Improved AOAC First Action 2011.08 for the Analysis of Vitamin B₁₂ in Infant Formula and Adult/Pediatric Formulas: First Action 2014.02.

This report documents improvement and single-laboratory validation performed on AOAC First Action Method 2011.08 for vitamin B12 in infant formula and adult/pediatric nutritional formula. The original validation study included a range of fortified products, from infant formulas to breakfast cereals or beverages. Extended validation data, including additional infant formulas and adult/pediatric nutritionals, has now been produced. In addition, the method has been modified to use ultra-HPLC and the calibration range extended in a multilevel calibration curve. Detection and quantification limits were also improved by increasing the sample weight used for analysis and the reconstitution rate adapted to the requirements. The Stakeholder Panel on Infant Formula and Adult Nutritionals Test Material Kit, designed to represent a large range of products within the category (infant formula and adult nutritionals made from any combination of milk, soy, rice, whey, hydrolyzed protein, starch, and amino acids, with and without intact protein), was used to determine performance characteristics of the method. The modifications included allow now full compliance with standard method performance requirements established for vitamin B12 (SMPR 2011.005). LOQ was ≤0.01 μg/100 g, working range between 0.01 and 5.0 μg/100 g, repeatability ≤7%, and recovery in the range 90-110%. The method was granted AOAC First Action status 2014.02.