Ultra-Sensitive Quantitation of Exenatide with Micro-Flow LC Trap-and-Elute and High Resolution and Triple Quadrupole Mass Spectrometry Workflow

摘要

Comparing with the published best results where 100 pg/mL was determined as the LLOQ, 10 fold sensitivity improvement was observed on this? UHPLC-MSHRAM platform configured with nanoLC 425 and TripleTOF 5600. The LLOQ of this method was determined at 10 pg/ml with 50uL injection. Wide linear dynamic range was observed from 10 to 2,000 pg/mL with excellent precision (%CV < 15% throughout the linear range), and accuracy within 89.5% to 112%. The ultra-sensitive quantitation workflow does not suffer from throughput and the total run time for each sample is 10 minutes. Carryover was a known issue for exenatide quantitation. And in this study, trifluoroethanol was introduced to effectively remove residue exenatide from sampling path as well as from separation column to maintain low system background and to ensure low level carryover. Compared with regular wash, the wash mechanism utilized in this method could reduce carryover by 87.5% and the observed carryover was at 0.17%.