AOAC SPIFAN ERP Meeting Book-March 16, 2016

VitK-01 w/SLV FOR ERP USE ONLY DO NOT DISTRIBUTE

Silliker Chem, Res. Center Crete, IL – Validation of a LC-MS/MS Method for Vitamin K Analysis

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

Total (trans + cis)

Paramtrs

Trans

Cis

Sample ID

mcg/100g of RF sample as is or a powder sample reconstituted in to a liquid

Mean

10.3

1.3 0.1

11.6

0.5 5.0

0.6 5.5

Stdev

SPIFAN Sample #6

11.4

% RSD

3

3

3

n

Mean

2.4 0.1 5.8 6.8 0.3 3.8 2

1.1 0.0 1.7 1.2 0.1 7.4 2

3.5 0.2 4.5 8.0 0.3 4.3 2

Stdev

SPIFAN Sample #7

% RSD

n

Mean Stdev

SPIFAN Sample #8

% RSD

3

3

3

n

Silliker Laboratories, Chemistry Research Center, Date: 1/30/15

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