Evaluation of Different RP-HPLC Methods for Lactoferrin Quantification in Goat Milk
Keywords:
Goat milk, heparin, lactoferrin quantification, resin size, sample preparationAbstract
Objective: This study aimed to develop and validate a reliable method for the quantification of lactoferrin in goat milk. It also addressed the challenges associated with applying standard heparin-based methods, originally developed for cow’s milk, to goat milk due to differences in fat and casein composition.
Materials and Methods: Different Reverse-phase High-performance Liquid Chromatography (RP-HPLC) methods for lactoferrin quantification were evaluated and validated. Goat milk samples were prepared using an optimized protocol that included two rounds of centrifugation at 4000 rpm, followed by filtration and high-speed centrifugation at 10,000 rpm. Defatting was identified as a critical step in sample preparation. In addition, the study investigated the use of cation-exchange resin with larger bead size as an alternative to conventional heparin-based beads for lactoferrin isolation.
Results: The optimized sample preparation method improved the clarity and suitability of goat milk samples for analysis. The use of larger cation-exchange resin beads facilitated better sample flow through the column and prevented clogging, which is a common issue with heparin-based columns. The validated RP-HPLC method showed good accuracy and robustness for lactoferrin quantification in goat milk.
Conclusion: A robust and accurate method for lactoferrin determination in goat milk was successfully developed. The combination of optimized sample preparation and the use of larger cation-exchange resin beads improve analytical performance and overcomes limitations of conventional heparin-based methods. This approach provides a reliable alternative for industrial and research applications involving goat milk.
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