Analysis of Six Human Milk Oligosaccharides (HMO) in Infant Formula and Adult Nutritionals by 2AB Labeling and Quantification with HILIC-FLD: First Action 2022.02.

Abstract

Human milk oligosaccharides (HMO) function as a prebiotic, enhance immune functions, and support brain development for infants when fed mother's milk. These are added to infant formula and adult nutritionals in order provide these same benefits. To develop and validate a method that can meet the AOAC Standard Method Performance Requirements (SMPR®) outlined by the AOAC INTERNATIONAL Stakeholder Panel for Infant Formula and Adult Nutritionals (SPIFAN) through a single-laboratory validation (SLV). This work describes a method that can analyze six different HMOs that include 2'-fucosyllactose, 3-fucosyllactose, 3'-sialyllactose, 6'-sialyllactose, lacto-N-tetraose, and lacto-N-neotetraose. The method utilizes a derivatization procedure that labels the HMO with the fluorescent compound 2-aminobenzamide. The method was optimized to provide a non-toxic derivatization procedure, automate the removal of excess derivatization reagent, and provide a chromatographic separation that can analyze multiple HMOs in a single profile. A summary from the SLV is provided. The SLV was reviewed by the AOAC SPIFAN Expert Review Panel, and determined the method met the SMPR requirements for six HMO. The method was granted First Action Official MethodsSM status.