Hae-In Choi (한국생명공학연구원 (KRIBB))

Kyeong-Ryoon Lee (한국생명공학연구원 (KRIBB))

Hae-In Choi (한국생명공학연구원 (KRIBB))

Hyeon-Cheol Jeong (한국생명공학연구원 (KRIBB))

Kyeong-Ryoon Lee (한국생명공학연구원 (KRIBB))

Tirzepatide is a novel dual GIP/GLP-1 receptor agonist with promising therapeutic potential for diabetes and obesity. Given the growing clinical interest in peptide-based therapeutics, the development of bioanalytical methods is important for pharmacokinetic evaluation. In this study, a sensitive and rapid LC-MS/MS method was developed and validated for quantifying tirzepatide in rat plasma. Sample preparation was performed using methanol-induced protein precipitation. Chromatographic separation was achieved using a C18 column with gradient elution, and detection was performed using multiple reaction monitoring mode with m/z transitions of 1204.4→1473.6 for tirzepatide and 1029.4→1238.4 for internal standard. The method showed good linearity (r² > 0.99) over the range of 1–1000 ng/mL. Accuracy and precision met regulatory criteria at all QC levels, and other validation parameters also complied with regulatory guidelines. The validated method was successfully applied to pharmacokinetic studies in Sprague-Dawley rats following single subcutaneous and intravenous administration of 0.3 mg/kg tirzepatide. Observed pharmacokinetic parameters were consistent with previous findings, supporting the reliability of the analytical method. This robust and reliable analytical method provides a solid foundation for the preclinical pharmacokinetic or further investigations of tirzepatide, and can also be expected to apply to the bioanalysis of emerging peptide therapeutics.

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