Identification of insulin variants using Raman spectroscopy.
Drop coating deposition Raman (DCDR) spectroscopy is used to obtain high-quality normal Raman spectra from small volumes (10 microl) of dilute insulin solutions (3-400 microM) for spectral identification and chromatographic detection. The results are used to demonstrate the spectroscopic classification (identification) of three natural insulin variants-human, bovine, and porcine-that differ by between one and three amino acid residues. DCDR measurements were performed on solutions obtained from reverse phase high-performance liquid chromatography (RP-HPLC) eluent fractions, either before or after lyophilization. Classification is demonstrated using replicate DCDR measurements, followed by normalized Savitsky-Golay second derivative preprocessing and partial least squares training with either leave-one-out or batch-to-batch testing.
Ortiz
Corasi
C
Department of Chemistry, Purdue University, West Lafayette, IN 47907, USA.
Zhang
Dongmao
D
Xie
Yong
Y
Davisson
V Jo
VJ
Ben-Amotz
Dor
D
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D014644:Genetic Variation; D006801:Humans; D007328:Insulin / Q000031:analogs & derivatives* / Q000032:analysis* / Q000737:chemistry; D008969:Molecular Sequence Data; D013059:Spectrum Analysis, Raman; D013552:Swine; D013696:Temperature
D007328:Insulin