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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