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Silliker

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

31

Spike Recovery Evaluation in SPIFAN samples at different spike levels:

Two beverages prepared from the respective powdered samples and three RTF samples were analyzed

for all the targeted folate compounds with and without spike with respective folate compounds. The

samples were spiked with the folate compound at three spike levels . The results are presented in Table

21.

Table 21: % Spike Recovery of folate compounds in SPIFAN samples spiked with folate

compounds at three different spike levels. The results are for total folate (sum of folate

compounds after adjustment to folic acid).

Sample#

Sample

Lot#

Spike

Level

Spike of

individual

folates

compounds

as folic acid

Spike of total

folate

calculated as

folic acid

Average %

Spike

Recovery of

total folate

(n=3)

% RSD 0f the

total folate

spike recovery

replicates

(n=3)

mcg/100g

1

Child Formula

Powder

00411RF00

REC'D

12/4/15

I

70-80

500.0

97.8

0.7

II

20

120.0

99.7

0.7

III

10

60.0

103.5

1.8

2

Infant

Elemental

Powder

00402RF00

REC'D

12/4/15

I

60-70

500.0

98.1

1.2

II

20

120.0

98.8

1.4

III

10

60.0

99.1

1.5

3

Adlt

Nutritional

RTF, High

Protein

00821RF00

REC'D

12/4/15

I

60-70

500.0

93.1

0.03

II

20

120.0

95

0.6

III

10

60.0

95.1

3.9

4

Adult

Nutritional

RTF, High Fat

00820RF00

REC'D

12/4/15

I

60-70

500.0

94.3

1.0

II

20

120.0

93.5

0.9

III

10

60.0

98.8

3.5

5

Infant Formula

RTF Milk Based

(SPIFAN Blank

Milk Form)

EV4H2Q

REC'D

12/4/15

I

60-70

500.0

104.4

3.5

II

20

120.0

107.2

1.0

III

10

60.0

111.5

0.9

* Samples were spiked with each targeted folate compound at three different spike levls. Spike recovery of total

Folate calculated based on sum of the folate compounds as folic acid. Each spike recovery value is an average of

three independent analysis.

Results in the above Table 21 for % spike recovery of folic acid at varied spike levels analyzed in

SPIFAN matrices ranged from 93 to 107%. One of the samples at one of the spike levels provided a

slightly higher recovery of 111.5%. All of the other samples at every spike level provided spike

recovery within the recovery range 90-110% as required by the Folate SMPR. Folate % spike

recovery analysis was satisfactory in all of the studied SPIFAN matrices at investigated spike levels

except one of the sample at the lowest spike level of 60 mcg/100g providing about 1.5% higher

recovery than the SMPR upper range.

The repeatability of % spike recovery analysis in multiple replicate was evaluated as % RSD value. %

RSD of % spike recovery analysis of total folic acid at varied spike levels in replicates of SPIFAN

matrices ranged from 0.03 % to 3.5 %. The repeatability of folate recovery analysis was found to be

2011.06/Fol-22 (February 2016)

FOR ERP USE ONLY

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