Glioblastoma is the most aggressive solid tumor of the brain. Prior studies from our lab found that carmofur, an acid ceramidase inhibitor has shown promising results in suppressing and killing tumor cell lines compared to the FDA-approved drug temozolomide. In addition to glioblastoma, carmofur has also gained attention as a potential lead compound in a variety of diseases such as colorectal cancer, acute lung injury, and SARS-CoV-2 (COVID-19). Therefore, a sensitive bio-analytical method is required in order to quantify the drug in biological matrices for further preclinical and clinical studies. To the best of our knowledge, there is no liquid chromatography-tandem mass spectrometric (LC-MS/MS) method available for carmofur analysis. Therefore, a rapid, sensitive, and accurate LC-MS/MS method was developed for quantitative bioanalysis of carmofur in rat plasma. The method was validated for specificity, linearity, carry-over, accuracy, precision, dilution integrity, and stability according to the FDA guideline for bioanalytical method validation. The analyte was extracted employing liquid-liquid extraction using ethyl acetate and 2-propanol at 85:15 (v:v) ratio. Carmofur and internal standard (5-chlorouracil) were separated on an XBridge BEH C18 XP column (100 mm × 3 mm, 2.5 µm) column with a runtime of eight minutes. The multiple reaction monitoring (MRM) transitions were m/z 256.25 to m/z 129.00 for carmofur and m/z 145.10 to m/z 42.00 for 5-chlorouracil in negative electrospray ionization. Carmofur showed good linearity over the range of 5-1,000 ng/mL. The three quality control samples low (5 ng/mL), medium (100 ng/mL), and high (1000 ng/mL) showed acceptable recovery, matrix effect, accuracy, precision, and stability. This validated method can be applied for pharmacokinetic studies in rat plasma. Application of this method to other types of matrices is currently underway.

Development and validation of LC-MS/MS method for quantitative bioanalysis of carmofur in rat plasma

ACS Spring Meeting 2021

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.