Study of SI-traceable purity assessment of bovine lactoferrin using mass balance approach and amino acid-based isotope dilution liquid chromatography-mass spectrometry.

The accurate measurement of bovine lactoferrin (bLF) attracts wide attention in food and nutraceutical applications as its important physiological and nutritional functions. We present SI traceable procedures for assessing bLF purity using mass balance method and amino acid (AA)-based isotope dilution mass spectrometry (IDMS). The mass balance method was revealed with a purity of 0.938 ± 0.011 g/g by deducting all aspects of impurities, including related structure impurities of 4.60%, ignition residue of 0.28%, Cl of 1.10%, SO of 0.13%, and moisture of 0.17%. The AA-based IDMS quantitative result was 0.937 ± 0.027 g/g. Hydrolysis conditions were optimized and methodology validation including, accuracy, precision, were studied. Good consistency was achieved between the two independent strategies and bLF purity assigned via the weighted mean value of their results was 0.938 ± 0.015 g/g. These analyses are expected to be applicable to proteins quantification and development of LF certified reference materials.