Validation of a Fast and Simple HPLC-UV Method for the Quantification of Adenosine Phosphates in Human Bronchial Epithelial Cells
A new HPLC method for the simultaneous quantitative analysis of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) was developed and validated. ATP, ADP, and AMP were extracted from human bronchial epithelial cells with a rapid extraction procedure and separ...
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MDPI AG
2021-10-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/26/20/6324 |
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| author | Ana Teresa Juarez-Facio Violaine Martin de Lagarde Christelle Monteil Jean-Marie Vaugeois Cécile Corbiere Tiphaine Rogez-Florent |
| author_facet | Ana Teresa Juarez-Facio Violaine Martin de Lagarde Christelle Monteil Jean-Marie Vaugeois Cécile Corbiere Tiphaine Rogez-Florent |
| author_sort | Ana Teresa Juarez-Facio |
| collection | DOAJ |
| description | A new HPLC method for the simultaneous quantitative analysis of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) was developed and validated. ATP, ADP, and AMP were extracted from human bronchial epithelial cells with a rapid extraction procedure and separated with a C18 column (3 × 150 mm, 2.7 µm) using isocratic elution with a mobile phase consisting of 50 mM of potassium hydrogen phosphate (pH 6.80). The absorbance was monitored at 254 nm. The calibration curves were linear in 0.2 to 10 µM, selective, precise, and accurate. This method allowed us to quantify the nucleotides from two cell models: differentiated NHBE primary cells grown at the air–liquid interface (ALI) and BEAS-2B cell line. Our study highlighted the development of a sensitive, simple, and green analytical method that is faster and less expensive than other existing methods to measure ATP, ADP, and AMP and can be carried out on 2D and 3D cell models. |
| first_indexed | 2024-03-10T06:20:37Z |
| format | Article |
| id | doaj.art-ed974ecde59c478fa5c57ed890343735 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T06:20:37Z |
| publishDate | 2021-10-01 |
| publisher | MDPI AG |
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| series | Molecules |
| spelling | doaj.art-ed974ecde59c478fa5c57ed8903437352023-11-22T19:21:31ZengMDPI AGMolecules1420-30492021-10-012620632410.3390/molecules26206324Validation of a Fast and Simple HPLC-UV Method for the Quantification of Adenosine Phosphates in Human Bronchial Epithelial CellsAna Teresa Juarez-Facio0Violaine Martin de Lagarde1Christelle Monteil2Jean-Marie Vaugeois3Cécile Corbiere4Tiphaine Rogez-Florent5Normandie Univ, UNIROUEN, UNICAEN, ABTE, 76000 Rouen, FranceNormandie Univ, UNIROUEN, UNICAEN, ABTE, 76000 Rouen, FranceNormandie Univ, UNIROUEN, UNICAEN, ABTE, 76000 Rouen, FranceNormandie Univ, UNIROUEN, UNICAEN, ABTE, 76000 Rouen, FranceNormandie Univ, UNIROUEN, UNICAEN, ABTE, 76000 Rouen, FranceNormandie Univ, UNIROUEN, UNICAEN, ABTE, 76000 Rouen, FranceA new HPLC method for the simultaneous quantitative analysis of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) was developed and validated. ATP, ADP, and AMP were extracted from human bronchial epithelial cells with a rapid extraction procedure and separated with a C18 column (3 × 150 mm, 2.7 µm) using isocratic elution with a mobile phase consisting of 50 mM of potassium hydrogen phosphate (pH 6.80). The absorbance was monitored at 254 nm. The calibration curves were linear in 0.2 to 10 µM, selective, precise, and accurate. This method allowed us to quantify the nucleotides from two cell models: differentiated NHBE primary cells grown at the air–liquid interface (ALI) and BEAS-2B cell line. Our study highlighted the development of a sensitive, simple, and green analytical method that is faster and less expensive than other existing methods to measure ATP, ADP, and AMP and can be carried out on 2D and 3D cell models.https://www.mdpi.com/1420-3049/26/20/6324HPLC-UVBEAS-2B cellsNHBE cellsadenosine phosphatescellular extraction |
| spellingShingle | Ana Teresa Juarez-Facio Violaine Martin de Lagarde Christelle Monteil Jean-Marie Vaugeois Cécile Corbiere Tiphaine Rogez-Florent Validation of a Fast and Simple HPLC-UV Method for the Quantification of Adenosine Phosphates in Human Bronchial Epithelial Cells Molecules HPLC-UV BEAS-2B cells NHBE cells adenosine phosphates cellular extraction |
| title | Validation of a Fast and Simple HPLC-UV Method for the Quantification of Adenosine Phosphates in Human Bronchial Epithelial Cells |
| title_full | Validation of a Fast and Simple HPLC-UV Method for the Quantification of Adenosine Phosphates in Human Bronchial Epithelial Cells |
| title_fullStr | Validation of a Fast and Simple HPLC-UV Method for the Quantification of Adenosine Phosphates in Human Bronchial Epithelial Cells |
| title_full_unstemmed | Validation of a Fast and Simple HPLC-UV Method for the Quantification of Adenosine Phosphates in Human Bronchial Epithelial Cells |
| title_short | Validation of a Fast and Simple HPLC-UV Method for the Quantification of Adenosine Phosphates in Human Bronchial Epithelial Cells |
| title_sort | validation of a fast and simple hplc uv method for the quantification of adenosine phosphates in human bronchial epithelial cells |
| topic | HPLC-UV BEAS-2B cells NHBE cells adenosine phosphates cellular extraction |
| url | https://www.mdpi.com/1420-3049/26/20/6324 |
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