Evaluation of Lactoferrin Transport In Vitro Using the Everted Intestinal Sac Method and Its In Vivo Effects on Growth and Liver Color (L*) in Rats

Faculty of Agricultural Technology, University of 17 Agustus 1945 Semarang, Semarang, Indonesia

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Abstract

This study aimed to evaluate the absorption and transport of lactoferrin in vitro using the everted intestinal sac method and to determine its in vivo effects on growth performance and liver color (L*) in rats. The research material included bovine lactoferrin, FeSO₄·7H₂O, 0.9% NaCl solution and male albino rats (Rattus norvegicus) aged 4-5 months with body weights ranging from 200–350 g (in vitro) and male or female rats 7 day aged with body weights 10-15 g (in vivo) among other materials. The in vitro experiment demonstrated that lactoferrin absorption increased and reached a plateau at 60–70 mg/80 mL of 0.9% NaCl solution (14.53-14.92%), declined at higher concentrations, and was highest in the duodenal segment (15.38%), followed by the jejunum (11.59%) and ileum (6.89%). Time-course analysis revealed a progressive increase in absorption up to 75 minutes (17.66%) followed by a slight decline, indicating an active and saturable absorption mechanism. In the in vivo, rats receiving a combination of FeSO₄·7H₂O and lactoferrin showed the greatest body weight gain (1.61 g) and darker liver coloration (L* = 34.41), reflecting enhanced iron utilization and storage. These findings indicate that lactoferrin enhances intestinal iron absorption and improves physiological responses when combined with FeSO₄·7H₂O. The integration of in vitro and in vivo results highlights lactoferrin’s potential as a functional bioactive protein for improving iron bioavailability and promoting growth.