WOLFAPPLE (SOLANUM LYCOCARPUM ST. HILL) STARCH: CHEMICAL AND PHYSICAL PROPERTIES FRUTA-DO-LOBO (SOLANUM LYCOCARPUM ST. HILL): PROPRIEDADES QUÍMICAS E FÍSICAS DO AMIDO.
Adriana dos Santos Fernandes1, Diego Palmiro Ramirez Ascheri1, Marcos Paulo Batista2, Jocielle Conceição Oliveira Cardoso3, Jacqueline Nascimento Gomes3, Yago Silva de Sousa1.
Abstract: In this study, starch from S. informações sobre sua caracterização. Para lycocarpum fruit (wolfapple) was extracted to isto, realizou-se extração do amido e o cálculo determine physical and chemical properties do seu rendimento. Determinou-se também sua and provide more information and further composição química (cinzas, extrato etéreo, evidence on its characteristics. Starch yield fibras, proteínas e teor de amilose); forma e was calculated. The chemical composition of tamanho dos grânulos; análise de viscosidade the wolfapple starch was determined for ash, (RVA); análise calorimétrica (DSC) e análise ether extract, fiber, protein and amylose cristalográfica. A extração do amido da fruta content. The shape and size of the granules de S. lycocarpum teve um rendimento de were assessed, as well as rapid visco analysis 27,9% e este se caracterizou por apresentar (RVA); differential scanning calorimetry 0,19% ±0,01 de cinzas, 0,07% ± 0,00 de (DSC); and crystallographic analysis. The extrato etéreo, 0,32% ± 0,01de proteínas, yield of the wolfapple starch was 27.9% and 0,03% ± 0,01 de fibras e 29,16% ± 0,09 de the chemical composition was characterized amilose. A análise de imagem do amido by the presence of 0.19% ± 0.01 ash, 0.07% ± mostrou grânulos com formato cônico e 0.00 ether extract, 0.32 % ± 0.01% proteins, diâmetros médios e respectivos desvios padrão 0.03% ± 0.01 fibers, and 29.16% ± 0.09 de 16,59±3,68 e 16,52±3,36 μm, para diâmetro amylose. Starch image analysis presented maior e menor, respectivamente. As análises conical shaped granules with average da viscosidade de pasta; calorimétrica e diameters and respective standard deviations cristalográfica revelou que o amido da fruta de of 16.59 ± 3.68 and 16.52 ± 3.36 μm, for the S. lycocarpum mostrou perfil com largest and smallest diameter, respectively. características desejáveis, como boa Paste viscosity, calorimetric, and estabilidade a alta temperatura, o que o elege crystallographic analyses showed that the como um bom ingrediente a ser usado em starch from wolfapple has desirable diversos setores alimentícios e industriais. characteristics such as stability at high temperature, making it a promising material in PALAVRAS-CHAVE: Amilose, gelatinização, various food and industrial sectors. grânulos de amido.
KEYWORDS: Amylose, gelatinization, 1Universidade Estadual de Goiás, Campus CET starch granule. - Henrique Santillo. [email protected];
2Universidade Federal de Goiás, Campus Resumo: O objetivo deste trabalho foi extrair Samambaia. e determinar as propriedades físicas e químicas 3Instituto Federal de Goiás. do amido da fruta de S. lycocarpum (fruta-do- lobo) visando obter e disponibilizar mais
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INTRODUCTION consumed by humans (SPIER, 2010). Starch is a plant-produced polymer Starch can be extracted from cereal grains, stored in small and semicrystalline granules vegetables and tubers, and has a variety of composed by two main components: food and non-food applications amylose and amylopectin. Amylose (SCHIRMER et al., 2013; FERNANDES et predominantly consists of linear chains, al., 2019). typically consisting of α (1-4) D-glucose The starch industry extracts starch units, while amylopectin has large and from various sources and processes it into a branched chains formed by α (1-4) D- wide variety of products, such as native glucose units in the rectilinear portions and starches, glucose, maltose and fructose α bonds ( 1-6) in the branches (BENINCA syrups, dextrose and maltodextrins et al., 2013). It is the main source of reserve (CEREDA, 2002). Figure 1 shows some carbohydrate in plants and an important applications of starch, both in natural and source of energy for many organisms, modified forms. providing between 70 to 80% of the calories
Figure 1. Main applications of starch in Brazil. Source: Adapted from Beninca et al, 2013.
Among other usages in the food used due to its viscosity, gelling power, industries, starch can facilitate food adhesion, tendency to retrograde, among processing and is used as thickener in soups, other properties affected by the broths, and meat sauces. It provides amylose/amylopectin ratio as well as the suspended solids and texture, gelling agents shape and size of granules (CIACCO; in cured processed meat, stabilizers in salad CRUZ, 1987). dressings, and increase the shelf life of In the textile industry, starch is used products (CEREDA, 2002). Starch is also for gumming threads to facilitate weaving 2 Revista Agrotecnologia, Ipameri, v.11, n.2, p.1-13, 2020
and in papermaking to produce papers of The most prominent starchy species different strengths and high print quality in the world market are corn, potato, and (BENINCA, et al., 2013). In glue and cassava. However, recent studies have adhesive manufacturing, in general, it is searched for new sources of starch with used for providing more viscous products, potential to be used in various sectors for its easy preparation and for combining (DHANAPAL et al., 2012; KASIRAJAN; with various resins and synthetic emulsions NGOUAJIO, 2012). Industrial applications (CIACCO; CRUZ, 1987). It can be used in consider availability and physicochemical the composition of biodegradable characteristics of the starch, which may packaging (FERNANDES et al., 2019). In vary according to its origin. Thus, every culture media, starch can be useful as an natural or native starch is considered unique inductor of the production of enzymes and (MATSUGUMA et al., 2009). microbial biomass. In addition, it can be The Cerrado region in Brazil has a hydrolyzed and the sugar produced from variety of plants with very specific hydrolysis can be fermented to produce characteristics due to their adaptations to ethanol used for food, beverage, the climate and soil. These plants are still pharmacological and laboratory purposes little explored and could be used as a source (AJIBOLA et al., 2012). The glucose of starch. Solanum lycocarpum St. Hill , obtained from starch is also widely used in commonly known as wolfapple, belongs to the manufacture of organic acids such as the family Solanaceae, the most common citric acid, glutamic acid, lactic acid, and genus found in the Brazilian Cerrado amino acids such as lysine, or (Figure 2). It can produce from 40 to 100 commercialized as crystalline glucose. fruits, of masses varying between 400 to Therefore, it is very important to have 900g, with good potential for starch different starch sources available for this extraction (CAMPOS, 1994; very wide market (AQUINO et al., 2015). FERNANDES et al.; 2019).
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Figure 2. S. lycocarpum a) plant; b) flower; c) and d) fruit cross section. Source: Adapted from Campos (1994).
In the present study, starch from S. properties and provide more information lycocarpum fruit (wolfapple) was extracted and further evidence on its characteristics. to determine physical and chemical
MATERIAL AND METHODS washed under running water for starch S. lycocarpum fruits were collected in separation and decantation in plastic the rural area of the municipality of containers. Following decantation, the Anápolis-GO. Pulp samples were collected material was washed with absolute alcohol in paper bags and stored in a refrigerator for to remove resins and vacuum filtered. After starch extraction. purification, the starch was vacuum dried at To obtain the starch, the pulp of the 40°C to constant weight. The dehydrated peeled fruits was cut into small pieces with starch was kept in a desiccator containing a stainless steel knife and stored in plastic silica gel for 12h and reduced to powder buckets containing a 5 g.L-1 sodium using mortar and pestle. Finally, the metabisulphite solution to prevent material was sieved to 48 mesh to obtain the browning. dry sample for analysis. The pulp was milled in a The yield was calculated as the ratio “CROTON” MA580 knife mill between the weight of the peeled pulp and (MARCONI, Piracicaba, Brazil) and sieved the starch produced. The material was to 10 mesh (1700 µm). The pulp was again characterized considering ash, fiber, ether sieved (150 to 400 mesh), and repeatedly extract, and protein content (AOAC, 2000). 4 Revista Agrotecnologia, Ipameri, v.11, n.2, p.1-13, 2020
Amylose content was determined according Calorimetric analysis was performed to the methodology proposed by Williams on a DSC Q200 differential scanning et al. (1970). calorimeter (TA Instruments, New Castle, Image analysis was used to evaluate USA), using indium calibration. To the shape and size of the isolated starch determine the gelatinization temperature, granules. The samples were collected with approximately 5.0 mg of the sample (at platinum wire on a glass slide, mixed with a known moisture) was placed into an airtight drop of distilled water, and covered with a aluminum container. The scanning profile coverslip. The slides were examined with a used was balanced at 5ºC, heating at 110 LEICA DME optical microscope (Wetzlar, ºC, heating rate of 10ºC.min-1, and flow of Germany), and the selected images were 50 mL nitrogen per minute. Gelatinization analyzed with the software LAS EZ. enthalpy was determined using the software The methodology described by Universal Analysis version 4.3A. Ascheri et al. (2006) was used for viscosity Crystallographic analysis was carried analysis, using the Rapid Visco Analyzer out with the samples fixed in an aluminum (RVA). Starch suspensions (2.5 g in 25 mL support and assessed at room temperature, of water) were corrected to 14% moisture using a Rigaku X-ray diffractometer (model and assessed using the following RU 200 R, Osaka, Japan), operating with a time/temperature regime: one minute at monochromatic filter, copper Ka radiation, 50oC , heating from 50 to 95°C at a rate of 0.8 kW power, 50 mA current, 40 kV 6ºC.min-1, holding at 95°C for 5 minutes voltage and wavelength of 1.54 A. The and cooling from 95 to 50°C at a rate of analyses were performed between 2 = 5º 6ºC.min-1. Viscosity was expressed as cP. and 2 = 40º, with scanning speed of Based on the graph obtained, the following 2º(2).min-1, and intensity expressed as parameters were evaluated: pasting peak count per second. temperature, maximum viscosity (peak), viscosity decrease (difference between RESULTS AND DISCUSSIONS maximum viscosity and viscosity at 95°C Yield of starch extracted from S. for 5 min), final viscosity, and lycocarpum was 27.9% on a dry matter retrogradation tendency (difference basis. Although this value is lower than that between final viscosity and viscosity at found by Di-Medeiros et al. (2014) for the 95°C for 5 min). same botanical source (51%), still this yield is considered good when compared with 5 Revista Agrotecnologia, Ipameri, v.11, n.2, p.1-13, 2020
other starch sources such as the commercial industrial fields, like production of films, starch from cassava roots, which can range biomembranes of low water permeability, between 21-33% (dry basis) (CEREDA, food additives, or pharmaceutical coating 2002). According to Lima et al. (2012), materials (Di-Medeiros et al, 2014). easiness to extract starch depends on Table 1. Chemical composition of S. lycocarpum fruit starch. granule size and other factors. S. lycocarpum fruit starch has large granules Mean ± Standard Deviation that facilitate decantation and extraction Constituent (% dry matter efficiency. basis) Difference between the yields Ash 0.19 ± 0.01 reported are due the methodologies used in Ether Extrato 0.07 ± 0.00 the extraction processes. Protein 0.32 ± 0.01 The results of centesimal composition Fiber 0.03 ± 0.01 (Table 1) were within the required by the Amylose 29.16 ± 0.09 Brazilian legislation for starches extracted Each value represents the mean and from natural sources. They indicate a high standard deviation of three repetitions. purity level of the S. lycocarpum fruit starch, with low contents of ash, ether The image analysis (Figure 3) showed extract, fiber, and protein. conical shaped granules, mean diameters Table 1 indicates that the starch from and standard deviations of 16.59 ± 3.68 and S. lycocarpum fruit has 29.16% of amylose 16.52 ± 3.36 µm, for larger and smaller in its content. This value is considered high diameters, respectively. and suggests it can be used in various
Figure 3. (A) Optical photomicrograph (x100) and (B) Scanning electron microscope (SEM) (x2000) granule diameters of S. lycocarpum fruit starch .
Eliasson (1996) classified starch small granules for sizes ranging from 1 to granules of different botanical sources as 10 µm and large granules for sizes ranging 6 Revista Agrotecnologia, Ipameri, v.11, n.2, p.1-13, 2020
from 10 to 35 µm. Di-medeiros et al. addition, it is an important parameter to (2014), studying the rheological and define the steps in the extraction process of biochemical properties of S. lycocarpum this polysaccharide. The extraction process starch also found conical shaped granules, with large granules, for example, is more but with average sizes from 10 to 14.4 μm. efficient due to the easiness of decantation This variation can occur because the and extraction. granule size and amylose content of The viscosity profile of the starch starches of the same species may vary due paste from S. lycocarpum is shown in to the environmental conditions of plant Figure 4. Paste temperature and the growth (CIACCO; CRUZ, 1987). viscosity peaked at 69.5°C and 4,461 cP, at Eliasson (1996) discussed that the the time of 2.6 min and 5.7 min, size of granules and their distribution are respectively. Viscosity was stable at hot among the factors that influence the temperatures with a slight decrease after functional properties of starches. In reaching the maximum viscosity peak.
Figure 4. Viscosity profile of the starch paste from S. lycocarpum fruit.
The viscosity peak of S. lycocarpum 923.5 cP, reaching final viscosity of 4438 starch showed a round shape, indicating cP. heterogeneous starch granules. After Retrogradation or recrystallization is viscosity rupture and subsequent cooling, a process that occurs after solubilization and there was a retrogradation tendency of during gelatinization. The amylose chains aggregate faster than the amylopectin and 7 Revista Agrotecnologia, Ipameri, v.11, n.2, p.1-13, 2020
form crystalline double helices stabilized by for the processed food industry, as well as hydrogen bonds. Upon cooling and/or in the adhesive industry (CEREDA 2002; staling processes, these helices form highly DI-MEDEIROS et al. 2014). stabilized three-dimensional crystalline Considering the calorimetric analysis structures (ELIASSON, 1996). of starches, Di-Medeiros et al. (2014) stated The viscosity profile in Figure 4 that this property is very important to shows that the starch from S. lycocarpum determine the functionality and utilization fruit has desirable features for the of starch in the industry. The gelatinization production of biodegradable films: good properties of the starch from S. lycocarpum stability at high temperature and and other botanical sources are presented in mechanical stability (FERNANDES et al., Table 2. 2019). In addition, it is also a good material
Table 2. Starch gelatinization properties of wolfapple (S. lycocarpum) and other botanical sources. Gelatinization
-1 To (ºC) Tp (ºC) Tc (ºC) ∆H (J.g ) Wolfapple (S. 63.68± 0.27 67.20 ± 0.06 77.81 ± 0.28 13.08 ± 0.72 lycocarpum) Cassava (Manihot 61.15 ± 0.18 67.73 ± 0.35 73.95 ± 0.17 13.73 ± 1.33 esculenta)* Edible Canas (Canna 65.44 ± 0.05 70.08 ± 0.01 74.84 ± 0.09 14.24 ± 0.72 edulis)* Turmeric (Curcuma 78.78 ± 0.18 82.68 ± 0.25 89.04 ± 0.55 13.73 ± 0.24 longa)* Ginger (Zingiber 81.77 ± 0.28 87.41 ± 0.35 93.28 ± 0.56 20.23 ± 1.30 officinale)*
* Source: Peroni (2003), T0 = initial temperature, Tp = peak temperature, Tc = conclusion temperature and ∆H = enthalpy variation.
Comparison of the initial was found by Peroni (2003) for cassava gelatinization temperatures (T0) obtained (Manihot esculenta), Edible Cana (Canna from DSC with the paste temperatures of edulis), turmeric (Curcuma longa), and RVA (Figure 4) shows that the initial ginger (Zingiber officinale) starches, which gelatinization temperature of the S. present higher values than those obtained by lycocarpum starch is lower than that of the DSC. According to Cereda (2002), the paste paste temperature (69.5°C). The same result temperature obtained by RVA was higher
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because it is sensitive to the first increases that amylose is not complexed with lipids, in the starch paste viscosity, which is since, if present, they could fuse and different from the initial gelatinization produce an endotherm within the temperature detected when the first temperature range of 91-100 °C. This is in granules begin to disrupt. DSC values are agreement with the findings of this study more accurate, while RVA values can show showing low fat content (0.07%) in S. temperature ranges. lycocarpum starch (Table 1). The value of Yoo and Jane (2002) showed that a this endotherm was 13.08 J.g-1 (Figure 5). single endotherm found for starches means
Figure 5. DSC thermogram for S. lycocarpum starch.
The peak in the DSC curve pointing ΔH gel may be correlated with both down in Figure 5 is a representation of the amylopectin crystallinity and the strength endothermic gelatinization reaction that with which the double helices formed by involves fusion of starch granules. their chains are associated with the starch
According Cereda (2002), T0, Tp, and Tc granule. Thus, the higher the ΔH gel, the are influenced by the molecular structure of greater the force required to break the the crystalline region, which corresponds to granule structure, resulting in the distribution of short chains of gelatinization. In thie study, S. Lycocarpum amylopectin. starch required 13.08J.g-1, which is
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considered high and indicates a strong confirm its potential in the production of association of the amylopectin. biodegradable films, having potential for A good gelatinization temperature coating of agricultural seeds that require range (R = (Tc - To)) was found for starch rapid germination (FERNANDES et al., of S. lycocarpum fruit, suggesting the 2019). presence of crystals with different stability Cereda (2002) and Ciacco and Cruz within the crystalline zone, which are (1987) found that crystallinity patterns are broken at different temperatures (Cereda, defined based on the interplanar spaces (d) 2002). and the relative intensity of the X-ray Abraharm and Simi (2008) discussed diffraction lines. The starch from S. that starch gel gelatinization is a response to lycocarpum produced six main peaks the nature of the granules, regarding their around the diffraction angles 4.2°, 6.5°, amylose and amylopectin contents, the 14.8°, 17.2°, 21.5° and 23.8° (Figure 6). spatial arrangement of both polysaccharides These peaks are typical of granules with in the internal structure of the granule, and type B structure, presenting few branched the degree of compaction. The starch chains and numerous long chains. gelatinization properties of S. lycocarpum
Figure 6. X-ray diffractogram of S. lycocarpum starch.
Table 3 shows the results for the angle 2, the interplanar space, and intensity of S. lycocarpum and Arracacia xanthorrhiza starches.
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Table 3. Main x-ray diffraction intensity peaks of starch from S. lycocarpum and A. Xanthorrhiza. Solanum lycocarpum Arracacia Xanthorrhiza* 2 Intensity d** 2 Intensity d** 6.55 192 13.48 5.55 146 15.90 14.8 1272 5.98 14.9 266 5.94 17.15 2253 5.16 17.02 635 5.20 19.1 1239 4.64 19.27 240 4.60 21.45 1219 4.14 22.03 331 4.03 23.75 1479 3.74 24.0 256 3.70 * Source: VIEIRA (2004) ** Calculated with Bragg's law.
In the characterization by x-ray However, further and more detailed diffraction of Arracacia xanthorrhiza research is needed to evaluate the presence starch, Vieira (2004) found similar values of substances unsuitable for food for the six main peaks around the diffraction consumption (LAJOLO; MENEZES, angles, showing that the starch is also type 2006). B, but the intensities found for the respective peaks were much lower. CONCLUSIONS Type B crystal starch has an open and S. lycocarpum fruits have potential highly hydrated structure with double as a raw material for starch extraction as helices arranged in a hexagonal fashion they produce yields comparable to that of (CEREDA, 2002). It is found in the starch raw materials from commercial starch from tubers, which are varieties with high sources. amylose content and retrograded starches. Chemical composition, paste This is in agreement with the high amylose viscosity, calorimetric and crystallographic content found for S. lycocarpum (Table 1) analyses showed that starch from S. in the present study. Lajolo and Menezes lycocarpum has desirable characteristics: (2006) also agree that because they are rich good stability at high temperature and in amylose, these types of starch present mechanical stability, which makes it a similar shapes and sizes and are resistant to promising material for several food and both enzymatic and acid hydrolysis, which industrial applications. makes them comparable to dietary fibers. This characteristic increases the potential REFERENCES use of this raw material in the food industry. ABRAHAM T E AND SIMI C K. Physicochemical Rheological and 11 Revista Agrotecnologia, Ipameri, v.11, n.2, p.1-13, 2020
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