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Silliker

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

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folate recovery analysis was found to be satisfactory in all of the studied SPIFAN matrices.

The grand average of % spike recovery of folate based on all of the SPIFAN samples was found to be

99.3% with a % RSD value of 5.4% (n=45), thus a satisfactory overall spike recovery was obtained

with a good precision of the replicate analysis .

LOD and LOQ of the Method:

The LOD and LOQ of the method were calculated based on replicate (n=7) analysis of one of the

SPIFAN reconstituted beverage sample with very low total folate levels (near detection). Total folate

compounds in the sample were analyzed and total folate calculated (mcg/100g) as sum of the detectable

folate compounds as folic acid. Standard deviation (Stdev) of the replicate analysis of total folate in the

sample was calculated. The LOD was calculated as 3 X Stdev and LOQ as 10X Stdev. The calculated

LOD and LOQ of the method are provided in the following Table 23.

Table 23: LOD and LOQ of the Method

Parameters

Analyte

Values as mcg/100g of

reconstituted

liquid/RTF

LOD

Total Folate

0.127

LOQ

0.425

Values based on replicate (n=7) analysis of samples with folate at

very low level (near detection level). Stdev of the replicate analysis

was calculated. LOD = 3xStdev. LOQ = 10xStdev.

LOD and LOQ required by the folate SMPR are < 0.1 and <0.5 mcg/100g respectively. The LOD and

and LOQ obtained for the method are within the requirements of the folate SMPR.

2011.06/Fol-22 (February 2016)

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