Application of ITC in foods: A powerful tool for understanding the gastrointestinal fate of lipophilic compounds

doi:10.1016/j.bbagen.2015.10.001

Biochimica et Biophysica Acta (BBA) - General Subjects

Volume 1860, Issue 5, May 2016, Pages 1026-1035

https://doi.org/10.1016/j.bbagen.2015.10.001 Get rights and content

Highlights

•
ITC is a powerful tool for characterizing interactions involving food components.
•
Food components under complex interactions within the gastrointestinal tract.
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ITC can characterize interactions such as binding, hydrolysis, and micelle formation.
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ITC may provide useful qualitative or quantitative information.

Abstract

Background

Isothermal titration calorimetry (ITC) is a biophysical technique widely used to study molecular interactions in biological and non-biological systems. It can provide important information about molecular interactions (such as binding constant, number of binding sites, free energy, enthalpy, and entropy) simply by measuring the heat absorbed or released during an interaction between two liquid solutions.

Scope of the review

In this review, we present an overview of ITC applications in food science, with particular focus on understanding the fate of lipids within the human gastrointestinal tract. In this area, ITC can be used to study micellization of bile salts, inclusion complex formation, the interaction of surface-active molecules with proteins, carbohydrates and lipids, and the interactions of lipid droplets.

Major conclusions

ITC is an extremely powerful tool for measuring molecular interactions in food systems, and can provide valuable information about many types of interactions involving food components such as proteins, carbohydrates, lipids, surfactants, and minerals. For systems at equilibrium, ITC can provide fundamental thermodynamic parameters that can be used to establish the physiochemical origin of molecular interactions.

General significance

It is expected that ITC will continue to be utilized as a means of providing fundamental information about complex materials such as those found in foods. This knowledge may be used to create functional foods designed to behave in the gastrointestinal tract in a manner that will improve human health and well-being. This article is part of a Special Issue entitled Microcalorimetry in the BioSciences — Principles and Applications, edited by Fadi Bou-Abdallah.

Introduction

Isothermal titration calorimetry (ITC) is an extremely powerful tool that can be used to provide valuable information in many areas of food science. In this article, we focus on a particular application of ITC to highlight its great potential: the gastrointestinal fate of ingested lipids. Understanding the behavior of lipids within the gastrointestinal tract (GIT) has recently become an important area of research within the food industry due to the focus on designing foods to increase human health and wellness. For example, functional foods are being designed to improve the bioavailability profile or control the delivery of hydrophobic nutraceuticals, nutrients, and vitamins [1], [2], [3]. A number of bioactive hydrophobic components that might be incorporated into food and beverages are listed in Table 1.

The gastrointestinal fate of food lipids within the human body is a complex process that depends on many factors, including the nature of the delivery system used, food matrix effects, and the concentration and nature of the lipids [4]. A number of physicochemical and physiological processes influence the absorption of dietary lipids. Triacylglycerols (TAGs) undergo digestion within the stomach and small intestine due to the presence of gastric and pancreatic lipases, which leads to the formation of free fatty acids (FFAs) and monoacylglycerols (MAGs). These surface-active lipid digestion products interact with endogenous surface-active lipids (bile salts and phospholipids) to form micelles, vesicles, and other colloidal structures [5]. These “mixed micelles” are capable of solubilizing lipophilic bioactive components and transporting them to the epithelium cells where they are absorbed. Lipid digestion is therefore a critical step in ensuring good absorption of ingested lipophilic bioactives [6]. The GIT fate of lipids is influenced by their interaction with other species within the food matrix or within the gastrointestinal fluids, and therefore it is important to understand the nature of these interactions and their impact on lipid digestion and absorption [7].

Section snippets

Lipid bioavailability

Oral bioavailability can be defined as the fraction of an ingested component (or its metabolic products) that eventually ends up in the systemic circulation [8], [9]. For lipophilic components, the overall bioavailability (F) can be defined as [9]: ${F = F}_{B} {\times F}_{A} {\times F}_{T} .$

Here, F_B is the bioaccessibility coefficient or the fraction of lipophilic components released from the food matrix into the GIT fluids; F_A is the absorption coefficient or the fraction of bioaccessible lipophilic components absorbed by the

Characterizing interactions amongst food components using ITC

The gastrointestinal fate of lipids is influenced by their interactions with various kinds of molecular species within the GIT. In this section, some of the applications of ITC for studying the interactions of both polar lipids (such as surfactants) and non-polar lipids (such as TAGs) with molecular species that may be found in the human GIT are discussed.

Conclusions/summary

ITC is an extremely powerful tool for characterizing molecular interactions in food systems, and can provide valuable information about many types of physicochemical phenomena involving food components such as proteins, carbohydrates, lipids, surfactants, and minerals. For systems at equilibrium, ITC can provide fundamental thermodynamic parameters (such as binding constants, number of binding sites, and free energy, enthalpy, and entropy changes) that can be used to establish the

Conflict of interest

The authors declare no competing financial interest.

Transparency document

Transparency document.

Acknowledgments

This material was partly based upon the work supported by the Cooperative State Research, Extension, Education Service, USDA, Massachusetts Agricultural Experiment Station (Project No. 831) and USDA, NRI Grants (2011-03539, 2013-03795, 2011-67021, and 2014-67021). I. A.-M. thanks the CONACYT, Mexico (Registration no. 208139) for financial support.

References (87)

D.J. McClements et al.
Physicochemical and structural aspects of lipid digestion
E. Fernandez-Garcia et al.
In vitro bioaccessibility assessment as a prediction tool of nutritional efficiency
Nutr. Res.
(2009)
B. Holst et al.
Nutrients and phytochemicals: from bioavailability to bioefficacy beyond antioxidants
Curr. Opin. Biotechnol.
(2008)
P. Reis et al.
Lipases at interfaces: a review
Adv. Colloid Interf. Sci.
(2009)
D.J. McClements et al.
Structured emulsion-based delivery systems: controlling the digestion and release of lipophilic food components
Adv. Colloid Interf. Sci.
(2010)
W.H. De Jong et al.
Particle size-dependent organ distribution of gold nanoparticles after intravenous administration
Biomaterials
(2008)
C. Aguirre et al.
Isothermal titration calorimetry determination of individual rate constants of trypsin catalytic activity
Anal. Biochem.
(2015)
K. Maximova et al.
Kinetics of trypsin-catalyzed hydrolysis determined by isothermal titration calorimetry
Anal. Biochem.
(2015)
M.K. Transtrum et al.
Enzyme kinetics determined by single-injection isothermal titration calorimetry
Methods
(2015)
P. Mukerjee
Micellar properties of drugs: micellar and nonmicellar patterns of self-association of hydrophobic solutes of different molecular structures — monomer fraction, availability, and misuses of micellar hypothesis
J. Pharm. Sci.
(1974)

L. Chiappisi et al.

An improved method for analyzing isothermal titration calorimetry data from oppositely charged surfactant polyelectrolyte mixtures

J. Chem. Thermodyn.

(2014)

S.D. Clas

Quaternized colestipol, an improved bile salt adsorbent: in vitro studies

J. Pharm. Sci.

(1991)

J.A. Monro et al.

Bile acid activity in the presence of dietary fibres, casein, calcium, phospholipid, fatty acid and cholesterol: factorial experiments in vitro

Food Chem.

(1992)

M. Thongngam et al.

Isothermal titration calorimetry study of the interactions between chitosan and a bile salt (sodium taurocholate)

Food Hydrocoll.

(2005)

C.G. Biliaderis et al.

Physicochemical properties of commercial starch hydrolyzates in the frozen state

Food Chem.

(1999)

M.G. Semenova et al.

Effect of maltodextrins on the surface activity of small molecule surfactants

Colloids Surf. B: Biointerfaces

(2003)

A. Wangsakan et al.

Isothermal titration calorimetry study of the influence of temperature, pH and salt on maltodextrin–anionic surfactant interactions

Food Hydrocoll.

(2006)

E.M.M. Del Valle

Cyclodextrins and their uses: a review

Process Biochem.

(2004)

S. Mun et al.

Study of inclusion complexes of cycloamylose with surfactants by isothermal titration calorimetry

Carbohydr. Polym.

(2009)

S.J. Hur et al.

In vitro human digestion models for food applications

Food Chem.

(2011)

Y. Li et al.

Modulating lipid droplet intestinal lipolysis by electrostatic complexation with anionic polysaccharides: influence of cosurfactants

Food Hydrocoll.

(2014)

O.K. Simo et al.

Novel strategies for fabricating reduced fat foods: heteroaggregation of lipid droplets with polysaccharides

Food Res. Int.

(2012)

A. Ye et al.

Effect of calcium on the kinetics of free fatty acid release during in vitro lipid digestion in model emulsions

Food Chem.

(2013)

A. Wangsakan et al.

Isothermal titration calorimetry study of the influence of temperature, pH and salt on maltodextrin–anionic surfactant interactions

Food Hydrocoll.

(2006)

D. Guzey et al.

Characterization of β-lactoglobulin–chitosan interactions in aqueous solutions: a calorimetry, light scattering, electrophoretic mobility and solubility study

Food Hydrocoll.

(2006)

Y. Chang et al.

Interaction of cationic antimicrobial (ɛ-polylysine) with food-grade biopolymers: dextran, chitosan, carrageenan, alginate, and pectin

Food Res. Int.

(2014)

D.J. McClements et al.

Structural design principles for delivery of bioactive components in nutraceuticals and functional foods

Crit. Rev. Food Sci. Nutr.

(2009)

D.J. McClements et al.

Emulsion-based delivery systems for lipophilic bioactive components

J. Food Sci.

(2007)

K.J. Yeum et al.

Carotenoid bioavailability and bioconversion

Annu. Rev. Nutr.

(2002)

G. Kossena et al.

Separation and characterization of the colloidal phases produced on digestion of common formulation lipids and assessment of their impact on the apparent solubility of selected poorly water-soluble drugs

J. Pharm. Sci.

(2002)

H.C.F.a.R.M. Clark

Intestinal absortion and tissue distribution of carotenoids.pdf

J. Nutr. Biochem.

(1997)

C. Versantvoort et al.

Development and applicability of an in vitro digestion model in assessing the bioaccessibility of contaminants from food.pdf

D.J. McClements et al.

Review of in vitro digestion models for rapid screening of emulsion-based systems

Food Funct.

(2010)

A.M.l. et al.

A dynamic in vitro lipolysis model 1 controlling the rate of lipopysis by continous addition of calcium.pdf

Eur. J. Pharm. Sci.

(2001)

P. Reis et al.

Lipases at interfaces: unique interfacial properties as globular proteins

Langmuir

(2008)

S. Mun et al.

Influence of interfacial composition on in vitro digestibility of emulsified lipids: potential mechanism for chitosan's ability to inhibit fat digestion

Food Biophys.

(2006)

G.A. van Aken

Modelling texture perception by soft epithelial surfaces

Soft Matter

(2010)

F. Kong et al.

Disintegration of solid foods in human stomach

J. Food Sci.

(2008)

M.J. Ferrua et al.

Modeling the fluid dynamics in a human stomach to gain insight of food digestion

J. Food Sci.

(2010)

E. Roger et al.

Biopharmaceutical parameters to consider in order to alter the fate of nanocarriers after oral delivery

Nanomedicine

(2010)

L.R. Johnson

Gastrointestinal physiology

(2001)

D.J. McClements, E.A. Decker, Y. Park, Physicochemical and structural aspects of lipid digestion., in: D.J. McClements...

R. Ghai et al.

Applications of isothermal titration calorimetry in pure and applied research—survey of the literature from 2010

J. Mol. Recognit.

(2012)

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^☆: This article is part of a Special Issue entitled Microcalorimetry in the BioSciences — Principles and Applications, edited by Fadi Bou-Abdallah.

¹: Present address: Departamento de Procesos y Tecnología, Universidad Autónoma Metropolitana-Cuajimalpa, Cuajimalpa, D.F. 05300, Mexico.

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Application of ITC in foods: A powerful tool for understanding the gastrointestinal fate of lipophilic compounds☆

Highlights

Abstract

Background

Scope of the review

Major conclusions

General significance

Introduction

Section snippets

Lipid bioavailability

Characterizing interactions amongst food components using ITC

Conclusions/summary

Conflict of interest

Transparency document

Acknowledgments

Nutr. Res.

Curr. Opin. Biotechnol.

Adv. Colloid Interf. Sci.

Adv. Colloid Interf. Sci.

Biomaterials

Anal. Biochem.

Anal. Biochem.

Methods

J. Pharm. Sci.

J. Chem. Thermodyn.

J. Pharm. Sci.

Food Chem.

Food Hydrocoll.

Food Chem.

Colloids Surf. B: Biointerfaces

Food Hydrocoll.

Process Biochem.

Carbohydr. Polym.

Food Chem.

Food Hydrocoll.

Food Res. Int.

Food Chem.

Food Hydrocoll.

Food Hydrocoll.

Food Res. Int.

Structural design principles for delivery of bioactive components in nutraceuticals and functional foods

Crit. Rev. Food Sci. Nutr.

Emulsion-based delivery systems for lipophilic bioactive components

J. Food Sci.

Carotenoid bioavailability and bioconversion

Annu. Rev. Nutr.

Separation and characterization of the colloidal phases produced on digestion of common formulation lipids and assessment of their impact on the apparent solubility of selected poorly water-soluble drugs

J. Pharm. Sci.

Intestinal absortion and tissue distribution of carotenoids.pdf

J. Nutr. Biochem.

Development and applicability of an in vitro digestion model in assessing the bioaccessibility of contaminants from food.pdf

Review of in vitro digestion models for rapid screening of emulsion-based systems

Food Funct.

A dynamic in vitro lipolysis model 1 controlling the rate of lipopysis by continous addition of calcium.pdf

Eur. J. Pharm. Sci.

Lipases at interfaces: unique interfacial properties as globular proteins

Langmuir

Influence of interfacial composition on in vitro digestibility of emulsified lipids: potential mechanism for chitosan's ability to inhibit fat digestion

Food Biophys.

Modelling texture perception by soft epithelial surfaces

Soft Matter

Disintegration of solid foods in human stomach

J. Food Sci.

Modeling the fluid dynamics in a human stomach to gain insight of food digestion

J. Food Sci.

Biopharmaceutical parameters to consider in order to alter the fate of nanocarriers after oral delivery

Nanomedicine

Gastrointestinal physiology

Applications of isothermal titration calorimetry in pure and applied research—survey of the literature from 2010

J. Mol. Recognit.