Our aim was to develop an accurate, precise, and reliable RP-HPLC method for estimating ferrous fumarate and ascorbic acid in pro-liposomes. The aim was to make the analytical method simple, environmentally friendly, and accurate. To measure ferrous fumarate (FF) and ascorbic acid (AA), an isocratic method was used in an HPLC, with 220 and 245 nm detection, using phosphate buffer (pH 3.0) and methanol (95:5). It was found that AA and FF were retained for 6.4 and 8.3 min, respectively. A linear relationship was observed between 0.1 and 50 μg/mL for both ferrous fumarate (r [2] = 0.994) and ascorbic acid (r [2] = 0.999). The resolution between the two peaks ≥2, tailing factor (TF) ≤2, and theoretical plate count >2000 were evaluated and recorded as system suitability parameters. The LOD and LOQ for FF were calculated to be 0.52 and 1.58 μg/mL, respectively, whereas the same for AA were calculated to be 0.17 and 0.53 μg/mL, respectively. The validation method was tested under various conditions, including flow rate, injection volume, column temperature, and mobile phase ratio. Additionally, the optimized method was successfully used to estimate the entrapment efficiency for FF and AA in the developed pro-liposomes, which were found to be 92 and 65%, respectively.

The global prevalence of iron (Fe) deficiency anemia (IDA) continues to pose major public health challenges, necessitating the development of effective fortification strategies. Dairy-based systems are promising vehicles for Fe fortification and delivery due to their widespread consumption and nutritional value, although their complex matrix presents significant challenges for maintaining Fe bioavailability and product quality. This review critically evaluates current knowledge on Fe fortification in dairy systems, integrating evidence on Fe forms, physicochemical interactions between Fe and the matrix, and their consequent effects on Fe bioavailability and physical functionality. Emerging fortification strategies are highlighted. Conventional Fe salts, including ferrous sulfate, ferrous fumarate, and ferric pyrophosphate, have varying degrees of solubility and absorption but are often associated with adverse sensory changes, lipid oxidation, and reduced product stability. Fe-ligand complexes, including protein-, carbohydrate-, and Maillard reaction product (MRP)-based systems, can impart improved stability to the fortified product and bioavailability by minimizing interactions with inhibitors of Fe absorption. The incorporation of Fe into dairy matrices influences product pH and structural and functional properties while also accelerating lipid oxidation and sensory deterioration when inorganic salts are used. Strategies such as Fe chelation, microencapsulation, and co-fortification with enhancers have shown potential to mitigate these limitations. In addition, the role of prebiotics, probiotics, and MRPs in modulating Fe absorption remains complex and context dependent. Despite promising advancements, gaps remain in understanding interactions between Fe, proteins, carbohydrates and MRP and their implications for Fe fortification of dairy systems. Future research should focus on developing integrated fortification approaches enabling the design of next-generation functional dairy foods to effectively combat global Fe deficiency.

Iron deficiency remains a global nutritional challenge, requiring stable and efficient delivery systems for fortification. This study aimed to develop a novel carrier by encapsulating ferrous fumarate-loaded niosomes within a hydrogel matrix composed of agar and chia seed gum (CSG). The niosomes exhibited a uniformity (PDI < 0.22) and negative surface charge (<-23 mV), ensuring electrostatic stability. FTIR indicated that the characteristic peak of ferrous was not present in the spectrum of hybrid systems. XRD showed that successful transition of crystalline iron into an amorphous state within the carriers. Rheological assessment revealed that the agar enhanced the viscoelastic moduli (G' > G'') and mechanical strength. FESEM demonstrated non-porous surface morphology in the dual-hydrogel system, which improved thermal stability (Tm > 300 °C). Release studies highlighted that while CSG were susceptible to rapid destabilization, the dual-hydrogel (≅42.6%) matrix provided superior protection. Overall, the agar-CSG-niosome composite represents a robust platform for the fortification of functional foods.

Iron is a normal component in human milk. Daily oral iron intake from prenatal vitamins or other multimineral supplements does not affect milk iron levels. Higher daily oral iron dosing has a minimal effect on milk iron levels and is not expected to cause harm to the breastfed infant if needed to treat the mother’s anemia, but it is not an adequate substitute for direct infant iron supplementation to prevent or treat infant anemia.

Background: Oral iron supplementation or food fortification is essential for managing or preventing iron deficiency but often causes gastrointestinal side effects. While solubility has traditionally been considered a requirement for iron uptake via the DMT1 transporter, recent evidence shows that insoluble iron can also be absorbed through endocytosis, raising questions about particle size and epithelial responses. Methods: Human intestinal cell lines (Hutu-80 and Caco-2) were exposed to physiologically relevant but elevated iron levels (0.5 mM Fe, 48 h) as ferric pyrophosphate, ferrous fumarate (both prone to precipitation), and soluble ferric EDTA. Cell survival and COX-2 protein were quantified by ELISA, solubility by ICP-OES, and particle size in cell culture medium by dynamic light scattering analyses. Results: Ferric pyrophosphate (0.62-3.8 μm) markedly increased COX-2 expression in Hutu-80 cells (254% ± 37%, n = 3, p = 4.11 × 10-5) and in Caco-2 cells (78% ± 8%, n = 3, p = 0.01) compared to the control. Ferrous fumarate (237-866 nm) also induced COX-2, but only in Hutu-80 cells (62% ± 11%, n = 3, p = 0.04), whereas ferric EDTA showed no effect in either cell line. COX-2 induction was associated with larger particles in the medium (≥237 nm), whereas smaller particles (<146 nm) were not. Conclusions: Particle size appears to be a critical determinant of cell survival and iron-induced epithelial COX-2 expression. Iron compounds that present as both soluble and particulate forms may optimize bioavailability, but controlling aggregate size (<146 nm) could reduce inflammatory signaling. These findings may have important implications for cell culture systems and warrant in vivo validation in iron supplemental studies.