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

Quantification of HMO in Infant Formula

Reference

Technique HPAEC-PAD & HILIC-FLD

Analyte(s) Comments

Austin et al. Molecules (2018) 23 : 2650

2’FL LNnT

HPAEC-PAD delivered results below the expected values on pilot plant produced products • Selectivity in bovine milk samples (not per se in HM) • DFL selectivity? • Enzymes not commercially available

Seydametova et al. Anal. Biochem. (2019) 582 : 113358

Enzymatic

2’FL

LOQ 1.75 P g/ml (approx. 0.0002%)

Prieto et al. J. Food Chem. Nutr. (2019) 7(1)

HPAEC-PAD

2’FL

Nijman et al. J. Agric. Food Chem . (2018) 66 : 6851

nano-LC/Q-TOF MS various

HPAEC-PAD also for selected HMO in milk samples Endogenous acidic BMO in infant formula from China & Malaysia

Ma et al. Int. Dairy J. (2019) 94 : 1-6

LC-MS

3’SL 6’SL 6’SLN DSL

Eurofins Food Feed & Water testing Nederland (2018)

HPAEC-PAD

2’FL

LOQ 0.002% and 0.01%

Quantification of HMO in Infant Formula Challenges Next to the challenges in relation to HMO analysis in milk as discussed above • No official/standard method available • Lack of standards of sufficient (and known) purity î Presence of other components (e.g. 2’FL/DFL, 3’SL/6’SL) î Moisture content • Interferences with other ingredients in IF (e.g. lactose, GOS, FOS, polydextrose, maltodextrin) • Good separation between analytes (e.g. 2’FL and 2’-fucosyllactulose) • Laborious sample preparation (lactose, fat, protein, non-HMO carbohydrate removal)