Estimation of Carmofur and 5-Fluorouracil in Plasma and Other Biological Tissues Using Mass Spectrometry

Mentor 1

Shama Mirza

Start Date

10-5-2022 10:00 AM

Description

Carmofur, also known as 1-hexylcarbanoyl 5-fluorouracil (HCFU) is a drug for colorectal cancer and has been in use for over four decades in Japan, China, Korea and Finland. In addition to colorectal cancer, it has been found effective against other cancers such as head and neck, breast, pancreatic, gastrointestinal, and solid tumors of the brain. Several recent studies also found carmofur beneficial against the novel coronavirus disease 2019, Krabbe disease, and lipopolysaccharide-induced acute lung injury etc. To date, carmofur does not have any LC-MS/MS or MALDI-MS based method for quantitative or qualitative bioanalysis to support further pre-clinical and clinical trials. Herein, we worked on developing and validating tandem mass spectrometric methods for analyzing carmofur and its active metabolite 5-fluorouracil in mouse plasma and other tissues such as brain, kidney and liver. Qualitative analysis was performed by MALDI imaging MS on cryo-sliced brain tissue sections using Shimadzu MALDI 7090 TOF/TOF MS instrument with negative ionization in reflectron mode. Collected MS data was analyzed using IonView software to generate images of the analytes in the brain tissue. Quantitative analysis was performed on Shimadzu LCMS 8040 triple quadrupole mass spectrometer. Carmofur was administered at a dose of 10 mg/kg intraperitoneally. Plasma and other tissues were collected over a period of 4 hours. Collected samples underwent liquid-liquid extraction and the extracted analytes were ionized by negative electrospray ionization and the quantification was achieved in multiple reaction monitoring (MRM) mode with transitions m/z 256.25 to m/z 129.00 for carmofur, m/z 129.15 to m/z 42.00 for 5-fluorouracil. Three quality control samples (low/medium/high) showed acceptable recovery, matrix effect, accuracy, precision, and stability. Finally, we tested the applicability of our method in female Swiss Webster mice for a pharmacokinetic and drug distribution study.

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May 10th, 10:00 AM

Estimation of Carmofur and 5-Fluorouracil in Plasma and Other Biological Tissues Using Mass Spectrometry

Carmofur, also known as 1-hexylcarbanoyl 5-fluorouracil (HCFU) is a drug for colorectal cancer and has been in use for over four decades in Japan, China, Korea and Finland. In addition to colorectal cancer, it has been found effective against other cancers such as head and neck, breast, pancreatic, gastrointestinal, and solid tumors of the brain. Several recent studies also found carmofur beneficial against the novel coronavirus disease 2019, Krabbe disease, and lipopolysaccharide-induced acute lung injury etc. To date, carmofur does not have any LC-MS/MS or MALDI-MS based method for quantitative or qualitative bioanalysis to support further pre-clinical and clinical trials. Herein, we worked on developing and validating tandem mass spectrometric methods for analyzing carmofur and its active metabolite 5-fluorouracil in mouse plasma and other tissues such as brain, kidney and liver. Qualitative analysis was performed by MALDI imaging MS on cryo-sliced brain tissue sections using Shimadzu MALDI 7090 TOF/TOF MS instrument with negative ionization in reflectron mode. Collected MS data was analyzed using IonView software to generate images of the analytes in the brain tissue. Quantitative analysis was performed on Shimadzu LCMS 8040 triple quadrupole mass spectrometer. Carmofur was administered at a dose of 10 mg/kg intraperitoneally. Plasma and other tissues were collected over a period of 4 hours. Collected samples underwent liquid-liquid extraction and the extracted analytes were ionized by negative electrospray ionization and the quantification was achieved in multiple reaction monitoring (MRM) mode with transitions m/z 256.25 to m/z 129.00 for carmofur, m/z 129.15 to m/z 42.00 for 5-fluorouracil. Three quality control samples (low/medium/high) showed acceptable recovery, matrix effect, accuracy, precision, and stability. Finally, we tested the applicability of our method in female Swiss Webster mice for a pharmacokinetic and drug distribution study.