Adaption and Optimization of the Ferric-nitrilotriacetate Spectrophotometric Method for Quantification of Lactoferrin in Raw Milk
Keywords:Lactoferrine, optimising, spectrophotometric, isolation, estimation
AbstractThis study aimed at adapting and optimising the Fe3+-nitrilotriacetate (Fe3+-NTA) spectrophotometric method for determining lactoferrin (LF) content in raw milk and milk fractions. Isolation of LF from acid whey was achieved by using cellulose phosphate resin. LF was quantified from the standard curve based on spectrophotometric reading at 465nm. The purity of LF isolates was established based on lactoperoxidase (LP) activity. The adopted method indicated under estimation of LF content. However, this was modified by incorporation of NaHCO3 and Fe3+-NTA in both the blank for the standard and test samples, which improved the performance of the method. Results generally indicated that modified Fe3+-nitrilotriacetate (Fe3+-NTA) method is a simple, quick and reproducible method that can give a better quantitative estimation of LF. A batch procedure using cellulose phosphate resin was highly effective for the isolation and purification of LF as demonstrated with less contamination of LP.