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IMPROVEMENTS MADE TO DERIVATIZATION  PROCEDURE • A protein precipitation step with acetonitrile was included. • Reagents were prepared according to Miwa et al. (2000), J. Chrom. A 881 365 which reduces the number of steps involved in the , , derivatization process. Ibid. for reaction conditions (80 °C, 5 min) which reduces overall sample processing time. • Excess reagent was not removed. There was no indication that the presence of excess reagent had any adverse effect on the analysis or the instrumentation. • One single reversed phase SPE step was used for solvent exchange and 5-fold concentration of extract. • Tandem mass spectrometry (MS/MS) was used which adds another level of selectivity compared to single stage MS. • Stable isotope labeled internal standard was used for accurate quantitation.

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CONCLUSIONS

Direct Analysis Method • A simple and high throughput LC-MS method has been developed for nutritional products analysis of MFA • Specificity and quantitation in matrix demonstrated • 30 ppb quantitation limit in powders • Implemented for routine analysis at a third part laboratory for samples analysis Derivatization Method • A streamlined derivatization method has been developed for nutritional products analysis of MFA

• Specificity and quantitation in matrix demonstrated • 10 ppb quantitation limit in powders • Confirmatory method

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