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Chem, Res. Center Crete, IL – Report of A Validation of LC-MS/MS Method for Folate Analysis
38
The SPIFAN samples were spiked with each targeted folate compound (at equal level of each of seven
targeted compound) and total spike level as folic acid calculated as sum of concentration of all folate
compounds as folic acid. Results in the above Table % spike recovery of folic acid at varied spike
levels analyzed in SPIFAN matrices ranged from 88 to 106%. Only one of the samples at one of the
spike level provided 88% spike recovery. All other samples at every spike level provided spike
recovery within the recovery range 90-110% stipulated by the SMPR. Folate recovery analysis was
satisfactory in all of the studied SPIFAN matrices at investigated spike levels.
The repeatability of % spike recovery analysis in multiple replicate was evaluated as % RSD value. %
RSD of % spike recovery analysis of folic acid at varied levels in replicates analyzed in SPIFAN
matrices ranged from 0.009 % to 7.2 %. The spike recovery of folic acid at different spike levels
analyzed in SPIFAN matrices was within range mentioned in the SMPR. The repeatability of folate
recovery analysis was found to be satisfactory in all of the studied SPIFAN matrices at investigated
spike levels.
The results of the spike recovery evaluation are presented in simplified form in the following Table. All
SPIFAN samples were spiked in triplicates with targeted folate compounds at three different levels
(spike Level I, II and III) were evaluated for spike recovery of total folate (sum of spiked/analyzed
folate compounds as folic acid) on different days. Averages of per cent spike recovery of total folate in
replicates of all the samples calculated irrespective of their level of the spike. This provides a
comprehensive and simpler evaluation of the spike recovery in the reconstituted and RTF SPIFAN
samples.
Table 29: % Spike recovery of total folate in samples spiked with targeted folate irrespective of
the level of the spike
Sample
#
Sample
Lot #/ID
Matrix
Analyzed
as
% Spike
Recovery
Parameters
Average
(%) Spike
Recovery
in All
Spike
samples*
1
Child
Formual
Powder
004011RF00
Powder
Reconst.
Beverage
Av % Recov
103.9
SD
2.1
% RSD
2.1
2
Infant
Formula
Powder
00402RF00
Powder
Reconst.
Beverage
Av % Recov
98.6
SD
2.4
% RSD
2.4
3
Adult
Nutritional
RTF High
Protein
00415RF00
Beverage
As is
Av % Recov
92.4
SD
1.6
% RSD
1.7
4
Adult
Nutritional
RTF High Fat
00407RF00
Beverage
As is
Av % Recov
92.6
SD
6.2
% RSD
6.7
5
Infant
Formula
Powder Milk
Based,
SPIFAN
00394RF00
Beverage
As is
Av % Recov
101.2
SD
2.4
% RSD
2.3
2011.06 (Fol-22) w/SLV
FOR ERP USE ONLY
DO NOT DISTRIBUTE