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5841 v2
Page 8 of 8
AsureQuality Laboratory Services
Table 4: Repeatability
Average
CLC10-b
197.42 201.01 196.08 196.28 197.93 201.54 198.54 198.39 199.34 198.81 197.00 197.30 198.30 0.86
11750017V3 12.77 12.73 12.67 12.77 12.69 12.69 12.53 12.73 12.61 12.62 12.65 12.70 12.68 0.55
1172572116 34.68 34.50 34.20 35.24 35.17 35.13 36.05 35.09 36.19 36.97 35.19 35.02 35.29 2.19
117257651Z 39.76 39.99 39.98 39.16 40.12 40.50 39.37 40.77 40.87 40.96 39.50 40.70 40.14 1.54
00394RF00 248.67 249.85 254.88 237.61 258.75 261.19 256.65 259.83 255.74 259.20 254.28 256.30 254.41 2.55
00412RF00 91.08 90.66 88.99 89.01 89.70 91.71 89.92 90.81 89.56 90.20 90.69 90.72 90.25 0.93
00403RF00 69.17 69.30 70.41 69.88 71.88 72.28 71.35 71.45 72.14 71.83 70.52 71.03 70.94 1.52
D04HTCVV 35.48 35.24 34.23 33.92 33.36 34.28 33.01 33.17 32.94 32.96 31.71 32.94 33.60 3.19
E29JVLV
43.30 43.25 45.40 44.72 45.02 44.38 45.29 44.85 45.15 44.78 44.76 45.08 44.67 1.58
Control Milk F 5.45 5.47 5.36 5.34 5.40 5.44 5.38 5.43 5.20 5.31 5.15 5.37 5.36 1.83
00414RF00 54.02 55.42 55.50 55.16 52.36 52.07 53.67 53.78 53.23 53.91 55.48 54.21 54.07 2.16
00406RF00 68.88 65.17 63.77 64.85 66.78 67.48 65.51 66.54 65.64 65.36 63.92 62.13 65.50 2.74
SRM 1849a
198.46 0.63
Biotin (µg/100g)
RSD
Day 1
Day 2
Day 3
Day 4
Day 5
Day 6
199.97
199.65
196.50
197.87
198.37
198.42
SPIFAN SLV
Samples
6. SUMMARY AND CONCLUSION
AsureQuality Auckland Laboratory has initiated a method to facilitate a specific, precise,
accurate and robust procedure for the analysis of biotin from Infant Formula and Adult /
Pediatric Nutritional Formulas.
The method involves immunoaffinity column (R-Biopharm Rhone, EASI-EXTRACT biotin
column) clean-up and extraction followed by liquid chromatography with UV detection.
The proposed method exceeds all the SPIFAN SMPR for biotin. The method also has an
assured limit of quantification of 0.1µg/100g (1ppb) based on a simple mathematical
relationship between lowest standard and dilution. This method is found to be suitable for
routine testing of biotin in infant formula and adult nutritionals. Comparison of SMPR for
biotin and the proposed method performance are given in Table 5.
Table 5: Performance of the proposed method (Bio -02) compared to SMPR 2014.005
Parameters
SMPR 2014.005
Proposed Method
Comments
Analytical Range
0.1 - 150 µg/100g
0.1 - 300 µg/100g
Analytical range of the proposed method is
wider based on the sample weight taken
and loading volume for sample extraction.
Limit of Quantitation
≤
0.1 µg/100g
≤
0.1 µg/100g
The proposed method meets the LOQ of
0.1 µg/100g as required by AOAC SMPR
2014.005.
Repeatability (RSD
r
)
>1 µg/100g:
≤
6%
>1 µg/100g:
≤
4%
Repeatability of the proposed method is
better than the SMPR 2014.005.
Recovery
(>1µg/100g)
90 to 110%
95 to 105%
All the spiked recoveries by the proposed
method are better than the recovery range
specified by AOAC SMPR 2014.005.
NIST SRM 1849a
199 ± 13
198 ± 3
NIST 1949a results are at the certified
value.
Bio-02 w/SLV
FOR ERP USE ONLY
DO NOT DISTRIBUTE