SPIFAN Nutrients ERP Book_9-29-15

Carn-06 (Choline Validation Report) FOR ERP USE ONLY DO NOT DISTRIBUTE

Robustness:

The robustness of the method has been evaluated on the following critical pa- rameters; extraction time and saponification time. Extraction time: The robustness test has been performed on a milk powder and a pet food con- taining choline. 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

Choline:

Ekstraktion 37°C

2500

2000

1500

1000

Pet food

Milk powder

500

Choline (mg/kg)

0

Time (hour)

From the data (see appendix 4) it is seen that an extraction time of 20 hours is sufficient to extract all the choline 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 choline. 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.