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Contents lists available at ScienceDirect Colloids and Surfaces A journal homepage: www.elsevier.com/locate/colsurfa Ultrasound-assisted extraction of starch nanoparticles from breadfruit (Artocarpus altilis (Parkinson) Fosberg) Ivo H.P. Andradea, Caio G. Otonib, Thaís S. Amorima, Geany P. Camillotoc, Renato S. Cruzc,* aGraduate Program in Food Science, School of Pharmacy, Federal University of Bahia, Rua Barão de Jeremoabo, s/n, Salvador, BA, 40170-115, Brazil b Institute of Chemistry, University of Campinas, Rua Josué de Castro, 126, Campinas, SP, 13083-861, Brazil c Department of Technology, State University of Feira de Santana, Av. Transnordestina, s/n, Feira de Santana, BA, 44036-900, Brazil A R T I C L E I N F O Keywords: Biocolloid Nanostarch Mechanochemistry Ultrasonic homogenization Cavitation A B S T R A C T In this contribution, we report on the characterization of starch nanoparticles (SNP) extracted from breadfruit by high-intensity ultrasound. These biocolloids were characterized regarding their spectroscopic, morphological, rheological, and thermal properties in addition to ζ potential, light transmission, and X-ray diffraction (XRD) measurements. After submitting a 0.5 % (w/v) aqueous suspension to 75min of ultrasound treatment, particles of ca. 145 nm in diameter were obtained. The relatively low ζ potential (ca. −17mV) did not prevent SNP aggregation. This was corroborated by transmission electron microscopy images. The viscosity of SNP suspen- sion (2.7mPa s) was lower than that of native breadfruit starch (4.2mPa s). Thermogravimetry indicated that SNP were more thermally unstable than native starch. Infrared spectroscopy indicated that CeO groups were the most weakened by the ultrasound treatment. XRD revealed the rupture of the crystalline structure of native starch, providing SNP with an amorphous character. Altogether, this set of characterization techniques de- monstrates the feasibility of producing SNP in a rapid fashion and in the absence of chemical modifications, characteristics that are in line with the contemporary trends towards greener products and processes. The SNP produced herein through a mechanochemical approach have enormous potential for colloidal-related applica- tions, including food, cosmetic, pharmaceutical, and biomedical systems. 1. Introduction Starch is an abundant naturally occurring, rapidly renewable, and biodegradable polysaccharide [1–4]. Starch is produced during photo- synthesis as semi-crystalline granules ranging in diameter from 1 to 100 μm [5,6]. Starch granules are hierarchically structured into alter- nate and concentric crystalline and amorphous layers [2]. Corn, potato, wheat, and cassava stand out as the most common sources of starch [7]. Breadfruit (Artocarpus altilis (Parkinson) Fosberg) – a fruit originated from Indonesia and Malaysia and favorably grown in warm and humid regions – denotes an alternative amylaceous source because of its high starch contents, typically between 53 and 76 % [8]. In addition to micro-sized granules, starch may fall within the col- loidal regime after downsizing native granules into starch nanoparticles (SNP). SNP feature the intrinsic characteristics of native starch (i.e., biodegradability, non-toxicity, and biocompatibility) as well as the advantage of having large specific surface areas (surface area-to-weight ratio), the latter providing SNP with unique properties such as increased adsorption capacity and decreased diffusion-related limita- tions. Furthermore, the production of SNP often involves low operating costs [9–11]. Commercial applications of SNP include – but are not limited to – pharmaceuticals, for controlled drug release [12,13]; dentistry, in formulations of materials with improved physical proper- ties [14]; polymer-based materials, for mechanical reinforcement pur- poses [15,16]; and foods [17]. The extraction of SNP from native granules typically involves the combination of high energy inputs and complex chemical modifica- tions. Ultrasound, however, is a top-down method that allows obtaining SNP with high yields, in short periods, and without the need for addi- tional chemical treatments, e.g. acid hydrolysis, which requires long processing periods and provides low yields [18]. Ultrasound is a top- down isolation approach of SNP and conveys energy through acoustic waves as frequencies typically higher than the human threshold of hearing, i.e. above 20 kHz. Ultrasonic waves have been demonstrated to cause physicochemical transformations in matter by means of acoustic cavitation, which is the collapse of bubbles of gases that propagate as a https://doi.org/10.1016/j.colsurfa.2019.124277 Received 10 October 2019; Received in revised form 22 November 2019; Accepted 24 November 2019 ⁎ Corresponding author. E-mail addresses: ivo_henriquee@hotmail.com (I.H.P. Andrade), cgotoni@gmail.com (C.G. Otoni), thaisouzamorim@gmail.com (T.S. Amorim), geanyperuch@yahoo.com.br (G.P. Camilloto), cruz.rs@uefs.br (R.S. Cruz). Colloids and Surfaces A 586 (2020) 124277 Available online 25 November 2019 0927-7757/ © 2019 Elsevier B.V. All rights reserved. T http://www.sciencedirect.com/science/journal/09277757 https://www.elsevier.com/locate/colsurfa https://doi.org/10.1016/j.colsurfa.2019.124277 https://doi.org/10.1016/j.colsurfa.2019.124277 mailto:ivo_henriquee@hotmail.com mailto:cgotoni@gmail.com mailto:thaisouzamorim@gmail.com mailto:geanyperuch@yahoo.com.br mailto:cruz.rs@uefs.br https://doi.org/10.1016/j.colsurfa.2019.124277 http://crossmark.crossref.org/dialog/?doi=10.1016/j.colsurfa.2019.124277&domain=pdf sound wave passes in a suspension medium [14,19,20]. To date, no previous study has focused on the exploitation of breadfruit as a botanical source of SNP. In this sense, this contribution set out not only to debut the production and characterization of SNP from breadfruit, but also to shed light on a mechanochemical strategy for achieving such an outcome through an efficient and sustainable protocol. An extensive characterization of this biocolloid was aimed in order to allow establishing correlations among its chemical structure, extraction protocol, and final properties in an effort to pave the route for commercial applications of SNP. 2. Materials and methods 2.1. Source of biological macromolecule Breadfruit starch of the apyrena variety (50 ± 5 % amylose con- tent; 1.47 ± 0.03 (g/ml) density; 0.030 ± 0.001 % phosphorus con- tent), extracted according to Adebowale et al. [21], was kindly pro- vided by the State University of Feira de Santana (UEFS). 2.2. Preparation of breadfruit starch nanoparticles Breadfruit SNP were prepared according with Bel Haaj et al. [22], with some modifications. Briefly, 40ml of aqueous breadfruit starch suspensions at 0.5 % (w/v) were submitted to ultrasound treatment at a frequency of 20 KHz using a 50-W probe ultrasonic processor, model Q55 (QSonica, LLC, USA) operating at 1.0 % power for 75min. Ultra- sound treated SNP were either kept in suspension or frozen and freeze- dried. The dried SNP were used for thermogravimetry, X-ray diffrac- tion, and infrared spectroscopy analyses. 2.3. Particle size and ζ potential Never-dried SNP suspensions at 0.5 % were analyzed by dynamic light scattering on a Zetasizer Nano ZS (Malvern Instruments Ltd., UK) at 25 °C using a light dispersion angle of 173°. From these data, average particle size and particle size distribution, the latter indicated by polydispersity index (PDI), were calculated. The same equipment was used for electrophoretic mobility measurements to determine ζ poten- tial. All analyses were carried out in triplicates. 2.4. Light transmission The transmittances of the SNP suspensions were measured at wa- velengths ranging from 350 to 550 nm on a ultraviolet-visible spec- trophotometer, model SP-220 (Biospectro, Brazil). This analysis was performed 24 h after preparation and rest at room temperature. 2.5. Morphology Native starch granules were imaged by scanning electron micro- scopy (SEM) on a JSM-6610LV (JEOL, Japan) microscope operating at 30 kV. Samples were previously dried and coated by a gold layer for ca. 2min on a Desk V (Denton Vacuum, USA) thin film deposition system. SNP were imaged through transmission electron microscopy (TEM) on a JEM-1230 (JEOL, Japan) operating at 80 kV. Prior to TEM imaging, SNP were diluted up to 1:500 (volume) in distilled water and a drop of the diluted suspension was deposited onto a copper grid, stained with a 2 % aqueous dispersion of uranyl acetate and finally dried at room temperature for 24 h. 2.6. Rheological measurements The steady shear viscosities of the native starch and the breadfruit SNP suspensions (same suspensions used for the ultrasound treatment) were determined at 25 °C on a DV-II + Pro viscometer (Brookfield, USA) equipped with a #61 spindle rotating at 150 rpm. The measure- ments were repeated five times. 2.7. Thermogravimetry Native starch and SNP samples of approximately 5mg were con- tinuously weighed while being heated from 25 to 600 °C at 10 °C/min on a thermal analyzer, model Pyris 1 TGA (PerkinElmer, USA), to in- vestigate the thermal degradation profile by means of thermogravi- metric (TG) and derivative TG (dTG) curves. 2.8. Attenuated total reflectance Fourier-transform infrared spectroscopy (ATR-FTIR) Infrared spectra of freeze-dried native starch and SNP were obtained on a vibration spectrometer, model Paragon 1000 (PerkinElmer, USA) operating in the infrared region with Fourier transform and equipped with attenuated total reflectance (ATR) module. Spectra were acquired at wavenumbers between 4000 and 500 cm−1 using a resolution of 2 cm−1. 2.9. X-ray diffraction (XRD) Freeze-dried native starch and SNP samples were analyzed on an X- ray diffractometer, model 6000 (Shimadzu Co.) with a CuKα radiation, and tension and current were 30 kV and 30mA. XRD patterns were recorded at Bragg angles (2θ) varying from 4 to 35° at 2.0° min−1. 2.10. Statistical analysis SNP suspensions were obtained in three repetitions. Particle size, ζ potential, PDI, and viscosity values are reported as mean values ± standard deviations. Particle size and steady shear viscosity were fur- ther submitted to analysis of variance (ANOVA) at a significance level of 5 %, followed by t-test at the same level. The OriginPro software, version 8.0724 (OriginLab, USA) was used for the statistical treatment of the obtained data. 3. Results and discussion 3.1. Size and stability of SNP The obtained SNP presented a polymodal size distribution, as shown in Fig. 1. The first population of particles (from left to right) shows a size distribution range from 8.7–37.8 nm and corresponds to 35.2 % of the total population. The second population corresponds to 43.8–64.0 nm and 23.3 % of the population. Finally, the third Fig. 1. Particle size distribution of breadfruit SNP. I.H.P. Andrade, et al. Colloids and Surfaces A 586 (2020) 124277 2 population ranges in diameter from 190.0–955.0 nm and comprises 38.1 % of the particles, therefore showing the highest intensity. A fourth population was also observed, with particles showing an average size of 4,836.7 nm, but this represents only 3.4 % of the total particles. Hebeish et al. [23] pointed out that nanoparticles are likely to ag- glomerate in an aqueous medium during size measurements. Conse- quently, the measured size represents that of the particle aggregates instead of the individual particles. Either particle aggregation or any possible contamination could justify the fourth population. However, the herein obtained SNP had an average diameter of 146 ± 51 nm. The studied SNP presented a PDI value of 0.46 ± 0.06. PDI is a measurement of the width of the distribution of particle sizes within a population. It ranges from 0 to 1, values close to zero indicating a homogeneous distribution, whereas values greater than 0.5 indicate high heterogeneity [23,24]. As mentioned before, the SNP produced in this study ranged in size from 43 to 955 nm and presented three main populations, each of which comprised between 20 and 40 % of the total population, therefore denoting a nonhomogeneous distribution and justifying the encountered PDI value. Bel Haaj et al. [22] pointed out that too intense ultrasound treat- ments, i.e. above 80 % power, the strong cavitation tends to increase the coalescence between the bubbles, which in turn decreases the me- chanochemical effects provided by such a treatment. In other studies, only part of the total power was actually used [25–28]. However, in our preliminary studies, we could only obtain a translucent suspension – characteristic of nanoparticle suspensions - using 1.0 % power. Other parameters that can interfere in the size of nanoparticles obtained through ultrasound-assisted procedures are processing time [29] and the size of the native starch granules, which depends on the botanical origin [30]. Bravo et al. [31] produced maize SNP larger than 400 nm. Silva et al. [28] obtained cassava SNP with 93.1 % of the particles having an average diameter of 77.51 nm. In both studies, the ultrasound apparatus and the sonication time and power were identical to those used in the present study (75min and 50W). Even using the same amylaceous source, different sizes of SNP can be obtained. For instance, Bel Haaj et al. [32] as well as Bravo et al. [31] produced maize SNP with average diameters of 37 and>400 nm using ultrasound powers of 400 and 50W, respectively. Concerning particle stability in suspension, the average ζ potential of the produced SNP was -16.9 ± 0.5mV. High nominal ζ potential values (higher than +30mV or lower than −30mV) indicate stable systems due to electrostatic repulsion between charged groups, which prevent particle aggregation and other destabilization mechanisms [33]. The negative surface charge of SNP arises from hydroxyl radicals that are deprotonated in aqueous media [18,23]. In addition to the negative character, the obtained ζ potential value indicates insufficient electrostatic repulsion among SNP, possibly leading to aggregation [32]. This observation is in line with DLS data, which suggest SNP aggregation. Similar outcomes with regards to SNP stability were ob- served in other investigations on ultrasound-assisted SNP extraction. Bravo et al. [31] produced cassava and maize SNP with ζ potential values equal to -8.70 and -1.90 mV, respectively. Silva et al. [28] ob- tained cassava SNP with a ζ potential of -8.67mV. 3.2. Morphology of SNP SEM and TEM images of breadfruit native starch and SNP, respec- tively, are presented in Figs. 2 and 3. Native starch granules were mostly quasi-spherical, but few polyhedral granules with irregular surfaces were also observed (Fig. 2A and B). Rincón and Padilla [34] also found these characteristics in breadfruit starch granules and at- tributed such irregularities to possible indentations caused by the compression of small starch granules during their development and extraction. Indentation arises from endosperm volume reduction and wrinkling during plant maturation [35]. Breadfruit SNP also presented a round shape (Fig. 3A and B), with some stretched, quasi-cylindrical exceptions (Fig. 3C and D). These structures were similar to those obtained by Wei et al. [36]. The pre- sence of some SNP clusters (as in Fig. 3E) corroborates the aggregation allowed by the relatively low ζ potential value. García et al. [37] and Lamanna et al. [38] obtained similarly aggregated structures and at- tributed it to the abundance of hydroxyl groups on SNP surface, creating strongly associate interactions that, in addition to a low surface charge, favor inter-SNP association. Finally, the darker SNP may be related to particle overlapping during fragmentation, as suggested by Hebeish et al. [23]. 3.3. Light transmission through SNP suspensions Pictures of SNP suspensions are presented in Fig. 4. Changes in the turbidity of SNP suspensions were clearly observed when comparing samples sonicated from 15 to 25min, with suspensions becoming vir- tually transparent at the end of this interval – indeed, after 25min of sonication, the average SNP diameter was 228 ± 71 nm, i.e., smaller than the wavelength of visible light, avoiding its scattering. Because we aimed to evaluate the influence of sonication time on suspension transparency, different sonication periods were also evaluated (Fig. 4A). As mentioned before, light scattering is directly related to the size of particles in suspension. When aiming light beams directly at colloidal suspension, small particles – as in the case of this study – are compar- able in size with the lengths of the incident waves. The scattering is observed when each particle in the light path behaves as a source of secondary light [22,39]. As colloidal particles are known to scatter visible radiation, the path of the beam that is passing through the Fig. 2. SEM micrographs of native breadfruit starch (A, 1000X; B, 3000X). I.H.P. Andrade, et al. Colloids and Surfaces A 586 (2020) 124277 3 suspension can be seen by naked eye. This phenomenon, so called Tyndall effect [40], was observed in the SNP suspensions upon aiming a light beam towards them. Suspensions that experienced shorter soni- cation times presented a lower transmittance as well as a higher dis- persed light intensity (Fig. 4B and C). According to Craig et al. [39], larger particles scatter more light. Thus, one can establish a direct relationship between sonication time, particle diameter, and light transmission. Suspension transparency therefore denotes a parameter that confirms particle fragmentation up to the nanoscale, which was also observed by Boufi et al. [41]. Light scattering by SNP was also observed when aiming the light beam at the suspension sonicated for 75min (Fig. 4D, left). In the case of water (Fig. 4D, right), this phenomenon was not observed because of the absence of colloidal particles in suspension. The transmittance curves (Fig. 5) clearly corroborate the direct re- lationship among suspension transparency and sonication time, with 75min of ultrasound treatment leading to a 96.6 % transmittance at 550 nm. It is worth mentioning that SNP concentration in all suspen- sions was the same. A remarkable change in transmittance at 550 nm was observed among suspensions sonicated for 15 (14.6 %), 20 (71.0 %), and 25min (95.8 %). 3.4. Viscosity of SNP suspensions The steady shear viscosity of breadfruit starch suspension was 4.2 ± 0.3mPa s, whereas for SNP suspension at the same solid content this parameter was 2.7 ± 0.1mPa s. The lower viscosity of the SNP suspension is caused by the reduced molecular weight of the granule due to the breakdown of the glycosidic bonds caused by the ultrasonic waves [42]. Reduced particle size is related to a lower capacity of water retention by the swollen granules. This leads to a lower viscosity of SNP suspensions in comparison with its native counterpart [42,43]. Other studies have also reported a reduction in the viscosity of starch sus- pensions submitted to ultrasound [19,29]. The decreased size of breadfruit SNP as well as the reduced viscosity of SNP suspensions make them potential candidates as binders in pa- permaking, to increase the binding capacity among starch and cellu- lose. Decreased viscosity is also beneficial for nanocomposites from the processing standpoint, wherein solid contents in suspensions should be as high as possible in order to avoid excessive dilution [44]. 3.5. Thermal properties of SNP The thermogravimetric (TG) and derivative TG (dTG) curves of breadfruit native starch and SNP are shown in Fig. 6. The TG curves of both starch particles presented the same pattern, including two stages of weight loss, behavior which were also reported by Bravo et al. [31], Lamanna et al. [38], and Le Corre et al. [45]. According to Hornung et al. [46], the first weight loss is related to water evaporation and release of volatile compounds. The second weight loss stage is attrib- uted to the degradation of the organic matter. The dTG curves allow for a better visualization of the weight loss events through their decomposition rate as a function of temperature. The first event in native starch was as evidenced by the peak centered at 65.41 °C, being the offset of this weight loss stage at 140.78 °C. SNP, in turn, had the maximum weight loss rate at 50.56 °C, and the process continued up to 127.75 °C. The second event was observed between 311.86 and 412.73 °C (with the maximum decomposition rate occur- ring at 371.96 °C) in native starch and from 294.88 to 426.79 °C (maximum weight loss rate at 369.65 °C) for SNP. Native starch was hence demonstrated to have a higher thermal stability than SNP. A similar behavior has also been observed for SNP elsewhere [28,38,47]. The higher thermal stability of native starch compared to SNP can also be observed by comparing their rates of maximum thermal decomposition, with native starch presenting a rate (in modulus) of 0.83mg/min while SNP 0.62mg/min. Native starch has more compact surface and inner structures than the nanoparticles de- rived from it. Denser structures require more energy for the phase Fig. 3. TEM micrographs of SNP (A, B, C, & D, scale bar: 200 nm; E, scale bar: 100 nm). I.H.P. Andrade, et al. Colloids and Surfaces A 586 (2020) 124277 4 transitions and thermal decompositions [48]. Due to its lower degree of compaction, SNP have a high number of OH groups on its surface, through which thermal degradation occurs in advance due to the higher hydrophilicity. Thus, weight losses generally occur earlier in SNP when compared to native starch [38,49]. 3.6. Spectroscopic characteristics of SNP FTIR spectra of native starch and breadfruit SNP (Fig. 7) were re- markably similar. This behavior was also documented in other studies on nanostarch production [9,50]. Some bands in SNP spectrum pre- sented different intensities in relation to native starch, in particular those within the fingerprint region (below 800 cm−1 and between 800 and 1500 cm−1), where the characteristic peaks of starch are observed [51]. Fig. 4. Tyndall effect on SNP suspensions. Fig. 5. UV–vis Transmittance curves of SNP suspensions. Fig. 6. TG (A) and dTG (B) of native starch and starch nanoparticles. Fig. 7. Spectra of native starch and starch nanoparticles. I.H.P. Andrade, et al. Colloids and Surfaces A 586 (2020) 124277 5 The most intense peak was observed at 1003 cm−1 and corresponds to the stretching vibrations of CeO groups, as well as the peak at 1079 cm-1 [52]. Since the band 1003 cm−1 was the most accentuated of the SNP, the CeO groups were the most affected by the ultrasonic waves, which caused the weakening and rupture of the chemical con- nections. The broad peak at 3290 cm−1 also presented accentuation in relation to native starch, but in a low intensity. This band is related to the stretching of the OeH hydrogen bonds. The region is characterized by the interval between 3000 and 3600 cm-1 [53,54]. The accentuation of such peak is related to the weakening of the hydrogen bond network, probably due to a possible reduction of the crystalline structure of the starch caused by the ultrasound [55]. Still in relation to the SNP spectrum, the peak at 1350 cm-1 suffered a slight accentuation com- pared to native starch. According to Pavlovic and Brandão [56], it is related to the deformations in the CeOeH groups. Some specific peaks that did not suffer alterations by the ultrasonic treatment were observed in both spectra, such as the one at 1640 cm-1, attributed to water mo- lecules adsorbed by the starch, at 2930 cm-1, related to the vibrations of the CeH bonds [51]. The breakdown of starch glycosidic bonds is the factor responsible for the formation of SNP. According to Chang et al. [20], the break- down of covalent bonds of polymeric materials is caused by the emis- sion of ultrasonic waves into aqueous suspension. Ultrasonic waves emitted in starch suspensions may form air bubbles that collapse and generate shear forces on starch granules. This energy leads to the breakdown of native starch chains as well as to fragmented particles, potentially within the nanoscale [14,20]. 3.7. Crystalline structure of SNP The XRD patterns of native starch and breadfruit SNP are presented in Fig. 8. It was observed that the native starch presented the char- acteristic peaks of B-type starch at Bragg angles (2θ) close to 5.6°, 15°, 17°, 22°, and 24° [57]. The presence of the characteristic V-type peak was also observed at ca. 20.6°, being associated with complexes formed by simple amyloid propellers and lipids [58]. The B-type pattern is typical of starches with high amylose contents (60–73 %), such as the breadfruit starch studied here [30]. The results are in agreement with recent studies on breadfruit starch, in which B-type crystalline patterns were also observed [59,60]. In relation to the SNP diffractogram, the diffraction peaks dis- appeared completely after the 75min of sonication, suggesting that the energy input arising from the ultrasound waves disrupted the original crystalline arrangement of starch. In other words, native starch un- derwent an amorphization process when converted into SNP. Liu [61] pointed out that the crystalline decomposition of starch can be attrib- uted to the decrease in particle size. This, in turn, results in the con- version of the ordered structure into fractions that are extremely small. The deformation of the crystalline region of different starches by ul- trasound treatment resulting in amorphous nanoparticles was also ob- served elsewhere [22,28,32,40]. Bernardo et al. [62] also observed a reduction in diffraction peaks and the appearance of fissures on the surface of some starch granules after applying ultrasound. They at- tributed this observation to cavitation-induced damage to the crystal- line structure of yam starch. However, the energy was not sufficient to modify the crystallinity pattern of starch, which remained unchanged. When used for reinforcement purposes in polymer-based film ma- trices, SNP and nanocrystals have different characteristics. Crystalline particles feature organized polymer chains in ordered arrays, forming a network that more effectively diffuses water and improves polymer matrix stiffness when compared to amorphous SNP, thus denoting better reinforcing agents [32]. Since amorphous particles do not feature such a degree of organization, the SNP produced in this study are not envisaged for reinforcement purposes in composite materials. Their poor dispersion and propensity to agglomeration also lead to a low reinforcing effect. 4. Conclusions In summary, ultrasound was herein demonstrated as a purely phy- sical means of isolating starch nanoparticles from breadfruit in a fast, straightforward fashion. The produced SNP presented an average size of 145.65 nm after 75min of sonication. The surface charge in SNP was not sufficient to prevent aggregation, which was further boosted by the high occurrence of hydroxyl groups, leading to SNP aggregates. The nanoparticles in suspension had a lower viscosity when compared to the native starch due to their lower capacity of water retention and less intermolecular interaction between the polymeric chains. The thermogravimetry indicated a lower thermal stability of the SNP compared to the native starch with a loss of volatiles and a de- gradation of the organic matter at lower temperatures. The chemical groups C–O were the most weakened by the ultrasonic waves during the process. The analysis of the XRD indicated the rupture of the crystalline region of the nanoparticles caused by the ultrasonic treatment and that led to the formation of amorphous particles. Thinking of future applications, the breadfruit SNP produced in this study have potential for a range of colloidal-related systems, including food formulations and as binders. Due to the small size of the breadfruit SNP produced here and also because they were produced through a purely physical procedure, they can directly contact foodstuffs without major concern regarding toxicity, therefore opening the possibility of being applied as fat mimetics – compounds that can mimic the sensory and physical properties of triglycerides, though not totally replacing fat in a gram-for-gram basis [63–66]. The combination of SNP with other food components may lead to a cream-like blend featuring similar properties to fats. An advantage of the use of SNP as mimetic fats is the reduction of calories [14]. Another potential food application of breadfruit SNP would be in shelf life extension of bakery and con- fectionery products, as modified starches are known to slow down the processes through which their moisture contents are reduced and their hardness is increased, therefore leading to impaired palatability [67]. CRediT authorship contribution statement Ivo H.P. Andrade: Conceptualization, Data curation, Writing - original draft. Caio G. Otoni: Data curation, Writing - review & editing. Thaís S. Amorim: Data curation. Geany P. Camilloto: Supervision, Validation. Renato S. Cruz: Conceptualization, Methodology. Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influ- ence the work reported in this paper.Fig. 8. XRD patterns for native breadfruit starch and starch nanoparticles. I.H.P. Andrade, et al. 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