RP-HPLC peptide profiling of cheese extracts: A study of sources of variation, repeatability and reproducibility

Peptide profiling of cheese extracts by RP-HPLC is widely used to study cheese quality and authenticity. Repeatability, reproducibility and the contribution of differences in sample/treatment, extraction, and the effect of the chromatographic run was studied. Chromatograms (108) of pH 4.6-soluble extracts from 180 d Cheddar cheeses were used to evaluate the precision of the method and to quantify the effect of treatment and replicate cheese trial. Repeatability and reproducibility coefficient of variations of peaks/RT classes varied widely and no clear correlation was found between peak size and reproducibility. ANOVA of Principal Component Scores identified significant differences amongst cheeses and a trial effect. Partial Least Square Discriminant Analysis identified variables related to cheese treatment. Statistical process control methods were used to study the variability of replicate injections over the life of a single column showing the discriminatory power of RP-HPLC and giving possibilities to improve both process control and process optimisation.