During stability studies at high temperature (70 degrees C) and low relative humidity ( approximately 0%), the recovery of an asparagine containing hexapeptide (VYPNGA) and its known deamidation products from solid polyvinylpyrrolidone (PVP) matrices was incomplete. To determine the causes of this mass loss, formulations were prepared by lyophilizing solutions containing PVP, glycerol, and the Asn-hexapeptide in pH 7.5 phosphate buffer, followed by storage at 70 degrees C and 0% relative humidity. Asn-hexapeptide loss was mono-exponential and reached a plateau at about 30% remaining. Total recovery of the peptide and its known deamidation products was approximately 30% of peptide load. Size exclusion chromatography with fluorescence detection indicated the formation of a PVP-peptide adduct that was stable in the presence of 6 M guanidine hydrochloride. Similar stability studies using N-acetyl phenylalanine, phenylalanine ethyl ester, and N-acetyl tyrosine ethyl ester demonstrated that the reaction involves the peptide N-terminus. The adduct was disrupted in the presence of carboxypeptidase-A, suggesting the formation of an amide bond between the peptide and PVP. (15)N solid-state nuclear magnetic resonance spectroscopy using (15)N-labeled valine as a model of the peptide N-terminus showed different populations of (15)N, suggesting that noncovalent peptide-polymer interactions precede amide bond formation.