Ion-Exchange Chromatographic Method for the Determination of the Free Amino Acid Composition of Cheese and Other Dairy Products : an Inter-Laboratory Validation Study

Although free amino acids (FAAs) represent a significant component of ripened cheeses and can provide useful information for their characterization, no inter-laboratory validated analytical method exists which allows a reliable comparison of data obtained by different laboratories and the adoption of quality control schemes based on FAA pattern. The objective of the present work was to test the effectiveness of an analytical protocol for the determination of the FAA composition of cheese and to verify the adequateness of this type of analysis for quality control procedures of Grana Padano PDO cheese as well as for research purposes. After an initial test to compare performances of ion-exchange chromatography (IEC) and HPLC techniques, an inter-laboratory collaborative study (seven laboratories, four samples) was organized to validate an IEC method with post-column ninhydrin derivatization and using l-norleucine as an internal standard. Determined amounts of individual FAA ranged from 8 to over 1380 mg/100 g cheese, with relative standard deviation of repeatability (RSDr) ranging from 0.5 to 4.6%, and relative standard deviation of reproducibility (RSDR) ranging from 1.3 to 9.9% for FAA concentrations over 100 mg/100 g. For lower concentrations, RSDr and RSDR were about thrice as high. On the basis of the results of this investigation, at present, the validated method is adopted as the official method for the determination of FAA patterns in the quality control of Grana Padano PDO cheese.