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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
17
SPIFAN
Sample
#15
Mean
7.5
1.3
8.8
Stdev
0.7
0.0
0.7
% RSD
8.8
3.0
7.8
n
3
3
3
SPIFAN
Sample
#16
Mean
6.4
2.3
8.7
Stdev
0.0
0.2
0.2
% RSD
0.4
7.3
2.2
n
2
2
2
SPIFAN
Sample
#17
Mean
7.4
2.7
10.1
Stdev
0.3
0.1
0.3
% RSD
3.7
3.7
2.9
n
3
3
3
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 2.7 – 12.0 µg/100g in
the analyzed samples. 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 slightly higher values (9.5% and 11.5%) in their replicate
analysis. Both of the samples with concentration of Vit K1 higher than 10 µg/100g provided % RSD
values under 5% (2.4 and 2.9%) as stipulated in the SMPR.
Cis –vitamin K1 was always found at relatively lower levels (0.3-2.7 µg/100g) in the samples with a%
RSD of the replicates in the range of 3.0-13.1%. 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 2.3 – 10.5 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%. Some of the samples however gave % RSD values some what higher values (10.9%-
11.0%) in their replicate analysis.
The sample with concentration of trans-Vit K1 higher than 10
µg/100g (10.5 µg/100g) did provide % RSD values under 5% (2.0%) as stipulated in the SMPR.
Table 5b: Analysis of different SPIFAN samples in replicates on
multiple days Day 2 Results
Day 2 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
Mean
9.4
1.3
10.7
VitK-01 w/SLV
FOR ERP USE ONLY
DO NOT DISTRIBUTE