<abstract xmlns="http://www.w3.org/1999/xhtml">Theophylline has been used in the treatment of bronchial asthma and chronic obstructive pulmonary disease (COPD) for over 70 years. In order to maximize the effectiveness and safety of theophylline therapy it is important to individualize the dosage of the drug. In our study we focused on determination of theophylline concentrations in guinea pig plasma. A rapid, specific, and reliable LC-MS/MS-based method was developed and validated according to European Medicine Agency (EMA) guidelines. A hydrophilic interaction liquid chromatography (HILIC) separation mode for reduction time of sample preparation was used. The analysed sample was quantified in a positive ionization mode. Multiple reaction monitoring (MRM) using transition m/z 181.06→124.06 and m/z 187.17→127.06 was performed to quantify theophylline with deuterated internal standard ([2H6]-theophylline), respectively. Modification of collision energies was performed in parallel with chromatographic separation to further eliminate interference from the matrix. The method was validated for a range of 0.5 to 30 μg/mL of plasma sample. The intra-day and inter-day precision and accuracy of the quality control samples at low, me dium, and high concentration levels exhibited relative standard deviations (RSD) of less than 10 %. The method was successfully applied for the quantitation of theophylline in guinea pig plasma for better understanding its effects in a model of ovalbumin-induced allergic inflammation.</abstract>

Theophylline has been used in the treatment of bronchial asthma and chronic obstructive pulmonary disease (COPD) for over 70 years. In order to maximize the effectiveness and safety of theophylline therapy it is important to individualize the dosage of the drug. In our study we focused on determination of theophylline concentrations in guinea pig plasma. A rapid, specific, and reliable LC-MS/MS-based method was developed and validated according to European Medicine Agency (EMA) guidelines. A hydrophilic interaction liquid chromatography (HILIC) separation mode for reduction time of sample preparation was used. The analysed sample was quantified in a positive ionization mode. Multiple reaction monitoring (MRM) using transition m/z 181.06→124.06 and m/z 187.17→127.06 was performed to quantify theophylline with deuterated internal standard ([2H6]-theophylline), respectively. Modification of collision energies was performed in parallel with chromatographic separation to further eliminate interference from the matrix. The method was validated for a range of 0.5 to 30 μg/mL of plasma sample. The intra-day and inter-day precision and accuracy of the quality control samples at low, me dium, and high concentration levels exhibited relative standard deviations (RSD) of less than 10 %. The method was successfully applied for the quantitation of theophylline in guinea pig plasma for better understanding its effects in a model of ovalbumin-induced allergic inflammation.

Quantification of Theophylline in Guinea Pig Plasma by LC-MS/MS Using Hydrophilic Interaction Liquid Chromatography Stationary Phase: Method Development, Validation, and Application in Study

Department of Pharmacology, Jessenius Faculty of Medicine in Martin

Biomedical Center Martin, Jessenius Faculty of Medicine in Martin

Acta Medica Martiniana

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Theophylline has been used in the treatment of bronchial asthma and chronic obstructive pulmonary disease (COPD) for over 70 years. In order to maximize the...

Quantification of Theophylline in Guinea Pig Plasma by LC-MS/MS Using Hydrophilic Interaction Liquid Chromatography Stationary Phase: Method Development, Validation, and Application in Study

Publicado en línea: 20 feb 2018

Páginas: 5 - 15

Recibido: 18 oct 2017

Aceptado: 30 oct 2017

DOI: https://doi.org/10.1515/acm-2017-0012

Palabras clave
theophylline, HILIC, liquid chromatography, mass spectrometry, guinea pig, plasma

© 2017 Kertys M et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Quantification of Theophylline in Guinea Pig Plasma by LC-MS/MS Using Hydrophilic Interaction Liquid Chromatography Stationary Phase: Method Development, Validation, and Application in Study

Publicado en línea: 20 feb 2018

Páginas: 5 - 15

Recibido: 18 oct 2017

Aceptado: 30 oct 2017

DOI: https://doi.org/10.1515/acm-2017-0012

Palabras clavetheophylline, HILIC, liquid chromatography, mass spectrometry, guinea pig, plasma

© 2017 Kertys M et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Palabras clave
theophylline, HILIC, liquid chromatography, mass spectrometry, guinea pig, plasma