Chemical Segregation and Reduction of Raman Background Interference Using Drop Coating Deposition

Authors: Zhang, Dongmao¹; Mrozek, Melissa F.¹; Xie, Yong¹; Ben-Amotz, Dor¹

Source: Applied Spectroscopy, Volume 58, Issue 8, Pages 220A-244A and 887-1022 (August 2004) , pp. 929-933(5)

Publisher: Society for Applied Spectroscopy

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Abstract:

A new application of the recently described drop coating deposition Raman (DCDR) method facilitates the segregation and independent spectral characterization of mixture components. The quality of the normal (un-enhanced) Raman spectra are significantly improved as a result of reduced spectral interference from fluorescent impurities and buffer compounds. Fluorescence of commercial amino acid (O-phospho-L-serine) and protein (myoglobin) samples is reduced by over an order of magnitude using DCDR, more effectively than prolonged photo-bleaching. Furthermore, DCDR is used to obtain high-quality Raman spectra of proteins, lysozyme, and insulin, derived from solutions with up to 1000-fold excess buffer concentration. Possible thermodynamic and kinetic contributions to the observed segregation phenomena are discussed.

Keywords: RAMAN; PROTEIN; PEPTIDE; AMINO ACID; BUFFER; FLUORESCENCE; SEGREGATION; PURIFICATION

Document Type: Research article

DOI: 10.1366/0003702041655430

Affiliations: 1: Purdue University, Department of Chemistry, West Lafayette, Indiana 47907

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