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Validation and evaluation of four sample preparation methods for the quantification of intracellular tacrolimus in peripheral blood mononuclear cells by UHPLC-MS/MS.
- Source :
-
Clinica chimica acta; international journal of clinical chemistry [Clin Chim Acta] 2020 Apr; Vol. 503, pp. 210-217. Date of Electronic Publication: 2019 Nov 30. - Publication Year :
- 2020
-
Abstract
- Rejection and toxicity occur despite monitoring of tacrolimus blood levels during clinical routine. The intracellular concentration in lymphocytes could be a better reflection of the tacrolimus exposure. Four extraction methods for tacrolimus in peripheral blood mononuclear cells were validated and evaluated with UHPLC-MS/MS. Methods based on protein precipitation (method 1), solid phase extraction (method 2), phospholipids and proteins removal (method 3) and liquid-liquid extraction (method 4) were evaluated on linearity, lower limit of quantification (LLOQ), imprecision and bias. Validation was completed for the methods within these requirements, adding matrix effect and recovery. Linearity was 0.126 (LLOQ)-15 µg/L, 0.504 (LLOQ)-15 µg/L and 0.298 (LLOQ)-15 µg/L with method 1, 2 and 3, respectively. With method 4 non-linearity and a LLOQ higher than 0.504 µg/L were observed. Inter-day imprecision and bias were ≤4.6%, ≤10.9%; ≤6.8%, ≤-11.2%; ≤9.4%, ≤10.3% and ≤44.6%, ≤23.1%, respectively, with methods 1, 2, 3 and 4. Validation was completed for method 1 and 3 adding matrix effect (7.6%; 15.0%) and recovery (8.9%; 10.8%), respectively. The most suitable UHPLC-MS/MS method for quantification of intracellular tacrolimus was protein precipitation due to the best performance characteristics and the least time-consuming rate and complexity.<br />Competing Interests: Declaration of Competing Interest The authors declared that there is no conflict of interest.<br /> (Copyright © 2019 Elsevier B.V. All rights reserved.)
- Subjects :
- Chemical Precipitation
Chromatography, High Pressure Liquid methods
Humans
Lipids isolation & purification
Liquid-Liquid Extraction standards
Proteins isolation & purification
Solid Phase Extraction standards
Specimen Handling standards
Tandem Mass Spectrometry methods
Leukocytes, Mononuclear chemistry
Specimen Handling methods
Tacrolimus analysis
Subjects
Details
- Language :
- English
- ISSN :
- 1873-3492
- Volume :
- 503
- Database :
- MEDLINE
- Journal :
- Clinica chimica acta; international journal of clinical chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 31794770
- Full Text :
- https://doi.org/10.1016/j.cca.2019.11.033