Structural characteristics and in vitro starch digestibility of oil-modified cooked rice with varied addition manipulations.

Lipid has crucial applications in improving the quality of starchy products during heat processing. Herein, the influence of lipid modification and thermal treatment on the physicochemical properties and starch digestibility of cooked rice prepared with varied addition manipulations was investigated. Rice bran oil (RO) and medium chain triglyceride oil (MO) manipulations were performed either before (BC) or after cooking (AC). GC-MS was applied to determine the fatty acid profiles. Nutritional quality was analyzed by quantifying total phenolics, atherogenic, and thrombogenic indices. All complexes exhibited higher surface firmness, a soft core, and less adhesive. FTIR spectrum demonstrated that the guest component affected some of the dense structural attributes of V-amylose. The kinetic constant was in the range between 0.47 and 0.86 min-1 wherein before mode presented a higher value. The lowest glucose release was observed in the RO_BC sample, whereas the highest complexing index was observed in the RO_AC sample, indicating that the dense molecular configuration of complexes that could resist enzymatic digestion was more critical than the quantity of complex formation. Despite the damage caused by mass and heat transfer, physical barrier, intact granule forms, and strengthened dense structure were the central contributors affecting the digestion characteristics of lipid-starch complexes.

Effect of starch and peanut oil on physicochemical and gel properties of myofibrillar protein: Amylose content and addition form.

Starch and peanut oil (PO) were widely used to improve the gel properties of surimi, however, the impact mechanism of addition forms on the denaturation and aggregation behavior of myofibrillar protein (MP) is not clear. Therefore, the effect of starch, PO, starch/PO mixture, and starch-based emulsion on the physicochemical and gel properties of MP was investigated. The results showed that amylose could accelerate the aggregation of MP, while amylopectin was conducive to the improvement of gel properties. The addition of PO, starch/PO mixture, or starch-based emulsion increased the turbidity, solubility, sulfhydryl content of MP, and improved the gel strength, whiteness, and texture of MP gel. However, compared with starch/PO mixture group, the gel strength of MP with waxy, normal and high amylose corn starch-based emulsion increased by 22.68 %, 10.27 %, and 32.89 %, respectively. The MP containing emulsion had higher storage modulus than MP with starch/PO mixture under the same amylose content. CLSM results indicated that the oil droplets aggregated in PO or starch/PO mixture group, while emulsified oil droplets filled the protein gel network more homogeneously. Therefore, the addition of starch and PO in the form of emulsion could effectively play the filling role to improve the gel properties of MP.

Hot air-assisted radio frequency drying of corn kernels: the effect on structure and functionality properties of corn starch.

Hot air (HA) drying caused quality damage of grains with long treatment time. Radio frequency (RF) heating as an emerging technology was applied to improve drying quality of cereals effectively. The effects of HA-RF drying (50 °C, 70 °C, 90 °C) of corn kernels on the morphology, structure, and physicochemical properties of starch were investigated and compared with HA drying. The surface of treated starch became rough, along with fragments and pores. Drying treatments increased the amylose content from 10.59 % to 23.88 % and the residual protein content of starch from 0.58 % to 1.23 %, and reduced the crystallinity from 31.95 % to 17.15 % and short-range order structures of starch from 0.918 to 0.868. The change of structures in turn resulted in the increase of pasting viscosity, gelatinization temperature, storage modulus and loss modulus. Furthermore, the HA-RF dried starch displayed stronger thermal stability, higher gelatinization degree and better gelation properties than the HA-treated starch at the same temperature. The data proved that the synergistic effects of HA and RF were more effective in modulating the starch structure and improving the functional characteristics of corn starch. This paper would like to provide potential reference for better application of HA-RF technologies to corn.

Physicochemical and digestive properties of corn starch nanoparticles incorporated different polyphenols.

The corn starch nanoparticles were prepared by incorporating three kinds of polyphenols, including quercetin, proanthocyanidins and tannin acid. The physicochemical and digestive properties of corn starch nanoparticles were researched. The quercetin showed a higher complexation index than proanthocyanidins and tannin acid when they complexed with corn starch. The mean size of corn starch quercetin, proanthocyanidins and tannin acid were 168.5 nm, 179.1 nm and 188.6 nm, respectively. XRD results indicated that all the corn starch-polyphenols complex showed V-type crystalline structure, the crystallinity of corn starch-quercetin complex was 19.31 %, which showed more formation of amylose-quercetin single helical formed than the other two starch-polyphenol complexes. In vitro digestion revealed that polyphenols could resist digestion and quercetin increased the content of resistant starch from 23.32 % to 35.24 % and polyphenols can form complexes with starch through hydrophobic interactions or hydrogen bonding. This study indicated the hydrophobic polyphenols had a more significant effect on the digestibility of corn starch. And the cell toxicity assessments demonstrated that all nanoparticles were nontoxic and biocompatible.

Insights into formation and stability mechanism of V7-type short amylose-resveratrol complex using molecular dynamics simulation and molecular docking.

Pre-formed V-type amylose as a kind of wall material has been reported to carry polyphenols, while the interaction mechanism between V-type amylose and polyphenol is still elusive. In this work, the formation and stability mechanism of a V7-type short amylose-resveratrol complex was investigated via isothermal titration calorimetry, molecular dynamics, and molecular docking. The results presented that two stoichiometric ratios of resveratrol to short amylose were calculated to 0.120 and 0.800, and the corresponding main driving force was hydrogen bonding and hydrophobic interaction, respectively. The folding and unfolding conformation of V7-type short amylose chains appeared alternately during the simulation. Resveratrol tended to be bound in the short amylose helix between 40 ns and 80 ns to form a more stable complex. Hydrogen bonds between resveratrol molecule and O6 at the 22nd glucose molecule/O2 at the 24th glucose molecules and hydrophobic interaction between resveratrol molecule and glucose molecules (19th, 20th, 21st and 23rd) could be found.

Multiscale structure changes and mechanism of polyphenol-amylose complexes modulated by polyphenolic structures.

This study was designed to investigate the effect of polyphenolic structure on the interaction strength and process between polyphenols (gallic acid (GA), epigallocatechin gallate (EGCG) and tannic acid (TA)) and amylose (AM). The results of Fourier transform infrared spectroscopy, isothermal titration calorimetry, X-ray photoelectron spectroscopy and molecular dynamic simulation (MD) suggested that the interactions between the three polyphenols and AM were noncovalent, spontaneous, low-energy and driven by enthalpy, which would be enhanced with increasing amounts of pyrogallol groups in the polyphenols. The results of turbidity, particle size and appearance of the complex solution showed that the interaction process between polyphenols and AM could be divided into three steps and would be advanced by increasing the number of pyrogallol groups in the polyphenols. At the same time, MD was intuitively employed to exhibit the interaction process between amylose and polyphenols, and it revealed that the interaction induced the aggregation of amylose and that the agglomeration degree of amylose increased with increasing number of pyrogallol groups at polyphenols. Last, the SEM and TGA results showed that TA/AM complexes had the tightest structure and the highest thermal stability (TA/AM˃EGCG/AM˃GA/AM), which could be attributed to TA having five pyrogallol groups.

Effects of microwave radiation on the physicochemical properties, structure, and digestibility of the synthesized different crystal forms of malic acid starch ester.

The effects of microwave combined with L-malic acid treatment on the degree of substitution (DS), structure, physicochemical properties, and digestibility of sweet potato starch (A-type), potato starch (B-type), and pea starch (C-type) were evaluated. The order of DS obtained was: DSM-POS > DSM-SPS > DSM-PES. Fourier transform-infrared spectroscopy (FT-IR) showed that the obtained modified starch produced a new absorption band at 1735 cm-1. Scanning electron microscopy (SEM) and polarized light microscopy indicated that different types of native starches exhibited different granular morphologies and appeared to have different degrees of damage, but still had polarized crosses after modification. Sweet potato starch had the smallest particle size, while potato starch had the largest. X-ray diffractometry (XRD) showed that the modified starches still retained the same crystal structure as the native starches, but the relative crystallinity decreased. The apparent viscosity and swelling power of modified starches dropped, but their water/oil holding capacity, amylose content, and resistant starch content all increased. The results demonstrate that the degree of influence on the structure, physicochemical properties, and digestibility of different starches varies under the same modification conditions.

Amylose content controls the V-type structural formation and in vitro digestibility of maize starch-resveratrol complexes and their effect on human gut microbiota.

The chain structure of starch affects its interaction with polyphenol molecules which in turn determines the nutritional function of starch. In this study, starch with different amylose content including waxy maize starch (WMS), normal maize starch (NMS) and G50 high-amylose maize starch (G50) were selected to complex with resveratrol (RA) in high-pressure homogenization (HPH) environment, and structural changes of the complexes, together with their effects on in vitro digestibility and gut microbiota were discussed. The results showed that with increasing amylose content, RA could form more inclusion complex with starch through non-covalent bonds accompanied by the increased single helix structure, V-type crystalline structure, compact nano-aggregates and total ordered structure content, which thus endowed the complex lower digestibility and intestinal probiotic function. Notably, when RA addition reached 3 %, the resistant starch (RS) content of HP-G50-3 % rose to 29.2 %, correspondingly increased the relative abundance of beneficial gut microbiota such as Megamonas and Bifidobacterium, as well as the total short-chain fatty acids (SCFAs) content. Correlation analysis showed that V-type crystalline structure positively correlated with the growth of Pediococcu and Blautia (p < 0.05) for producing SCFAs. These findings provided feasible ideas for the development of personalized nutritional starch-based foods.

Inhibition of α-amylase digestion by a Lonicera caerulea berry polyphenol starch complex revealed via multi-spectroscopic and molecular dynamics analyses.

Polyphenol-starch complexes exhibit synergistic and beneficial effects on both polyphenols and resistant starches. This study evaluates the inhibitory effects and mechanisms of α-amylase on a Lonicera caerulea berry polyphenol-wheat starch (LPWS) complex following high hydrostatic pressure treatments of 400 MPa for 30 min and 600 MPa for 30 min. The IC50 values for α-amylase inhibition by the complex were 3.61 ± 0.10 mg/mL and 3.42 ± 0.08 mg/mL at a 10 % (w/w) polyphenol content. This interaction was further supported by Fourier-transform infrared spectroscopy and circular dichroism, which confirmed that the alpha helix component of the secondary structure of α-amylase was reduced due to the complex. Multifluorescence spectroscopy revealed that the complex induces changes in the microenvironment of fluorophores surrounding the α-amylase active site. Molecular dynamics simulations and molecular docking revealed that the active site of amylose within the complex becomes enveloped in polyphenol clusters. This wrapping effect reduced the hydrogen bonds between amylose and α-amylase, decreasing from 16 groups to just one group. In summary, the LPWS complex represents a low-digestible carbohydrate food source, thus laying the groundwork for the research and development of functional foods aimed at postprandial hypoglycemic effects.

Gallic acid forms V-amylose complex structure with starch through hydrophobic interaction.

This study aimed to investigate the role of amylose and amylopectin in the formation of starch-polyphenol complex and elucidate the interaction mechanisms. Gallic acid (GA) was used to complex with maize starch with various amylose contents. Results showed GA formed V-type crystals with normal maize starch (NMS) and high amylose maize starch (HAMS), while higher relative crystallinity was exhibited in HAMS-GA complexes than NMS counterparts. Molecular structure analysis revealed more amylose in GA-starch complexes than in treated starch counterparts without GA, and this was more apparent in HAMS than NMS, implying amylose is preferred to complex with GA than amylopectin. FTIR detected higher R1047/1022 value in starch-GA complexes than their starch counterparts without GA, suggesting increased short-range ordered structrure of complexes. Typical signatures of hydrophobic interactions were further revealed by isothermal titration calorimetry, indicating the complexation of GA to starch is mainly through hydrophobic bonds. More binding sites were observed for HAMS (72.50) than NMS (11.33), which proves the preferences of amylose to bind with GA. Molecular dynamics simulated the complexation of GA to amylose, and confirmed hydrophobic bond is the main interaction force. These findings would provide guidance for precise design and utilization of starch-polyphenol complexes in functional foods.

Multi-scale structural disruption induced by radio frequency air cold plasma accelerates enzymatic hydrolysis/ hydroxypropylation of tapioca starch.

This study investigated the effects of radio frequency air cold plasma (RFACP) pretreatment on the multi-scale structures, physicochemical properties, enzymatic hydrolysis, and hydroxypropylation of tapioca starch. The results showed that cold plasma (CP) made starch granules rough on the surface and disrupted long- and short-range ordered structures, reducing relative crystallinity from 43.8 % to 37.4 % and R1047/1022 value from 0.992 to 0.934. Meanwhile, the starch molecules were depolymerized and oxidized by CP, reducing weight-average molecular weight from 9.64 × 107 to 2.17 × 107 g/mol, while increasing carbonyl and carboxyl groups by up to 118 % and 53 %. Additionally, CP-treated starches exhibited higher solubility and swelling power, along with lower gelatinization enthalpy. Short-time CP pretreatment (10 min) promoted the hydroxypropylation of starch and increased the molar substitution (0.081-0.112). Also, CP pretreatment accelerated enzymatic hydrolysis of starch, as indicated by the increase in hydrolysis rate (1.846 × 10-3-2.033 × 10-3 min-1) and degree of hydrolysis (51.45 % - 59.92 %). Overall, the multi-scale structural disruption induced by CP treatment facilitated the accessibility of enzymes/chemical reagents into starch granules and glucan chains. This study suggested that RFACP could be used for starch pretreatment to increase production efficiency in modified starch production, as well as in brewing and fermentation industries.

The selective adsorption mechanism of V-type starches for key off-odors of sea cucumber intestinal peptides.

The off-odors of sea cucumber intestinal peptide (SCIP) severely limit its application. In this study, the V-type starches were derived from high amylose maize starch to adsorb odors of SCIP and the adsorption mechanism was explored. The inclusion complexes formed by V-type starches and volatile compounds of SCIP were characterized by X-ray diffraction and Fourier transform infrared spectroscopy. The electronic nose results revealed a decreasing trend in response values of SCIP, with significant differences before and after deodorization (p < 0.05). Furthermore, 82 volatiles were identified from SCIP, and six were determined as key volatiles using gas chromatography-mass spectrometry. The V6- and V7-type starches with smaller cavity sizes selectively adsorb butyric acid, isobutyric acid and nonaldehyde, and V8-type starches with a larger cavity size selectively adsorb trimethylamine. This study proved that using V-type starches for deodorization could effectively improve SCIP flavor.

Beneficial glycaemic effects of high-amylose barley bread compared to wheat bread in type 2 diabetes.

BACKGROUND: Cereals foods with a high content of dietary fibres or amylose have potential to lower postprandial glucose levels. Optimisation of cereal foods may improve management of type 2 diabetes (T2D). METHODS: We investigated the impact on 4 h postprandial glucose responses given as incremental area under curve (iAUC) of bread made of either 50% RNAi-based (genetically modified) amylose-only barley flour (AmOn) (and 50% wheat flour), 50% hulless barley flour (and 50% wheat flour) or 75% hulless barley (and 25% wheat flour) in subjects with T2D compared with 100% wheat flour bread. DESIGN: Twenty adults with T2D were randomly allocated to one of four breads at four separate visits. We measured fasting and 4 h postprandial responses of glucose, insulin, glucagon, triacylglycerol (TG), free fatty acids (FFA), glucagon-like peptide-1 (GLP-1) and gastric inhibitory polypeptide (GIP). Mixed model ANOVA was used to examine the differences. RESULTS: Bread made from 50% AmOn lowered the 4 h postprandial glucose by 34%, 27%, 23% (P < 0.05) compared with 100% wheat, 50% or 75% hulless barley, respectively. Bread made from 75% hulless barley reduced the postprandial glucose response (iAUC) by 11% (P < 0.05) compared to 100% wheat bread. Postprandial insulin responses (iAUC) were reduced for 50% AmOn compared with 100% wheat and 50% hulless barley and for 75% hulless compared to 50% hulless barley bread (P < 0.05). 4 h postprandial glucagon (tAUC) did not differ between the four bread types (P > 0.05). Lower postprandial GIP (iAUC) was observed after all barley breads compared to 100% wheat (P < 0.05), whereas no difference was seen in postprandial GLP-1. Postprandial TG and FFA (tAUC) were difficult to judge due to differences in fasting values. CONCLUSIONS: Bread made by replacing wheat flour with either 50% high-amylose or 75% hulless barley flour lowered postprandial glucose responses compared to 100% wheat bread indicating a beneficial impact on glucose regulation in T2D subjects. This trial was registered at clinicaltrials.gov as NCT04646746.

Fabrication and release property of self-assembled garlic essential oil-amylose inclusion complex by pre-gelatinization coupling with high-speed shear.

Our aim was to investigate the preparation of self-assembled garlic essential oil-amylose inclusion complexes (SGAs) using garlic essential oil (GEO) and corn starch (CS), and evaluated their release properties. SGAs were fabricated by pre-gelatinization coupling with high-speed shear at different GEO-CS mass ratios. When the mass ratio of GEO to pre-gelatinized corn starch was set at 15 % (SGA-15 %), with a fixed shear rate of 9000 rpm and a shear time of 30 min, the allicin content was 0.573 ± 0.023 mg/g. X-ray diffraction (XRD) results revealed a starch V-type crystalline structure in SGAs with peaks at 13.0°, 18.0°, and 20.0° (2θ). Fourier Transform Infrared (FTIR) spectra of SGAs displayed a shift in the characteristic peak of diallyl trisulfide from 987.51 cm-1 to 991.45 cm-1. Scanning electron microscope (SEM) images revealed that SGAs exhibited lamellar structures covered with small granules. SGAs exhibited higher residual mass (approximately 12 %) than other samples. The resistant starch content of SGAs increased from 10.1 % to 18.4 % as GEO contents varied from 5 % to 15 %. In vitro digestion tests showed that about 53.21 % of allicin remained in SGA-15 % after 8 h. Therefore, this dual treatment can be a new method for fabricating controlled-release inclusion complexes of guest molecules.

A co-carrier for plasmid DNA and curcumin delivery to treat pancreatic cancer via dendritic poly(l-lysine) modified amylose.

Pancreatic cancer is one of the most lethal malignancies in the world and remains one of the leading causes of cancer related death. For its treatment, a lot of investigations have dealt not only with individual chemotherapy by using polymeric carriers to deliver anticancer drugs, but also with individual gene therapy by using polymeric carriers to deliver nucleic acids such as small interfering RNA (siRNA) and plasmid DNA. However, relatively few studies have been focused on the co-delivery of gene and anticancer drug by multifunctional polymeric carriers for its synergistic therapy. In this work, a DPLL-functionalized amylose (ADP) was prepared by the click reaction between azidized amylose and propargyl focal point poly(l-lysine) dendrons, and then used to co-deliver plasmid pIRES2-EGFP-TNFα and curcumin for pancreatic cancer treatment. Due to the internal hydrophobic cavity of amylose component, ADP could load efficiently curcumin with anticancer activity and showed a sustained release behavior. Moreover, the curcumin-loaded ADP could form colloidally stable nanocomplexes with plasmid DNA in aqueous system due to the existence of cationic poly(l-lysine) dendrons and exhibited high gene transfection efficiency. The in vitro and in vivo tests confirmed the effectiveness of using ADP to co-deliver plasmid pIRES2-EGFP-TNFα and curcumin for synergistic therapy of pancreatic cancer.

Kinetic Study on the Base-Catalyzed Imine-Enamine Tautomerism of a Chiral Biologically Active Isoxazoline Derivative by HPLC on Amylose Tris(3,5-dimethylphenylcarbamate) Chiral Stationary Phase.

Isoxazoline is a nitrogen- and oxygen-containing five-membered heterocyclic scaffold with diverse biological profiles such as antimicrobial, fungicidal, anticancer, antiviral, analgesic and anti-inflammatory activity. Accordingly, the use of this peculiar structural framework in drug discovery is a successful strategy for the development of new drug candidates. Here, a chiral saccharin/isoxazoline hybrid was considered to investigate the tendency of the imine moiety of the heterocyclic ring to tautomerize to the enamine form in the presence of a basic catalyst. The pseudo-first-order rate constants for the base-catalyzed tautomerization process were measured in different solvents and at different temperatures by off-column kinetic experiments based on the amylose (3,5-dimethylphenylcarbamate)-type chiral stationary phase. The kinetic results obtained in this study may be a useful aid in the perspective of designing experimental conditions to control the stereointegrity of these types of pharmacologically active compounds and drive their synthesis toward the preferred, imine or enamine, tautomer.

Effects of different content of EGCG or caffeic acid addition on the structure, cooking, antioxidant characteristics and in vitro starch digestibility of extruded buckwheat noodles.

The effects of different types and content of polyphenol addition on the structure, cooking, antioxidant characteristics and in vitro starch digestibility of extruded buckwheat noodles were investigated in this study. The result showed epigallocatechin-3-gallate (EGCG) was more easily combined with starch to form complex than caffeic acid, and amylose tended to be combined with polyphenols to form more complex. Amylose had a protective effect on polyphenols during extrusion process, which led to a significant increase of polyphenol content and antioxidant activity of extruded noodles. The addition of polyphenol and high amylose corn starch (HACS) improved the cooking quality of extruded buckwheat noodles. The extruded buckwheat noodles with 20 % HACS and 1 % EGCG had the lowest cooking loss of 6.08 %. The addition of EGCG and HACS increased the content of resistant starch and reduced predicted glycemic index (pGI). The noodles with 20 % HACS and 3 % EGCG had the lowest pGI (63.38) and the highest resistant starch (RS) content (61.60 %). This study provides a theoretical basis for the development of low pGI extruded buckwheat noodles.

The interaction and physicochemical properties of the starch-polyphenol complex: Polymeric proanthocyanidins and maize starch with different amylose/amylopectin ratios.

This study investigated the impact of polymeric proanthocyanidins (PPC) on the physicochemical characteristics of maize starch with varying amylose content, and their potential interaction mechanism. PPC with a lower content (1 %) reduced the viscoelasticity of the high amylose maize starch (HAM) system, inhibited amylose rearrangement, and enhanced its fluidity. However, excessive PPC restrained the interaction between PPC and amylose. In contrast to HAM, PPC improved the gelation ability of waxy maize starch (WAM) as PPC concentration was raised. PPC suppressed the recrystallization of starch during storage, and PPC had a superior inhibition influence on the retrogradation of WAM in comparison to HAM. This indicated that amylopectin was more likely to interact with PPC than amylose. Hydrogen bonds were the main driving force between PPC and starch chains, which was clarified by Fourier transform-infrared, nuclear magnetic resonance, X-ray diffraction, iodine bonding reaction, and dynamic light scattering data. Additionally, the mechanism of interaction between PPC and the two starch components may be similar, and variance in physicochemical attributes can be primarily credited to the percentage of amylose to amylopectin in starch.

Fermentation of resistant starch from the starch-ferulic acid inclusion complex compared with high-amylose corn starch.

Fermentation of resistant starch from the starch-ferulic acid inclusion complex, one representative of the starch-polyphenol inclusion complex, was investigated in this study. It was found that this complex-based resistant starch, high-amylose corn starch and the mixture of ferulic acid and high-amylose corn starch were mainly utilized at the initial 6 h as indicated by the gas production and pH. Besides, the supplement of high-amylose corn starch, the mixture and the complex promoted production of short-chain fatty acids (SCFAs), reduced the ratio of Firmicutes/Bacteroidetes (F/B) and selectively stimulated the proliferation of some beneficial bacteria. Specifically, the production of SCFAs in the control and high-amylose starch, mixture and complex groups was 29.33 mM, 140.82 mM, 144.12 mM, and 167.4 mM after fermentation for 48 h, respectively. Moreover, the F/B ratio of those groups was 1.78, 0.78, 0.8 and 0.69, respectively. These results suggested that the supplement of the complex-based resistant starch led to the most SCFAs and the lowest F/B ratio (P < 0.05). Moreover, the complex group had the largest abundance of beneficial bacteria, including Bacteroides, Bifidobacterium and Lachnospiraceae_UCG-001 (P < 0.05). In summary, the resistant starch from the starch-ferulic acid inclusion complex exhibited stronger prebiotic activity than high-amylose corn starch and the mixture.

Solid state cationization reaction of microporous starch with betaine hydrochloride under repeated heating/cooling cycles: Design of a green approach for corn starch modification.

In this research, the cationization process of microporous starch with betaine hydrochloride (BHC) in the presence of H3PO4 (as a catalyst) under heating/cooling cycles was reported for the first time. Granular microporous starch was initially prepared from normal corn starch (NS) through amyloglucosidase treatment. Then, solid state cationization reaction of microporous starch (MS) with betaine hydrochloride (BHC) was performed under repeated dry-heat modification. The cationic microporous starch showed higher substitution degree (0.031) and reaction efficiency (89.1 %) in comparison with cationic starch based on NS (0.021, 60.3 %), which this can be attributed to the increased probability of effective collision between BHC molecules and starch granules after enzymatic treatment. The analysis of cationic starches by FTIR and 13C NMR confirmed the presence of cationic functional groups on starch chains. Further examinations on the modified starches by single and dual treatments were accomplished with respect to morphology, particle size distribution, X-ray powder diffraction (XRD), colour parameters, zeta potential, amylose content, viscosity, solubility, and swelling power. The greenness of the suggested dual treatment (score: 82) in this work was evaluated and compared to a conventional method reported in literature (score: 67) on the preparation of cationic starches.