Journal
ANALYTICAL CHEMISTRY
Volume 93, Issue 2, Pages 920-927Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.0c03567
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Funding
- EPSRC [EP/T014318/1, EP/T014245/1]
- STFC
- EPSRC [EP/T014318/1, EP/T014245/1] Funding Source: UKRI
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Glycine is used as a model system to evaluate the sensitivity of 2D-IR spectroscopy in detecting low-molecular-weight protein components in blood serum. By combining data acquisition schemes and peak pattern analysis, the spectral signature of Gly can be separated from dominant protein fractions, with a detection limit of around 3 mg/mL. The potential for 2D-IR to complement IR absorption methods for measuring concentrations of amino acids, peptides, and low-molecular-weight proteins in serum samples is highlighted.
Glycine (Gly) is used as a model system to evaluate the ability of ultrafast two-dimensional infrared (2D-IR) spectroscopy to detect and quantify the low-molecular-weight proteinaceous components of blood serum. Combining data acquisition schemes to suppress absorption bands of H2O that overlap with the protein amide I band with analysis of peak patterns appearing in the off-diagonal region of the 2D-IR spectrum allows separation of the Gly spectral signature from that of the dominant protein fraction of serum in a transmission-mode 2D-IR measurement without any sample manipulation, e.g., filtration or drying. 2D-IR spectra of blood serum samples supplemented with varying concentrations of Gly were obtained, and a range of data analysis methods compared, leading to a detection limit of similar to 3 mg/mL for Gly. The reported methodology provides a platform for a critical assessment of the sensitivity of 2D-IR for measuring the concentrations of amino acids, peptides, and low-molecular-weight proteins present in serum samples. We conclude that, in the case of several clinically relevant diagnostic molecules and their combinations, the potential exists for 2D-IR to complement IR absorption methods as the benefits of the second frequency dimension offered by 2D-IR spectroscopy outweigh the added technical complexity of the measurement.
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