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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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range stipulated by the SMPR. The precision of the spike recovery evaluation in replicates as % RSD
was calculated. The values obtained were quite satisfactory and in the range of 1.1-6.0%
An average summary of the % spike recovery in replicates on two separate days for vitamin K2-7 is
provided in Table 9c.
Table 9c: Vitamin K2-4 Spike recovery in SPIFAN matrices on multiple days
VIT K2-4 Spike Recovery Parameters
% Recovery
Day 1
Day 2
average:
97.3
100.1
Stdev:
5.0
3.7
%RSD:
5.1
3.7
n:
30
15
Spike Recovery Parameters
Average of both days for all
replicates
average:
98.2
Stdev:
4.7
%RSD:
4.8
n:
45
An average of the spike recovery for vitamin K2-4 obtained in replicate analysis of the analyzed spiked
and non-spiked samples on two separate days at all the investigated spike levels is presented in Table
9c. An average spike recovery of 98.2% was obtained and is within the range stipulated by the SMPR.
The precision of the spike recovery evaluation in replicates as % RSD was calculated. The values
obtained were quite satisfactory and in the range of 3.7-5.1%.
The method LOD and LOQ:
The LOD and LOQ of the method were calculated based on replicate analysis of one of the SPIFAN
sample (Sample # 10, low in vitamin K1 concentration). Vitamin K1 in the sample was analyzed in
replicates (n=3) and its total vitamin K1 (cis + trans) concentration (mcg/100g of the reconstituted
sample) in the sample calculated. The LOD was calculated as 3 X of the standard deviation and LOQ
as 10X of the standard deviation of the replicates of the analysis.
Sample# 10 Vitamin K1 mcg/100g RTF
Trans Cis Total
Mean 2.27 0.29 2.56
SD 0.05 0.01 0.06
N 3 3 3
VitK-01 w/SLV
FOR ERP USE ONLY
DO NOT DISTRIBUTE