An Algorithm for the Reproducible Spectral Subtraction of Water from the FT-IR Spectra of Proteins in Dilute Solutions and Adsorbed Monolayers

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Spectral subtraction and baseline correction via interactive graphics programs have been extensively used for the analysis of weak IR absorption signals which are under strong bands, such as occur in aqueous protein solutions. However, personal bias and uncertainty can result in significant differences in the spectra of the same protein substracted and baseline corrected by different persons. This is demonstrated in the case of albumin adsorbed on the surface of a germanium ATR crystal in contact within pure water. A simple computer algorithm to automatically subtract the water spectrum and remove any baseline tilt or curvature, and thus remove this individual bias and uncertainty, is presented, with a comparison of the results between different persons and the algorithm.

Keywords: Baseline correction; Computer alagorithm; FT-IR; Spectral subtraction

Document Type: Research Article


Affiliations: National Center for Biomedical Infrared Spectroscopy, Battelle's Columbus Laboratories, 505 King Avenue, Columbus, Ohio 43201-2693

Publication date: March 1, 1986

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