Development of a standardised human in vitro digestion protocol based on macronutrient digestion using response...

Area of Science:

• Food Science
• Gastroenterology
• Toxicology

Background:

• Bioaccessibility studies are crucial for assessing physiological effects of ingested compounds.
• Existing in vitro digestion protocols lack optimization, leading to variable results.
• A standardized, optimized protocol is needed for reliable bioaccessibility assessments.

Purpose of the Study:

• To optimize an in vitro digestion protocol for simulating human digestion.
• To evaluate the influence of pH, incubation time, and enzyme concentrations on macronutrient digestion.
• To establish a validated in vitro digestion model for pre-colonic steps.

Main Methods:

• Response surface methodology with central composite designs was used to model digestion parameters.
• The study investigated the digestion of starch, albumin, and triolein under varying conditions.
• Optimization focused on salivary, gastric, and abiotic duodenal digestion steps.

Main Results:

• An optimized in vitro digestion protocol was successfully established.
• Specific conditions for salivary (pH 6.9, 5 min, 3.9 U α-amylase/mL), gastric (pH 2, 90 min, 71.2 U pepsin/mL), and duodenal (pH 7, 150 min, 9.2 mg pancreatin, 55.2 mg bile extract/mL) steps were determined.
• The model effectively simulates macronutrient digestion.

Conclusions:

• The developed in vitro digestion protocol provides a validated method for bioaccessibility studies.
• This optimized model can enhance the evaluation of physiological effects of ingested substances.
• The protocol offers a reliable tool for research in food science and toxicology.