Bioanalytical method validation and sample analysis for nirmatrelvir in dried blood collected using the Tasso-M20...

Area of Science:

• Pharmacology
• Analytical Chemistry
• Biomedical Science

Background:

• Nirmatrelvir is a critical antiviral medication.
• Accurate quantification in biological matrices is essential for therapeutic drug monitoring and clinical trials.
• Dried blood spot (DBS) collection offers a minimally invasive alternative to traditional blood draws.

Purpose of the Study:

• To develop and validate a sensitive and selective analytical method for determining nirmatrelvir concentrations.
• To utilize the Tasso-M20 device for collecting dried human blood samples for nirmatrelvir analysis.
• To establish a robust assay suitable for large-scale clinical applications.

Main Methods:

• Development of a high-performance liquid chromatography coupled with tandem mass spectrometric detection (HPLC-MS/MS) method.
• Isolation of nirmatrelvir and a stable-labeled internal standard from dried blood spots (DBS) collected using the Tasso-M20 device.
• Extraction of analytes from approximately 20 μL of blood using methanol, followed by supernatant dilution.

Main Results:

• The developed method demonstrated high sensitivity and selectivity for nirmatrelvir quantification.
• The assay validation confirmed accuracy and precision across the concentration range of 20.0 to 20,000 ng/mL.
• Hematocrit levels were found to have no significant impact on assay accuracy.
• Comprehensive stability studies were performed, covering various drying and storage conditions.

Conclusions:

• A fully validated, sensitive, and selective method for nirmatrelvir determination in Tasso-M20 collected DBS was successfully established.
• The method is robust, unaffected by hematocrit variations, and demonstrates excellent stability.
• The validated assay has been successfully implemented in multiple clinical studies, showcasing its practical utility and reliable performance.