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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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Conclusions
1.
A LC-MS/MS method was developed and validated for analysis of different folate
vitamers (Folic acid + 5- CH3-THF +
5-CHO-THF + THF + 10-CH3-FA + 10-CHO-FA +
Methenyl-THF). The
sum of these compounds (as folic acid) was calculated to
represent total folic acid.
2.
HPLC conditions and mass spec parameters were optimized.
3.
The composition of extraction buffer and analysis buffer were adjusted to provide
the best analytical response of the analytes.
4.
SPE conditions and elution solvents were selected to provide the best recovery of
the analytes.
5.
The method accuracy was evaluated and established by analysis of various
reference materials.
6.
The method employed to analyze a series of the SPIFAN II samples. The powder
samples were reconstituted to the liquid form and RTF samples were analyzed as
is. The results expressed as mcg/100g of total folate as folic acid (sum of the
detected folate compounds after calculation as folic acid).
7.
The method precision was evaluated for folate analysis in the SPIFAN samples
and was found satisfactory and within the folate SMPR specifications.
8.
Results of selected folate vitamers are presented for certain reference materials.
9.
Five SPIFAN samples (two powders and three RTF) were spiked with targeted
folate compounds at three different levels (spike Level I, II and III). % spike
recovery of folates as folic acids at the evaluated levels was within the recovery
range 90-110% stipulated by the SMPR except in one of the sample at lowest spike
level spike level where recovery was slightly higher (1.5%) than the SMPR upper
range. Folate spike recovery when calculated as average of all the levels in every
sample was found to be in the range of 94-108% and with in the SMPR range. %
Spike recovery of total folate was found to be satisfactory in all of the studied
SPIFAN matrices.
10. The LOD of the method was calculated to be 0.1 mcg/100g in reconstituted liquid
and the RTF samples. The LOQ of method was calculated to be 0.4 mcg/100g.
LOD and LOQ of the method are within the folate SMPR required range.
References:
AOAC 2011.06
Folate SMPR AOAC 2011.006
2011.06/Fol-22 (February 2016)
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