Quantitative analysis of peptides with NMR spectroscopy

Cynthia K. Larive, Dimuthu Jayawickrama, L. Őrfi

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

The determination of peptide concentration with 1H nuclear magnetic resonance (NMR) spectroscopy using an internal standard or an external standard in a sealed glass capillary was investigated for three tyrosine-containing tripeptides. Trimethylsilylpropionic acid (TSP) and maleic acid were tested as external standards for quantitation by proton NMR. Although comparable results were obtained for either standard, the performance of maleic acid was found to be superior because of its better long-term stability in the sealed capillary. Loss of TSP from solution occurred over time due to adsorption onto the walls of the capillary, necessitating frequent recalibration against the primary standard, potassium acid phthalate (KHP). The peptide contents of solid peptides determined with 1H NMR are compared with those obtained from ultraviolet (UV) absorbance measurements of the tyrosine chromophore. The versatility of NMR for the quantitative analysis of peptides that do not contain an appropriate UV chromophore make it well-suited for the determination of peptide concentration in aggregation studies or for the preparation of solutions for high-throughput screening of biological activity.

Original languageEnglish
Pages (from-to)1531-1536
Number of pages6
JournalApplied Spectroscopy
Volume51
Issue number10
Publication statusPublished - Oct 1997

Fingerprint

magnetic resonance spectroscopy
Peptides
quantitative analysis
Nuclear magnetic resonance spectroscopy
peptides
nuclear magnetic resonance
acids
Nuclear magnetic resonance
Acids
Chemical analysis
tyrosine
Chromophores
chromophores
Tyrosine
phthalates
proton magnetic resonance
versatility
activity (biology)
Bioactivity
Potassium

Keywords

  • External standards
  • NMR
  • Peptide concentration
  • Quantitative analysis

ASJC Scopus subject areas

  • Spectroscopy
  • Instrumentation

Cite this

Larive, C. K., Jayawickrama, D., & Őrfi, L. (1997). Quantitative analysis of peptides with NMR spectroscopy. Applied Spectroscopy, 51(10), 1531-1536.

Quantitative analysis of peptides with NMR spectroscopy. / Larive, Cynthia K.; Jayawickrama, Dimuthu; Őrfi, L.

In: Applied Spectroscopy, Vol. 51, No. 10, 10.1997, p. 1531-1536.

Research output: Contribution to journalArticle

Larive, CK, Jayawickrama, D & Őrfi, L 1997, 'Quantitative analysis of peptides with NMR spectroscopy', Applied Spectroscopy, vol. 51, no. 10, pp. 1531-1536.
Larive CK, Jayawickrama D, Őrfi L. Quantitative analysis of peptides with NMR spectroscopy. Applied Spectroscopy. 1997 Oct;51(10):1531-1536.
Larive, Cynthia K. ; Jayawickrama, Dimuthu ; Őrfi, L. / Quantitative analysis of peptides with NMR spectroscopy. In: Applied Spectroscopy. 1997 ; Vol. 51, No. 10. pp. 1531-1536.
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