AOAC SPIFAN ERP Meeting Book-March 16, 2016

Bio-02 w/SLV FOR ERP USE ONLY DO NOT DISTRIBUTE

The placebos or unfortified SPIFAN matrices were spiked at 50% and 150% of the typical target level. In absence of a placebo, fully fortified product was over-spiked at 50% and 100%. The biotin results in both SRM 1849a and IRM were close to 100% of the respective target values confirming the accuracy of the method (Table 2). All the biotin spiked recoveries were within 95% to 105% of added value confirming accuracy of the method (Table 3). The recoveries well exceeds the SMPR 2014.005 requirements of 90% to 110% in all instances.

Table 2: NIST SRM 1849a and IRM Reference Materials Limits

Biotin (µg/100g)

Day 1 199.97

Day 2 199.65

Day 3 196.50

Day 4

Day 5 Day 6

NIST SRM 1849a

199 ± 13

197.87 198.37 198.42

IRM

26.55 ± 7.7

26.12

27.58

25.36

25.61

25.13

25.39

Table 3: Spiked recovery

Recovery (%)

Spiked Levels

Placebo

Day 1

Day 2

Day 3

Average

50%

99.18 99.40 99.42 94.91 97.98 95.41 97.15 94.87

99.66 98.89 99.94 94.38 98.56 95.25 97.73 95.13

101.00 100.95 99.39 99.72

99.98 98.71 99.63 94.81 98.23 95.40 97.32 95.20

Child Formula Powder

150%

99.28

98.05 98.90 95.33 97.58 94.61 97.50 95.86

99.97 96.65 99.91 99.14 94.47 94.47 99.21 97.85 97.35 94.66 97.18 97.85 95.06 94.77

50%

100.44

Infant Elemental Powder

150%

95.28 98.17 95.11 96.51 95.51

50%

Adult Nutritional RTF, High Fat

150%

50%

Infant Formula RTF, Milk Based

150%

*Infant Formula RTF, Milk Based

50%

97.49

98.07

93.52

94.50

97.51 98.19

96.55

Spiked Levels

Sample

Day 1

Day 2

Day 3

Average

10 3.0 0 96. 25

103. 98 97.6 5

50%

103.50

105.21

103.42 103. 01

103.69

**Adult Nutritional RTF High Protein

95.2 4

100% 96.73 * Calibration forced through origin as the 50% levels were lower than the lowest calibration. **Placebo for Adult Nutritional RTF, High Protein Lot#/ID 00415RF00 could not be used for spiked recovery studies due to the bacterial fermentation of milk. Instead the fully fortified sample of the same matrix Lot#/ID00414RF00 was used for over spiking. 96.95 97.04 97.24

5.6 Precision (Repeatability)

Repeatability was determined by analysing the 12 different matrices in the SLV test materials kit in duplicate on each of the 6 days by two different analysts. In addition two different liquid chromatography equipment were utilized during the course of validation. Precision data for all the samples can be found in Table 4. Coefficient of variation (%RSD) was calculated on all the 12 data points on all the 12 SLV matrices. The %RSD was not more than 3.2% and is well below the set limit of 6% as per SMPR 2014.005.

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