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Silliker
Chem, Res. Center Crete, IL – Validation of a LC-MS/MS Method for Vitamin K Analysis
Silliker Laboratories, Chemistry Research Center, Date: 1/30/15
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All powder samples reconstituted before the analysis (About 25 in
200 mL water. RF samples analyzed as such.
Concentration of Vitamin K1 (Total, trans + cis) was found to be in the range of 2.6 – 10.7 µg/100g in
the analyzed samples when analyzed second time in replicates of three. The precision of the vitamin K1
analysis in terms of the % RSD was estimated to be equal or under 8% for most of the analyzed
samples with concentration of total (trans + cis) vitamin K1 within the range of 1 -10 mcg/100g as
stipulated in the Vitamin K SMPR 2014.001. Two of the samples however gave % RSD values higher
values (10.2% and 15.7%) in their replicate analysis. One of the samples with concentration of Vit K1
higher than 10 µg/100g provided % RSD values under 5% (2.6%) as stipulated in the SMPR but the
value provide by the replicates of the other one were slightly higher (6.2%).
Cis –vitamin K1 was always found at relatively lower levels (0.3-2.8 µg/100g) in the samples with a%
RSD of the replicates in the range of 0.0-18.9%. The lower relative level of the analyte in samples may
be associated with the observed higher variance in the analysis of the replicates.
Trans –vitamin K1 was found to be in the range of 1.6 – 9.4 µg/100g in the analyzed samples. % RSD
of the trans-vitamin K1 analysis was estimated to be under 8% in replicates of most of the analyzed
samples. % RSD in the analysis of the replicates of the samples was found to be in the range of 0.4-
11%. Two of the samples however gave higher % RSD values (10.3% and 15.8%) in their replicate
analysis.
Table 5c: Analysis of different SPIFAN samples in replicates on
multiple days Day 3 Results
Day 3 of Multiple Day Replicate analysis
Sample ID
Paramtrs
Trans
Cis
Total
(trans +
cis)
mcg/100g of RF sample as is or a powder
sample reconstituted in to a liquid
SPIFAN
Sample #6
Mean
10.3
1.3
11.6
Stdev
0.5
0.1
0.6
% RSD
5.0
11.4
5.5
n
3
3
3
SPIFAN
Sample #7
Mean
2.4
1.1
3.5
Stdev
0.1
0.0
0.2
% RSD
5.8
1.7
4.5
n
2
2
2
SPIFAN
Sample #8
Mean
6.8
1.2
8.0
Stdev
0.3
0.1
0.3
% RSD
3.8
7.4
4.3
n
3
3
3
VitK-01 w/SLV
FOR ERP USE ONLY
DO NOT DISTRIBUTE