Beyond Fourier Transform Infrared Spectroscopy: External Cavity Quantum Cascade Laser-Based Mid-infrared Transmission Spectroscopy of Proteins in the Amide I and Amide II Region

Analytical Chemistry
Andreas SchwaighoferBernhard Lendl

Abstract

In this work, we present a setup for mid-IR measurements of the protein amide I and amide II bands in aqueous solution. Employing a latest generation external cavity-quantum cascade laser (EC-QCL) at room temperature in pulsed operation mode allowed implementing a high optical path length of 31 μm that ensures robust sample handling. By application of a data processing routine, which removes occasionally deviating EC-QCL scans, the noise level could be lowered by a factor of 4. The thereby accomplished signal-to-noise ratio is better by a factor of approximately 2 compared to research-grade Fourier transform infrared (FT-IR) spectrometers at equal acquisition times. Employing this setup, characteristic spectral features of three representative proteins with different secondary structures could be measured at concentrations as low as 1 mg mL-1. Mathematical evaluation of the spectral overlap confirms excellent agreement of the quantum cascade laser infrared spectroscropy (QCL-IR) transmission measurements with protein spectra acquired by FT-IR spectroscopy. The presented setup combines performance surpassing FT-IR spectroscopy with large applicable optical paths and coverage of the relevant spectral range for protein analysis. T...Continue Reading

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Citations

Mar 3, 2020·Expert Opinion on Drug Delivery·Yijie SongNing Zhang
Apr 3, 2020·Journal of Physics. Condensed Matter : an Institute of Physics Journal·Christoph KratzKarsten Hinrichs
Jan 3, 2020·Nature·Ioachim PupezaFerenc Krausz
Sep 29, 2018·Applied Spectroscopy·Susanne PahlowRohit Bhargava
Mar 24, 2020·Chemical Reviews·Victor A Lorenz-Fonfria
Feb 24, 2021·Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy·Andreas SchwaighoferBernhard Lendl
Apr 7, 2021·Optics Express·Yohan YoonR Andrew McGill
Jul 3, 2021·Pharmaceutics·Vladimir Villanueva-LópezSamuel P Hernandez-Rivera

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