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FOS-04

Determination of Fructans in Infant and Adult/Pediatric Nutritional Formula as well as

ingredient commodities

High-Performance Anion-Exchange Chromatography with Pulsed Amperometric Detection

Principle

The basic principle of this method is as follows. A two part analysis is performed. Part 1 is the qualitative classification

of the fructan present in the sample, if not already known. This classification is based on rules related to

presence/absence of GF

3

(Nystose) and GF

4

(fructofuranosylnystose), specifically, and higher oligomers, generally. It

groups the fructans into one of three DP ranges and allows selection of an appropriate commodity factor for converting

fructose content to total fructan content. Part 2 is the quantitative determination of total fructan in the sample. The

sample to be analyzed is weighed and diluted with lab water, as appropriate. Part 1 of the analysis is then performed. Part 2 can

be run in parallel, whereby an aliquot of the diluted product is treated with sucrase to hydrolyze any sucrose present. Glucose

and fructose released by the enzymatic hydrolysis, as well as inherent glucose and fructose, are then reduced to sugar alcohols by

the addition of sodium borohydride. Excess borohydride is neutralized by the addition of acetic acid. Lastly an aliquot of

internal standard (glucoheptose) and an aliquot of fructanase are added to the sample solution. After the fructanase hydrolysis is

completed the samples are analyzed for fructose on HPAEC-PAD instrumentation. The total fructan content is calculated from

the fructose, adjusted by a commodity factor (

CF, determined by part 1

).

Part 1

Qualitative Fructan ID

Materials

Fructans:

1.

scFOS: Ingredion Incorporated (Nutraflora), vendor lot # 7642339101 used

2.

oligofructose + inulin: Beneo Orafti Synergy 1, vendor lot # YAGKL7HAF8 used

3.

enriched inulin: Beneo Orafti HP, vendor lot # HPQEK7CEK7 used

Product Matrices:

1.

Powdered pediatric formula (milk based with oligofructose)

2.

Powdered infant formula (soy based with scFOS)

3.

Powdered adult product (milk based with Synergy 1)

4.

Powdered infant formula (low lactose, milk based with GOS): Spiked in lab with inulin (HP)

5.

SPIFAN material

a)

Placebo Child Formula Powder (00847RF00)

b)

Toddler Formula Powder Milk-Based (4052755861)

c)

Infant Formula Powder Milk Based (4044755861)

d)

Child Formula Powder (00866RF00)

e)

Infant Formula Powder FOS/GOS Based (50350017W1)

f)

Infant Elemental Powder (00796RF)

g)

Adult Nutritional RTF, High Protein (00821RF00)

h)

Adult Nutritional RTF, High Fat (00820RF00)

i)

Infant Formula RTF, Milk based (EV4H2Q)

j)

Infant Formula Powder Partially Hydrolyzed Milk Based (410057652Z)

k)

Infant Formula Powder Partially Hydrolyzed Soy Based (410457651Z): Spiked in lab with inulin (HP)

l)

Adult Nutritional Powder Low Fat (00859RF00)

m)

Infant Formula Powder Soy Based (E10NWZC)

n)

Infant Formula RTF Milk Based (EV4H2R)

o)

Adult Nutritional RTF High Protein (00730RF00)

Fos-04 (February 2016)

FOR ERP USE ONLY

DO NOT DISTRIBUTE