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Development and validation of an analytical method using UPLC–MS/MS to quantify everolimus in dried blood spots in the oncology setting

Development and validation of an analytical method using UPLC–MS/MS to quantify everolimus in dried blood spots in the oncology setting
Development and validation of an analytical method using UPLC–MS/MS to quantify everolimus in dried blood spots in the oncology setting

While the therapeutic drug monitoring (TDM) of everolimus has been routinely performed for over 10 years in solid organ transplantation medicine, in order to optimize the balance between effectiveness and toxicity, it is yet uncommon in the treatment of malignancies. The aim of this study was to develop and validate a bioanalytical method to quantify everolimus in dried blood spots (DBS) to facilitate TDM for the oncology outpatient setting. The hematocrit effect of everolimus was investigated. An 7.5 mm disk from the central part of the DBS was punched, followed by the extraction of everolimus from the DBS by methanol/acetonitrile (80/20%) spiked with deuterium-labelled everolimus as internal standard. Subsequently, everolimus was separated and analyzed using ultra performance liquid chromatography-tandem mass spectrometry (UPLC–MS/MS). The UPLC–MS/MS method was validated according to the European Medicine Agency (EMA) guideline. Everolimus concentrations could be quantified over the range of 3–75 μg/L. The intra- and inter-assay precision and accuracy of the method were shown to be acceptable (coefficient of variation ≤10.7% and relative error ≤4.4%, respectively). The matrix effects appeared to be influenced by the hematocrit effect. The hematocrit effect was tested in a range of 0.20–0.50 L/L, at which hematocrit accuracy and precision were satisfactory at values ≥0.25 L/L. However, at 0.20 L/L hematocrit in combination with high everolimus concentrations of 20 and 40 μg/L, the precision was adequate (≤7.4%), but the accuracy was >15% of the nominal concentration. Everolimus was stable in DBS for at least 80 days at 2–8 °C. Given these results, the everolimus DBS method has been successfully developed and validated. Special attention is necessary for cancer patients with both a 0.20 L/L hematocrit in combination with everolimus concentrations ≥20 μg/L. A clinical validation for the use of everolimus DBS in cancer patients is currently being undertaken.

Analytical method, Dried blood spot testing, Everolimus, Hematocrit, Oncology, UPLC–MS/MS
0731-7085
106-113
Knapen, Lotte M.
a2c486f6-09fa-4b02-82b8-7658d206db69
de Beer, Yvo
24e2c303-d5cc-4e78-8264-a16cf73d03d5
Brüggemann, Roger J.M.
0d16fa0b-dc4e-4ca1-88d0-74ecd073b62d
Stolk, Leo M.
162c1cbf-0e75-4770-9946-b793e31a5129
de Vries, Frank
10245a32-6083-4feb-9d20-7e7db0f358b1
Tjan-Heijnen, Vivianne C.G.
66e662b5-75f1-474a-85bf-6c4f9ddbd811
Erp, Nielka P.van
e1342d63-a3f7-4459-a400-6821556c52ce
Croes, Sander
2cf9e562-32ac-43ba-ba14-4523a6ca4120
Knapen, Lotte M.
a2c486f6-09fa-4b02-82b8-7658d206db69
de Beer, Yvo
24e2c303-d5cc-4e78-8264-a16cf73d03d5
Brüggemann, Roger J.M.
0d16fa0b-dc4e-4ca1-88d0-74ecd073b62d
Stolk, Leo M.
162c1cbf-0e75-4770-9946-b793e31a5129
de Vries, Frank
10245a32-6083-4feb-9d20-7e7db0f358b1
Tjan-Heijnen, Vivianne C.G.
66e662b5-75f1-474a-85bf-6c4f9ddbd811
Erp, Nielka P.van
e1342d63-a3f7-4459-a400-6821556c52ce
Croes, Sander
2cf9e562-32ac-43ba-ba14-4523a6ca4120

Knapen, Lotte M., de Beer, Yvo, Brüggemann, Roger J.M., Stolk, Leo M., de Vries, Frank, Tjan-Heijnen, Vivianne C.G., Erp, Nielka P.van and Croes, Sander (2018) Development and validation of an analytical method using UPLC–MS/MS to quantify everolimus in dried blood spots in the oncology setting Journal of Pharmaceutical and Biomedical Analysis, 149, pp. 106-113. (doi:10.1016/j.jpba.2017.10.039).

Record type: Article

Abstract

While the therapeutic drug monitoring (TDM) of everolimus has been routinely performed for over 10 years in solid organ transplantation medicine, in order to optimize the balance between effectiveness and toxicity, it is yet uncommon in the treatment of malignancies. The aim of this study was to develop and validate a bioanalytical method to quantify everolimus in dried blood spots (DBS) to facilitate TDM for the oncology outpatient setting. The hematocrit effect of everolimus was investigated. An 7.5 mm disk from the central part of the DBS was punched, followed by the extraction of everolimus from the DBS by methanol/acetonitrile (80/20%) spiked with deuterium-labelled everolimus as internal standard. Subsequently, everolimus was separated and analyzed using ultra performance liquid chromatography-tandem mass spectrometry (UPLC–MS/MS). The UPLC–MS/MS method was validated according to the European Medicine Agency (EMA) guideline. Everolimus concentrations could be quantified over the range of 3–75 μg/L. The intra- and inter-assay precision and accuracy of the method were shown to be acceptable (coefficient of variation ≤10.7% and relative error ≤4.4%, respectively). The matrix effects appeared to be influenced by the hematocrit effect. The hematocrit effect was tested in a range of 0.20–0.50 L/L, at which hematocrit accuracy and precision were satisfactory at values ≥0.25 L/L. However, at 0.20 L/L hematocrit in combination with high everolimus concentrations of 20 and 40 μg/L, the precision was adequate (≤7.4%), but the accuracy was >15% of the nominal concentration. Everolimus was stable in DBS for at least 80 days at 2–8 °C. Given these results, the everolimus DBS method has been successfully developed and validated. Special attention is necessary for cancer patients with both a 0.20 L/L hematocrit in combination with everolimus concentrations ≥20 μg/L. A clinical validation for the use of everolimus DBS in cancer patients is currently being undertaken.

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More information

Accepted/In Press date: 28 October 2017
e-pub ahead of print date: 29 October 2017
Published date: 5 February 2018
Keywords: Analytical method, Dried blood spot testing, Everolimus, Hematocrit, Oncology, UPLC–MS/MS

Identifiers

Local EPrints ID: 417453
URI: https://eprints.soton.ac.uk/id/eprint/417453
ISSN: 0731-7085
PURE UUID: bec325e0-9403-4289-a90a-79e2c31ddc26

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Date deposited: 31 Jan 2018 17:30
Last modified: 31 Jan 2018 17:30

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Contributors

Author: Lotte M. Knapen
Author: Yvo de Beer
Author: Roger J.M. Brüggemann
Author: Leo M. Stolk
Author: Frank de Vries
Author: Vivianne C.G. Tjan-Heijnen
Author: Nielka P.van Erp
Author: Sander Croes

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