Identification of insulin variants using Raman spectroscopy
Identification of insulin variants using Raman spectroscopy
15 September 2004
Received 9 February 2004. Available online 24 July 2004.
Corasi Ortiza, Dongmao Zhanga, Yong Xiea, V. Jo Davissonb and Dor Ben-Amotza,
Analytical Biochemistry
Volume 332, Issue 2
ScienceDirect
aDepartment of Chemistry, Purdue University, West Lafayette, IN 47907, USA
bDepartment of Medicinal Chemistry and Molecular Pharmacology, Purdue University, West Lafayette, IN 47907, USA
Abstract
Drop coating deposition Raman (DCDR) spectroscopy is used to obtain high-quality normal Raman spectra from small volumes (10 ?l) of dilute insulin solutions (3?400 ?M) 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.
Keywords: Insulin; Variants; Raman; Classification
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