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Chem, Res. Center Crete, IL – Report of A Validation of LC-MS/MS Method for Folate Analys
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about 10% of the total folates in the samples. One other sample, infant formula powder based on
partially hydrolyzed milk was found to contain a very small amount of minor folate which contributed
to 6.5% of total folate in the sample.
% Spike Recovery of total folate (sum analyzed folates as folic acid) after spike with the targeted
folate compounds and their analysis in SPIFAN II matrices:
Details of Reconstitution of the samples before the analysis:
All of the 5 samples received from SPIFAN for spiking purposes 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 20.
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.
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 employing trienzyme extraction
including conjugase on different days. The samples were spiked with each targeted folate compound
and their sum as folic acid calculated as total folate spike level (folic acid). Spike levels are calculated
as mcg/100g in the reconstituted liquid and as is the RTF samples.
Table 20: Details of the spike levels and reconstitution of the SPIFAN samples processed for %
spike recovery evaluation
Matrix
Lot #/ID
Reconstitution of Powder in
to water
Analyzed as
Spike levels
(mcg/100g)*
Total Folate (T),
Individual Folate
Compound (I)
Powder (g)
In to water (g)
Level
1
Level
2
Level
3
Child Formula Powder
004011RF00
Recd 12/4/15
25.0083
200
Reconst.
Beverage
T=
500
I = 60-
80
T=120
I=20
T=60
I=10
Infant Formula Powder
00402RF00
Recd 12/4/15
25.0026
200
Reconst.
Beverage
Adult Nutritional RTF High
Protein
00821RF00
Recd 12/4/15
Recd as Beverage
As is
Adult Nutritional RTF High
Fat
00820RF00
Recd 12/4/15
Recd as Beverage
As is
Infant Formula Powder
Milk Based, SPIFAN Blank
Milk Form
EV4H2Q Recd
12/4/15
Recd as Beverage
As is
* Samples were spiked with each targeted folate compound. Total Folic calculated as sum of the folate
compounds after adjusting their values to folic acid.
2011.06/Fol-22 (February 2016)
FOR ERP USE ONLY
DO NOT DISTRIBUTE