AOAC SPIFAN Stakeholder Panel Meeting Book (September 7, 2019)

Analytical Techniques: HPLC-based methodology

By http://en.wikipedia.org/wiki/User:Kommando - Own work. Originally uploaded at http://en.wikipedia.org/wiki/File:HPLC.jpg, CC BY-SA 3.0, https://commons.wikimedia.org/w/index.php?curid=15808573

HPLC-UV Methods

X Most analytical applications are reverse phase (RP). C4 column commonly used X Most targeted to in-tact protein analysis

X Raw material: Typically UV at 280nm (GB >95%)

X Many possible interferences (BSA, lactalbumin noted in lit) X Sufficient characterization of raw material?

X Finished product or milk analysis usually requires a cleanup step X Heparin column – affinity chromatography. Multiple references X Cation exchange cleanup. Multiple references X Cleanups offer additionaly selectivity for non-denatured protein

Novel cation exchange chromatography

X

X Billakanti, J., McRae, J., Mayr, M., & Johnson, K. (2019). Advanced analytical tools for bovine lactoferrin identification and quantification in raw skim milk to finished lactoferrin powders. International Dairy Journal , 104546. https://doi.org/10.1016/j.idairyj.2019.104546