Seungman Park (Presenter)
GreenCross Laboratories
Bio: MD, Laboratory medicine (Clinical pathology). Director of department of special chemistry in GreenCross Laboratories. In PhD. course in Seoul National University
Authorship: Seungman(1,2), Juhee Park(1), Jungsun Han(1), Songhyeon Yang(1), Euna Park(1), Eunhee Lee(1)
Green Cross Laboratories, South Korea(1) Seoul National University (2)
| Short Abstract We developed and validated lap developed test for simultaneous determination of sirolimus and everolimus using liquid chromatography tandem mass spectrometry (LC-MS/MS). The accuracy, limit of quantification, precision, linearity, and carry-over were evaluated. The accuracy, precision, and linearity were excellent. The limit of quantification and carry-over were acceptable. But, result of proficiency test was unacceptable. After calibrator change, the concentrations of PT materials were within acceptable range. Although the LC-MS/MS assay shows excellent analytical performance, attending PT program is helpful for harmonization with peer group. |
Long Abstract
Introduction: Measuring the concentration of immunosuppressive agent is essential for patients with organ transplantation. We developed and validated lap developed test for simultaneous determination of sirolimus and everolimus using liquid chromatography tandem mass spectrometry (LC-MS/MS).
Methods: The accuracy and limit of quantification were evaluated with commercially available standard materials (Sigma-Aldrich, US) mixed with blank whole blood. The precision, linearity, and carry-over were evaluated with ClinChek control materials (Recipe, Germany).
Results: The recoveries of LC-MS/MS method for standard material were 92.7-105.2% over concentration of 1-80 ng/mL for sirolimus and 92.8-101.2% over 1-80 ng/mL for everolimus. The limit of quantitation was 1 ng/mL for both assays. The imprecision (CVs) of LC-MS/MS for sirolimus were 6.4%, 4.9%, and 3.9% for low (4.18 ng/mL), mid (12.6 ng/mL), and high (21.0 ng/mL) concentration. The imprecision (CVs) for everolimus were 7.3%, 3.8%, and 3.6% for low (3.89 ng/mL), mid (12.3 ng/mL), and high (20.3 ng/mL) concentration. The sirolimus assay was linear (R2=0.9997) over concentration of 1.62-52.9 ng/mL and the everolimus assay was linear (R2=0.9997) over concentration of 1.45-49.4 ng/mL. The carry-over was estimated less than 1% for both assays. Since the report of College of American Pathology (CAP) proficiency test (PT) showed significantly higher result than peer group, we reviewed procedure, reagent, calibrator and LC-MS/MS equipment. Since the assays were performed with ClinCal¢ç Calibrator (Recipe, Germany), we tried MassCheck¢ç Immunosuppressants Whole Blood Calibrator (Chromsystems, Germany). After calibrator change, the concentrations of CAP PT materials were within acceptable range.
Conclusions: The LC-MS/MS assay for simultaneous quantification of sirolimus and everolimus showed excellent analytical performance. But, attending PT program is helpful for harmonization with peer group.
References & Acknowledgements:
| Description | Y/N | Source |
| Grants | no | |
| Salary | no | |
| Board Member | no | |
| Stock | no | |
| Expenses | no |
IP Royalty: no
| Planning to mention or discuss specific products or technology of the company(ies) listed above: | no |