Contribution of minor folates to the total folate content of infant formula and
adult/pediatric nutritionals / F. Martin (NRC, AS)
03 Feb 2016
CONFIDENTIAL
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B A C K G R O U N D A N D O B J E C T I V E S
The Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN) of the AOAC has identified
vitamin B
9
as one of the nutritional parameters which needs new analytical methodology for its
determination. The Expert Review Panel (ERP) reviewed and granted First Action status to two
methods: AOAC Official Method 2013.13 (Nestlé’s method) and AOAC Official Method 2011.06
(Silliker’s method). The two methods differ on several points, which are summarized in the following
table:
Parameter
AOAC 2013.13
AOAC 2011.06
Compounds targeted
Folic acid, 5ͲmethylͲ
tetrahydrofolate
Folic acid, 5ͲmethylͲtetrahydrofolate,
5ͲformylͲtetrahydrofolate,
tetrahydrofolate, 5,10 methenylͲ
tetrahydrofolate, 10ͲmethylͲfolic
acid, 10ͲformylͲfolic acid
Extraction procedure
DualͲenzyme (amylase,
protease)
Trienzyme (amylase, protease,
deconjugase)
SPE cleanͲup
Strong anion exchange (SAX)
Weak anion exchange (WAX)
It is worth noting that the SMPR (Standard Method Performance Requirements), issued by the
SPIFAN, target folic acid and 5ͲmethylͲtetrahydrofolate (including the 5ͲmethylͲtetrahydrofolate
polyglutamates).
During method development of AOAC 2013.13, experiments showed that other minor folates
potentially present in infant formula and adult nutritionals represented a minor contribution to the
total vitamin B
9
content. Moreover, trials with deconjugase showed only a minor liberation of
polyglutamate folates.
The main objective of this note is to assess the content of minor folates
1
in products belonging to the
SPIFAN II kit with and without inclusion of the polyglutamate forms.
1
Minor folates targeted are: 5ͲmethylͲtetrahydrofolate (5ͲMe THF), 5,10 methenylͲtetrahydrofolate (5Ͳ
10MylͲTHF), tetrahydrofolate (THF), 5ͲformylͲtetrahydrofolate (5CHOͲTHF), 10ͲformylͲfolic acid (10CHOͲFA)
and 10ͲmethylͲfolic acid (10MeͲFA).
2013.13 (Fol-21) / (February 2016) - NOTES
FOR ERP USE ONLY
DO NOT DISTRIBUTE