Determination of Bovine Lactoferrin in Food by HPLC with a Heparin Affinity Column for Sample Preparation

An HPLC method was developed for the quantitative determination of bovine lactoferrin (bLF) in sterilized milk, modified milk, fermented milk, infant formula, adult formula, rice cereal, vitamin function drink, and protein powder products. bLF was first extracted with a phosphate buffer (pH 8), underwent cleanup in a heparin affinity column, and was detected by HPLC with a C4 column and diode-array detector at a wavelength of 280 nm. The proposed method provided a linear detection range of 10.0-1000 μg/mL with an LOD of 0.6 mg/100 g in liquid samples and 3 mg/100 g in solid samples and an LOQ of 2 mg/100 g in liquid samples and 10 mg/100 g in solid samples. In addition, the method showed good recovery for various samples, ranging from 76 to 96%. The method had several remarkable advantages, including ease of handling, high sensitivity and accuracy, good reproducibility, and low-cost detection. Based on the distinctive properties presented here, we believe the proposed HPLC assay holds great promise for the oversight and detection of bLF in testing organizations, dairy enterprises, and regulatory authorities.