Development of UHPLC–MS/MS method for simultaneous determination of tacrolimus and sirolimus in human whole blood and comparisons with two immunoassays

Abstract

Tacrolimus (TAC) and sirolimus (SIR) antirejection medications are widely used in organ transplantation. We aimed to develop an ultra‐high performance liquid chromatography tandem mass spectrometry (UHPLC–MS/MS) assay for quantifying TAC and SIR simultaneously and evaluating agreement with chemiluminescence microparticle immunoassay (CMIA) and electrochemiluminescence immunoassay (ECLIA). Whole blood samples collected from 209 TAC and 208 SIR patients were assessed by UHPLC–MS/MS, CMIA and ECLIA. The agreement of the three techniques was assessed using the Bland–Altman plot. The UHPLC–MS/MS assay had a calibration range of 1–100 ng/ml for TAC and SIR. The accuracy and precision were −2.73–4.32% and <4.71% for TAC, respectively, and 0.07–4.84% and <6.5% for SIR, respectively. The three methods had good correlation. In comparison with UHPLC–MS/MS, two immunoassays showed a slight deviation in proportion. An UHPLC–MS/MS method for simultaneously detecting TAC and SIR in human whole blood was developed, validated and comparatively analyzed with CMIA and ECLIA. For determining TAC and SIR, immunoassays displayed acceptable analytical performances in terms of precision and correlation compared with UHPLC–MS/MS. However, further investigation is warranted to examine the novel method.

A gold-based immunochromatographic strip for the specific detection of tacrolimus in whole blood

“…29 The immunoassay is simple to perform and low cost, but it has the disadvantages of low sensitivity and lack of specificity. 30…”

“…29 The immunoassay is simple to perform and low cost, but it has the disadvantages of low sensitivity and lack of specificity. 30 In recent years, the colloidal gold-based immunochromatographic strip (CG-ICS), as a low-cost, high-throughput, timesaving, easy-to-operate and portable biosensor, has been widely applied in areas of food safety, 31 environmental pollution, 32 COVID-19 control 33 and clinical research. 34 The CG-ICS visualizes the specific combination of antigen and antibody through the optical properties of colloidal gold nanoparticles.…”