AOAC SPIFAN Nutrients ERP-Final Review (Jan. 2019)

2015.10 (Jan. 2019) Carn-07 MLT Report FOR ERP USE ONLY DO NOT DISTRIBUTE

choline were determined from a common “free” or “total” extract, and the data is presented in this manner. Individual laboratory results for blind duplicate analysis of each sample are presented in Tables 1.1 and 1.2 for free carnitine and choline analysis, respectively. No outliers were removed for free carnitine statistical analysis within the SMPR concentration requirements for carnitine. Two samples were reported as below the method LOQ for free carnitine analysis. No outliers were identified within the SMPR concentration requirements for free choline analysis. Three samples for free choline from one laboratory were determined to be statistical outliers for free choline analysis, but all samples with outlier results were less than the SMPR LOQ requirement of 2.00 mg/100g. An investigation into these sample results from the one laboratory did not identify any issue with the analyses. Statistical analysis of the free carnitine and choline analysis is presented in Tables 1.3 and 1.4. Repeatibility (RSD r ) for free carnitine analysis was 1.6-3.9% against an SMPR requirement of 8%. For free carnitine analysis, reproducibility (RSD R ) was 2.6-6.3%, versus a target of ≤15%. HorRat ratios for free carnitine analysis in all matrices were all <1, indicating good performance against predicted RSD R . All blinded SRM values were within the certified range for free carnitine. For free choline analysis of samples >2.00mg/100g “as fed”, repeatability was 1.7-3.7% across these samples. The range of RSD R for these samples was 4.4-9.1% across these same samples, versus a target RSD R of 10-15% for total choline, depending on level. For free choline analysis of samples <2.00 mg/100g and outside of the SMPR scope, the repeatability was 0.8-3.0%. Reproducibility of these 3 same samples was 5.7-10.6% with outliers excluded. HorRat ratios for free choline analysis ranged from 0.49 to 1.44. Samples with free choline results under 2.00 mg/100g also exhibited good performance against the predicted RSD R values, showing good performance in this concentration range but with one laboratory having outlier results. Overall, the free method extraction showed good reproducibility for carnitine and choline in all matrices tested. All analysis results met the SMPR requirements as listed in those documents for free carnitine and choline analysis. Individual laboratory results are presented in Tables 1.5 and 1.6 for total carnitine and choline analysis, respectively. Statistics for this portion of the analysis are presented in Tables 1.7 - 1.8. Two outlier values were identified for total choline analysis. The outlier results were excluded from the statistical analysis. For total carnitine analysis, RSD r ranged from 1.8-3.9% across the 8 samples with detectable levels of carnitine. 2 samples had results below the method LOQ. RSD R for total carnitine in these same samples was 3.7-5.8%, easily meeting the SMPR requirement of 15%. HorRat values were 0.35 -0.74 for the samples with detectable amounts of total carnitine, indicating good reproducibility compared to predicted RSD R . Total choline analysis gave RSD r values ranging from 1.3-3.0%. RSD R values ranged from 5.2%-11.4%. Results obtained for choline content of the samples were higher than the free choline content, as expected. HorRat values for total choline analysis ranged from 0.82-1.84.

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