Ultra-sensitive LC-MRM analysis for trastuzumab-emtansine quantification in rat plasma


Featuring the SCIEX Triple Quad™ 7500 LC-MS/MS System – QTRAP® Ready, powered by SCIEX OS Software

Zuzana Demianova and Lei Xiong
SCIEX, USA 

Abstract


With the adoption of immunoaffinity based sample preparation, an ultra-low LLOQ of 1 ng/mL was achieved. The assay shows high precision, accuracy, and good linearity, demonstrating the robustness and performance of the developed method.

Quantification of protein therapeutics in biological matrices, play a critical role in the drug discovery and development process. LC-MS/MS has been routinely adopted for quantification of this molecular class in bioanalytical laboratories, serving as an alternative technology to the traditional ligand binding assays (LBAs). Although mass spectrometry offers higher specificity than LBA, there still remains a need for improved sensitivity for accurate quantification of low concentration analytes in complex matrices while still maintaining high analysis throughput, instrument easy-of-use, and robustness.

In this work, the SCIEX Triple Quad 7500 System coupled with an analytical flow HPLC system is employed to quantify trastuzumab-emtansine in rat plasma. Multiple hardware improvements on the ion source and the front end of the mass analyzer significantly boost the system sensitivity.1 

Key features of the peptide quantification workflow
 

  • Immunoaffinity-LC-MRM workflow2 offers solid quantification of trastuzumab-emtansine in rat plasma at 1 ng/mL, with high precision, accuracy, and linearity

  • Hardware improvements on the SCIEX 7500 System provide significant gains in sensitivity for peptide quantification: the OptiFlow® Pro Ion Source with E Lens™ Technology provides improvements in ion generation and the D Jet™ Ion Guide improves ion sampling

  • SCIEX OS Software—an easy to use, compliance ready, single platform for acquisition, processing and data management