Development and validation of a liquid chromatography‐tandem mass spectrometry method for the determination of temozolomide in mouse brain tissue

Abstract

AbstractTemozolomide is a Food and Drug Administration‐approved anticancer drug that has poor drug delivery via oral or intravenous routes. A potential strategy to combat this problem is investigating alternative routes of administration, requiring quantitation of the drug in the brain tissues by liquid chromatography‐mass spectrometry. However, current methods used to extract the drug from brain tissues resulted in poor recovery and substantial matrix effects. Herein, we reported a new two‐step extraction method that involves the use of Proteinase K to lyse tumor tissues to efficiently release the drug, followed by ethanol protein precipitation. The extracts were then separated on a C18 column and analyzed in positive electrospray ionization, a multiple reaction monitoring mode of the triple quadrupole. We found this new method led to a recovery of 82% with negligible matrix effects. The method has been validated in accordance with Food and Drug Administration guidance for linearity, specificity, selectivity, accuracy, precision, carry‐over, stability, and lower limit of quantitation. In conclusion, we have developed and validated a liquid chromatography‐mass spectrometry method with a novel sample preparation method that was able to efficiently extract temozolomide from mouse brain tissue with high recovery.