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

Common methods of HMO determination

HPAEC-PAD + High separation power + No need for derivatisation + Some selectivity from electrochemical detection + Identity from RT î Response factors variable between analytes î Connectivity to MS is tricky LC-FLD / LC-UV + Wide range of UV or fluorescent tags available + Some selectivity from derivatisation chemistry + Identity from RT + Response factors between OS are comparable

LC-MS + Separation in LC and in MS + No need for derivatisation + Some selectivity from MS detection + Identity from RT and MS data î Response factors variable between analytes CE-LIF + Some selectivity from derivatisation chemistry + Identity from RT + Response factors between OS are comparable + Possibility to multiplex î Connectivity to MS not straight forward î Need for derivatisation before analysis

+ Connectivity to MS straight forward î Need for derivatisation before analysis

Received from S. Austin

Quantification of HMO in Infant Formula Why? • R & D and product development • Confirmation of HMO level for safety trials , stability testing , efficacy trials , regulatory purposes • Quality control purposes to assure labeling compliance Are there methods available? • Methods developed for (academic) HMO analysis in milk could be applied • Small number of methods involving IF reported in literature • Methods not aligned as no official method is available