Research Article

Servizo de Toxicoloxía, Instituto de Ciencias Forenses, Universidade de Santiago de Compostela, Rúa San Francisco, s/n, 15782 Santiago de Compostela, Spain

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Elena Lendoiro

Servizo de Toxicoloxía, Instituto de Ciencias Forenses, Universidade de Santiago de Compostela, Rúa San Francisco, s/n, 15782 Santiago de Compostela, Spain

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Esteban Otero-Antón

Unidade de Transplante Abdominal, Complexo Hospitalario Universitario de Santiago de Compostela, Rúa da Choupana, s/n, 15706 Santiago de Compostela, Spain

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Esther Molina-Pérez

Unidade de Transplante Abdominal, Complexo Hospitalario Universitario de Santiago de Compostela, Rúa da Choupana, s/n, 15706 Santiago de Compostela, Spain

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Evaristo Varo-Pérez

Unidade de Transplante Abdominal, Complexo Hospitalario Universitario de Santiago de Compostela, Rúa da Choupana, s/n, 15706 Santiago de Compostela, Spain

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Manuel López-Rivadulla

Servizo de Toxicoloxía, Instituto de Ciencias Forenses, Universidade de Santiago de Compostela, Rúa San Francisco, s/n, 15782 Santiago de Compostela, Spain

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Angelines Cruz

Servizo de Toxicoloxía, Instituto de Ciencias Forenses, Universidade de Santiago de Compostela, Rúa San Francisco, s/n, 15782 Santiago de Compostela, Spain

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Ana de-Castro-Ríos

*Author for correspondence: Tel.: +34 88181 2446; Fax: +34 88181 2459;

E-mail Address: ana.decastro@usc.es

https://orcid.org/0000-0002-9832-012X

Servizo de Toxicoloxía, Instituto de Ciencias Forenses, Universidade de Santiago de Compostela, Rúa San Francisco, s/n, 15782 Santiago de Compostela, Spain

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Published Online:11 Sep 2019https://doi.org/10.4155/bio-2019-0143

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Abstract

Aim: This study aimed: to develop and validate an LC–MS/MS method for mycophenolic acid, tacrolimus, sirolimus, everolimus and cyclosporin A in oral fluid (OF), as an essential tool to study the usefulness of OF as an alternative matrix for immunossuppressants’ therapeutic drug monitoring; and to find the best OF collector for these analytes. Materials & Methods: Chromatographic separation was achieved using an XBridge^® Shield RP18 analytical column maintained at 65ºC, using 2 mM ammonium formate and 0.1% formic acid in water (A) and acetonitrile (B) as mobile phase. OF sample was extracted with solid phase extraction after sonication and protein precipitation. Results & Conclusions: Method validation met all the acceptance criteria. LODs were 0.05–1 ng/ml, and LOQs 0.1–5 ng/ml. Silanized tubes offered the best recoveries. The method was successfully applied to 31 OF specimens, describing everolimus detection in OF for the first time. Conclusion: The proposed method is sensitive enough for the detection of OF trough concentrations in patients receiving immunosuppressants when using an appropriate OF collector.

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Papers of special note have been highlighted as: • of interest; •• of considerable interest

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Vol. 11, No. 16

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History

Received 11 June 2019

Accepted 14 August 2019

Published online 11 September 2019

Published in print August 2019

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Acknowledgments

The authorss would like to thank the Xunta de Galicia (Galicia, Spain) for the Competitive Reference Groups Help (ED431C 2017/05). The authors sincerely thank the sanitary staff involved in the collection of the samples at the University Clinical Hospital Complex of Santiago de Compostela, Spain and all the patients who voluntarily participated in the study.

Financial & competing interests disclosure

L Paniagua-González and E Lendoiro thank the Consellería de Cultura, Educación e Ordenación Universitaria, Xunta de Galicia, for their predoctoral (ED481A-2018/059) and postdoctoral (ED481B2016/103-0) contracts, respectively. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval (Galician Ethics Committee, Spain; ref. number 2018/157), and procedures were in accordance with the tenets of the Declaration of Helsinki. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

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A multidrug LC–MS/MS method for the determination of five immunosuppressants in oral fluid

Abstract

Graphical abstract

References