Solubility and Impact of Ascorbic Acid on the in vitro Bioavailability of Two Casein-Based Iron Fortificants from Reconstituted Milk

Abstract Objectives The two objectives were 1) to evaluate the solubility of two iron casein complexes (ICCs) under a condition mimicking gastric pH, 2) to evaluate the impact of ascorbic acid (AA) on the in vitro iron absorption of ICCs after incorporation in reconstituted whole milk powder. Methods The in vitro solubility was determined over time after addition of diluted HCl (pH 1.7), ultracentrifugation and measurement of iron appearing in the supernatant by ICP-OES (n = 2). The impact of AA on iron uptake from the Fe compounds in reconstituted milk was determined using the in vitro digestion coupled with the Caco-2 cell model and the measurement of ferritin/total protein produced by the cells (n = 3). The molar ratio of AA to iron of 2 to 1 recommended by the WHO for iron absorption optimization has been tested with an iron level corresponding to 3.3 mg Fe/serving of milk. Ferrous sulfate (FeSO4), the reference compound for iron bioavailability and micronized ferric pyrophosphate (FePP), main salt used for milk fortification were used as references. Results The dissolution test showed a rapid solubilization of iron from the ICCs i.e., >75 ± 19.3% at 5 min and >89 ± 0.3% at 90 min. The kinetics of soluble iron from the complexes were like that from FeSO4. The solubility of FePP was only 37.6 ± 4.7% at 90 min. Without AA, the iron uptake from FeSO4 was lower than expected translating into a relative in vitro bioavailability (iRBA) of FePP and of the two ICCs to FeSO4 of 66, 169 and 215%. This might be explained by a rapid conversion of soluble iron from FeSO4 into Fe3+ and insoluble iron hydroxide when the pH increased from 2 to >7 during in vitro digestion. However, with the addition of AA in the milk, iron uptake by the cells was found to be increased to levels of 341.8 ± 8.9, 124 ± 12.2, 403.1 ± 117.8 and 362.9 ± 36.9 ng ferritin/mg protein for FeSO4, FePP and the two ICCs respectively. This translates into iRBAs to FeSO4 of 36% for FePP and of 118 and 106% for the two ICCs. Conclusions The solubility and the demonstrated impact of AA on Fe uptake suggest that ICCs are absorbed to a similar amount as FeSO4 and thus provide an excellent source of Fe. Funding Sources Société des Produits Nestlé, NPTC Konolfingen, Switzerland.