The Pennsylvania State University The Graduate School College of Agricultural Sciences USE OF STARCH INCLUSION COMPLEXES FOR IMPROVED DELIVERY OF DIETARY POLYPHENOLS TO THE ORAL CAVITY BY CHEWING GUM A Thesis in Food Science by Debie Wesley Blair © 2010 Debie Wesley Blair Submitted in Partial Fulfillment of the Requirements for the Degree of Master of Science December 2010 The thesis of Debie W. Blair was reviewed and approved* by the following: Gregory R. Ziegler Professor of Food Science Thesis Adviser Joshua D. Lambert Assistant Professor of Food Science John D. Floros Professor of Food Science Head of the Department of Food Science *Signatures are on file in the Graduate School. ii ABSTRACT Starch inclusion complexes can increase the stability, dispersibility, and quite possibly bioavailability, of bioactive guest compounds encapsulated within a starch helix. Bhosale and Ziegler (unpublished) successfully emulsified beta-carotene into a starch inclusion complex, which increased its dispersibility in water, protected the compound from degradative reactions, and also provided a means for controlled release of this sensitive hydrophobic compound. In this thesis, the overall effectivenesss of chewing gum as a delivery vehicle for bioactive polyphenols found in green tea was investigated. In order to fully understand this delivery method, the release of the catechins from three different gum treatments were measured by HPLC. The inhibitory effect that green tea catechins may have on amylase activity was tested to evaluate the effect of starch addition to the gum base. Lastly, the effect of the complex on bioavailability of bioactive components was investigated in-vivo by dosage in 72 male CF-1 mice and HPLC analysis of beta-carotene absorption in plasma and fecal samples. Overall, the results of these studies support the need for further investigation of the use of chewing gum and starch inclusion complexes as a delivery vehicle for bioactive components. There is no evidence of interference by tea polyphenols on the breakdown of starch by amylase and hence the release of the bioactive components into the body. Although the physical mixture of starch, tea polyphenols and ascorbyl palmitate into chewing gum possessed the highest Cmax, the mixture of starch-ascorbyl palmitate complex and tea polyphenols could be the most effective treatment in relation to both possible organoleptic properties as well as total release of bioactive components when compared to gum with tea polyphenols alone. iii TABLE OF CONTENTS List of tables.................................................................................................................................. vi List of figures ............................................................................................................................... vii Acknowledgements .................................................................................................................... viii Chapter 1: Introduction ...............................................................................................................1 1.1 Objectives ..............................................................................................................................4 Chapter 2: Literature Review ......................................................................................................6 2.1 Using chewing gum as a delivery vehicle of bioactive green tea polyphenols ................6 Growth of the gum industry ....................................................................................................8 Gum composition ....................................................................................................................9 Texture ..................................................................................................................................12 Encapsulation and coatings ...................................................................................................13 Encapsulation ..................................................................................................................13 Coatings ...........................................................................................................................15 Chewing gum as an oral delivery vehicle .............................................................................16 2.2 Inhibition of salivary and gastrointestinal enzymes by polyphenolic compounds ........16 Inhibition of lipase by polyphenols .......................................................................................18 Inhibitory effects on amylase and amyloglucosidase ............................................................18 Inhibitory effects of black tea compounds ............................................................................22 Overall inhibitory effect and product feasibility ...................................................................24 2.3 Bioavailability of nutrients in starch inclusion complexes and food matrices .............24 Lipid interactions affecting bioavailability ...........................................................................25 Starch interactions affecting bioavailability ..........................................................................27 Micronutrient availability ......................................................................................................30 Matrices and interactions with guar gum ..............................................................................32 Bioavailability and product feasibility ..................................................................................34 Chapter 3: Materials and methods ............................................................................................35 3.1 Chewing gum study ...........................................................................................................35 Chemicals and reagents.........................................................................................................35 Complexation of amylose and ascorbyl palmitate ................................................................35 Complexation of amylose, ascorbyl palmitate, and tea extract ............................................36 Ingredient preparation for chewing gum ...............................................................................36 Addition of complex to gum base and formation of chewing gum ......................................37 Experimental design..............................................................................................................37 iv Analysis of polyphenols in samples by HPLC .....................................................................38 3.2 Inhibition study ..................................................................................................................38 Chemicals and reagents.........................................................................................................38 Preparation of solutions ........................................................................................................39 Glucose production ...............................................................................................................39 3.3 Bioavailability study ..........................................................................................................40 Chemicals and reagents.........................................................................................................40 Inclusion complex .................................................................................................................40 Animals and treatment ..........................................................................................................41 Extraction of plasma .............................................................................................................41 Extraction of feces ................................................................................................................42 Analysis of beta-carotene in samples by HPLC ...................................................................42 Chapter 4: Results and discussion .............................................................................................43 4.1 Chewing gum study ...........................................................................................................43 4.2 Inhibition study ..................................................................................................................54 4.3 Bioavailability study ..........................................................................................................56 Chapter 5: Conclusions ..............................................................................................................62 5.1 Overall conclusions ............................................................................................................63 References .....................................................................................................................................65 v LIST OF TABLES Table 1. Chewing gum composition (D’Amelia et al. 1984) .......................................................10 Table 2. Formulations for gum treatments per 4.0 g and 5.15 g stick of gum. Values for complex mixture are based on the weights of A6P and starch used to make the inclusion complexes ......................................................................................................................................37