Abstract
Background. Although high-performance liquid chromatography (HPLC) is the method of choice for blood sirolimus determination, the microparticle enzyme immunoassay (MEIA) run on the IMx® analyser is widely used in therapeutic monitoring of this immunosuppressant agent. The aim of our study was to evaluate the possible determination of sirolimus using the fluorescence polarization immunoassay (FPIA) commercialized for everolimus quantification.Methods. Sirolimus concentrations were determined in whole-blood samples from liver and kidney transplant recipients using the Innofluor® Certican® FPIA (Seradyn Inc.) run on a TDx® analyser (Abbott Laboratories), Sirolimus MEIA run on an IMx® analyser (Abbott Laboratories), and HPLC (UV detection) methods.Results. The Innofluor® FPIA has a similar cross-reactivity with everolimus and sirolimus, and the within- and between-run coefficients of variation obtained for sirolimus determination were 2.7%–13.3%. In analysing different blood samples from liver and kidney transplant patients the linear regressions obtained were: FPIA = 1.12 HPLC + 0.43 (n=104, r=0.874), MEIA = 1.14 HPLC (n=146, r=0.892), and FPIA = 1.00 MEIA + 0.29 (n=106, r=0.941). Better correlation coefficients were obtained between the methods in the liver transplant samples (r≥0.900) than in the kidney transplant samples (r≥0.849). No significant effect was found for sirolimus clearance or the blood hematocrit on the relationship between the results produced by both immunoassays and HPLC.Conclusion. The Innofluor® FPIA is a valid alternative with an analogous performance to the MEIA for the therapeutic monitoring of sirolimus.
Highlights
High-performance liquid chromatography (HPLC) is the method of choice for blood sirolimus determination, the microparticle enzyme immunoassay (MEIA) run on the IMx† analyser is widely used in therapeutic monitoring of this immunosuppressant agent
The results obtained in the imprecision study for fluorescence polarization immunoassay (FPIA), MEIA, and high-performance liquid chromatography (HPLC) may be considered satisfactory
The FPIA imprecision was in general slightly greater than that given by the MEIA assay, and this fact may be due to the FPIA procedure for preparing blood samples, which requires methanol extraction prior to the addition of the precipitation reagent, whereas MEIA uses a simpler procedure with only one extraction/precipitation reagent
Summary
High-performance liquid chromatography (HPLC) is the method of choice for blood sirolimus determination, the microparticle enzyme immunoassay (MEIA) run on the IMx† analyser is widely used in therapeutic monitoring of this immunosuppressant agent. Sirolimus concentrations were determined in whole-blood samples from liver and kidney transplant recipients using the Innofluor† Certican† FPIA (Seradyn Inc.) run on a TDx† analyser (Abbott Laboratories), Sirolimus MEIA run on an IMx† analyser (Abbott Laboratories), and HPLC (UV detection) methods. The Innofluor† FPIA has a similar cross-reactivity with everolimus and sirolimus, and the within- and between-run coefficients of variation obtained for sirolimus determination were 2.7%Á13.3%. No significant effect was found for sirolimus clearance or the blood hematocrit on the relationship between the results produced by both immunoassays and HPLC. The Innofluor† FPIA is a valid alternative with an analogous performance to the MEIA for the therapeutic monitoring of sirolimus
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