Microflow UPLC and high-resolution MS as a sensitive and robust platform for quantitation of intact peptide hormones

Bioanalysis. 2019 Jun;11(13):1275-1289. doi: 10.4155/bio-2019-0081. Epub 2019 Jul 12.

Abstract

Aim: Recent advances in microflow ultra performance liquid chromatography (UPLC) systems offer higher sensitivity with robustness to meet the routine bioanalytical demands. Modern high-resolution mass spectrometers (HRMS) enable the development of highly selective methods with broad dynamic range. Results: The quantitative performances of tandem quadrupole MS and HRMS were comprehensively compared using seven intact peptide hormones up to 9.4 kDa. Results show comparable performance between two platforms in sensitivity, accuracy and linearity. For some peptides, HRMS provided lower background interference. The benefit of increased sensitivity using microflow UPLC was also demonstrated. Conclusion: HRMS is a versatile platform capable of both basic characterization and reliable quantitation in complex matrices. Microflow UPLC provides lower LLOQs than conventional flow systems, even with less sample volume injected.

Keywords: HRMS; TOF-MRM; large molecules; microflow-UPLC.

MeSH terms

  • Amino Acid Sequence
  • Chromatography, High Pressure Liquid / methods*
  • Chromatography, High Pressure Liquid / standards
  • Limit of Detection
  • Peptide Hormones / analysis*
  • Peptide Hormones / isolation & purification
  • Peptide Hormones / standards
  • Quality Control
  • Reproducibility of Results
  • Solid Phase Extraction
  • Tandem Mass Spectrometry / methods*
  • Tandem Mass Spectrometry / standards

Substances

  • Peptide Hormones