This method detects four BTK-related analytes in plasma/CSF with a 3-min run time, evaluation of hemolytic/lipemic matrices, and clinical validation, increasing throughput, reducing costs, and broadening applicability, providing preliminary feasibility data for clinical TDM.
Abstract
Background
Bruton's tyrosine kinase inhibitors (BTKis, ibrutinib, zanubrutinib, orelabrutinib) are key targeted therapies for B-cell lymphomas, but interindividual variability and adverse reactions limit their use. Metabolized by CYP3A, co-medications or hepatic dysfunction cause plasma fluctuations, necessitating therapeutic drug monitoring (TDM). Cerebrospinal fluid (CSF) distribution is also critical for CNS lymphoma treatment.
Objective
To develop and validate a UHPLC-MS/MS method for simultaneous quantification of three BTKis and one metabolite in human plasma and CSF to support TDM.
Methods
After protein precipitation with methanol, samples were analyzed on an ACQUITY HSS T3 column with a 3-min gradient at 0.3 mL/min and ESI (+) MRM. Mobile phase: water-acetonitrile-formic acid-ammonium acetate (1 M) (950:50:1:1, A) and (50:950:1:1, B).
Results
The method showed excellent selectivity and linearity (r > 0.997) over 1-200 ng/mL (ibrutinib, zanubrutinib, ibrutinib metabolite) and 5-1000 ng/mL (orelabrutinib) in both matrices. Recoveries were 89.4-103.8%, matrix factors 0.93-1.06, and precision CV ≤8.5%. Stability was confirmed. The method was applied to 34 clinical samples (25 plasma, 9 CSF) from 20 patients.
Conclusion
This method detects four BTK-related analytes in plasma/CSF with a 3-min run time, evaluation of hemolytic/lipemic matrices, and clinical validation, increasing throughput, reducing costs, and broadening applicability, providing preliminary feasibility data for clinical TDM.
Objectives: A high-performance liquid chromatography (HPLC) with mass spectrometry (MS) detection method was developed to quantify several tyrosine kinase inhibitors (TKIs) and their relevant metabolites. This method is suitable for therapeutic drug monitoring (TDM) of alectinib, brigatinib, dabrafenib, lenvatinib, lorlatinib, osimertinib and trametinib in patients with solid tumors using volumetric absorptive microsampling (VAMS®). Methods: The HPLC-MS system contained four pumps, a Turboflow HTLC CycloneTM 1.0 × 50 mm solid phase extraction column for analyte enrichment, and a Kinetex 2.6 µm C18 100Å, 100 × 3.0 mm column for analyte separation. An acetonitrile–water gradient was used for the separation, and the ions generated by ESI (+)-ionization were detected in single-ion mode. This method was validated according to recent European Medicines Agency (EMA) and Food and Drug Administration (FDA) guidelines. Results: The accuracy and precision shown during method validation were within the acceptable limits for all analytes in plasma and whole blood. All analytes showed acceptable stability in both matrices for at least 28 days when stored at −21 °C. So far, 100 venous plasma and 94 capillary blood samples have been collected and analyzed. Conclusions: We developed a reliable method to quantify several TKIs from plasma and capillary blood, which is intended for TDM purposes in clinical practice.
A reliable, sensitive, and selective liquid chromatography-tandem mass spectrometry method suitable for high-throughput pharmacokinetic studies and therapeutic drug monitoring of ALB in human plasma is developed and validated.
Venkateswararao Agraharapu, V. Vutla, Vidyadhara Suryadevara· Journal of Applied Pharmaceu...· 0 citations
Bosutinib is a second-generation tyrosine kinase inhibitor (TKI) primarily used for the treatment of chronic myeloid leukemia (CML). The aim of this study was to develop and validate a rapid and accurate ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous determination of bosutinib and its major metabolite M5. Chromatographic separation was performed on an Acquity UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) using a gradient elution with acetonitrile and 0.1% formic acid as the mobile phase, and gefitinib was used as the internal standard (IS). Biological analysis showed that both bosutinib (2-400 ng/mL) and its metabolite M5 (1-60 ng/mL) exhibited good linearity in their respective concentration ranges, with correlation coefficients (r2) of 0.996 and 0.993, respectively. Both of them exhibited intra- and inter-day precision (RSD%) below 15% and accuracy (RE%) within ± 15%. Recoveries were all within the range of 85.3-98.0%, and matrix effects were all within the range of 91.6-101.6%. Ultimately, human liver microsomes (HLM) was used to study in vitro metabolic stability, while in vivo pharmacokinetics were evaluated in rats. In vivo pharmacokinetic studies confirmed that this method could be successfully used for the analysis of bosutinib after oral administration of 50 mg/kg in rats. In vitro results showed that bosutinib had the intrinsic clearance (CLint) value of 0.03 mL/min/mg and the half-life (t1/2) value of 79.94 min. This study provides a potential reference for the clinical investigation of bosutinib.
Peiqi Wang, Yan Chen, Jun Wu et al.· Chemico-Biological Interacti...· 0 citations
The first validated liquid chromatography tandem mass spectrometry (LC-MS/MS) bioanalytical method for the quantification of atirmociclib in mouse plasma is reported, validated in compliance with international council for harmonisation-M10 (ICH-M10) guidelines.
Ashwani Gaur, U. Patel, Ashutosh Gavali et al.· Journal of chromatography. B...· 0 citations
As an antithrombotic agent, indobufen is widely used in China. However, the relationship between drug concentration and therapeutic efficacy in specific patient populations remains unclear. Therefore, it is crucial to establish a method for quantifying indobufen concentrations in human plasma to guide dosage recommendations. An electrospray ionization (ESI) power supply was utilized in conjunction with multireaction monitoring (MRM) in positive ion mode. Protein precipitation was employed to extract indobufen from the plasma sample. Separation was carried out using a ChromCORE‐C18 column with a gradient elution of the mobile phase, which was composed of 0.1% formic acid in water (Solvent A) and methanol (Solvent B) at a flow rate of 0.3 mL/min. The typical standard curve equation was Y = 0.763X + 0.0986 (r = 0.9993), demonstrating a good linear relationship within the range of 0.5–30 μg/mL. All methodological outcomes complied with regulatory guidelines. In patients with moderate renal insufficiency, it is advisable to reduce the dosage to 100 mg once daily. For patients with severe renal insufficiency, the dosage should be reduced to 1/3–1/4 of the standard dose. For patients undergoing continuous renal replacement therapy (CRRT), indobufen can be gradually eliminated, and most patients do not require dose adjustment.
Hongyan Ji, Haijun Qu, Qie Guo et al.· Journal of Analytical Method...· 0 citations
Aficamten (CK-3773274) is a next-in-class selective cardiac myosin inhibitor now approved for use for chronic oral therapy in hypertrophic cardiomyopathy (HCM). The present study established an innovative, rapid, and highly sensitive HPLC-MS/MS approach for quantifying aficamten concentrations in human plasma and urine. Samples were prepared using protein precipitation with acetonitrile containing CK-3942912 (D5-labelled aficamten) as the internal standard (IS). Sample separation was achieved using a C18 analytical column, with a mobile phase comprising acetonitrile and ammonium acetate. A Xevo TQS tandem mass spectrometer performed the detection in electrospray ionization (ESI) positive mode. The following MRM ion transitions were monitored: m/z 338.2 → 213.2 for CK-3773274 and m/z 343.2 → 218.2 for IS. This method was validated with excellent selectivity and sensitivity. Linear calibration curves (1-500 ng/mL) demonstrated excellent accuracy (RE% ±15%) and precision (RSD% ≤15%) for both intra- and inter-run analyses. Furthermore, critical validation parameters including matrix effects, extraction recovery, carryover, and stability were also assessed. The validated method was subsequently utilized to quantify aficamten concentrations in a clinical pharmacokinetic study involving healthy Chinese adults.
Xiao-Fei Wu, Aijing Liu, Adrienne Griffith et al.· Journal of chromatography. B...· 0 citations
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