SPIFAN Nutrients ERP Book_9-29-15

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

Eurofins Lab. LTD., Wolverhampton: SOP CHROM/341-Choline in pet foods by ion chromatography with conductivity detection.

The quantification by LC-MS/MS is based on the analytical concepts of the follow- ing two articles.

STARKEY et AL. J.AOAC INT. 91,1,2008,130-142.Single-Laboratory Validation of a Liquid chromatographic/Tandem Mass Spectrometric Method for the Deter- mination of Free and Total Carnitine in Infant Formula and Raw Ingredients.

Brief method description:

The sample is shaken in a mixture of water and methanol thereby extracting car- nitine including their phosphorylated forms. An aliquot is treated with potassium hydroxide solution at elevated temperature for 4 hours thereby converting the phosphorylated forms to free carnitine. The mixture is neutralised and deuterered internal standard is added. Quantification is done by LC-MS.

Statistics:

In the current validation report, a p-value < 0.05 is considered statistically signifi- cant. Consequently, confidence intervals (CI) are calculated at a confidence level at 95 %. Paired t-testing is performed on normal distributed raw data. Data which is not within the normal distribution is clearly marked within the text.

Matrices:

This method can be used for quantification of free and/or total carnitine in the fol- lowing matrices:

 Food and feed.  Supplements, premixes and raw products.  Milk powder and nutritional formulas.  Animal feed / pet food (dry and wet).

Specificity:

The specificity has been evaluated by examining chromatograms of natural sam-