Ann Lab Med.  2021 May;41(3):268-276. 10.3343/alm.2021.41.3.268.

Immunosuppressive Drug Measurement by Liquid Chromatography Coupled to Tandem Mass Spectrometry: Interlaboratory Comparison in the Korean Clinical Laboratories

• Kim HK
• Park HD
• Lee SG
• Chae H
• Song SH
• Lee YW
• Yun YM
• Ahn S
• Kim S
• Lee SM
• Lee SY
• Chun S
• on behalf of the Clinical Mass Spectrometry Research Committee of the Korean Society of Clinical Chemistry

• KMID: 2512684
• DOI: http://doi.org/10.3343/alm.2021.41.3.268

Abstract

Background
Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) is increasingly used for immunosuppressive drug tests. However, most LC-MS/MS tests are laboratory-developed and their agreement is unknown in different Korean laboratories. This interlaboratory comparison study evaluated test reproducibility and identified potential error sources.
Methods
Test samples containing three concentrations of tacrolimus, sirolimus, everolimus, cyclosporine, and mycophenolic acid were prepared by pooling surplus samples from patients undergoing routine therapeutic drug monitoring and tested in duplicate in the participating 10 clinical laboratories. Reconstitution and storage experiments were conducted for the commonly used commercial calibrator set. The robust estimators of reproducibility parameters were calculated. Spearman’s rank correlation coefficient (rho, ρ) was used to evaluate the correlation between drugs. Multiple linear regression was used to determine whether the experimental conditions alter the calibration curves.
Results
The reproducibility coefficient of variation exceeded 10% only for sirolimus concentrations 1 and 2 (10.8% and 12.5%, respectively) and everolimus concentrations 1 and 2 (12.3% and 11.4%, respectively). The percent difference values showed weak correlations between sirolimus and everolimus (ρ = 0.334, P = 0.175). The everolimus calibration curve slope was significantly altered after reconstitution following prolonged 5°C storage (P = 0.015 for 14 days; P = 0.025 for 28 days); the expected differences at 6 ng/mL were 0.598% for 14 days and 0.384% for 28 days.
Conclusions
LC-MS/MS test reproducibility for immunosuppressive drugs seems to be good in the Korean clinical laboratories. Continuous efforts are required to achieve test standardization and harmonization, especially for sirolimus and everolimus.

Keyword

Immunosuppressive drug; Therapeutic drug monitoring; Liquid chromatography coupled to tandem mass spectrometry; Interlaboratory comparison; Sirolimus; Everolimus; Reproducibility

Figure

• 

  Fig. 1 Correlation between the test samples. (A) The percent difference in individual laboratory results (D%) for each test sample. The cells are colored according to the D% values. The laboratory order is based on the average of the D% values, except for Lab E and Lab D, for pattern readability. (B) Correlation of D% values for test samples with similar concentrations for SIR and EVE. Test sample pairs are indicated by a triangle or circle, and the colors indicate the laboratory code. Abbreviations: CSA, cyclosporine; EVE, everolimus; NA, not available; SIR, sirolimus; TAC, tacrolimus.

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