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Silliker
Chem, Res. Center Crete, IL – Report of A Validation of LC-MS/MS Method for Folate Analysis
36
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
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 Formual Powder
004011RF00
25.0
200
Reconst.
Beverage
T=
492
I =
75
T=126
I=19
T=63
I=10
Infant Formula Powder
00402RF00
24.8
200
Reconst.
Beverage
Adult Nutritional RTF High
Protein
00415RF00
Recd as Beverage
As is
Adult Nutritional RTF High
Fat
00407RF00
Recd as Beverage
As is
Infant Formula Powder
Milk Based, SPIFAN Blank
Milk Form
00394RF00
Recd as Beverage
As is
* Samples were spiked with each targeted folate compound. Total Folic calcluted as sum of the folate
2011.06 (Fol-22) w/SLV
FOR ERP USE ONLY
DO NOT DISTRIBUTE