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

8

8

8

n

Mean

7.4 0.6 8.1

0.9 0.1 6.6 1.1 0.1 9.8 9

8.3 0.6 7.1

Stdev

SPIFAN Sample #12

% RSD

9

9

n

Mean

5.0 0.6

6.1 0.7

Stdev

SPIFAN Sample #13

11.4

11.0

% RSD

9

9

9

n

Mean

5.4 0.3 4.9 7.8 0.7 9.4 6.2 0.5 7.6 7.7 0.4 5.7 8 9 8

1.1 0.1

6.5 0.3 3.9 9.1 0.8 8.6 8.5 0.5 5.4 8 9

Stdev

SPIFAN Sample #14

10.7

% RSD

8

n

Mean Stdev

1.4 0.1 8.0

SPIFAN Sample #15

% RSD

9

n

Mean

2.3 0.3

Stdev

SPIFAN Sample #16

12.5

% RSD

8

8

n

Mean

2.8 0.2 5.4

10.5

0.5 5.0

Stdev

SPIFAN Sample #17

% RSD

n 8 All powder samples reconstituted before the analysis (About 25 in 200 mL water. RF samples analyzed as such. 8 8

Concentration of Vitamin K1 (Total, trans + cis) was found to be in the range of 2.6 – 11.5 µg/100g in the analyzed samples when analyzed third time in replicates. The precision of the vitamin K1 analysis in terms of the % RSD was estimated to be equal or under 8% in 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. Some samples however gave some what higher % RSD values (11 - 13%,) in their replicate analysis. The sample with concentration of Vit K1 higher than 10 µg/100g, provided % RSD values closer to 5%. 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 5.0 -12.5 %. 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 2.2 – 10.1 µg/100g in the analyzed samples. % RSD of the trans-vitamin K1 analysis was estimated to be under 8% in replicates of many of the analyzed samples. But % RSD value for trans-vitamin K1analysis of the replicates of some of the samples was some what higher (9.4-15.3%). The sample with concentration of Vit K1 higher than 10 µg/100g provided % RSD value of 6.2%. The precision of the method for vitamin K1 analysis can be stated based on the average % RSD value of all the 12 SPIFAN samples tested in replicates (usually 3) on three different days thus replicates of usually 8-9 for every sample. The precision of the vitamin K1method based on the average % RSD

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

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