The follow-up of patients under lifelong immunosuppressant therapy is pivotal to prevent allograft rejection after transplant. Part of the difficulties associated with routine monitoring of immunosuppressant concentrations can be alleviated by home sampling using dried blood spots (DBSs).


To evaluate the applicability of a DBS method for the determination of immunosuppressants in venous blood samples, making use of an automated extraction platform.


Paired venous DBSs and whole blood samples were analyzed for tacrolimus (n = 162), sirolimus (n = 47), everolimus (n = 45), and cyclosporin A (n = 61) with liquid chromatography coupled to tandem mass spectrometry, using fully automated extraction for DBSs. Agreement between the automated DBS and whole blood method was assessed by using Bland-Altman comparison. Both an analytical and a clinical acceptance limit were predefined at more than 67% of all paired samples within 20% of the mean of both samples and more than 80% of all paired samples within 20% of the whole blood concentration, respectively.


An impact of the hematocrit (hct) on DBS quantitation was observed for all analytes, which could be alleviated for all analytes by using a hct conversion formula based on a tacrolimus data subset: [DBScorrected] = [DBSmeasured]/(1.6305 – 1.559*hct). After correction, both analytical and clinical acceptance criteria were met for all analytes.


Automated DBS analysis shows great potential for routine therapeutic drug monitoring of immunosuppressants, avoiding any manual sample handling.

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Author notes

Two supplemental digital content files are available for this article. See text for hyperlinks.

This work was supported by the Research Foundation-Flanders (FWO) (grant No. 11F3121N).

The authors have no relevant financial interest in the products or companies described in this article.

Part of the results outlined in this manuscript have been presented at the following meetings: at the 7th European Bioanalysis Forum Young Scientist Symposium (YSS 2021) (and online); June 10, 2021; Ghent, Belgium; at the 19th international congress of Therapeutic Drug Monitoring and Clinical Toxicology (IATDMCT 2021); September 22, 2021; Rome, Italy; and at the Toxicological Society of Belgium and Luxembourg meeting (BLT 2021); October 8, 2021; Echternach, Luxembourg.

Supplementary data