Volumetric Absorptive Microsampling as a New Biosampling Tool for Monitoring of Tamoxifen, Endoxifen, 4-OH Tamoxifen and N-Desmethyltamoxifen in Breast Cancer Patients

• Published in: Drug design, development and therapy development and therapy, 2021-01, Vol.15, p.2417-2430
• Main Authors: Maggadani, Baitha Palanggatan, Harmita, Haryono, Samuel J, Rinaldi, Marcellino Ryan, Harahap, Yahdiana
• Format: Article
• Language: English
• Subjects: Absorptivity; Accuracy; Acetonitrile; Antineoplastic Agents, Hormonal - administration & dosage; Antineoplastic Agents, Hormonal - analysis; Antineoplastic Agents, Hormonal - pharmacokinetics; Breast cancer; Breast Neoplasms - drug therapy; Calibration; Cancer; Cancer patients; Care and treatment; Chromatography, High Pressure Liquid - methods; Desiccants; Drug Monitoring - methods; Drying; Estrogen; Estrogens; Female; Flow velocity; Formic acid; Humans; Metabolism; Metabolites; Monitoring; Oncology, Experimental; Organic acids; Original Research; Plasma; Polymorphism; Propranolol; Quality control; Reproducibility of Results; Room temperature; Sample preparation; Sonication; Stability analysis; Tamoxifen; Tamoxifen - administration & dosage; Tamoxifen - analogs & derivatives; Tamoxifen - analysis; Tamoxifen - pharmacokinetics; Tandem Mass Spectrometry - methods; Time Factors
• ISSN: 1177-8881; 1177-8881
• DOI: 10.2147/DDDT.S286409

In this research, we used a volumetric absorptive microsampling (VAMS) technique to collect blood samples from the patients. A rapid and simple sample preparation method and LC-MS.MS assay was then developed and validated for the simultaneous analysis of tamoxifen and its three active metabolites. VAMS extraction was performed in methanol by sonication-assisted extraction method for 25 min after 2 hof VAMS drying. Separation was carried out using Acquity UPLC BEH C column (2.1 x 100 mm; 1.7 µm), with a flow rate of 0.2 mL/min, and the mobile phase gradient of formic acid 0.1% and formic acid 0.1% in acetonitrile for 5 min. The multiple reaction monitoring (MRM) values were set at m/z 358.31>58.27 for -desmethyltamoxifen, m/z 372.33>72.28 for tamoxifen, m/z 388.22>72.28 for 4-hydroxytamoxifen, m/z 374.25>58.25 for endoxifen, and m/z 260.26>116.12 for propranolol. The lower limit of quantification value (LLOQ) was 2.50 ng/mL for tamoxifen, 2.50 ng/mL for endoxifen, 1.50 ng/mL for 4-hydroxitamoxifen, and 2.00 ng/mL for N-desmethyltamoxifen. Accuracy (%bias) and precision (%CV) were within 20% for LLOQ and 15% for other concentrations. There were no interference responses >20% of the LLOQ and 5% of the internal standard. The level of ion suppression in all analytes was less than 7%. The preparation system developed in this study successfully extracted more than 90% of analytes from the matrix with precision below 15%. Carryover was shown to be below 6% in all analytes. Stability of analytes in VAMS was demonstrated for up to 30 days, under room temperature storage in a sealed plastic bag with desiccant. This method was successfully applied to analyze tamoxifen and the metabolites level in 30 ER+ breast cancer patients.