NDLI: Validation and application of a fast semi-automated whole blood extraction for LC-MS/MS simultaneous quantification of cyclosporine A, tacrolimus, sirolimus and everolimus – application to high throughput routine therapeutic drug monitoring

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Validation and application of a fast semi-automated whole blood extraction for LC-MS/MS simultaneous quantification of cyclosporine A, tacrolimus, sirolimus and everolimus – application to high throughput routine therapeutic drug monitoring

Content Provider	Royal Society of Chemistry (RSC)
Author	Jolliet, Pascale Deslandes, Guillaume Dailly, Eric Bouquié, Régis Renaud, Christian
Copyright Year	2013
Abstract	Numerous laboratories carried out simultaneous whole blood quantitation of everolimus, sirolimus, tacrolimus and cyclosporine-A on a liquid chromatography-tandem mass-spectrometry (LC-MS/MS) system with online solid phase extraction (SPE) after manual deproteinization and centrifugation. This method had a relatively low sample throughput due to labor intensive and time-consuming sample preparation. We develop a fast and robust method of semi-automated whole blood sample pre-treatment able to support over 120 samples daily. Whole blood samples were mixed in a V-bottom plate with 2 volumes of ZnSO4/methanol containing deuterated internal standards. After centrifugation, the supernatant was loaded on an online-SPE before being transferred and monitored on the LC-MS/MS system. Assay performance included inter- and intra-day accuracy and imprecision, relative and absolute matrix effects, extraction recovery and selectivity. Intra- and inter-day imprecision was <15% for all tested concentrations. The lower limit of quantification of cyclosporine-A, everolimus, sirolimus and tacrolimus was respectively 26.6, 1.13, 1.57 and 1.28 ng mL−1, with an accuracy between −0.63% and 11.48% and a precision <12% of the nominal concentration. Standard line slope imprecision did not exceed 20% and ion enhancement or suppression was negligible. The comparison methods were performed by Passing and Bablok regression analysis: a high correlation was found between the two pre-treatment methods for tacrolimus, everolimus and sirolimus and between LC-MS/MS vs. radio-immuno-assay for cyclosporine-A. We developed a fast, sound and less misleading analytical method for high throughput (over 120 samples daily) therapeutic drug monitoring of multiple immunosuppressive drugs in routine practice.
Starting Page	5079
Ending Page	5088
Page Count	10
File Format	HTM / HTML PDF
ISSN	17599660
Volume Number	5
Issue Number	19
Journal	Analytical Methods
DOI	10.1039/c3ay40603b
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Centrifuge Whole blood Therapeutic drug monitoring Bablok Solid phase extraction Sirolimus Tacrolimus Ion Regression analysis Everolimus Methanol
Content Type	Text
Resource Type	Article
Subject	Analytical Chemistry Engineering Chemical Engineering

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Robust, high-throughput LC-MS/MS method for therapeutic drug monitoring of cyclosporine, tacrolimus, everolimus, and sirolimus in whole blood.

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Validation and application of a fast semi-automated whole blood extraction for LC-MS/MS simultaneous quantification of cyclosporine A, tacrolimus, sirolimus and everolimus – application to high throughput routine therapeutic drug monitoring

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Validation and application of a fast semi-automated whole blood extraction for LC-MS/MS simultaneous quantification of cyclosporine A, tacrolimus, sirolimus and everolimus – application to high throughput routine therapeutic drug monitoring