Food Hydrocolloids 97 (2019) 105212

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Food Hydrocolloids
journal homepage: www.elsevier.com/locate/foodhyd

Effect of amylose content in morphological, functional and emulsification T

properties of OSA modified corn starch
Madai Lopez-Silvaa, Luis A. Bello-Pereza,∗, Edith Agama-Acevedoa, Jose Alvarez-Ramirezb
a
Instituto Politécnico Nacional, CEPROBI, Km. 6.5 Carr. Yautepec-Jojutla Col. San Isidro, Calle CEPROBI No. 8, Yautepec, Morelos, Mexico
b
Departamento de Ingeniería de Procesos e Hidráulica, Universidad Autónoma Metropolitana-Iztapalapa, Apartado Postal 55-534, Ciudad de México, 09340, Mexico

A R T I C LE I N FO A B S T R A C T

Keywords: Octenyl succinic starch esters were prepared from three corn starches with different amylose content; namely,
Amylose content waxy (5.43%), normal (25.16%) and Hylon VII (65.84%). The degree of substitution varies from 0.0103 for
OSA starch waxy starch to 0.0125 for Hylon VII, meaning that the degree of substitution is positively correlated with the
FTIR amylose content. Scanning electron microscopy images showed that the esterification reaction affected only the
Emulsification
granule surface while leaving the integrity of the granules. Solubility and water retention capacity were in-
creased with the OSA modification, with the effect more pronounced for higher amylose contents. X-ray analysis
revealed that the crystallinity type was not affected by esterification. FTIR analysis corroborated the changes in
the hydrated and ordered structures of starch granules. Emulsification tests showed that octenyl succinic starch
ester was an effective emulsifier for stabilization of oil-in-water emulsions. In particular waxy and Hylon VII
starches induced high stabilization degrees as revealed by long-term stability of oil-in-water emulsions.

1. Introduction branching points of amylopectin molecules. Analysis techniques like X-

Esterification by octenylsuccinic anhydride (OSA) is an effective
method for modification of starch. The task is to improve the functional
properties of starch by adding partial hydrophobic character, such that
the esterified starch presents amphiphilic character. OSA-modified
starch has found sound applications in food products, including im-
provement of resistant starch fraction, stabilization of emulsions by the
“Pickering” mechanism, and stabilization of thin edible films
(Sweedman, Tizzotti, Schäfer, & Gilbert, 2013). It has been pointed out
that OSA esterification likely impairs the binding of α-amylase and
decrease the starch digestion (He, Song, Ruan, & Chen, 2006; Zhu, Xie,
Song, & Ren, 2011). Overall, it has been demonstrated that OSA-mod-
ified starch is a good derivative with important applications in the food
industry (Sweedman et al., 2013).
The preparation method and the physicochemical properties of OSA
starch esters have been studied at some detail. However, the mechan-
isms underlying the starch modification by OSA have not been com-
pletely elucidated (Bai, Shi, & Wetzel, 2009). The issue is important for
the design of proper procedures aimed to obtain starch esters with
tailored properties. It was postulated that OSA substitution is carried
out in the amorphous regions of starch (Bai, Kaufman, Wilson, & Shi,
2014), which are located in the periphery of the granules (Jane & Shen,
1993; Pan & Jane, 2000) and randomly interspersed among the
ray diffraction of wide-angle did not identify the OSA modification
since changes in the X-ray diffraction pattern were not observed. In
contrast, FTIR spectroscopy showed evidences of the esterification ef-
fects (Bai et al., 2009), suggesting that esterification takes place at lo-
cated functional groups of the starch chains. The distribution of OSA
groups within the starch structure has been explored in the recent
years. OSA groups appeared to be homogeneously distributed in waxy
corn starch where the amorphous regions are located in the branching
points of the amylopectin (Shogren, Viswanathan, Felker, & Gross,
2000), although further studies suggested that the distribution is un-
even (Bai et al., 2014). Potato and normal maize starch with similar
amylose content, showed more OSA groups in the inner region of the
granule than OSA potato starch. Also, a higher number of OSA groups in
the periphery than in the inner of both starch granules was reported
(Wang, Su, & Wang, 2010, 2013). Overall, these studies indicated that
the distribution of OSA groups determine largely the physiochemical
and functional properties of modified starches. OSA reaction on starch
granules is a complex process affected by amylose content, particle size,
and molecular structure. Several reports have focused on the role of
these factors on OSA starch reactions and emulsifying properties. The
report by Sweedman, Hasjim, Schäfer, and Gilbert (2014) showed that
high amylopectin starches with a higher degree of branching (DB) and
more rigid structure showed the best colloidal stability. Also, Wang,

∗
Corresponding author.
E-mail address: labellop@ipn.mx (L.A. Bello-Perez).

https://doi.org/10.1016/j.foodhyd.2019.105212
Received 20 February 2019; Received in revised form 3 June 2019; Accepted 3 July 2019
Available online 06 July 2019
0268-005X/ © 2019 Elsevier Ltd. All rights reserved.
M. Lopez-Silva, et al.
Tang, Fu, Huang, and Zhang (2016) showed that smaller granules with
larger specific surface area had higher degree of substitution when re-
acted with OSA and showed more uniform distribution of octe-
nylsuccinate substituents.
The effects of amylose content in the properties of OSA-modified
starch have been scarcely studied. The positive effect of amylose con-
tent on the OSA modification was characterized (He, Song, Ruan, &
Chen, 2006), showing that amylose content increased the degree of
substitution and modifies the pasting properties of OSA starches (Song,
Zhao, Li, Fu, & Dong, 2013). Starches with similar amylose content
(∼25%) showed different degree of esterification (Whitney, Reuhs,
Martinez, & Simsek, 2016). The results indicated that the OSA groups
were present only in the amylose and perhaps in the amylopectin that
was hydrolyzed during the chemical modification.
The results published to date have indicated that amylose is the
main receptor of OSA esterification. This suggests that the design of
starch esters with specific properties should rely on the accurate char-
acterization of the amylose content effects on esterification. In this re-
gard, the aim of this study was to compare the morphological proper-
ties, crystallinity, thermal and pasting properties of OSA starch with
different amylose/amylopectin ratio.
2. Materials and methods
2.1. Materials
Waxy corn (Amioca, CODE: 04401106), normal corn (Globe AA
CODE: 03401037), and high-amylose corn (Hylon VII) starches were
obtained from Ingredion®. All other chemicals and solvents were of
analytical grade.
2.2. Preparation of OSA-modified starch
The corn starch (30 g, dry weight) was dispersed in 100 mL of dis-
tilled water under gently stirring. The pH of the suspension was ad-
justed to 8.75 using a solution of NaOH (0.1 M). A weighed amount of
OSA (3.0% of the dried starch base) was added slowly over 2 h while
controlling the pH at 8.75. The reaction was allowed to proceed for a
total of 6 h at 25 °C. After the reaction, the pH was adjusted to 7 with a
solution of HCl (1 M). The mixture was centrifuged and washed three
times with distilled water and once with acetone. The OSA-modified
starch was dried in an oven at 35 °C for 24 h, ground and then passed
through a 100 mesh.
2.3. Determination of amylose
The amylose content was determined using an amylose/amylo-
pectin kit from Megazyme (Wicklow, Ireland).
2.4. Determination of degree of substitution (DS) and reaction efficiency
(RE)
The DS is the average number of hydroxyl groups substituted per
glucose unit. The DS of OSA starch was determined using a titration
method (Timgren, Rayner, Dejmek, Marku, & Sjöö, 2013). The OSA
starch (1.25 g) was dispersed by stirring for 30 min in 12.5 mL of a
0.1 M HCl solution. Subsequently, the sample was centrifuged at
3000×g for 10 min. The precipitate was washed with ethanol (90%)
and with distilled water. The starch was resuspended in 75 mL of dis-
tilled water and subjected to a boiling water bath for 10 min. It was
allowed to cool to room temperature. Then, it was titrated with a
standard solution of 0.1 N NaOH until pH 8.3 and as a reference it was
titled native starch. The DS was calculated using the following equa-
tion:
2
Food Hydrocolloids 97 (2019) 105212
0.162 × (A × M )/ W
DS =
(
1 − 0.21
A×M
W )
where A is the titration volume of NaOH solution (mL), M is the mo-
larity of NaOH solution, and W is the dry weight (g) of the modified
starch. The reaction efficiency (RE) was calculated as follows:
Measured DS
RE = × 100
Theoretical DS
The theoretical DS was calculated by assuming that all the added
OSA reacted with starch to form the ester derivative.
2.5. Polarized light microscopy
A polarized light microscope (Eclipse 80i, Nikon, Japan) equipped
with a 40x objective lens and a digital camera (Digital imaging Head,
DC330 camera MTI, Japan) was used. The dried starch was spread on a
slide and a coverslip was added to the slide. A drop of deionized water
was added to the edge of the coverslip and the images were captured
under polarized light.
2.6. Scanning electron microscopy (SEM)
Starch samples were fixed to conductive tape mounted on a brass
disc. Samples were then coated with gold using Polaron E5100 (Polaron
Equipment Ldt., Watford, UK). Images of starches were captured using a
scanning electronic microscope model JSM-5800LV (JEOL, Tokyo,
Japan).
2.7. Particle size distribution
The size distribution of the starch granules was determined by laser
diffraction analysis (Mastersizer 2000; Malvern Instruments Ltd.,
Malvern, United Kingdom). The samples were analyzed using the Hydro
2000S accessory. The powders were diluted in water to achieve a sa-
turation between 14 and 16% (concentration of ∼0.001%). The sam-
ples were sonicated (250 rpm for 2 min) during the analysis to avoid
aggregation of the starch granules. The particle size is expressed as the
mean diameter D [v, 0.5], which is defined as the diameter for which
50% of the particles by volume are larger and the cumulative volume
distribution.
2.8. Water retention capacity and solubility
The water retention capacity was determined according to the
Hallgren method (Hallgren, 1985). Briefly, 5 mL of water was added to
0.25 g of starch samples (native and modified) in pre-weighed cen-
trifuge tubes at room temperature and heated at different temperatures
(50–90 °C) for 15 min, with agitation at 5 and 10 min . The tubes were
centrifuged for 15 min at 1000×g, 10 min. The supernatant was dec-
anted and the tubes were allowed to drain for 10 min at a 45Z° angle.
The tubes were weighed and then the weight gain was used to calculate
the percentage of gain as the water holding capacity. The experiments
were performed in triplicate.
2.9. X-ray diffraction analysis
The X-ray diffraction patterns were obtained using a Rigaku dif-
fractometer, model MiniFlex600 (Rigaku, Corporation Japan). The
moisture content of the samples was adjusted to 86% by storing the
samples in a desiccator with a saturated potassium sulfate solution for
7 day at room temperature (Song and Jane, 2000). The range of the
scanned diffraction angle (2θ) was 2–60° and the scanning speed was
2°/min.
M. Lopez-Silva, et al. Food Hydrocolloids 97 (2019) 105212

2.10. Thermal properties Table 1

Amylose content (%) of native corn starches and modified with 3% OSA.
The thermal properties were determined using a differential scan- Starch Native (g/100 g) Modified (g/100 g) Change (%)
ning calorimeter (DSC) (Q10, TA Instruments, USA). 2 mg of starch was
placed in an aluminum tray at room temperature (25 °C) and 7 μL Waxy 5.43 ± 0.15 c
4.04 ± 0.64 c
−25.59 ± 1.63b
Normal 25.16 ± 0.51b 14.85 ± 0.45b −59.65 ± 1.87a
(7 mg) of deionized water was added to obtain a moisture content of
Hylon VII 65.84 ± 0.67a 54.12 ± 1.00a −17.82 ± 1.12c
about 70%. The trays were hermetically sealed and allowed to stand for
12 h at room temperature for even distribution of water. The sample The values are the average of six replicates ± standard deviation. Different
trays were heated at a speed of 10 °C/min from 20 to 140 °C. An empty letters in the same column mean statistically significant difference (p < 0.05).
tray was used as a reference. The start temperature, the maximum
temperature, the completion temperature and the enthalpy of gelati- Table 2
nization (ΔH) were recorded. Degrees of substitution and reaction efficiency of corn starch esterification with
3% OSA.
2.11. FTIR spectroscopy
Starch DS × 102 RE (%)

c
FTIR analysis was carried out in a Vertex 70 FT-IR (Bruker Optik Waxy 1.03 ± 0.3 49.43 ± 0.35a
GmbH., Ettlingen, Germany) spectrometer equipped with a mercury Normal 1.12 ± 0.1b 52.58 ± 0.47b
Hylon VII 1.25 ± 0.4a 58.38 ± 0.93c
cadmium telluride detector and KBr beam splitter using a Platinum
Diamond ATR accessory with a diamond crystal at an angle of incidence The values are the average of six replicates ± standard deviation.
of 45°. Spectra were collected using 256 scans at 4 cm−1 resolution over DS = degrees of substitution; RE (%) = Reaction efficiency. Different letters
the entire 4000-400 cm−1 wavenumber region. Two spectra were col- mean statistically significant difference (p < 0.05).
lected for each sample and averaged. The analysis was performed using
OPUS version 7.0 software.

2.12. Preparation of emulsions

Oil-in-water emulsions were prepared using the method of Timgren

et al. (2013) with some modifications. The continuous phase of the
emulsions was made with 5 mmol/L of phosphate butter at pH 7 and
0.2 mol/L of NaCl, and the dispersed phase was liquid paraffin. Four
milliliters of continuous phase, 2 mL of dispersed phase, 1 mg of Solvent
Red 26 oil-soluble dye and starch in variable amounts (0–550 mg of
starch/ml of oil) were emulsified in glass test tubes by means of a
mixer, high shear in a SilentChruser M mixer (Heidolph, Germany) at
25,000 rpm for 120 s. The emulsions were observed for seven days.

2.13. Average droplet size

The volume-weighted mean droplet diameter (d4,3) was determined

on fresh emulsions and after 3, 7, 14, 21 and 28 days of storage at room
temperature (∼20 °C) with a laser light diffraction particle size ana-
lyzer (Mastersizer 2000, Malvern Instruments, Ltd.,Malvern,
Worcestershire, UK) using deionized water as dispersant. A relative
refractive index of 1.103 (ratio of the refractive indices of oil and water
phases) was used for all measurements.

2.14. Creaming stability

A dispersion analyzer (LUMiSizer, L.U.M. GmbH, Berlin, Germany)
was used to determinate the creaming rate of the emulsions. It is a novel
instrument employing centrifugal sedimentation to accelerate the oc-
currence of instability phenomena such as sedimentation, flocculation
or creaming. Samples (0.4 mL) of emulsions were subjected to cen-
trifugation at 4000 rpm for about 127 min at 25 °C, and 255 measure-
ments were determined at time intervals of 30 s. The creaming rate was
estimated by the SEPView equipment software, and was correlated to
the emulsion stability; namely, the higher the creaming rate, the lower
the emulsion stability.
2.15. ζ-Potential
The ζ-Potential of the fresh emulsions was measured with
ZetasizerNano ZS equipment. Emulsions samples were diluted in deio-
nized water (ratio 1:100 v/v) and then filled into the test cell and
placed on the measuring equipment. The ζ-Potential was calculated by
instrument using the Smoluchowski model.
Fig. 1. Micrographs of polarized light of the granules of native and 3% of OSA
modified corn starches. Waxy (A, a), normal (B, b), and Hylon (C,c). Left panel –
native, and right panel - modified with OSA.
3. Results and discussion
3.1. Amylose content
Table 1 presents the amylose content of the three different un-
modified corn starches. As expected, waxy starch exhibited the lowest
amylose content (5.43%), while Hylon VII presented the highest value
(65.84%). The OSA esterification decreased the content of detectable
amylose (i.e., apparent amylose), with the highest decrease for the
normal corn starch (59.65%). The results in Table 1 suggest that OSA
esterification took place mainly on amylose chains, such that their
presence with standard amylose measurement methods was not de-
tected (Whitney et al., 2016). The issue is of importance given that

3
M. Lopez-Silva, et al.
Fig. 2. Scanning electron micrographs (SEM) of native and 3% of OSA-modified
corn starches. The magnification of images was 2500×. Uppercase letters in-
dicate native starches, lowercase letters indicate modified starches. Waxy (A, a)
the arrows signal the pores on the surface and cracks; normal (B, b) the arrows
signal the pores on the surface and hylon VII (C, c) the signal arrows agglom-
erated on the surface after being esterified.
amylose content has large effect in the starch functionality. In fact, even
small differences in amylose content can have large effects in the
functional properties and digestibility features of starch (Jane et al.,
1999). On the other hand, it has been pointed out that changes in ap-
parent amylose content could be attributed to OSA esterification or to
the mild alkaline treatment during the modification procedure (Simsek,
Ovando-Martinez, Marefati, & Rayner, 2015).
3.2. Degree of substitution and reaction efficiency
The degree of substitution (DS) is presented in the first column of
Table 2. The DS was positively correlated with the amylose content,
suggesting that OSA esterification takes place primordially in the
amorphous regions where amylose chains are concentrated. Similarly,
the reaction efficiency (RE) was positively correlated with the amylose
content, as shown by Table 2 where the RE increased with the amylose
content.
3.3. Polarized light microscopy
Fig. 1 presents images of polarized light microscopy for unmodified
and OSA-modified starches. Visible changes in the morphology of the
starch granules were not observed, indicating that OSA modification
did not affect the integrity of the granules. It is apparent that OSA es-
terification has the major effects in the fine structure of starch chains,
while leaving untouched the solid structure of starch granules.
4
Food Hydrocolloids 97 (2019) 105212
Fig. 3. Particle size distribution (PSD) of native (black continuous line)) and
OSA-modified (red dashed line) corn starches: (a) Waxy, (b) normal, and (c)
Hylum VII. (For interpretation of the references to colour in this figure legend,
the reader is referred to the Web version of this article.)
3.4. SEM microscopy
The left panel of Fig. 2 presents SEM images of the unmodified
starch granules. The surface is smooth, without cavities or fractures.
However, the OSA modification disrupted slightly the morphology of
the starch granule surface. In fact, some granules exhibited holes,
cavities and small fractures produced by the starch treatment. However,
it is not clear at all whether the surface modifications were provoked by
esterification reactions or by mild hydrolysis by the preliminary NaOH
treatment (Yan & Zhengbiao, 2010). It is noted that granules of OSA-
modified Hylon VII starch (Fig. 3c) presents some protrusions linked
probably to the aggregation of OSA-esterified amylose chains otherwise
concentrated on the granule surface. The large amylose content of
Hylon VII allowed the extensive action on OSA starch. In contrast, the
surface of the OSA-modified waxy starch (Fig. 3a) presented large
cavities, caused probably by erosion induced by alkaline reactions.
3.5. Particle size distribution
Fig. 3 presents the particle size distribution of unmodified and OSA-
modified starches. The mean diameter of waxy starch exhibited a slight
increase, from 18.23 μm to 19.35 μm (Fig. 3a). In contrast, the Hylon
VII mean diameter was unchanged. However, the mean diameter of the
normal corn starch exhibited a large increase, from 17.54 μm to
25.42 μm. The increase of the mean diameter could be due to ag-
gregation effects induced by changes in the amphiphilic nature of OSA-
modified starches.
M. Lopez-Silva, et al. Food Hydrocolloids 97 (2019) 105212

Fig. 4. Solubility of native (■) and OSA-modified (□) corn starches: (a) Waxy,
(b) normal, and (c) Hylum.
Fig. 6. X-ray diffraction pattern of native (black line) and OSA-modified (red
line) of corn starches: (a) Waxy, (b) normal, and (c) Hylum. (For interpretation
of the references to colour in this figure legend, the reader is referred to the
Web version of this article.)

Table 3
Gelatinization characteristics of native and 3% of OSA modified corn starches.
Starch T0 (°C) Tp (°C) Tc (°C) ΔH(J/g sample)

a a a
NW 63.20 ± 0.64 71.98 ± 0.86 79.76 ± 0.26 9.61 ± 0.88a
MW 63.18 ± 0.62a 71.67 ± 0.28a 77.88 ± 0.35b 6.55 ± 0.63b
NN 68.40 ± 0.42a 73.81 ± 0.37a 81.50 ± 0.29a 10.44 ± 0.37a
MN 67.72 ± 0.08a 72.97 ± 0.09a 79.39 ± 0.17a 6.94 ± 0.10b
NH 71.52 ± 0.42a 89.72 ± 0.20a 107.80 ± 0.15a 10.45 ± 0.14a
MH 72.59 ± 0.08a 83.16 ± 0.46b 100.48 ± 0.05b 5.48 ± 0.20b

NW: native waxy, MW: modified waxy, NN: native normal, MN: modified
normal, NH: native Hylon VII, MH: modified Hylon VII. T0, Tp and Tc indicate
the temperatures of the onset, peak and conclusion of gelatinization, respec-
tively. ΔH indicates the enthalpy change of gelatinization. All data reported on
dry basis and represent the mean of triplicates. Means inside the columns of NW
and MW; NN and MN; NH and MH with different letters are significantly sta-
tistically different (p < 0.05).

3.6. Solubility and water retention capacity

The solubility of the unmodified and OSA-modified starches is dis-

Fig. 5. Water retention capacity (WRC) of native (■) and OSA-modified (□)
corn starches: (a) Waxy, (b) normal, and (c) Hylum VII.
played in Fig. 4. The solubility as function of temperature increased
with the OSA treatment, although the increase was quite low (less than
15%) for waxy and normal corn starches. In contrast, the increase was
more important for Hylon VII, with increases of the order of 120% for
90 °C. The high content of amylose and leaching of amylose chains
could be behind the relatively high solubility of Hylon VII. The water
retention capacity (WRC) is presented in Fig. 5. The WRC of the waxy
starch was greatly increased after the OSA treatment. In contrast, Hylon
VII presented the smaller increase of the WRC. Temperature in WRC

5
M. Lopez-Silva, et al.
Fig. 7. FTIR diffraction pattern of native (black line) and OSA-modified (red
line) of corn starches: (a) Waxy, (b) normal, and (c) Hylon VII. (For inter-
pretation of the references to colour in this figure legend, the reader is referred
to the Web version of this article.)
exhibited a mixed effect in the increase of the WRC. Waxy corn starch
presented high WRC increase for low temperatures, while the opposite
behavior was exhibited by Hylon VII.
3.7. X-ray diffraction
Fig. 6 presents the XRD pattern of unmodified and OSA-modified
starches. Waxy corn starch (Fig. 6a) shows a typical A-type starch
crystallinity, with d-spacings at 5.8, 5.2 and 3.8 A (Vasanthan & Bhatty,
1996). Similar pattern was exhibited by the normal corn starch
(Fig. 6b). Hylon VII exhibited a different pattern (Fig. 5c), with a B-type
crystallinity pattern. As already reported (Bai et al., 2014), the OSA
modification did not change the crystallinity of the starch granules. This
is in line with the SEM image observations (Fig. 2), showing that the
OSA modification only affected the surface of the starch granules.
3.8. Thermal properties
The thermal properties, obtained via DSC analysis, are presented in
Table 3. The onset, peak and conclusion temperatures increased with
the amylose content, indicating that amylose is closely linked to the
internal ordering of starch chains within the granules. These tempera-
tures suffered a slight decrease, of about 2 °C, after the OSA treatment.
This suggests that the internal ordering of the starch granules was
slightly disrupted by the OSA treatment. However, the effect could be
linked more to the NaOH conditioning of starch granules, rather than to
the OSA reactions per se. The effect can be better assessed by the ge-
latinization enthalpy, as this parameter decreased about 30–40% after
treatment. The greater enthalpy decrease was exhibited by Hylon VII.
6
Food Hydrocolloids 97 (2019) 105212
Fig. 8. (a) Illustration of the deconvolution of the FTIR signal in the starch
fingerprint region. (b) Intensity ratio R1047/1022 reflecting the ratio between
ordered and amorphous structures. (c) Intensity ratio R995/1022 reflecting the
ratio between hydrated and amorphous structures.
The changes of the thermal properties in Table 2 suggest, as has been
already postulated by other researchers (Bai et al., 2014; Shogren et al.,
2000) that OSA treatment affects primordially the amylose chains.
However, the OSA treatment also affected the internal ordering (double
helices of amylopectin chains) of the starch granules.
3.9. FTIR analysis
Fig. 7 presents the FTIR spectra of the unmodified and OSA-mod-
ified starches. The interpretation of the FTIR spectrum follows the de-
scription by Simsek et al. (2015). The broad peak at 3500-3000 cm−1 is
linked to the presence of hydroxyl groups, reflecting the presence of
hydrating structures. The intensity of this peak increased after the OSA
treatment, and the increase was positively linked to the amylose con-
tent. In turn, the change in the intensity of the O–H stretching peak
reflects the increase in the water retention capacity exhibited in Fig. 5.
Peaks located at 2930 and 1650 cm−1 are attributed to the C–H
stretching vibration and bound water present in the starch, respec-
tively. The fingerprint region of starch is the large peak at 1100-
900 cm−1 (C–O bond stretching). Here, the fingerprint peak is com-
posed by individual overlapping peaks that can be obtained by de-
convolution methods. Fig. 8.a illustrates the deconvolution results for
the starch fingerprint region. The peak at about 1047 cm−1 has been
linked to ordered structures, and the peak at about 1022 cm−1 to
amorphous structures. On the other hand, the peak at 995 cm−1 re-
flected hydrated structures. The ratios R1047/1022 and R995/1022 have
been proposed as indicators of ordered-to-amorphous and hydrated-to-
amorphous starch structures (Van Soest, Tournois, de Wit, &
Vliegenthart, 1995). Fig. 8b and 8.c exhibits the estimated ratios R1047/
1022 and R995/1022 for the different starch samples. The waxy starch
exhibited the largest ratio R1047/1022, suggesting that the internal or-
dering is linked to the amylopectin structure. The value of this ratio
M. Lopez-Silva, et al. Food Hydrocolloids 97 (2019) 105212

Fig. 11. (a) Graphs of integrated transmission (%) versus time (s), and (b)
creaming velocity (μm/s) of emulsions.

Table 4
ζ-Potential of fresh emulsions.
Fig. 9. Photomicrographs of transmitted light (A) and polarized light (B) mi-
croscopy of the emulsions stabilized with native corn starches and OSA (3%) Emulsion ζ-Potential (mV)
modified starch at seven days. NW: native waxy, MW: modified waxy, NN:
MW −10.1 ± 0.2c
native normal, MN: modified normal, NH: native Hylon VII, MH: modified
MN −12.9 ± 0.5c
Hylon VII. Conditions: 550 mg/mL oil, 0.2 mol/L NaCl. Scale bar 10 μm. MH −93.7 ± 0.7a

Different alphabets indicate significant difference

(p < 0.05) between column values.

increased slightly after the OSA treatment, supporting the proposal that
OSA affects principally the amorphous regions of the starch granule.

3.10. Stability of emulsions

The emulsification capacity of OSA-modified starches is an im-

portant property for application in the food industry. Fig. 9 presents
images of the oil-in-water emulsion stabilized with unmodified and
OSA-modified starches. The stability of the emulsions is due to the
hydrophobic nature of starch particles, which migrate to the oil/water
interface to form a barrier against droplet coalescence and coagulation.
Images in the left panel of Fig. 9 illustrate the stability of the emulsions,
while the right panel uses light microscopy images for assessing the
morphology of droplets. OSA-modified starches led to emulsions with
improved stability, as showed by the absence of coalescence, creaming
and coagulation. In contrast, unmodified starches were unable to sta-
Fig. 10. Variation of mean droplet diameter (d4,3) of the emulsions monitored
bilize the emulsions. For instance, the normal unmodified starch gave
for 10 days of storage at room temperature.
the worst emulsifying properties as illustrates by the sharp separation of
phases. Emulsion made with unmodified starch exhibited coagulation
and sedimentation, while unmodified Hylon VII led to partial separa-
tion of phases and coagulation. It is apparent that the best emulsifying
properties were obtained for OSA-modified waxy starch since no phase
separation was observed.

7
M. Lopez-Silva, et al.
The results in Fig. 9 showed that OSA-modified starch has the ability
of forming stable emulsions. Fig. 10 presents the variation of the mean
droplet size for emulsions from 0 to 10 days. Emulsion made with MW
exhibited the smallest drop size (∼1.4 μm). In contrast, emulsion from
MH (ENWS) presented the highest drop size (∼3.0–5.0 μm). All emul-
sions were stable in terms of the droplet size, showing only slight in-
crease of about 10% in the first 10 days. The mean drop size of the
emulsion made with MH increased about 15%, to ∼4.8 μm, suggesting
that the emulsion was affected to some extent by drop coalescence. It is
apparent that drops tended to be aggregated for this emulsion, sug-
gesting that stability is not sufficient in terms of electrostatic repulsion
to maintain droplets distant from each other.
Creaming can be also present in the evolution of emulsions. The
tendency to creaming was assessed by accelerating conditions, and the
results are exhibited in Fig. 11.a. The integral transmission quantifies
the amount of transmitted light through the emulsion bulk, and is
considered as an index of creaming. In this way, the emulsion made
with MW presented the slowest creaming rate (Fig. 11b), with the other
two emulsions exhibiting similar values of the creaming rate
(∼90–110 μm/s). Some insights on the origin of creaming can be ob-
tained by analyzing the ζ-potential of the emulsions. The measurement
of the ζ-potential is based on the hydraulic mobility of particles in a
fluid. Since oil drops dispersed in the aqueous phase are relatively large
(> 1.0 μm), the charges carried out by the oil-water interface hardly
contributed to the measured electrostatic charge. Instead, the larger
contribution is made by the particles dispersed in the continuous phase.
Table 4 shows that the emulsion made with MW exhibited the smallest
ζ-potential value (−10.1 mV), while the emulsion made with MH ex-
hibited the highest value (−23.7 mV). In turn, the emulsion made with
MH presented high value of the creaming rate. High ζ-potential values
suggest that the continuous phase is a repulsive medium that probably
led aggregation of the dispersed phase. In contrast, a continuous
medium with relatively low ζ-potential allows the free (Brownian)
mobility of the dispersed phase.
4. Conclusions
The effects of amylose content (waxy: 5.43%, normal: 25.16%, and
Hylon VII: 65.84%) on the physicochemical, thermal and emulsification
properties of OSA corn starch esters have been studied. The amylose
content is positively correlated with the effect of the OSA treatment. In
fact, solubility, water retention capacity and emulsification ability were
improved by OSA treatment, and the increase was positively linked to
amylose content. In turn, this indicates that OSA reactions affected
extensively the amylose chains, and to a minor extent to amylopectin
chains. Besides, FTIR analysis showed that OSA modification increased
the short-range ordering, which is probably linked to the idea that es-
terification focuses on the amorphous regions of the starch granules.
On the other hand, stable emulsions were obtained for waxy starch,
while emulsions with limited stability were obtained for starches con-
taining a large fraction of amylose. It was suggested that the stabili-
zation of the oil-water interface was achieved by the formation of
complex layers attached onto the interface. ζ-potential measurements
indicated that OSA-modified starch formed repulsive medium that
likely led to creaming for long storage times. Overall, the results
showed that OSA-modified starch chains have the ability of stabilizing
oil-water interfaces, with amylopectin leading to more stable conditions
in the long term.
Conflicts of interest
The authors declare no conflict of interest.
Food Hydrocolloids 97 (2019) 105212
Acknowledgments
We appreciate the economic support from SIP, IPN, EDI-IPN
CONACYT-México and COFAA-IPN. MLS also acknowledges the scho-
larship from CONACYT-Mexico.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.foodhyd.2019.105212.
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