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taining both carnitine.

The sample is weighed into the extraction bottle, extraction solvents are added,

the bottle is closed and placed on the shaking table at 37°C. At the time t. the

sample is removed from the shaking table, centrifuged and placed on cold stor-

age until saponification.

t / hour

1 2 3 4 5 22

Carnitine:

From the data (see appendix 4) it is seen that an extraction time of 20 hours is

sufficient to extract all the carnitine from the sample.

Saponification time:

The robustness test has been performed on raw egg, pet food, milk powder (in-

fant formula), cornflakes and fish feed containing both carnitine.

An aliquot of the extraction solution is transferred to the saponification bottle, po-

tassium hydroxide and water is added and the mixture is saponifyed at a 100°C.

At the time t. the sample is removed from the oven and neutralized with hydro-

chloric acid.

t / min

0 (free)

30 60 90 120 150 180 240 300 360

From the data (see appendix 4) it is seen that a saponification time of 240 min is

sufficient to convert all bound carnitine to the free forms for all the tested ma-

trixes.

Carnitine:

0

50

100

150

200

250

300

350

400

Carnitine (mg/kg)

Time (hour)

Ekstraktion 37°C

Pet food

Milk powder

Carn-06 - Validation Report

FOR ERP USE ONLY

DO NOT DISTRIBUTE