This article describes the method development approaches for bioassay of substances containing unstable functional groups and forming unstable metabolites using the example of mycophenolic acid, methyldopa and mebeverine metabolite

Dzhurko, Yu. A.

Khokhlov, A. L.

Shitov, L. N.

Shitova, A. A.

Yaichkov, I. I.

Research Results in Pharmacology

Introduction: This article describes the method development approaches for bioassay of substances containing unstable functional groups and forming unstable metabolites using the example of mycophenolic acid, methyldopa and mebeverine metabolites.
        
          Materials and Methods: The concentration of mycophenolic acid, which contains one phenolic hydroxyl and forms glucuronides during metabolism, was measured in plasma using HPLC-MS/MS, HPLC-MS and GC-MS. The determination of methyldopa, containing two phenolic hydroxyls, in stabilised plasma was performed by HPLC-MS/MS in the range of 0.02-3.00 μg/ml. Desmethyl mebeverine acid, which contains one phenolic hydroxyl and is metabolised by forming phenolic glucuronide, was assayed simultaneously with mebeverine acid in the range of 10-2000 ng/ml.
        
          Results and Discussion: The selection of storage conditions of the samples containing unstable substances should begin with selecting an anticoagulant based on the study of its short-term stability and freeze/thaw stability. If an unacceptable result is obtained, a combination of the anticoagulant and a stabiliser solution, as well as a concentration of this solution and its volume ratio to the biological fluid should be titrated. After which, this method should be validated by using the selected anticoagulant or the combination of the anticoagulant and stabiliser solution.
        
          Conclusion: The application of this approach to developing a bioanalytical method for determination of unstable compounds makes it possible to avoid obtaining false assay results

Khokhlov, Alexander L.

Yaichkov, Ilya I.

Dzhurko, Yuriy A.

Shitov, Leonid N.

Shitova, Anastasia A.

ZENODO

Methodical approaches to bioassay of substances containing unstable functional groups

DSpace at Belgorod State University

Khokhlov,Alexander

Yaichkov,Ilya

Dzhurko,Yuriy

Shitov,Leonid

Shitova,Anastasia

ARPHA OAI-PMH Endpoint

ARPHA Preprints

Introduction: This article describes the method development approaches for bioassay of substances containing unstable functional groups and forming unstable metabolites using the example of mycophenolic acid, methyldopa and mebeverine metabolites.
          Materials and Methods: The concentration of mycophenolic acid, which contains one phenolic hydroxyl and forms glucuronides during metabolism, was measured in plasma using HPLC-MS/MS, HPLC-MS and GC-MS. The determination of methyldopa, containing two phenolic hydroxyls, in stabilised plasma was performed by HPLC-MS/MS in the range of 0.02-3.00 μg/ml. Desmethyl mebeverine acid, which contains one phenolic hydroxyl and is metabolised by forming phenolic glucuronide, was assayed simultaneously with mebeverine acid in the range of 10-2000 ng/ml.
          Results and Discussion: The selection of storage conditions of the samples containing unstable substances should begin with selecting an anticoagulant based on the study of its short-term stability and freeze/thaw stability. If an unacceptable result is obtained, a combination of the anticoagulant and a stabiliser solution, as well as a concentration of this solution and its volume ratio to the biological fluid should be titrated. After which, this method should be validated by using the selected anticoagulant or the combination of the anticoagulant and stabiliser solution.
          Conclusion: The application of this approach to developing a bioanalytical method for determination of unstable compounds makes it possible to avoid obtaining false assay results.</jats:p

Alexander L. Khokhlov

Ilya I. Yaichkov

Yuriy A. Dzhurko

Leonid N. Shitov

Anastasia A. Shitova

Crossref

NEUROSURGERY ENTHUSIASTIC WOMEN SOCIETY

http://dspace.bsu.edu.ru/bitstream/123456789/22853/1/Khokhlov_Methodical_18.pdf

Methodical approaches to bioassay of substances containing unstable functional groups

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ZENODO

DSpace at Belgorod State University

DSpace at Belgorod State University

ARPHA OAI-PMH Endpoint

ARPHA Preprints

Crossref

NEUROSURGERY ENTHUSIASTIC WOMEN SOCIETY