AOAC SPIFAN ERP Meeting Book-March 16, 2016

2011.06 (Fol-22) w/SLV FOR ERP USE ONLY DO NOT DISTRIBUTE

Silliker Chem, Res. Center Crete, IL – Report of A Validation of LC-MS/MS Method for Folate Analysis

The results of total folate as folic acid (sum of the detected folate compounds after calculation as folic acid) in the SPIFAN samples are presented in the above Table. The samples were analyzed in triplicates for three separate days. Averages and % RSD were calculated for all the replicates analyzed for the respective sample for all three days. Intermediate precision or interday precison as % RSD not clarified in the SMPR requirements. Interday precision as % RSD obtained values are well below the reproducibility requirement (as %RSD R ) of the SMPR (< 16%). The range of the inter-day precision (as % RSD) obtained by the method are 7 – 12.7% . % Spike Recovery of total folate (sum analyzed folates as folic acid) after spike with targeted folate compounds and their analysis in Different SPIFAN Matrices: Details of Reconstitution of the samples before the analysis: All the 5 samples received from SPIFAN were analyzed in replicates by the method for the targeted folate analytes with and without spike with the targeted folate compounds. The powder sample were reconstituted by taking approximately 25 g of the powder into 200 mL of water. Details of the samples and reconstitution are provided in the following Table 30. All samples were spiked in triplicates with targeted folate compounds at three different levels (spike Level I, II and III). Samples with different spike levels were analyzed on different days. The samples were spiked with each targeted folate compound and their sum as folic acid calculated as total spike level (folic acid). Spike levels are calculated as mcg/100g in the reconstituted liquid and the RTF SPIFAN samples (as received). There are total of five samples for spiking purposes out of which two are powders. All the powdered samples were reconstituted as ready to feed beverages. Table 27: Details of the spike levels and reconstitution of the SPIFAN samples processed for % spike recovery evaluation

Spike levels (mcg/100g)* (Total Folate (T)/Individual Folate Compound (I))

Reconstitution of Powder in to water

Matrix

Lot #/ID

Analyzed as

Level 1

Level 2

Level 3

Powder (g)

In to water (g)

Reconst. Beverage

25.0

200

Child Formual Powder

004011RF00

Reconst. Beverage

24.8

200

Infant Formula Powder

00402RF00

T= 492

Adult Nutritional RTF High Protein

Recd as Beverage

As is

T=126 I=19

T=63 I=10

00415RF00

I = 75

Adult Nutritional RTF High Fat

Recd as Beverage

As is

00407RF00

Infant Formula Powder Milk Based, SPIFAN Blank Milk Form

Recd as Beverage

As is

00394RF00

* Samples were spiked with each targeted folate compound. Total Folic calcluted as sum of the folate

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