Properties of octenyl succinic anhydride (OSA) modified starches and their application in low fat mayonnaise

Accepted Manuscript Properties of octenyl succinic anhydride (OSA) modified starches and their application in low fat mayonnaise

Ritika Bajaj, Narpinder Singh, Amritpal Kaur PII: DOI: Reference:

S0141-8130(18)36666-2 https://doi.org/10.1016/j.ijbiomac.2019.03.054 BIOMAC 11884

To appear in:

International Journal of Biological Macromolecules

Received date: Revised date: Accepted date:

1 December 2018 24 February 2019 7 March 2019

Please cite this article as: R. Bajaj, N. Singh and A. Kaur, Properties of octenyl succinic anhydride (OSA) modified starches and their application in low fat mayonnaise, International Journal of Biological Macromolecules, https://doi.org/10.1016/ j.ijbiomac.2019.03.054

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ACCEPTED MANUSCRIPT Properties of octenyl succinic anhydride (OSA) modified starches and their application in low fat mayonnaise. Ritika Bajaj, Narpinder Singh* and Amritpal Kaur Department of Food Science and Technology, Guru Nanak Dev University, Amritsar- 143005, Punjab, India

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*Corresponding author: Narpinder Singh, e-mail: [email protected]

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ABSTRACT

Wheat starch (WS), corn starch (CS), waxy corn starch (WCS), potato starch (PS), sweet

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potato (SP), rice starch (RS) and kidney bean starch (KB) were modified using octenyl succinic anhydride (OSA) and evaluated for various properties. Degree of substitution

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(DS) showed significant increase with increase in amylose (AM) content. OSA modified starches showed higher paste viscosities compared to their native counterparts. OSA

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groups acted majorly on the surface and caused some superficial pores, but crystalline pattern was not significantly altered for all starches. OSA modified starches were used in

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preparing low fat mayonnaise by substituting 75% fat. OSA modified starches enhanced the emulsifying properties of mayonnaise. Mayonnaises prepared using OSA modified

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starches were evaluated for phase separation brightness (L*), color index (dE), rheological parameters (G' and G''). Mayonnaises prepared using OSA modified starches

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showed higher G' and exhibited gel like structure. Fat substituted (FS) mayonnaise was preferred over full fat (FF) mayonnaise by the consumers. No significant effect of fat

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substitution was observed on particle size and phase separation for all mayonnaise samples.

KEYWORDS: Starch, octenyl succinic anhydride (OSA), mayonnaise, rheology, pasting properties. ABBREVIATIONS. WS, wheat starch; CS, corn starch; WCS, waxy corn starch; PS, potato starch; SP, sweet potato; KB, kidney bean; RS, rice starch; DS, degree of substitution; AM, amylose; OSA, Ocetyl Succinic Anhydride; FS, Fat-substituted; PV, peak viscosity; FV, final viscosity; BD, breakdown viscosity; SV, setback viscosity; PT, pasting temperature; To, onset transition temperature; Tp, peak transition temperature;

ACCEPTED MANUSCRIPT Tc, conclusion transition temperature; ΔHgel, enthalpy of gelatinization; RDS, rapidly digestible starch; SDS, slowly digestible starch INTRODUCTION Starch, a natural carbohydrate polymer acts as most useful ingredient in food industries.

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Wide ranging applications of starches were explored in different industries like fillers in

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paper industry, thickeners in soup industry. Along with applications, starches showed

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limitations like reduced solubility in water, higher retrogradation tendencies, poor

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functional properties and processing tolerances limiting their usage in food industries. Modification of starch refers to techniques used to create new products by modifying

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important characteristics to improve their functional properties which can help tailoring

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starch for certain applications in food industries (1). Modifications were classified as physical, chemical and enzymatic based on the type of technique required and properties

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to be altered. When using OSA for modification, hydrophilic character of native starch

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got altered to hydrophobic character by substitution of octenyl molecules, resulting in making the whole molecule amphiphilic. These modified polymers showed broad range applications,

particularly

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of

in

the

pharmaceuticals,

emulsification,

enamels,

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encapsulation, and gel production. OSA modified starches are not limited only to the food industries; instead, they can be utilized in any industry where stable emulsions are required (2). Applications of OSA modified starches in industries would be influenced by functional properties of polymer used for modification. During OSA modification, slightly basic conditions were required for maximum DS which would further contribute towards reduction in intermolecular

ACCEPTED MANUSCRIPT bonding between starch chains by replacing OH groups that favor swelling of granules and allowed penetration of water molecules within swollen starch granules. OSA modified starch performed better emulsification properties because of substitution of functional groups (ocetyl succinic groups) that behaved both as hydrophilic and

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hydrophobic character (3). Industrial applications of OSA modified starches includes in

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production of sauces, puddings, and baby foods. Various starch sources and techniques

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have been utilized to develop fat-mimetic. The chemical modifications help in reducing limitations of starch and enhancing properties including binding, thickening, gelling and

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dispersion. For instance, starch modification facilitates formation of gels which showed

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higher stability at low pH and also showed greater resistance towards shearing altering gelatinization temperatures making them softer or firmer as per constraint.

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US FDA has permitted OSA to be used in foods at a level of 3%. Substitution of octenyl

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group offers hydrophobic nature to starch which weakens internal bonding and holds starch granules together. The increasing demand for functional foods that were found

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beneficial for human health has been encouraged by the consumers (4).

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Demand for low-fat products has increased but development of such products with reduced fat is challenge as it adversely affects organoleptic properties like appearance,

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odor, taste, texture and mouth feel (5, 6). Mayonnaise is oil-in-water type emulsion, containing higher amount of fat (70-80%), egg yolk, additives. For preparation of FS mayonnaise, it becomes essential to reduce the discrete phase and also add to the water content. Modified starches mainly OSA modified are thus utilized as fat replacer due to their low cost, tasteless, and unique character which provides creamy texture and required mouth feel (7).

ACCEPTED MANUSCRIPT The study was designed to modify starches from various cereals, tubers and beans, evaluating their physico-chemical, morphological, pasting and thermal properties and comparing with native starches. These OSA modified starches were then utilized in preparation of FS mayonnaise by replacing fat up to 75% of the FF mayonnaise (FFM)

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and then evaluated for its rheological and sensory properties.

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MATERIALS AND METHODS

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Materials. Varieties of different botanical sources (WS, CS, SP, PS, RS, KB and WCS) commercially available were obtained from local market. Starches from different

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botanical sources were extracted using standard laboratory methods as described

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previously by Bajaj et al. (8).

Preparation of octenyl succinic anhydride (OSA) esterified starches. 20% (w/w)

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starch suspension was prepared with agitation for 10 min. pH of suspension was then

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altered to 4, 6 and 8 using 3% NaOH solution. 3% OSA (dwb) of total weight of suspension was added slowly during reaction time of 3 h. After 3 h, solution pH was

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reduced to 6.5 using 3% HCl solution and then it was centrifuged, rinsed two times with

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distilled water and then 70% aqueous alcohol, after which it was oven-dried at 40C for 24 h. The dried starch was then ground using pestle and mortar and stored in air tight

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containers.

Degree of substitution (DS) and amylose content (AM). Five grams (dwb) of OSA modified starch was suspended in 25 mL of HCl-IPA solution (2.5M) with intermittent stirring for 30 min followed by filtration using vacuum filter and washing with 90 % IPA solution was done until no Clˉ could be noticed by titrating with 0.1 M AgNO3 solution. The solution was then re-suspended in distilled water (300 mL) and heated for 20 min in

ACCEPTED MANUSCRIPT water bath at high temperature. The suspension was then titrated using 0.1 M NaOH solution using phenolphthalein as indicator. Native starch was treated same way and taken as control. DS was calculated using the following equation: DS = 0.162 x (AxM)/W 1 - [0.210x(A xM)/W]

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where A is the titration volume of NaOH (mL); M is the molarity of NaOH solution and

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W is the dry weight (g) of OSA starch.

AM of native and OSA modified starches were determined using method as described by

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Williams et al. (9).

Granule size distribution and morphology. Granular size distribution of native and

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OSA modified starches were evaluated using particle size analyzer (S3500, Microtrac

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Inc., USA).

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Morphological characteristics of starches were analyzed using digital scanning electron microscope, (Model EVOLS10, ZEISS, Oberkochen, Germany). 1% starch suspension

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was prepared in alcohol causing dehydration of starch before analysis. The dehydrated samples were then sprayed on metal stub covered with conductive adhesive tape and

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coated with 20 nm silver layer using Metalizer (Model SCD 050, Balzers, Liechtenstein).

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Crystalline structure. X-ray diffractograms of native and OSA modified starches were studied fully equilibrated at 25 C for 48 h before analysis using analytical diffractometer (Pan Analytical, Phillips, Holland), operating at 40 kV and 30 mA. Scanning speed of goniometer during analysis was adjusted to 4/min with step size of 0.02 from 4 to 40. Relative crystallinity was determined as the ratio of area of crystalline region to total area of diffractogram (2θ).

ACCEPTED MANUSCRIPT Thermal properties of starch. Starch (3-5 mg, dwb) was weighed into aluminum pan of quantity 40 μL and water was adjusted to sample so as to achieve 70% starch suspension (w/w). Pans were then sealed and kept for 1 h at room temperature to achieve uniform suspension before heating using heat flow DSC by Mettler Toledo 822e. The heating

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profile used was set from 40 to 110C at rate of 10C/min followed by cooling to 10C at

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the same rate. Onset temperature (To), peak temperature (Tp), conclusion temperature

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(Tc), and enthalpy of gelatinization (ΔHgel) were calculated using Stare Software for

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thermal analysis Ver. 8.10.

Pasting properties of starch. Pasting parameters were determined using dynamic

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rheometer (Anton Paar Rheo Plus/32 model MCR-301) as described by Bajaj et al. (8). Paste viscosities (PV, BV, FV and SV) and pasting temperature (PT) were recorded.

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In-vitro starch digestibility. RDS, SDS and resistant starch were determined using

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slight modification to the method of Englyst et al. (10).

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Preparation of low fat mayonnaise. Low fat mayonnaise was prepared by substituting oil (upto 75%) used in FF mayonnaise with gelatinized OSA modified starches. Firstly,

min.

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starch suspension 20% (w/w) was gelatinized in water bath maintained at 90°C for 30

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FF and FS mayonnaise (100 g) was prepared using ingredients as given in (Table 1). Firstly, powdered ingredients, egg white and starch paste (gelatinized) were added and mixed using high speed hand blender operating at speed 4 for 2 min., followed by addition of water and one third of vinegar. Further, oil was drizzled in fine stream to this mixture alternating with addition of remaining vinegar. Both vinegar and oil was added slowly with intermediate mixing over period of 5 min. until color of mayonnaise changed

ACCEPTED MANUSCRIPT from yellow to white upon mixing. FF mayonnaise was prepared using all ingredients except starch paste. The prepared mayonnaise was then kept under refrigerated temperature for 24 h before further analysis. Table 1. Ingredients for the preparation of 100 g of FF and FS mayonnaise. Full Fat (FF)

Fat Substituted (FS)

Oil

80

Vinegar

9

Water

3

Eggs

18

18

Salt

2

2

9.6

9.6

0.02

0.02

0.2

0.2

-

40

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Ingredients

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Sugar

Guar gum

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Citric acid

9 28

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Starch paste (20% w/w)

35

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Rheological properties of mayonnaise. Rheological properties were determined using

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strain sweep test (0.1-100%) operating using dynamic rheometer (Anton Paar Rheo Plus/32 model MCR-301) equipped with 40 mm parallel plates at fixed frequency of 5 Hz (11). Dynamic frequency sweep (0.1 - 100 Hz) operating at a constant strain of 0.5% was used to study viscoelastic behavior of prepared samples. Particle size measurement. 0.05 g of mayonnaise samples were dispersed with 0.1% sodium dodecyl sulfate solution (100 mL). The samples were gently mixed before

ACCEPTED MANUSCRIPT analysis and analyzed using particle size analyzer particle size analyzer (Microtrac S3500, Microtrac Ins. Ltd., USA). Color characteristics of mayonnaise. Color characteristics (L*, a*, b*) and color difference index (dE) for FF and FS mayonnaises was recorded using Ultra Scan VIS

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Hunter Lab (Hunter Associates laboratory Inc., Reston, VA, U.S.A.).

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Phase separation of mayonnaise. 20 g (F0) of mayonnaise was taken in pre-weighed

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centrifuge tubes (50 mL), sealed tightly and stored for 48 h at 50 C. After 48 h, samples

measured using following equation:

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Separated oil portion (%) = (F0- F1/F0) ×100

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were centrifuged at 3000 rpm/10 min. Separated oil phase (F1) was collected and

Sensory characteristics of mayonnaise. Sensory evaluation was done according to the

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methodology of Singh et al. (12). The panelists evaluated randomly presented

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9-point hedonic scale.

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mayonnaise for color, aroma, taste, texture, consistency and overall acceptability using a

Statistical analysis. All observations were taken in triplicate. Results were subjected to

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analysis of variance (ANOVA) and expressed as mean± standard deviation using Minitab Statistical Software (MINITAB 14.12.0, State College, PA).

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RESULTS AND DISCUSSION Degree of substitution (DS) and amylose content (AM) DS refers to substitution of OH groups per glucose molecule, which was found to be associated to degree of esterification. Starches from different botanical sources i.e. cereals, tubers and beans modified using OSA (3%, db) at different reaction pH were evaluated for DS analysis, results are summarized in Table 2a. The highest DS was observed for KB (0.060) at

ACCEPTED MANUSCRIPT pH 8 followed by PS (0.028) and SP (0.029) while WCS showed the lowest degree of esterification (0.018).Statistical analysis also revealed significant effect of reaction pH on DS and AM among starches from different botanical sources as reported in Table 2b. Sweedman et al. (13) suggested that varying degree of esterification might be due to changes in

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composition, morphology and structure. The distribution of hydroxyl groups was reported to

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be non- uniform among the granules, while OSA groups preferably got attached at the surface

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of granules. He et al. (14) reported that OSA esterification occurred majorly among amylose molecules as compared to amylopectin. Therefore, difference in structure between amylose

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and amylopectin was considered as the principal cause for different activities of OSA. The

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results were in accordance to AM content. DS increased with increase in AM content (Table 2a). KB showed highest AM content (38.43%) at pH 8 with highest DS of 0.060 while lowest

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DS was observed by WCS with minimum DS of 0.018. The substitution of OSA groups was

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also dependent upon the reaction conditions where pH of the reaction played a significant role. NaOH initiated reaction acted as a catalyst by formation of OH - ions along with starch

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polymer (15). However, DS decreased significantly for pH between <7.5 and >8. This might

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be due to the fact that at pH >8 side reactions were favored whereas when treated at pH <7.5, OH groups were not stimulated for nucleophilic attack. Thus pH 8.0 was found to be most

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suitable to have maximum substitution. The statistical analysis also revealed strong relationship among native and OSA modified starches for granule size, AM content and relative crystallinity. Average granule size and morphological characteristics Granule size of native and OSA modified starches are summarized in Table 2a. Starches from different sources showed wide variation in the distribution pattern and percent

ACCEPTED MANUSCRIPT volume of the granules. PS showed the highest granule size of 35.74 µm with wide granule size distribution compared to other starches whereas RS showed the smallest granule size of 5.41 µm with narrow range of granule size distribution. The statistical analysis also showed significant effect of OSA modification on average granule size of

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native and OSA modified starches. OSA modified starches showed decrease in granule

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size among all the starches as compared to their native counter parts except PS. PS

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showed significant increase in average granule size ranging from 35.35 µm at pH 4 to 36.11 µm at pH 8 whereas WS, RS, CS showed decrease in granule size as compared to

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their native counterparts. Both native and modified OSA starches were insoluble in water

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and thus, esterification becomes a heterogeneous process. OSA reacted on the surface of granules causing significant changes on the surface and thus size of the granules.

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Scanning Electron Microscopy (SEM) has been used extensively to study changes in

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structure caused during modifications. SEM of native and modified starches are represented in Figure 1. Most of the changes especially during chemical modifications

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like hydroxyl propylation took place at central region of starch granule (16, 17). SEM for

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OSA modified starches revealed that modification caused changes at structural level. OSA modified starch granules displayed rough surfaces and edges were lost upto some

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level with presence of superficial pores developed on the surface which were index of higher swelling power and water retention in the granules, thus improving its hydration characteristics and viscosity. OSA modified starches showed pronounced depressions on the surface for all starches (Figure 1) especially in starches with higher DS value (KB with 0.060). Crystalline structure

ACCEPTED MANUSCRIPT XRD patterns of both native and OSA modified starches are represented in Figure 2 and relative crystallinity is summarized in Table 2a. Starches exhibit semi crystalline structure and showed a typical diffraction pattern according to the botanical origin. Cereal starches (WS, CS, WCS and RS) showed typical A-type pattern with major

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diffraction peaks at 15, 17, 18 and 23 (2θ) while B-type diffraction pattern with

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strong diffractions at 2θ = 5.6, 14.5, 17.1, 22.3 and 24.2 was displayed by

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tubers (PS and SP). In addition, bean starches (KB) displayed C-type diffraction pattern

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which is mixture of both A and B type and also exhibited weak diffraction at 2θ= 20, which was indicative of V-shaped amylose-lipid complexes (18). Crystallinity of starches

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was the result of branched amylopectin structure located mainly in the crystalline domain within the granule, while amorphous region was mainly associated with amylose. No

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significant change was observed in the diffraction pattern for all starches mainly those

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with low DS whilst, when treated at lower pH these granules of modified starches were

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damaged to some extent by the modification process. Starches when treated at higher pH (pH 8) showed slight variation in intensities of diffraction peaks thus, caused differences

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in relative crystallinity, indicating little effect on crystalline structure of native starch.

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OSA modified KB starch with a high DS (0.060), showed decrease in relative crystallinity from 38.30 % to 18.74% while WCS with lowest DS of 0.018 showed significant increase in RC from 3.28% to 10.55 % OSA modification has been reported to occur in amorphous region causing minimal change in relative crystallinity (18). Results observed revealed that OSA modification caused disruptive effect on the surface with least effect on internal crystalline structure, thus, allowing water molecules to interact with starch granules, and increasing swelling power for all sources (20).

ACCEPTED MANUSCRIPT Thermal properties Thermal properties of both native and OSA modified starches are represented in Table 3a. Cereal starches (WS, CS, RS and WCS) were characterized by two distinct endotherms while tubers (PS and SP) and KB starch displayed one distinct endotherm in

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their respective thermograms. Transition temperatures (To, Tp and Tc) and enthalpy of

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gelatinization (Hgel) of OSA modified starches decreased with increase in reaction pH.

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To, Tp, Tc and Hgel for crystallite melting ranged from 54.38 to 72.57 C, from 60.63 to

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75.58 C, from 64.52 to 80.26 C and from 1.89 to 14.16 J/g respectively. Statistical analysis revealed significant differences in thermal properties of native and OSA

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modified starches as shown in Table 3b.OSA modified cereal starches, especially RS showed lower transition temperatures at pH 4 with lower DS as compared to pH 8. This

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reduction in transition temperatures especially Tp could be attributed to the insertion of

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octenyl succinic groups into much stabilized starches resulting into destabilization of

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starch chains causing linearity in the chains. Hgel also showed a significant decrease after OSA modification which showed that more energy for melting of crystalline

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structure of the modified starches was required. This could thus be concluded that OSA

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modification led to structural disruption and weakening of starch granules. While, OSA modified PS and SP starches showed much higher To and Tp than their native counterparts than observed for other sources. This may be contributed to the dense material present in the inner surface of PS and also ordered structure on outer surface which require higher temperatures for weakening of hydrogen bonds compared to other sources. Bulky groups (OSA) when incorporated into the backbone of starch enhanced

ACCEPTED MANUSCRIPT structural elasticity and contributed to reduction of transition temperature (21). Similar results have been reported for OSA modified WCS (19) and RS (22). Pasting properties Pasting parameters such as PV, BV, TV, FV, SV and PT of both native and OSA

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modified starches are listed in Table 4a. These properties were indicative of behavior of

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starches during hot or cold processing of food products. Also, relationships between structural and functional properties could be easily expressed through pasting properties.

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Compared to native starches, no significant difference was observed for PT among OSA modified starches, whereas, starches from different sources showed significant

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differences. Lower PT for tubers (PS and SP) compared to cereals (WS, RS and CS)

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indicated ease of gelatinization, however, higher gelatinization temperature contributes to better thermal stability. The statistical analysis also showed significant effect of OSA

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modification on pasting properties of native and OSA modified starches as reported in

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Table 4b. PV referred to as an important parameter of starch which represented maximum swelling of starch granules upon heating in starch suspension. Among all sources, PS and

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SP showed maximum PV which could be attributed to the presence of esterified

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phosphate group which got ionized upon heating opening branched amylopectin molecules and increased swelling (24). Substitution of bulky octenyl groups increased PV for all starches which may be attributed to declining effect of intra and intermolecular forces and thus restricted incorporation of water within the starch molecules. Hydrophobic character of OSA also contributed to increase in viscosity of starch. This attribute was found desirable during manufacture of mayonnaise with enhanced emulsion stability.SV represented retrogradation tendency of starch granules upon cooling. OSA

ACCEPTED MANUSCRIPT modified starches from different sources showed significantly lower SV than native starches which could be due to obscurity after modification in reassociation of amylose molecules due to large and bulky succinyl groups (7). These observations revealed that due to presence of bulky octenyl succinic ester, having characteristic hydrophobic

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properties could led to higher paste viscosities which acted as desirable property for

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products like mayonnaise.

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in -vitro starch digestibility

RDS, SDS and resistant starch of native and OSA-modified starches with different DS

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values are shown in Table 5a. Starch has been classified into three fractions according to

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Englyst and Englyst (25) on the basis of extent and rate of digestibility mainly RDS, SDS, and resistant starch. Variations in the digestibility of starches among different

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sources could be accredited to numerous factors, such as botanical origin, granular

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morphology, amylose-amylopectin ratio, crystallinity and molecular structure (26). As shown in Table 5a, subsequent reduction in RDS was observed with substantial increase

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in DS among OSA modified starches. RDS decreased from 85.92% (native) to 54.71%

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for modified starches with DS of 0.028. Comparatively, SDS increased from 1.22% (native) to 35.75%, and resistant starch increased from 2.53% to 26.76%. OSA-WCS

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showed decrease in RDS from 81.41% for native to 76.30% of modified starch with DS of 0.018 while SDS increased from 10.51% to 14.24%, and resistant starch from 8.57% to 9.46%. Han and Be Miller (28) reported significant increase in % SDS and resistant starch upon OSA modification. He et al. (14) also reported that OSA modified starches acted similarly to both amyloglucosidase and pancreatic -amylase, and delay was observed for the release of enzymes from the substrate due to physical obstruction or

ACCEPTED MANUSCRIPT amphiphilic character incorporated by bulky groups of OSA. The statistical analysis also revealed significant effect of modification on digestibility studies of native and OSA modified starches as reported in Table 5b. Rheological properties of mayonnaise

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Rheological properties of FF mayonnaise (control) and FS mayonnaise prepared using

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OSA modified starches are represented in Figure 4. Rheological parameters were

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conducted with controlled strain of 0.5% within the LVR range. It was observed that both FF and FS mayonnaise prepared using OSA modified starches were characterized with

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gel like structure as G′ was found to be significantly higher than G′′ in all the samples.

fat particles to form gel like network.

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This was mainly attributed to network development of egg proteins which incorporated

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All FS mayonnaises prepared using OSA modified starches showed less tan ẟ than FF

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mayonnaise, thus, showed less gel like structure compared to FF mayonnaise. This also proved that with increase in fat substitution, viscosity decreases giving liquid

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characteristic which may be due to higher water based continuous phase. Ma and

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Barbosa-Cánovas (29) reported that more the fat content, more G'. However, fat droplet size also found relevant effect on storage modulus (G′) resulting in more solid-like

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structure (30). Higher the fat substitution using OSA modified starches, less resisted would be the fat globules to force applied during the measurement. Thus, all mayonnaise samples showed shear thinning behavior however, FS mayonnaise prepared using OSA modified starches showed structure which could be easily disrupted or could not regain their original structure. This might be due to hydrophobic interactions and gel like network formation as micro-particulate gel in both FF and FS mayonnaise (31).

ACCEPTED MANUSCRIPT Particle size, color and phase separation measurement of FF and FS mayonnaise Images of FF and FS mayonnaise are represented in Figure 3. Particle size of fat molecules in FF and FS mayonnaise prepared using OSA modified starches was determined and summarized in Table 6. It was observed that fat particle size in FF

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mayonnaise was highest compared to FS mayonnaise. The lesser particle size of fat

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globules in FS mayonnaises could be attributed to the fact that OSA modified starch

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enhanced the emulsifier properties and also added to egg yolk proteins to stabilize the emulsion more than in FS mayonnaise causing smaller size fat globules.

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L* and a* values of both FF and FS mayonnaises prepared using OSA modified starches

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were determined and it was observed that all FS mayonnaises showed higher L* and a* values than FF samples. Generally, it was observed that samples with higher fat content

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and smaller fat droplet size contributes to higher L* and a* values resulting in good

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refraction of light (3). Thus, in case of FS mayonnaises, addition of OSA modified starch

FF mayonnaises.

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and smaller fat globules might have contributed higher L* and a* values as compared to

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Mayonnaise samples prepared using OSA modified starches did not show phase separation while FF mayonnaise showed clear phase separation. This might be due to

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emulsion stabilization effect of OSA and prevention of breaking of fat globules during centrifugation, thus, resulting in no phase separation. Sensory characteristics of FF and FS mayonnaise Color, aroma and taste differences between FF and FS mayonnaise prepared using OSA modified starches were not professed by the panelists as represented in Table 7. This showed that replacement of fat by OSA modified starches did not affect overall acceptability of

ACCEPTED MANUSCRIPT mayonnaise. However, consistency of FS-RS, FS-CS, FS-SP and FS-KB were more preferred over all other samples. Texture and mouth feel of FS mayonnaises prepared using OSA modified starches was preferred more over FF mayonnaise. Insignificant differences were observed for textural attributes which could be due to high consistency of OSA

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modified starches which obscured the negative attributes of FF mayonnaise. FS mayonnaise

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showed insignificant changes on thickness and spoon ability compared to FF mayonnaise. It

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was also observed that FS mayonnaise prepared by substituted with OSA modified starches showed higher viscosity which helped in masking the negative effects on textural

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characteristics (7).

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Secondly, FS mayonnaises with OSA starch substitution received higher scores for aroma. Acetic acid or vinegar added was perceived more for FS mayonnaises compared

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to FF mayonnaise showed preference of panelists for vinegar like aroma (7). In case of

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overall acceptability, least scores were received by FS- WCS and maximum was obtained for FS-CS whereas insignificant differences were observed for all other samples, which

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showed that starches from various cereals, tubers and beans could be used to prepare

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mayonnaises using OSA modified starches with lesser fat content. Thus, from all the observations, it could be concluded that OSA modified starches could successfully

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imitate structure, texture and flavor of a real mayonnaise. ACKNOWLEDGEMENT RB acknowledges grant fellowship from UPE scheme.

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(7) T.M. Ali, S. Waqar, S. Ali, S. Mehboob, A. Hasnain. Comparison of textural and

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sensory characteristics of low‐ fat mayonnaise prepared from octenyl succinic anhydride

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modified corn and white sorghum starches. Starch‐ Stärke, (2015), 67, 183-190. (8) R. Bajaj, N. Singh, A. Kaur, N. Inouchi. Structural, morphological, functional and digestibility properties of starches from cereals, tubers and legumes: a comparative study. J. Food Sci. Technol. (2018), DOI.org/10.1007/s13197-018-3342-4. (9) P.C. Williams, F.D. Kuzina, I. Hlynka (1970). A rapid colorimetric procedure for estimating the amylose content of starches and flours. Cereal Chemistry, 47, 411–420.

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(12) J. P. Singh, A. Kaur, K. Shevkani, N. Singh. Influence of jambolan (Syzygium cumini) and xanthan gum incorporation on the physicochemical, antioxidant and sensory

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properties of gluten-free eggless rice muffins. Int. J. Food Sci. Technol. (2015), 50, 1190-

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physicochemical properties of octenyl succinic anhydride modified starches: A

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review. Carbohydr Polym. (2013), 92, 905-920.

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(14) G.Q. He, X.Y. Song, H. Ruan, F. Chen. Octenyl succinic anhydride modified early indica rice starches differing in amylose content. J. Agric. Food Chem. (2006), 54, 2775-

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(15) U. Funke, M.G. Lindhauer. Effect of reaction conditions and alkyl chain lengths on the properties of hydroxyalkyl starch ethers. Starch‐ Stärke, (2001), 53, 547-554. (16)L. Kaur, N. Singh, J. Singh. Factors influencing the properties of hydroxypropylated potato starches. Carbohydr Polym. (2004), 55, 211-223. (17) H.R. Kim, A.M. Hermansson, C.E. Eriksson. Structural characteristics of hydroxypropyl

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substitution. Starch‐ Stärke, (1992), 44, 111-116.

depending

on

their

molar

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functional properties of waxy maize and amaranth starches. Carbohydr Polym.

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(2007), 68, 447-456.

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octenyl succinic anhydride modified potato starch. Food Chem. (2009), 114, 81-86. (21) O.S. Lawal. Succinyl and acetyl starch derivatives of a hybrid maize:

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Physicochemical characteristics and retrogradation properties monitored by differential

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scanning calorimetry. Carbohydr Res. (2004), 339, 2673-2682. (22) D. Thirathumthavorn, S. Charoenrein. Thermal and pasting properties of native and

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acid-treated starches derivatized by 1-octenyl succinic anhydride. Carbohydr Polym.

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(2006), 66, 258-265.

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succinylation on the structure and properties of high-amylose maize starch. Carbohydr

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Polym. (2011), 84, 1276-1281. (24) T. Noda, N.S. Kottearachchi, S. Tsuda, M. Mori, S. Takigawa, C. Matsuura-Endo, H. Yamauchi. Starch phosphorus content in potato (Solanum tuberosum L.) cultivars and its effect on other starch properties. Carbohydr Polym. (2007), 68, 793-796. (25) K.N. Englyst, H.N. Englyst. Carbohydrate bioavailability. British J Nutr. (2005), 94, 1-11.

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red gram. Int. J. Food Prop. (2014), 17, 1469-1481.

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(28) J.A. Han, J.N. Be Miller. Preparation and physical characteristics of slowly digesting

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modified food starches. Carbohydr Polym. (2007), 67, 366-374.

(29) L. Ma, G.V. Barbosa-Cánovas,. Rheological characterization of mayonnaise. Part II:

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Flow and viscoelastic properties at different oil and xanthan gum concentrations. J Food

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Eng. (1995) 25, 409-425.

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(30) M. Langton, E. Jordansson, A. Altskär, C. Sørensen, A.M. Hermansson.

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Microstructure and image analysis of mayonnaises. Food Hydrocoll.(1999)13, 113-125. (31) K. Maruyama, T. Sakashita, Y. Hagura, K. Suzuki. Relationship between rheology,

PT

particle size and texture of mayonnaise. Food Sci and Technol. Res. (2007), 13, 1-6.

CE

Legends to Figures and Tables

AC

Figure 1. SEM of native and OSA modified starches. Figure 2. X-ray diffractograms of native and OSA modified starches. Figure 3. Images of FF and FS OSA modified starch mayonnaises. Figure 4. Dynamic rheological spectra of FF and FS OSA modified mayonnaises. Table 1.

Ingredients for preparation of 100 g of full fat (FF) and fat substituted

mayonnaises.

ACCEPTED MANUSCRIPT Table 2. Physicochemical properties of native and OSA modified starches. Table 3. Thermal properties of native and OSA modified starches. Table 4a. Pasting properties of native and OSA modified starches from different

T

botanical sources

IP

Table 5a. in-vitro digestibility of native and OSA modified starches from different

CR

botanical sources

Table 6. Particle size, color characteristics and phase separation of FF and FS

US

mayonnaise

AN

Supplementary Table 2b. F values from ANOVA analysis of the data (botanical source

M

verses modification) reported in Table 2a

Supplementary Table 3b. F values from ANOVA analysis of the data (botanical source

ED

verses modification) reported in Table 3a

PT

Supplementary Table 4b. F values from ANOVA analysis of the data (botanical source

CE

verses modification) reported in Table 4a Supplementary Table 5b. F values from ANOVA analysis of the data (source verses

AC

modification) reported in Table 5a Supplementary Table 7. Sensory characteristics of FF and FS mayonnaise

ACCEPTED MANUSCRIPT Table 2. Physico chemical properties of native and OSA modified starches.

SP

KB

bc 0.020 ± 0.00 0.022b±0.00 0.028c±0.00 0.017b±0.00 0.019ab±0.00 0.023b±0.00 0.025c±0.00 0.027c±0.00 0.027c±0.00 0.019bc±0.00 0.021b±0.00 0.024b±0.00 0.010a±0.00 0.017a±0.00 0.018a±0.00 0.020bc±0.00 0.025e±0.00 0.029c±0.00 0.039d±0.00 0.053d±0.00 0.060d±0.00

37.57e±0.05 37.45f±0.03 32.60f±0.04 26.53e±0.05 37.52e±0.04 36.93e±0.01 30.57e±0.05 24.14d±0.04 34.28d±0.03 33.51d±0.01 28.12d±0.02 20.36c±0.02 32.64b±0.01 32.24c±0.02 26.19c±0.04 18.19b±0.03 29.21a±0.04 27.37a±0.05 19.43a±0.03 15.23a±0.05 33.12c±0.04 32.19c±0.04 26.47c±0.05 20.71c±0.22 38.30f±0.02 31.45b±0.03 24.32b±0.04 18.74b±0.03

CE

WCS

RC (%)

d

IP

T

16.25 ±0.24 17.0d±0.17 18.69d±0.20 23.02d±0.14 21.54e±0.21 22.41e±0.23 24.43e±0.17 26.35e±0.13 9.06b±0.28 8.84b±0.08 10.41b±0.15 14.47b±0.31 26.38f±0.10 28.34f±0.12 29.34g±0.13 30.41e±0.19 3.28a±0.12 4.54a±0.13 6.23a±0.07 10.55a±0.14 14.06c±0.04 14.33c±0.13 16.72c±0.07 20.22c±0.06 29.62g±0.16 30.96g±0.12 32.41g±0.16 38.43g±0.21

CR

35.74 ±0.12 36.35f±0.10 36.69f±0.13 36.11f±0.30 19.19c±0.16 23.07d±0.14 22.92d±0.11 22.55d±0.12 5.41a±0.02 4.86a±0.00 4.85a±0.01 4.83a±0.00 18.19b±0.12 17.40b±0.09 16.75b±0.08 16.64b±0.09 18.26b±0.16 17.92b±0.62 16.15b±0.11 16.67b±0.12 18.57bc±0.02 19.52c±0.03 19.73c±0.07 18.40c±0.01 25.50d±0.04 26.29e±0.05 27.68e±0.03 28.34e±0.04

US

CS

Amylose content (%)

e

AN

RS

Avg. size (µm)

M

WS

N 4 6 8 N 4 6 8 N 4 6 8 N 4 6 8 N 4 6 8 N 4 6 8 N 4 6 8

DS

ED

PS

pH

PT

Sample

*Values represent mean± standard deviation. Means with different superscripts in the same column are

AC

significantly different (P ≤ 0.05); the number of replications (n), n≥3.

ACCEPTED MANUSCRIPT Table 3. Thermal Properties of native and OSA modified starches. Peak I Sample PS

WS

RS

CS

WCS

SP

KB

Peak III

Peak II Hgel(J/g)

To (C)

Tp (C)

Tc (C)

Hgel(J/g)

To (C)

Tp (C)

Tc (C)

Hgel(J/g)

9.47 ±0.05 10.96f±0.09

NA NA

NA NA

NA NA

NA NA

NA NA

NA NA

NA NA

NA NA

71.03c±0.20 72.19c±0.08 67.24b±0.12 66.41b±0.08 66.33a±0.07 65.80a±0.15 72.46c±0.14 69.52c±0.05 68.43b±0.09 67.52b±0.10 77.85f±0.08

12.39d±0.14 14.16e±0.14 9.37c±0.24 6.78b±0.50 6.54b±0.10 5.49b±0.08 7.41a±0.12 4.09a±0.05 2.68a±0.19 1.89a±0.13 11.78e±0.56

NA NA 93.55a±0.13 92.97a±0.16 92.87a±0.08 89.34a±0.09 NA NA NA NA NA

NA NA 98.54a±0.17 96.86a ±1.66 95.81a ±0.17 92.49 a ±0.14 NA NA NA NA NA

NA NA 102.9a±0.06 100.19a±2.11 98.86a±0.19 96.46a±0.16 NA NA NA NA NA

NA NA 0.71a±0.18 0.45a±0.07 0.38a±0.04 0.38a±0.04 NA NA NA NA NA

NA NA 86.96a±0.28 84.18a±1.94 82.86a±0.19 82.13a±0.05 NA NA NA NA NA

NA NA 85.46a±0.08 82.30a±1.81 81.34a±0.15 80.13a±0.09 NA NA NA NA NA

NA NA 82.13a±1.85 78.96a±2.25 77.61a±0.06 75.83a±0.06 NA NA NA NA NA

NA NA 0.75a±1.25 0.82a±0.07 0.76a±0.12 0.62a±0.05 NA NA NA NA NA

73.27 ±0.20

f

77.35 ±0.28

e

10.01 ±0.04

NA

NA

NA

NA

NA

NA

NA

NA

72.55f±0.09 70.50e±0.36 69.35d±0.24 68.72d±0.09 68.77d±0.13 67.61c±0.21 70.48e±0.07 70.90e±0.07

75.34e±0.09 74.68d±0.12 73.52d±0.28 72.99d±0.06 73.21d±0.25 72.36c±0.06 76.36e±0.02 76.64e±0.09

7.92bc±0.55 6.77c±0.40 8.44b±0.24 7.46c±0.31 7.17b±0.56 5.29b±0.00 10.30d±0.06 10.34e±0.07

NA NA NA NA NA NA NA NA

NA NA NA NA NA NA NA NA

M

NA NA NA NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA NA NA NA NA NA NA

NA NA NA NA NA NA NA NA

66.48e±0.05

71.59e±0.03

76.59f±0.05

12.48d±0.05

NA

NA

NA

NA

NA

NA

NA

NA

f

f

f

e

NA NA NA NA NA

NA NA NA NA NA

NA NA NA NA NA

NA NA NA NA NA

NA NA NA NA NA

NA NA NA NA NA

NA NA NA NA NA

NA NA NA NA NA

pH

To (C)

Tp (C)

Tc (C)

N 4

a

57.72 ±0.10 57.68a±0.20

c

67.23 ±0.14 61.26a±0.09

a

64.52 ±0.16 65.44a±0.19

6 8 N 4 6 8 N 4 6 8 N

62.19c±0.05 63.28c±0.07 60.16c±0.06 58.35b±0.19 58.24b±0.05 57.31b±0.09 58.64b±0.26 58.73c±0.04 56.54a±0.13 54.38a±0.21 69.80f±0.28

66.42c±0.05 68.78d±0.05 63.31a±0.21 62.58b±0.27 62.45b±0.07 61.22b±0.11 64.29b±0.23 63.39c±0.05 61.67a±0.09 60.63a±0.17 74.12f±0.24

4

f

69.57 ±0.06

f

6 8 N 4 6 8 N 4

67.28f±0.43 65.53e±0.26 67.00e±0.09 64.74d±0.11 64.24d±0.32 63.82d±0.09 65.62d±0.04 65.59e±0.03

6 8 N 4 6 8

67.15 ±0.06 72.57g±0.26 70.07f±0.05 68.76g±0.16 63.12c±0.02

72.60 ±0.03 75.58g±0.07 74.81g±0.16 74.40g±0.05 68.37d±0.05

c

E C

T P

77.21 ±0.03 80.26g±0.06 79.16g±0.09 79.47g±0.05 75.70e±0.05

C A

D E

14.02 ±0.01 8.62b±0.02 8.16d±0.07 7.89c±0.02 7.23d ±0.16

C S

U N

A

I R

T P

*Values represent mean± standard deviation. Means with different superscripts in the same column are significantly different (P ≤ 0.05); the number of replications (n), n≥3. To, onset temperature; Tp, peak temperature; Tc, conclusion temperature and Hgel (enthalpy of gelatinization).

ACCEPTED MANUSCRIPT

PT(C)

PV(cP)

FV(cP)

N

c

68.62 ±0.10

f

6524 ±0.12

f

8742 ±23.9

682 ±18.2

3432d±25.42

4

69.88b±0.03

6427g±40.51

9528f±56.89

553a±9.29

3586f±64.09

6

70.33d±0.10

6742g±35.73

9766g±28.04

493a±6.24

3553f±25.79

8

70.46c±0.11

6841g±45.80

9781g±20.22

425a±7.02

3695e±25.48

N

68.50b±0.30

2110a±16.3

1758a±18.2

2347d±55.4

1056ab±10.5

4

67.05a±0.06

2108a±15.6

1860a±12.0

1362b±17.4

1117b±11.2

6

68.35b±0.12

2124a±12.49

1945a±28.31

1327b±9.29

1149b±10.02

8

66.89a±0.07

2324a±6.00

2140a±17.67

1199b±11.93

1416bc±10.41

N

68.05a±0.30

2326b±40.4

3951e±31.15

2745g±32.62

1084ab±9.46

4

67.25a±0.05

2230b±71.86

3807e±43.47

934a±11.72

6

66.09a±0.17

2469b±18.04

3331c±31.90

1775d±14.18

852a±8.50

8

67.43b±0.12

2478b±24.98

US

2502f±30.79

3543b±21.39

1627d±16.04

702a±7.51

N

76.72g±0.16

2362b±13.42

3508d±41.0

1979b±24.7

1019a±29.7

4

75.37e±0.48

2493c±36.02

3534c±32.52

2449e±38.07

1375d±19.43

6

76.49g±0.06

2708d±15.62

3875d±13.32

1440c±19.08

2639d±27.23

8

77.70g±0.14

3252e±14.98

4280d±11.14

1329c±9.54

2357d±11.72

N

70.52d±0.24

2432c±45.20

3118c±52.40

2464e±24.81

1158b±29.41

4

69.64b±0.13

2562d±18.15

3125b±54.78

2369d±21.55

1252c±29.01

69.51c±0.45

2666c±41.62

3263b±18.15

1945e±7.0

1372c±10.02

RS

T

IP

BV(cP)

8

71.47d±0.04

3089c±32.72

3995c±26.86

1775e±10.60

1473c±8.02

N

73.08e±0.72

5216e±36.30

2874b±36.3

2244c±21.1

3059c±26.5

4

73.52c±0.61

5228f±35.80

3842e±19.08

2131c±21.39

3000c±15.01

6

73.47f±0.06

5272f±14.98

4174e±17.79

2016f±10.69

3110e±10.02

8

73.35e±0.12

6067f±21.39

4658f±24.58

1624d±8.72

3573e±7.57

N

75.42f±0.05

2876d±26.42

3542d±15.62

2568f±24.63

1016a±18.62

4

74.76d±0.18

2981e±22.48

3637d±14.74

2461e±27.23

1026b±26.00

6

72.35e±0.07

3066e±14.00

4270f±34.87

2250g±28.35

1147b±16.09

8

74.36f±0.06

3161d±22.03

4344e±18.88

1805f±6.43

1353b±11.37

WCS

AC

CE

6

SP

a

PT

CS

SV(cP)

CR

WS

AN

PS

pH

ED

Source

M

Table 4. Pasting properties of native and OSA modified starches.

KB

ACCEPTED MANUSCRIPT *Values represent mean± standard deviation. Means with different superscripts in the same column are significantly different (P ≤ 0.05); the number of replications (n), n≥3. PT(Pasting Temperature);PV(Peak Viscosity);FV(Final Viscosity);SV(Setback Viscosity);BV(Breakdown Viscosity). Table 5. In-vitro digestibility of native and OSA modified starches. Source

pH

RDS(%)

SDS(%)

a

g

RS(%)

AC

CE

PT

ED

M

AN

US

CR

IP

T

N 62.32 ±0.86 31.30 ±0.80 6.38b±0.89 4 61.17a±1.95 31.70f±0.75 7.13b±1.21 PS a g 6 60.12 ±3.28 31.97 ±2.13 7.91c±1.15 a e 8 54.71 ±1.93 35.75 ±0.56 9.55b±2.49 N 80.36d±0.73 13.53e±0.95 6.11b±0.27 cd d 4 81.30 ±0.40 12.70 ±0.20 6.0d±0.10 WS 6 79.69d±0.59 13.96d±0.38 6.36b±0.29 c d 8 72.71 ±2.23 18.04 ±1.93 9.25b±0.32 e f N 81.25 ±0.08 16.28 ±0.04 2.53a±0.09 4 81.10c±0.21 16.17e±0.11 2.75a±0.29 RS c f 6 78.26 ±2.72 17.25 ±1.07 4.48a±1.85 e d 8 74.38 ±2.40 17.19 ±2.70 8.43a±0.80 c d N 79.25 ±0.12 11.90 ±0.33 8.87c±0.44 c c 4 81.08 ±1.68 11.70 ±0.36 7.22b±1.67 CS c e 6 78.09 ±1.55 14.61 ±1.19 7.30c±1.69 d d 8 73.46 ±2.89 17.18 ±2.52 9.36b±0.92 e c N 81.41 ±0.72 10.51 ±0.64 8.57c±0.15 c e 4 81.10 ±0.67 11.08 ±0.57 7.98c±1.02 WCS d c 6 79.01 ±0.63 12.33 ±0.16 8.66d±0.53 f c 8 76.30 ±0.39 14.24 ±0.37 9.46b±0.70 b a N 74.61 ±0.63 1.22 ±0.15 24.17e±0.70 b a 4 74.58 ±0.05 1.33 ±0.16 24.09f±0.19 SP b a 6 72.21 ±0.31 2.60 ±0.17 25.19f±0.15 b a 8 68.64 ±0.61 4.60 ±0.30 26.76d±0.80 f b N 85.92 ±0.26 2.67 ±0.08 11.42d±0.20 4 85.93d±0.18 2.67b±0.01 11.41e±0.18 KB e b 6 83.40 ±0.09 4.44 ±0.17 11.90e±0.38 g b 8 80.47 ±1.12 6.25 ±0.93 13.28c±1.87 *Values represent mean± standard deviation. Means with different superscripts in the same column are significantly different (P ≤ 0.05); the number of replications (n), n≥3. RDS (Rapidly Digestible Starch); SDS (Slowly Digestible Starch); RS(Resistant Starch).

ACCEPTED MANUSCRIPT

Table 6. Particle size, color characteristics and phase separation of FF and FS mayonnaise. Sample

Particle size (µm)

L

a

Full Fat (FF) FS-WS

12.36f±0.26 8.62e±0.08

72.76a±0.02 83.19c±0.08

-1.20b±0.05 -1.16b±0.00

7.41c±0.10 6.96b±0.02

77.84a±0.12 86.61c±0.05

-1.30a±0.00 -1.20b±0.01

FS-PS

6.24c±0.56

82.32b±0.06

-1.00c±0.01

7.05b±0.02

85.89b±0.06

-1.04c±0.01

d

d

d

d

e

b

d

L*

dE

Phase separation (%)

8.70c±0.06 7.73b±0.04

78.33a±0.08 86.96b±0.01

64.20a±0.52 ND

7.87b±0.00

86.26b±0.06

ND

8.64 ±0.01 6.89a±0.04 7.00a±0.04 8.48c±0.01

d

88.10 ±0.08 86.26b±0.05 88.97d±0.01 87.49c±0.03

ND ND ND ND

7.95b±0.05

86.54b±0.01

ND

a*

FS-KB FS-CS FS-SP FS-WCS

8.24 ±0.21 6.18c±0.27 7.61d±0.84 2.54a±0.24

84.48 ±0.04 82.43b±0.06 85.74e±0.12 83.75c±0.12

-0.95 ±0.01 -1.04c±0.02 -1.30a±0.01 -1.31a±0.00

7.78 ±0.04 6.21a±0.03 6.40ab±0.02 7.62d±0.08

87.67 ±0.06 85.98b±0.10 88.69e±0.08 87.06d±0.03

-0.98 ±0.01 -1.08c±0.00 -0.85d±0.05 -1.35a±0.06

FS-RS

4.48b±0.32

82.66b±0.03

-1.05c±0.00

7.14bc±0.05

86.17c±0.05

-1.08c±0.01

b*

I R

C S

U N

T P

c

*Values represent mean± standard deviation. Means with different superscripts in the same column are significantly different (P ≤ 0.05); the number of

A

replications (n), n≥3.

D E

T P

C A

E C

M

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Starches from various cereals, tubers and beans were subjected to OSA esterification under different reaction pH of 4, 6 and 8. DS showed increase with increase in AM content. OSA acted majorly on the surface of starch granules and caused superficial pores. OSA modification showed increase in paste viscosities without altering crystalline structure. FS mayonnaise prepared using OSA modified starches showed 75% reduction in fat content.

AC

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Figure 1

Figure 2

Figure 3

Figure 4