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Single Lab Validation Report for GOS in Infant Formula and Adult Nutritionals

13

5

Appendices

5.1

Detailed Method Description

Determination of trans-galacto-oligosaccharides in infant, adult and pediatric

formula

Ion-Exchange Chromatography

Principle:

Galacto-oligosaccharides (GOS) also known as oligogalactosyllactose, oligogalactose, or transgalacto-

oligosaccharides is produced by transgalactosylation of lactose. This method has been set up to

determine the GOS content in infant, adult and pediatric formula.

Currently, the standard method for analyzing GOS in food products is AOAC 2001.02 (De Slegte, 2002).

GOS and lactose in the sample are hydrolyzed with an enzyme to galactose and glucose. GOS content is

calculated from the difference in galactose content of a hydrolyzed and non-hydrolyzed sample.

The disadvantage of the AOAC 2001.02 method for IFT is that the high lactose content in IFT results in a

high quantity of galactose formed after hydrolysis. The resulting galactose quantity is very high

compared to the galactose quantity released from GOS. For the GOS calculation two large numbers are

subtracted, which leads to an estimate for GOS content that is not precise. Precipitation with cold

ethanol can be applied to remove part of the lactose. However, a considerable amount of lactose

remains and part of GOS may be removed as well.

In the currently proposed method, which is developed in collaboration with Danone Nutricia Research,

lactose is removed with a chromatographic preparative step before hydrolysis. The method consists of

the following steps:

1) preparative separation

2) SPE purification and concentration

3) enzymatic hydrolysis

4) HPAEC-PAD analysis

The GOS concentration is expressed as g GOS in 100 g reconstituted product.

GOS-02 SLV

FOR ERP USE ONLY

DO NOT DISTRIBUTE