Quality assessment of some nutraceuticals derived from milk / Hadeer Gamaleldin Mohamed ; Supervised Hala E. Zaazaa , Medhat A. Alghobashy , Mervat A. Fouad

Thesis (M.Sc.) - Cairo University - Faculty of Pharmacy - Department of Analytical Chemistry

In recent years, Lactoferrin has received great attention for its nutritional and immunological uses.Nowadays it has approved its great role as an antiviral agent in recent treatment protocols for COVID-19.As a minor element, its inclusion in milk with other milk proteins hindered its direct determination as it has a great ability to form a complex with some milk proteins e.g. Caseins. So downstream purification of Lactoferrin from skimmed milk was optimized using a vacuum-driven approach in the presence of phosphate buffer at pH 6.5 relative to the isoelectric point of Lactoferrin (pI 7.8-8.7) using stepwise increase of salt concentration from 0.5M to 1M.The purity of Lactoferrin (obtained from purified fraction after downstream system)was monitored using gel electrophoresis along with validated RP-HPLC and SE-HPLC assays. On the other hand, total protein content was determined using ordinary spectrophotometric assays as direct assay at 214nm and 280nm and Bradford assay at 595nm.SE-HPLC and RP-HPLC were also employed to study factors affecting Lactoferrin during milk processing, handling, and storage which shows its greatly unstable to different factors e.g. temperature, oxidation, acidic and alkaline conditions, and repeated freeze and thaw.These methods were used for investigation for the presence of Lactoferrin in variousnutraceutical products in the Egyptian market which assessed to withdraw our attention towards careful monitoring to track concentration, purity and stability of Lactoferrin during in-process quality control which showed to be affected by many deleterious reactions during processing causing significant denaturation of the milk proteins

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