Clinica chimica acta; international journal of clinical chemistry · 2026
High-throughput quantification of tacrolimus in whole blood by acoustic ejection mass spectrometry: analytical validation and comparative assessment against conventional LC-MS/MS for therapeutic drug monitoring.
СтатьяСравнительное исследованиеВалидационное исследованиеНазвание и аннотация: Перевод в очереди
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Wu Xin-Yu, Fu Xiao-Kang, Li Ao, Zhang Wei, Jiang Han-Peng, An Na, Zhu Quan-Fei, Wang Shao-Ting, Feng Yu-Qi
Аннотация
BACKGROUND: Therapeutic drug monitoring (TDM) of tacrolimus requires accurate, high-throughput methods, but conventional liquid chromatography-tandem mass spectrometry (LC-MS/MS) is limited by lengthy run times and complex sample preparation.
METHODS: We developed a chromatography-free assay for quantification of tacrolimus in whole blood by integrating cold-induced phase separation with acoustic ejection mass spectrometry (AEMS). This workflow only requires less than 10 μL of whole blood, eliminates centrifugation, and enables direct nanoliter-scale injection into the mass spectrometer. The AEMS assay performance was validated in accordance with CLSI C62-A and ICH M10 guidelines using 62 patient samples and matrix-matched calibrators, demonstrating its suitability for high-throughput therapeutic drug monitoring.
RESULTS: The AEMS assay exhibited excellent linearity (0.5-50 μg/L; R2 = 0.9987), with a lower limit of quantification of 0.5 μg/L, which was well below clinically relevant trough concentrations. Accuracy (93.3%-100.1%) and precision (intra-day RSD: 3.7%-11.3%; inter-day RSD: 2.3%-12.4%) met regulatory acceptance criteria. Results showed strong concordance with routine LC-MS/MS (Spearman ρ = 0.972; Passing-Bablok slope = 1.100 [95% CI: 1.026-1.177]). The total analysis cycle time was less than 3 s per sample, enabling a throughput exceeding 1000 samples per hour theoretically.
CONCLUSION: This study establishes the first chromatography-free mass spectrometry platform validated for clinical TDM of TAC. By integrating ultra-high throughput capability, minimal sample consumption, and robust analytical performance, the AEMS workflow offers a transformative analytical solution for high-demand clinical environments, including transplant centers and pediatric monitoring settings.
METHODS: We developed a chromatography-free assay for quantification of tacrolimus in whole blood by integrating cold-induced phase separation with acoustic ejection mass spectrometry (AEMS). This workflow only requires less than 10 μL of whole blood, eliminates centrifugation, and enables direct nanoliter-scale injection into the mass spectrometer. The AEMS assay performance was validated in accordance with CLSI C62-A and ICH M10 guidelines using 62 patient samples and matrix-matched calibrators, demonstrating its suitability for high-throughput therapeutic drug monitoring.
RESULTS: The AEMS assay exhibited excellent linearity (0.5-50 μg/L; R2 = 0.9987), with a lower limit of quantification of 0.5 μg/L, which was well below clinically relevant trough concentrations. Accuracy (93.3%-100.1%) and precision (intra-day RSD: 3.7%-11.3%; inter-day RSD: 2.3%-12.4%) met regulatory acceptance criteria. Results showed strong concordance with routine LC-MS/MS (Spearman ρ = 0.972; Passing-Bablok slope = 1.100 [95% CI: 1.026-1.177]). The total analysis cycle time was less than 3 s per sample, enabling a throughput exceeding 1000 samples per hour theoretically.
CONCLUSION: This study establishes the first chromatography-free mass spectrometry platform validated for clinical TDM of TAC. By integrating ultra-high throughput capability, minimal sample consumption, and robust analytical performance, the AEMS workflow offers a transformative analytical solution for high-demand clinical environments, including transplant centers and pediatric monitoring settings.