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Silliker
Chem, Res. Center Crete, IL – Report of A Validation of LC-MS/MS Method for Folate Analys
15
Method Validation Results:
The Evaluation of Accuracy of the Method by Analysis of Reference Materials :
The method accuracy was evaluated by analysis of a variety of Reference materials. The reference
values for total folate or some of the selected folate vitamers is available for some of the Reference
materials. The Results are presented in Table 7.
Average Folic Acid value obtained by the method in the NIST SRM 1849a matched closely with its
Certified value. The average value of 5-methyl-THF obtained by our method was almost twice of
reported by NIST. The NIST value for this vitamer is based on the NIST’s in house analysis by a LC-
MS/MS method and not a consensus value. It’s a reference value and carries relatively less confidence
in its accuracy than the certified value. Thus the comparison of our results of 5-CH3-THF with that of
NIST reference value may be a comparison of performance of two methods for the analysis of this
analyte. Total folate as folic acid in this reference material was found to be close to that obtained using
the microbiological analysis.
Intensity,
cps
Max. 4.7e4
4.0
4.2
4.4
4.6
4.8
5.0
5.2
5.4
5.6
5.8
6.0
6.2
6.4
6.6
6.8
7.0
7.2
7.4
7.6
7.8
8.0
Time, min
0.0
1.0e4
2.0e4
3.0e4
4.0e4
5.0e4
6.0e4
7.0e4
8.0e4
9.0e4
1.0e5
1.1e5
1.2e5
1.3e5
1.4e5
6.14
Figure 5: Folate Chromatograms with sodium ascorbate.
2011.06/Fol-22 (February 2016)
FOR ERP USE ONLY
DO NOT DISTRIBUTE