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

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

% Recovery

VIT K2-4 Spike Recovery Parameters

Day 1

Day 2 100.1

97.3

average:

5.0 5.1

3.7 3.7

Stdev: %RSD:

30

15

n:

Average of both days for all replicates

Spike Recovery Parameters

average:

98.2

Stdev: %RSD:

4.7 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

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

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