Monitoring caffeine in human saliva using a newly developed ELISA
- Caffeine is a useful indicator to quickly assess liver function. High-throughput tests are needed for single-point caffeine measurements, with low cross-reactivity toward its major metabolite, paraxanthine. A newly developed ELISA was compared with an LC-MS/MS reference method, using 60 saliva samples from 10 individuals, before and after caffeine intake. Bland-Altman plot, Student t-test and F-test were used to compare the two methods. Proteins were precipitated using organic solvent and the caffeine recoveries compared with those obtained using sample microfiltration. The antibody, with a low cross-reactivity toward paraxanthine (0.08%), allows quantification of caffeine in saliva samples from 2.5 µg/L to 125 µg/L with high precision and the ELISA shows comparable results to those obtained by LC-MS/MS. A one-step protein precipitation using an organic solvent provides comparable results to a more costly and time-consuming microfiltration pre-treatment of samples. The new ELISA is aCaffeine is a useful indicator to quickly assess liver function. High-throughput tests are needed for single-point caffeine measurements, with low cross-reactivity toward its major metabolite, paraxanthine. A newly developed ELISA was compared with an LC-MS/MS reference method, using 60 saliva samples from 10 individuals, before and after caffeine intake. Bland-Altman plot, Student t-test and F-test were used to compare the two methods. Proteins were precipitated using organic solvent and the caffeine recoveries compared with those obtained using sample microfiltration. The antibody, with a low cross-reactivity toward paraxanthine (0.08%), allows quantification of caffeine in saliva samples from 2.5 µg/L to 125 µg/L with high precision and the ELISA shows comparable results to those obtained by LC-MS/MS. A one-step protein precipitation using an organic solvent provides comparable results to a more costly and time-consuming microfiltration pre-treatment of samples. The new ELISA is a fit-for-purpose method to accurately and precisely determine caffeine in saliva samples.…
Autor*innen: | José Joao Carvalho, Michael G. WellerORCiD, Ulrich PanneORCiD, Rudolf SchneiderORCiD |
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Dokumenttyp: | Zeitschriftenartikel |
Veröffentlichungsform: | Verlagsliteratur |
Sprache: | Englisch |
Titel des übergeordneten Werkes (Englisch): | Analytical letters |
Jahr der Erstveröffentlichung: | 2012 |
Verlag: | Taylor & Francis |
Verlagsort: | Philadelphia, Pa. |
Jahrgang/Band: | 45 |
Ausgabe/Heft: | 17 |
Erste Seite: | 2549 |
Letzte Seite: | 2561 |
Freie Schlagwörter: | Caffeine; Cytochrome P450 CYP1A2; Enzyme-linked immunosorbent assay; LC-MS/MS; Liver function tests; Saliva |
DOI: | 10.1080/00032719.2012.696226 |
ISSN: | 0003-2719 |
ISSN: | 1532-236X |
Verfügbarkeit des Dokuments: | Physisches Exemplar in der Bibliothek der BAM vorhanden ("Hardcopy Access") |
Bibliotheksstandort: | Sonderstandort: Publica-Schrank |
Datum der Freischaltung: | 20.02.2016 |
Referierte Publikation: | Ja |
Datum der Eintragung als referierte Publikation: | 05.12.2012 |