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Item Insights into wheat grain microstructure and composition for the development of novel flour with slow digestion properties and enhanced functional characteristics : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Food Technology at Massey University, Palmerston North, New Zealand(Massey University, 2025) AbhilashaWheat has been consumed as whole grains, broken grains, flattened format, and puffed format other than the flour format, which has a wide application in different types of food preparations. Wheat flour possesses a unique ability to form a cohesive dough that has viscoelastic properties. A range of products with wheat as their major ingredient are high glycaemic index (GI) foods as wheat flour contains highly digestible starch. However, the consumption of high GI foods is associated with chronic diseases such as diabetes, coronary heart disease, and obesity due to a rapid increase in blood glucose levels and secretion of insulin. The major objective of the research studies of this thesis included creating slowly digestible flour with improved functionality using slowly digested starch sources and non-starch components. Modifying wheat grain through different processing techniques alters the microstructure, and therefore, starch digestibility is impacted. Microstructure modification through various processing techniques, which can control the access of digestive enzymes to starch, could help develop products with controlled starch digestibility. To advance the understanding of the impact of wheat grain microstructure on starch hydrolysis, Chapter 3 explored a study on whole wheat grain in different commercially available forms (kibbled, cut grains, and flour) to understand the influence of microstructural changes on in vitro starch digestibility. The process of size reduction from raw intact grains to kibbled grains and flour caused an increase in overall starch hydrolysis (%) during simulated digestion in the order of flour>kibbled>cut>intact whole wheat grains. Cooking of these formats further increased their starch hydrolysis. However, both cooked cut and intact grains were low glycaemic with the expected glycaemic indices (eGI) of values of 54.08±0.03 and 41.98±0.04, respectively, revealing the role of intact microstructure in starch hydrolysis of wheat grains. Based on the role of intact microstructure, Chapter 4 investigated the possibility of reducing the starch hydrolysis in wheat grain formats (whole, flakes, and flour) by hydrothermal treatment and low-temperature storage of whole wheat grains. The extent of starch hydrolysis after oral-gastro-small intestinal digestion in vitro was significantly lower (p<0.05) in intact grains, flakes, and flours from the cold-stored grains than their non-cold-stored counterparts. In this study, scanning electron micrographs, pasting properties, water retention capacities, and relative crystallinity of the resulting flours revealed an enhanced degree of gelatinisation with the treatment temperature; however, cold-storage of treated grains resulted in a change in these properties due to the retrogradation of the starch. This study indicates that hydrothermal pre-treatment of grains followed by low-temperature storage for prolonged periods might help to reduce the starch digestibility of wheat grains and their resulting products and could be an effective strategy in developing reduced glycaemic impact grain products. However, in our preliminary trials, the flours from hydrothermally treated and low-temperature stored grains resulted in doughs of inferior viscoelastic properties. Furthermore, intending to create slowly digestible flour, Chapter 5 employed two approaches to modify a resistant starch: one involving soluble extracts from wheat flour and vital gluten (water solubles, salt-assisted water-solubles, and acid-solubles) and the other utilising hydrocolloids (guar gum, xanthan gum, locust bean gum, and carboxymethyl cellulose). Modifications from both approaches resulted in modified starch morphology with the formation of starch clusters mimicking the wheat flour. Moreover, the modification with hydrocolloids resulted in an improved pasting profile. Furthermore, in vitro digestion studies revealed that the starch hydrolysis rate was decreased for most of the cooked modified starches with wheat solubles and a slower starch hydrolysis profile until 60 min of simulated small intestinal digestion for most of the hydrocolloids used, carboxymethyl cellulose being the least effective in slowing the starch hydrolysis rate. Additionally, Chapter 6 evaluates the functionality and starch digestibility of a wheat flour system (dough and flatbread-chapatti) by utilising the modified starches created in Chapter 5 as low glycaemic ingredients. The interaction of the modified starches with vital gluten and wheat flour components resulted in improved viscosity of the functional flour. The microstructure of the functional flour dough indicated that the modified starches with wheat solubles (soluble extracts from wheat flour and vital gluten) and hydrocolloids improved the starch-protein matrix and gluten network. Furthermore, the in vitro digestion study revealed the overall starch hydrolysis of chapattis from all the functional flour formulations was significantly lower than the wheat flour chapatti. In conclusion, structural modifications of wheat grain could help reduce the overall starch hydrolysis of wheat grain products. Moreover, the wheat grain components have the potential to modify resistant starch sources to improve their functionality while retaining their slow digestion property. Also, utilising hydrocolloids to modify resistant starch sources could be an effective strategy to enhance the functionality of resistant starches in wheat-based systems. Modified resistant starches created using wheat solubles (soluble extracts from wheat flour and vital gluten) and hydrocolloids have potential applications with slow digestibility and improved functionality in wheat-based products.Item Starch-Tannin Interactions: Influence of Grape Tannins on Structure, Texture, and Digestibility of Starches from Different Botanical Sources(Elsevier Ltd, United Kingdom, 2025-05) Kaur H; Mehta A; Kumar LThis study investigated the effect of grape seed (GSd) and grape skin (GSk) tannins on the physicochemical, rheological properties and in-vitro digestibility of starches (corn, pea and wheat) derived from three different botanical sources. Quantification of bound and unbound tannins using MCP and HPLC analysis demonstrated that majority of the tannins were bound to starch molecules. The results of particle size distribution, starch-iodine binding and FTIR studies indicated the development of inclusion complexes through hydrophobic interactions with tannins in pea starch, while other two starches prominently formed non-inclusion complexes via hydrogen bonding. Back extrusion analysis of textural properties indicated that wheat starch-tannin complexes resulted in firmer starch-tannin gels compared to other two starches. Rheological studies revealed an increase in the viscoelastic modulus (G’ and G”) with improved elastic behavior for all starch-tannin gels. Starches complexed with tannins demonstrated strong antioxidant properties and in-vitro starch digestion studies revealed significant reductions in rapidly digestible starch (RDS) and slowly digestible starch (SDS), along with an increase in resistant starch (RS), particularly in pea starch complexed with GSd tannins. This study enhanced our understanding of how GSd and GSk tannins influence the properties of starches from various botanical origins, helping in understanding starch-tannin interactions and enabling the creation of foods with improved texture and digestibility.Item Cooked Rice-Based and Wheat-Based Food Structure Influenced Digestion Kinetics and Glycemic Response in Growing Pigs(Elsevier Inc on behalf of American Society for Nutrition, 2023-05-03) Nadia J; Olenskyj AG; Stroebinger N; Hodgkinson SM; Estevez TG; Subramanian P; Singh H; Singh RP; Bornhorst GMBACKGROUND: How starch-based food structure can affect the rate and extent of digestion in the small intestine and resulting glycemic response is not properly understood. One possible explanation is that food structure influences gastric digestion, which subsequently determines digestion kinetics in the small intestine and glucose absorption. However, this possibility has not been investigated in detail. OBJECTIVES: Using growing pigs as a digestion model for adult humans, this study aimed to investigate how physical structure of starch-rich foods affects small intestinal digestion and glycemic response. METHODS: Male growing pigs (21.7 ± 1.8 kg, Large White × Landrace) were fed one of the 6 cooked diets (250-g starch equivalent) with varying initial structures (rice grain, semolina porridge, wheat or rice couscous, or wheat or rice noodle). The glycemic response, small intestinal content particle size and hydrolyzed starch content, ileal starch digestibility, and portal vein plasma glucose were measured. Glycemic response was measured as plasma glucose concentration collected from an in-dwelling jugular vein catheter for up to 390 min postprandial. Portal vein blood samples and small intestinal content were measured after sedation and euthanasia of the pigs at 30, 60, 120, or 240 min postprandial. Data were analyzed with a mixed-model ANOVA. RESULTS: The plasma glucose Δmaxoverall and iAUCoverall for couscous and porridge diets (smaller-sized diets) were higher than that of intact grain and noodle diets (larger-sized diets): 29.0 ± 3.2 compared with 21.7 ± 2.6 mg/dL and 5659 ± 727 compared with 2704 ± 521 mg/dL⋅min, for the smaller-sized and larger-sized diets, respectively (P < 0.05). Ileal starch digestibility was not significantly different between the diets (P ≥ 0.05). The iAUCoverall was inversely related to the starch gastric emptying half-time of the diets (r = -0.90, P = 0.015). CONCLUSIONS: Starch-based food structure affected the glycemic response and starch digestion kinetics in the small intestine of growing pigs.Item Influence of food macrostructure on the kinetics of acidification in the pig stomach after the consumption of rice- and wheat-based foods: Implications for starch hydrolysis and starch emptying rate(Elsevier Ltd, 2022-11-15) Nadia J; Olenskyj AG; Subramanian P; Hodgkinson S; Stroebinger N; Estevez TG; Singh RP; Singh H; Bornhorst GMHow the stomach can serve as a biochemical environment for starch digestion and the implications on starch emptying are not well-understood. Biochemical changes during gastric digestion of cooked wheat- and rice-based diets of varying particle size and microstructure were investigated using a growing pig model. In larger-particle size diets (rice grain, rice noodle, pasta), pH >3 was maintained in the proximal stomach digesta even until 240 min digestion, resulting in extended remaining amylase activity and accumulation of maltose from starch hydrolysis in the stomach. In smaller-particle size diets (couscous, rice couscous, semolina porridge), gastric acidification occurred faster to produce homogeneous intragastric pH and deactivated amylase. The hypothesis of the study was that food macrostructure would impact gastric acidification kinetics, and the resulting biochemical environment for starch hydrolysis in the stomach may further affect the mechanisms of food breakdown in the stomach and gastric emptying of starch.Item Contribution of the proximal and distal gastric phases to the breakdown of cooked starch-rich solid foods during static in vitro gastric digestion.(Elsevier, 2022-07) Nadia J; Bronlund JE; Singh H; Singh RP; Bornhorst GMIn vitro gastric digestion studies commonly focus on the acidic environment of the stomach (the distal phase), neglecting that the contact time between food and salivary amylase can be extended during bolus' temporary storage in the proximal stomach (the proximal phase). Consequently, the role of the proximal phase of gastric digestion on the breakdown of solid starch-based foods is not well understood. This study aimed to address this question using a static in vitro digestion approach. Cooked starch-rich foods of different physical structures (wheat couscous, wheat pasta, rice couscous, rice noodle, and rice grain) were subjected to 30 s oral phase digestion, followed by prolonged incubation of the oral phase mixture (pH 7) for up to 30 min representing different proximal phase digestion times. Each proximal phase sample was sequentially incubated in excess simulated gastric fluid (distal phase, pH 2) for up to an additional 180 min. The proximal phase aided solid food breakdown through starch hydrolysis that caused leaching of particles <2 mm. The distal phase led to softening of food particles, but the softening process was not enhanced with longer proximal phase. In foods with smaller initial size (couscous and rice couscous), a proximal phase of 15 min or longer followed by 180-min distal phase increased starch hydrolysis in the liquid and suspended solid fractions of the digesta, indicating the influence of food structure on acid hydrolysis during in vitro gastric digestion.Item Probing the Double-Layered Cotyledon Cell Structure of Navy Beans: Barrier Effect of the Protein Matrix on In Vitro Starch Digestion(MDPI (Basel, Switzerland), 2023-01) Do DT; Singh J; Johnson S; Singh H; Bordoni AThe microstructure of legumes plays a crucial role in regulating starch digestion and postprandial glycemic responses. Starch granules are double encapsulated within the outer cell wall and the inner protein matrix of legume cotyledon cells. Despite progress in understanding the role of cell walls in delaying starch digestion, the role of the protein matrix has received little research attention. The aim of this study was to evaluate if the protein matrix and cell wall may present combined physical barriers retarding enzyme hydrolysis of intracellular starch. Intact cotyledon cells were isolated from navy beans and used to assess the barrier effect of the protein matrix on the digestion of starch under conditions simulating the upper gastrointestinal tract. The cells were pretreated with pepsin at 37 °C and pH 2.0 for 1, 4, or 24 h and without pepsin for 24 h (control) to facilitate removal of the intracellular protein matrix prior to cooking and simulated in vitro digestion. A longer pretreatment time resulted in a lower protein content of the cells and a higher initial rate and extent of starch hydrolysis. We suggest that in addition to the primary cell wall barrier, the protein matrix provides a secondary barrier restricting the accessibility of α-amylase to starch. This study provides a new fundamental understanding of the relationship between the structural organization of legume cotyledon cells and starch digestion that could inform the design of novel low glycemic index foods.Item Glycaemic potency reduction by coarse grain structure in breads is largely eliminated during normal ingestion(Cambridge University Press on behalf of The Nutrition Society, 2022-05-28) Srv A; Mishra S; Hardacre A; Matia-Merino L; Goh K; Warren FJ; Monro JAThe hypothesis that coarse grain particles in breads reduce glycaemic response only if the particles remain intact during ingestion was tested. Three breads were formulated: (1) White bread (WB - reference), (2) 75 % of kibbled purple wheat in 25 % white bread matrix (PB) and (3) a 1:1 mixture of 37·5 % kibbled soya beans and 37·5 % of kibble purple wheat in 25 % white bread matrix (SPB). Each bread was ingested in three forms: unchewed (U), as customarily consumed (C) and homogenised (H). Twelve participants ingested 40 g available carbohydrate portions of each bread in each form, with post-prandial blood glucose measured over 120 min. Glycaemic responses to WB were the same regardless of its form when ingested. Unchewed PB had significantly less glycaemic effect than WB, whereas the C and H forms were similar to WB. Based on a glycaemic index (GI) of 70 for WB, the GI values for the C, U and H breads, respectively, were WB: 70·0, 70 and 70, PB: 75, 42 and 61, SPB: 57, 48 and 55 (%) (Least significant difference = 17·43, P < 0·05, bold numbers significantly different from WB). The similar glycaemic response to the H and C forms of the breads, and their difference from the U form, showed that the glycaemia-moderating effect of grain structure on starch digestion was lost during customary ingestion of bread. We conclude that the kibbled-grain structure may not effectively retard starch digestion in breads as normally consumed because it is largely eliminated by ingestive processes including chewing.Item Characterisation of de-structured starch and its interactions in whey protein isolate gels : a thesis submitted in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Food Technology at Massey University, Palmerston North, New Zealand(Massey University, 2022) Ang, Cai LingStarch serves as an important additive to enhance the physico-chemical properties of many food products. With the increased pursuit of natural products, there is an increasing demand for “clean-label” starches. In this study, waxy potato starch was physically-modified at elevated temperatures of 120–150 °C for 30 min at 300 rpm, in a pressurised reactor. The treatment converted native starch granules into their macromolecular chains (denoted as de-structured waxy potato starch, DWPS). This doctoral thesis presents the: (i) method of modifying starch (i.e., the de-structuring process), (ii) the mechanism of starch de-structuring, (iii) the rheological changes in DWPS samples and the shear-thickening mechanism, and (iv) the interactions of these DWPS ingredients with whey protein isolate (WPI) in a protein-based gel system, at different pH and ionic strength. The molar mass (Mᵥᵥ), particle size, rheological properties, degree of branching (DB) and side-chain length distribution of DWPS samples were characterised to elucidate the starch de-structuring mechanism. DWPS treated at 120 °C DWPS showed similar Mᵥᵥ (~3.6 × 10⁸ Da) as its native form (~3.7 × 10⁸ Da) indicating that the treatment at 120 °C resulted in the disassembly of starch granules into their macromolecular chains. Reduction in viscosity, Mᵥᵥ and particle size was observed with an increase in temperature from 120 to 150 °C, suggesting a cleavage in amylopectin chains. The DB and side-chain distribution data suggest that the reduction in Mᵥᵥ is likely due to the cleavage at α-1,4 linkages near the middle of the main amylopectin backbone. Particle size analysis by laser diffraction measurements revealed the presence of large fragment particles (> 1 µm) in DWPS samples, indicating that the starch de-structuring process into its macromolecules was incomplete even at 150 °C for 30 min. The DWPS (5% w/w) samples were found to exhibit a wide range of rheological properties—Newtonian, shear-thinning, shear-thickening and anti-thixotropy behaviours—depending on their treatment temperature (120–150 °C). In particular, 120 °C DWPS exhibited interesting shear-thickening, anti-thixotropy and shear-induced gelation. These rheological properties are different from the shear-thinning and thixotropy behaviours observed in most conventionally gelatinised waxy potato starches treated at 95 °C. The complex shear-induced structures of 120 °C DWPS were attributed to a two-step process: (i) upon shear at the critical shear rate (~10–20 s⁻¹), the shear stress caused a size reduction in the starch fragments and (ii) the increased number of small fragments together with the amylopectin chains in very close proximity could lead to the formation of a complex network probably consisting of amylopectin chains and a large number of fragments (2–20 μm). Shear thickening properties were attributed largely to these soft fragment particles colliding and sliding past each other during shear. The data from this study has also shown that the hydrogen bonding, electrostatic, hydrophobic interactions, or the combination of these interactions did not cause the shear-thickening behaviour. The influence of 4% w/w DWPS on 13% w/w WPI gels was studied by characterising the phase stability of the liquid mixtures, and mechanical properties, microstructure, and water-immersion stability of fine-stranded polymeric and coarse-stranded particulate protein gels at pH 7 and pH 5, respectively. At neutral pH, synergistic gel hardness of WPI was obtained with the incorporation of 140 °C DWPS. The increased gel strength was attributed to the enhanced density of a very fine-stranded gel network. The ability of the gel to retain its shape when immersed in water for 40 h was most noticeable for the composite gels containing either gelatinised starch or DWPS samples (swollen gels but with intact shape). In contrast, pure WPI gel and composite gel containing maltodextrin turned into very weak fluid-like and disintegrated gels, respectively. At pH 5, WPI formed particulate gels. The addition of gelatinised starch or DWPS weakened the particulate protein gels, likely due to phase separation and interrupted protein network with starch polymers acting as inactive fillers. The effects of NaCl and CaCl₂ (i.e., type of salts and ionic strength) on the mechanical and microstructural properties of composite gels containing 13% w/w WPI and 4% w/w 140 °C DWPS were also evaluated. Thermodynamic incompatibility between WPI and 140 °C DWPS was observed upon the addition of NaCl (~75 mM) or CaCl₂ (10–75 mM). The combined effects of such thermodynamic incompatibility with the changes in protein connectivity induced by varied ionic strength led to the formation of distinctive gel structures (inhomogeneous self-supporting gels with a liquid centre and weak gels with paste-like consistency) that were different from thermodynamic compatible homogeneous self-supporting gels (pure WPI and WPI + maltodextrin gels). At ≥ 250 mM NaCl, instead of a paste-like texture, a recovered soft self-supporting gel structure was observed when using 140 °C DWPS. The ability to generate a range of textures in WPI gelation-based foods by using 140 °C DWPS under different ionic conditions, is a feasible strategy for structuring high-protein foods for dysphagia—aimed to be either thickened fluids or soft solids. Additionally, this acquired knowledge is also relevant when formulating food gels for 3-D printing. The desirable rheological properties of DWPS samples and their ability to alter WPI gel structure signify the potential of DWPS as a clean-label ingredient to structure foods of specific needs (e.g., whipping cream for enhanced structure upon shear and high-protein foods for dysphagia sufferers).Item Starch retrogradation in tuber : mechanisms and its implications on microstructure and glycaemic features of potatoes : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in School of Food and Advanced Technology at Massey University, Palmerston North, Manawatū, New Zealand(Massey University, 2020) Chen, Yu-Fan NicoleAn increase in the occurrence of diabetes mellitus, cardiovascular disease and obesity in recent years led to the project “Starch retrogradation in tuber: mechanisms and its implications on microstructure and glycaemic features of potatoes”. Potato products can play a role in mitigating these hyperglycaemic events, if starch in these processed products is slowly digested and/or starch-derived glucose is released into the circulation in a slower and more attenuated manner. Three stages were envisaged for the project with an aim to create slowly digestible starch in whole potato tuber (in tuber) through starch retrogradation. Plant-based whole food systems, such as potato tubers encompass different cell compartments, (e.g. cell wall, vacuole, cytoplasm and intracellular spaces) within which starch gelatinisation and retrogradation occur, subject to local interactions of other cell components and water availability. Structural changes of potato starch during retrogradation in tuber and its resulting digestibility were studied. Different water pools in a cooked whole tuber were discerned by the low-field NMR (LF-NMR), having relaxation times T20 at <1 ms, T21 at 10-15 ms, T22 at 70–200ms, and T23 at > 400 ms. A significant reduction in eGI was observed after cooling and storage compared to freshly cooked tubers. Reheating of retrograded tuber restored some of the susceptibility to enzymatic hydrolysis and internal water mobility. Longer chilled storage (7 days) yet improved the stability of retrograded tuber against reheating effects (at 90 °C). Realignment of the gelatinised amylose and amylopectin changed the distribution of crystalline and amorphous regions during refrigerated storage and subsequent reheating, resulting in starch digestibility varying with treatment combination. Several, but not all, of time-temperature cycle processes were observed to induce stepwise nucleation and propagation, facilitating starch retrogradation in tuber more than did storage fixed at 4 °C. Sous vide processing (at 55 and 65°C), akin to annealing, combined with starch retrogradation in tuber, resulted in potatoes with intermediate eGI (40-72). After reheating at 60°C, the eGI of sous vide cooked-chill potatoes increased moderately, displaying a mixture of partially gelatinised starch and swollen granules. Food processing, i.e. optimum TTC process or sous vide process might facilitate the formation of retrograded starch in tuber, resulting in a reduced eGI (than freshly cooked tubers). To retain the resistance to digestive enzymes in retrograded starch in tuber, reheating at low temperatures (50-60°C) were needed.