Large-molecule quantification: sensitivity and selectivity head-to-head comparison of triple quadrupole with Q-TOF.

BACKGROUND: Bioanalysts are continuously looking for innovative ideas or instruments to increase the sensitivity and selectivity of their assays. Research for better mass spectrometers is becoming crucial with the emerging trend of large-molecule quantification. This study lists the different advantages of high-resolution MS (HRMS) over standard triple quadrupole instruments and proposes basic guidelines on how to use HRMS for large-molecule quantification in a regulated environment. RESULTS: A direct comparison between HRMS and triple quadrupole instruments for the quantification of six different model peptides (desmopressin, calcitonin, enfuvirtide, exenatide, glucagon and somatostatin) was completed. The HRMS instrument, when used specifically for targeted quantification ('quant/quant'), showed equivalent or better sensitivity for all compounds tested. CONCLUSION: This paper demonstrates that the use of a HRMS instrument in a regulated environment is a viable technique for quantification of large molecules. The latter was able to allow flexibility and selectivity to adapt the specificity of each assay with sensitivity comparable to the triple quadrupole instrument.

An alternative solution to overcome carryover issues in bioanalysis.

LC-MS/MS instruments are considered as the analytical technique of choice for bioanalytical assays. The LC-MS/MS technique acquired this status due to its robustness, specificity, sensitivity and good precision and accuracy. Nonetheless, with the use of these instruments, users also have to deal with some complex analytical problems such as carryover. Even if carryover is well known and widely discussed, it still remains a common issue in bioanalytical work. Although new autosampler instruments manufactured are specifically designed to fix the problem, the issue is still present because methods developed by the industry also change and evolve with the instrument. Requests for wider dynamic ranges and the use of a very sensitive detector that can reach very low LOQ are among the reasons. This article reviews the causes of carryover and proposes an autosampler needle seat backflush as a novel solution to address the issue.

Reliable procedures to evaluate and repair crosstalk for bioanalytical MS/MS assays.

Louis-Philippe Morin is a senior instrument application specialist at Algorithme Pharma, a CRO located in Laval, Canada. He has been working in the bioanalysis industry for the past 10 years where he became a subject matter expert in analytical instrumentation, especially in the MS field. His responsibilities in his current position are to optimize the workflow of the laboratory and to find new procedures, or approaches, to fix complex analytical problems. Louis-Philippe's expertise acquired over the years has led him to multiple publications regarding instrumentation. LC-MS/MS is the analytical technique of choice for the quantification of drugs in biological fluids. In recent years, MS/MS detection has been impacted by the rapid evolution of bioanalysis industry requirements. The availability of fast chromatographic systems, the demand for wider dynamic ranges and the extensive use of stable isotope-labeled internal standards in bioanalysis has pushed some triple quadrupole detectors to their limits of operation. Consequently, this situation has led to a re-evaluation of the problem of crosstalk as a potential cause of issues in bioanalysis. In this article, the importance of crosstalk verification on the MS/MS instrument will be demonstrated. Additionally, procedures to identify, evaluate and fix possible crosstalk issues during bioanalytical assays on MS/MS instruments are proposed.