Quantification of lactoferrin in dietary supplements using a Heparin affinity clean-up column by HPLC PDA detector

Abstract

The use of dietary supplements containing lactoferrin is a growing trend. As a result, there is a need for a simple and reliable analytical method to determine the content of lactoferrin in these supplements to ensure food quality. In this study, an HPLC method was developed and validated for determining lactoferrin in dietary supplements using a HiTrap Heparin clean-up column in combination with HPLC. The study involved selecting optimal conditions for extracting lactoferrin from the sample matrix, followed by chromatography conditions using a C4 column (150 mm × 4.6 mm, 3.5 µm). The limit of detection (LOD) and limit of quantification (LOQ) were determined, with the achieved values being 1.2 mg/100g and 4.0 mg/100g, respectively. The method has shown good recovery (99.5–103.4%) and precision (RSD 1.76–3.44%), meeting the method performance requirements outlined in AOAC SMPR 2020.005. The validated method was applied to determine the contents of lactoferrin in dietary supplement samples.