of c-GMP in the tissues of laboratory animals is an urgent task of experimental pharmacology and physiology.Purpose of the study. Development of a method for the quantitative determination of cyclic guanosine monophosphate in various tissues of rats using high performance liquid chromatography with mass spectrometric detection.Methods. The biomaterial was homogenized with deionized water. Extraction of c-GMP from homogenates was performed with methanol, acyclovir was used as an internal standard. Detection of c-GMP and acyclovir was performed using a Sciex QTrap 3200MD mass spectrometer, chromatographic separation was performed using an Agilent Technologies 1260 Infinity II HPLC. The mobile phase was methanol and deionized water.Results. Detection of c-GMP was performed by MRM transitions m/z 346.2/152.1; 346.2/135.1, chromatographic determination of c-GMP was performed in reverse phase mode on an Agilent InfinityLab Poroshell 120 EC-C18 4.6×100 mm, 2.7 µm column. The retention time of c-GMP and acyclovir was 7.85 and 7.45 minutes, respectively, the total duration of the chromatographic analysis was 12 minutes. The analytical range of the procedure for determining c-GMP in homogenates was 0.5–1000.0 pmol/ml. The content of c-GMP in the tissues of intact Wistar rats was analyzed using the developed method.Conclusion. The developed bioanalytical HPLC-MS/MS method for the quantitative determination of c-GMP fully complies with the validation requirements. The metrological characteristics of the method make it possible to estimate the content of c-GMP in various tissues of rats with high accuracy.
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institution Kabale University
issn 2587-7836
2686-8830
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publishDate 2023-11-01
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series Фармакокинетика и Фармакодинамика
spelling doaj-art-15ad24806c7b4a8fb047fc84bf6db7922025-08-20T03:57:48ZrusLLC “Publisher OKI”Фармакокинетика и Фармакодинамика2587-78362686-88302023-11-0103283810.37489/2587-7836-2023-3-28-38342Quantitative determination of cyclic guanosine monoposphate (c-GMP) in rat tissues using liquid chromatography and tandem mass spectrometryN. S. Popov0V. Yu. Balabanyan1N. Yu. Kolgina2G. A. Petrov3S. A. Donskov4I. B. Atadzhanov5FSBEI HE Tver SMU MOH RussiaLomonosov Moscow State University,FSBEI HE Tver SMU MOH RussiaFSBEI HE Tver SMU MOH RussiaFSBEI HE Tver SMU MOH RussiaFSBEI HE Tver SMU MOH RussiaRelevance. Cyclic 3′,5′-guanosine monophosphate is a secondary intracellular messenger that plays a key role in many physiological processes.Quantitative determination of the level of c-GMP in the tissues of laboratory animals is an urgent task of experimental pharmacology and physiology.Purpose of the study. Development of a method for the quantitative determination of cyclic guanosine monophosphate in various tissues of rats using high performance liquid chromatography with mass spectrometric detection.Methods. The biomaterial was homogenized with deionized water. Extraction of c-GMP from homogenates was performed with methanol, acyclovir was used as an internal standard. Detection of c-GMP and acyclovir was performed using a Sciex QTrap 3200MD mass spectrometer, chromatographic separation was performed using an Agilent Technologies 1260 Infinity II HPLC. The mobile phase was methanol and deionized water.Results. Detection of c-GMP was performed by MRM transitions m/z 346.2/152.1; 346.2/135.1, chromatographic determination of c-GMP was performed in reverse phase mode on an Agilent InfinityLab Poroshell 120 EC-C18 4.6×100 mm, 2.7 µm column. The retention time of c-GMP and acyclovir was 7.85 and 7.45 minutes, respectively, the total duration of the chromatographic analysis was 12 minutes. The analytical range of the procedure for determining c-GMP in homogenates was 0.5–1000.0 pmol/ml. The content of c-GMP in the tissues of intact Wistar rats was analyzed using the developed method.Conclusion. The developed bioanalytical HPLC-MS/MS method for the quantitative determination of c-GMP fully complies with the validation requirements. The metrological characteristics of the method make it possible to estimate the content of c-GMP in various tissues of rats with high accuracy.https://www.pharmacokinetica.ru/jour/article/view/379hplc-ms/mstandem mass spectrometryguanosine monophosphatec-gmp
spellingShingle N. S. Popov
V. Yu. Balabanyan
N. Yu. Kolgina
G. A. Petrov
S. A. Donskov
I. B. Atadzhanov
Quantitative determination of cyclic guanosine monoposphate (c-GMP) in rat tissues using liquid chromatography and tandem mass spectrometry
Фармакокинетика и Фармакодинамика
hplc-ms/ms
tandem mass spectrometry
guanosine monophosphate
c-gmp
title Quantitative determination of cyclic guanosine monoposphate (c-GMP) in rat tissues using liquid chromatography and tandem mass spectrometry
title_full Quantitative determination of cyclic guanosine monoposphate (c-GMP) in rat tissues using liquid chromatography and tandem mass spectrometry
title_fullStr Quantitative determination of cyclic guanosine monoposphate (c-GMP) in rat tissues using liquid chromatography and tandem mass spectrometry
title_full_unstemmed Quantitative determination of cyclic guanosine monoposphate (c-GMP) in rat tissues using liquid chromatography and tandem mass spectrometry
title_short Quantitative determination of cyclic guanosine monoposphate (c-GMP) in rat tissues using liquid chromatography and tandem mass spectrometry
title_sort quantitative determination of cyclic guanosine monoposphate c gmp in rat tissues using liquid chromatography and tandem mass spectrometry
topic hplc-ms/ms
tandem mass spectrometry
guanosine monophosphate
c-gmp
url https://www.pharmacokinetica.ru/jour/article/view/379
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