# Impact of reduced pancreatin and bile on Fe and Zn bioaccessibility assessment using the INFOGEST in vitro digestion method with stable isotopic labelling

**Authors:** Alexandre Minami Fioroto, Molly Muleya, Lolita Wilson, Kaja Kristensen, Ruth Price, David A. Gray, Eduardo Purgatto, Elizabeth H. Bailey

PMC · DOI: 10.1016/j.fochx.2026.103483 · 2026-01-02

## TL;DR

This study examines how reducing pancreatin and bile in digestion experiments affects the measurement of iron and zinc availability in different foods.

## Contribution

The study reveals that reducing pancreatin and bile impacts mineral solubility and digestion, highlighting the need for standardized methods in mineral bioaccessibility assessments.

## Key findings

- Fe solubility increased in cereals, beans, and nuts but decreased in sweet potato.
- Zn solubility increased in oats, nuts, and leafy vegetables but decreased in potatoes and beans.
- Standard INFOGEST conditions generally showed higher Fe and Zn solubility.

## Abstract

In a previously proposed stable isotope approach based on the INFOGEST protocol, pancreatin and bile amounts were reduced due to their high mineral content, which interferes with Fe and Zn measurements. The present study examined the impact of reagent reduction on macronutrient digestibility and, consequently, mineral release in ten food samples. Macronutrient digestibility was higher under INFOGEST conditions. However, Fe and Zn solubility were not consistently modified across matrices, although, a general trend of increased solubility was observed in the standard INFOGEST method. Fe solubility increased by 64–91 % in cereals, beans, and nuts but decreased by 48 % in sweet potato. Similarly, Zn solubility increased by 11–75 % in oats, nuts, and leafy vegetables, while it decreased by 18–64 % in potatoes and beans. Overall, the influence of enzymatic action and background minerals could not be isolated, thus standardisation of the INFOGEST method for mineral bioaccessibility assessment is urgently needed.

•High mineral background in fluids interferes with Fe and Zn bioaccessibility measurements.•Reducing the amounts of pancreatin and bile in the fluids can impair protein and starch hydrolysis.•Fe and Zn solubility varied across food matrices and in vitro digestion methods.•Fe and Zn solubility tends to be higher under standard INFOGEST conditions.

High mineral background in fluids interferes with Fe and Zn bioaccessibility measurements.

Reducing the amounts of pancreatin and bile in the fluids can impair protein and starch hydrolysis.

Fe and Zn solubility varied across food matrices and in vitro digestion methods.

Fe and Zn solubility tends to be higher under standard INFOGEST conditions.

## Linked entities

- **Chemicals:** Fe (PubChem CID 23925), Zn (PubChem CID 23994)

## Full-text entities

- **Chemicals:** Zn (MESH:D015032), Fe (MESH:D007501)
- **Species:** Solanum tuberosum (potatoes, species) [taxon 4113]

## Figures

5 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12814079/full.md

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Source: https://tomesphere.com/paper/PMC12814079