Accepted Manuscript

Physicochemical and functional properties of Thai organic rice flour

Supaluck Kraithong, Suyong Lee, Saroat Rawdkuen

PII: S0733-5210(17)30623-9
DOI: 10.1016/j.jcs.2017.10.015
Reference: YJCRS 2468

To appear in: Journal of Cereal Science

Received Date: 13 August 2017

Revised Date: 16 October 2017
Accepted Date: 18 October 2017

Please cite this article as: Kraithong, S., Lee, S., Rawdkuen, S., Physicochemical and functional
properties of Thai organic rice flour, Journal of Cereal Science (2017), doi: 10.1016/j.jcs.2017.10.015.

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2 Physicochemical and Functional Properties of Thai Organic Rice Flour
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9 To be submitted to
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11 Journal of Cereal Science
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18 Supaluck Kraithong,1 Suyong Lee,2 & Saroat Rawdkuen1*
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25 Innovative Food Packaing and Biomaterials Unit, School of Agro-Industry, Mae Fah Luang
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26 University, Muang, Chiang Rai, 57100 Thailand

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28 Carbohydrate Bioproduct Research Center, Department of Food Science & Technology,
29 Sejong University, Seoul 143-747, South Korea
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38 *To whom correspondence should be addressed. Tel: +66-53-916752.
39 Fax: +66-53-916739. E-mail: saroat@mfu.ac.th
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45 Abstract

46 Rice flour can be applied into many kinds of foods, but it can have an effect on the final product

47 quality. This study aimed to assess the properties of four different types of flours, namely Thai

48 organic white (Phitsanulok ;PRF), colored (Riceberry ; RBF), brown Jasmine (BJF), and red

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49 Jasmine (RJF). The chemical composition (moisture, ash, crude fat, protein, fiber, and carbohydrate

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50 contents) of all samples were significantly different (p<0.05). PRF contained the highest amylose

51 content (33.80%) (p<0.05). The highest peak (4,067.33 cP) and breakdown viscosities (2,077.33 cP)

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52 were observed in BJF (p<0.05) whereas the highest setback (1,551.33 cP) was found in PRF

53 (p<0.05). The onset, peak, and conclusion temperatures were significantly different (p<0.05) while

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gelatinization enthalpy were not significantly different (p>0.05). The highest water absorption (7.14
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55 g/g) and water solubility indexes (7.05%) were found in colored rice flour (p>0.05) (RJF and BJF,
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56 respectively). Ultimately, higher amylose content resulted in lower gelatinization temperature and in

57 percentage of crystallinity, but there was an increasing setback value for PRF. Meanwhile, low
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58 amylose content in colored rice flour promoted high values of gelatinization temperature, pasting
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59 (peak and breakdown viscosities), and functional properties (water solubility index).

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61 Keywords: organic rice flours; physicochemical; functional properties; proximate, colored rice

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68 1. Introduction

69 Thailand is a major rice producing country. In 2017, the total volume of Thai rice exports was

70 7,395,579 metric tons (US$ 3, 324.03 million) (Thai Rice Exporters Association, 2017). Thai rice

71 can be classified into 2 types: non-pigmented (or white rice) and pigmented rice (or colored rice).

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72 White rice is obtained by removing the husk and outer layer from the entire grain while colored rice

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73 is achieved by eliminating the husk and only a small amount of the outer layer that contains some

74 pigment. Different pigments that include anthocyanin, proanthocyanidins, and carotenoids produce

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75 differences in rice grain color that range in black, blue, red, and brown (Pereira-Caro et al., 2013).

76 Even though white rice has long been a staple food in Asia, few people know that pigmented rice

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actually contains higher health benefits (Sampaothong, 2016). Recently, pigmented rice has become
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78 a popular alternative for health conscious consumers.

79 Rice flour (obtained from rice grain) is widely used as an ingredient in many food products,
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80 including beverages, meat products, puddings, salad dressings, and gluten-free diets (Falade and
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81 Christopher, 2015). Organic rice flour has also become a popular raw material in many health food
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82 products because of consumers’ health concerns. It is considered a healthier and safer product than

83 non-organic flour due to the wide use of irradiation, pesticides, herbicides, and chemical fertilizers.
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84 Variations in properties of rice flour have significant effects on the overall quality of the product.

85 The high ability of flour to absorb water is required for processing foods with soft and smooth
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86 textures (Shafi et al., 2016). Oil absorption capacity is related with rancidity that naturally occurs in
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87 foods, which is a major predictor of shelf-life (Tharise et al., 2014). Fine and massive textures in

88 food products are promoted by rice flour that is a small particle size(Chung et al., 2011). High

89 viscosity rice flour can be properly employed into high viscosity food as a thickening agent (Ye et

90 al., 2016). Processed foods with rigid structures contribute to high setback rice flour (Hsu et al.,

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91 2015). The presence of low gelatinization temperature promotes complete gelatinization upon

92 cooking or steaming rice flour (Oka and Ugwa, 2011). The thermal properties of rice flour are

93 related to the presence of crystallinity (Zeng et al., 2015). The various values of pasting, thermal,

94 and functional properties in rice flour are influenced by their chemical composition (e.g. amylose,

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95 lipid, protein contents) (Prasad et al., 2012). The presenting color in pigmented rice flour contributes

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96 to higher health benefits, such as reducing and preventing the risk of chronic diseases due to its

97 phenolic compounds (Anggraini et al., 2015). Moreover, color is one important factor that affects

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98 consumer’s purchase decision-making.

99 Accordingly, physicochemical and functional properties of rice flour need to be determined.

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Many studies about the qualities of non-organic rice flour have been reported while the study of
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101 organic rice flour properties have been less reported. Therefore, this study aims to determine

102 physicochemical and functional properties of four organic rice flours (white and colored rice flours)
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103 in Thailand.
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104
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105 2. Materials and Methods

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107 2.1. Raw materials

108 Four varieties of Thai organic rice (Oryza sativa) were used in this study. They are white
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109 (Phitsanulok) and pigmented rices (Riceberry, brown Jasmine (Khao Dawk Mali 105) and red
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110 Jasmine (Hom Mali Dang). Phitsanulok rice flour (PRF) was provided by Urmatt Ltd. (Chiang Rai,

111 Thailand). Riceberry flour (RBF) was purchased from Boonleang Farm (Phetchabun, Thailand).

112 Brown and red Jasmine rice grains were purchased from Siam Organic Food Products Co., Ltd.

113 (Bangkok, Thailand).

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114

115 2.2. Rice flour preparation

116 The brown and red Jasmine rice flours (BJF and RJF) were prepared by grinding their grains into

117 powders with a hammermill (CMC-20, Thailand). The rice powders was passed through a 60 mesh

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118 sieve (250 µm) and then vacuum-packed in polyethylene bags. Four Thai organic rice flours were

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119 stored at 4°C before further analysis.

120

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121 2.3. Chemical properties

122 2.3.1. Proximate composition

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The contents of moisture (method 934.01), ash (method 945.46), crude protein (N ×5.95) (method
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124 992.15 (39.1.16)), crude fat (method 954.02), and crude fiber (method 978.10) of the rice flours

125 were determined according to the standard methods of AOAC (2000). Total carbohydrate content
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126 was estimated by the deducting the total percentage of the other components from one hundred.
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127
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128 2.3.2. Amylose content

129 Amylose content was examined by the colorimetry method described in Juliano (1971). The
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130 samples (100 mg) were mixed with 1 mL of 95% ethanol and 9 mL of 2 M NaOH. After that, the

131 mixtures were adjusted to 100 mL with distilled water. The solutions were added with 2.0 mL of
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132 0.2% iodine solution and the absorbance was measured at 620 nm (Genesy 10S UV-Vis
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133 spectrophotometer, Thermoscientific, USA). The amylose content was estimated by referring to a

134 calibration curve. The amylose from potato starch was used as a standard.

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137 2.4. Physical properties

138 2.4.1. Color attributes

139 The color was determined by using a colorimeter (Miniscan EZ, USA). The color parameters were

140 L* (L* = 0, black and L* = 100, white), a* (−a* = greenness and +a* = redness), and b* (−b* =

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141 blueness and +b* = yellowness). The colorimeter was calibrated with a white calibration tile.

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142

143 2.4.2. Particle size distributions

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144 The size of the organic rice flours was observed using a particle size analyzer (Mastersize, Malvern

145 2000, England). The measurement was carried out with a dry dispersion module under vacuum

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(Scirocco 2000) with 2-3 bars of pressure and 50-90% of the feed rate. After that, the particle size
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147 distribution was recorded.

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149 2.5. Pasting properties

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150 A rapid visco analyser (RVA 4500, Perten Instruments, Sweden ) was used for determining the
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151 pasting properties. The specimens (3 g of rice flours) were placed in a canister and added with

152 distilled water (approximately 25 mL, 14% moisture basis). The RVA profile was obtained under
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153 specific conditions. The temperature was held at 50°C for 1 min and then raised up to 95°C in 3.8

154 min (held for 2.5 min). After that, it was cooled to 50°C within 3.8 min and held for 1.4 min.
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155 The RVA parameters including the peak viscosity (PV), trough viscosity (TR), breakdown (BD),
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156 final viscosity (FV), setback (SB), and pasting temperature (PT) were all recorded.

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158 2.6. Thermal properties

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159 Thermal properties were determined with a differential scanning calorimetry (DSC8000,

160 PerkinElmer, USA) based on Thumrongchote et al. (2012). The specimens (4 mg) with 16 mg of

161 distilled water were accurately weighed into stainless steel pans and hermetically sealed. After that,

162 the samples were allowed to stand for 12 h at room temperature and then scanned over the range of

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163 0-100°C with a heating rate of 10°C/min in a nitrogen atmosphere (20 mL/min). The DSC

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164 parameters including the onset (To), peak (Tp), conclusion temperatures (Tc), and gelatinization

165 enthalpy (∆Hg) were recorded.

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167 2.7. Functional properties

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2.7.1. Water absorbtion Index (WAI) and water solubility Index (WSI)
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169 The WAI and WSI were determined following the method of Kadan et al. (2008). The rice flours (1

170 g) were suspended in 10 mL of distilled water and mixed with a vortex mixer for 1 min. The
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171 suspensions were heated in a water bath at 30°C for 30 min with gentle stirring. They were then
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172 centrifuged at 3,000 rpm (AVANTI j-30I, Beckman, Germany) for 10 min. The supernatants were
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173 carefully poured into an aluminum moisture dish before being dried at 105°C overnight. The

174 sediments were collected and weighed. The WAI and WSI were calculated using the following
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175 formulas:

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Weight of wet sediment (g)

177 WAI (g/g) =
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Dry weight of flour (g)

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Weight of dried supernatan t (g)

179 WSI (%) = × 100
Dry weight of flour (g)

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181 2.7.2. Bulk density

182 Bulk density was determined by following the method of Falade and Christopher (2015). The rice

183 flours (50 g) were weighed into a 100 mL graduated cylinder. After that, the samples were tapped

184 30 times. The volumes of the samples were recorded. The bulk density was calculated by using the

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185 following formula:

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186 Weight of sample (g)
Bulk density (g/cm 3 ) =
Volume of sample after tapp ing (cm 3 )

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187 2.7.3. Oil Absorption Index (OAI)

188 The OAI was examined based on the protocol developed by Malomo et al. (2012). The samples (1

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189 g) were mixed with 10 mL of soybean oil. The mixtures were centrifuged at 4,000 rpm for 20 min.
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190 After that, the surplus oil was decanted while the residues (weight of oil absorbed) were weighed.

191 The calculation of OAI was as follows:

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Weight of oil absorbed (g)

192 OAI (g/g) =
Weight of sample (g)
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193 2.8. Morphological properties

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194 2.8.1. Isolation of rice starch

195 The rice starch was isolated by using an alkali extraction method in accordance with the method
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196 described in Lumdubwong and Seibi (2000). The rice starch was steeped in 0.05 M NaOH solution

197 for 5 h at room temperature in order to remove the protein that was adhered to the fiber and lipid in
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198 the endosperm. The starch slurry was centrifuged at 3000 rpm at room temperature for 20 min. The
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199 sediments were washed until obtaining white color starches. The obtained starch was then dried

200 with a hot air oven at 40ºC for 48 h. The starches were ground with a blender and stored in an air-

201 tight container at 4ºC before use.

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203 2.8.2. Determination of starch morphology

204 The rice starch was coated with gold under 20 mA of plasma current by a sputter coater (SC7620,

205 Polaron, UK). After that, the morphologies of the samples were examined under a scanning electron

206 microscope (SEM, model 1450 VP, LEO, England) with a magnification 2,500X at an accelerating

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207 voltage of 15kV.

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209 2.8.3. X-ray diffraction pattern and crystallinity

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210 The X-ray diffraction pattern was carried out with an X-ray diffractometer (XRD) (X'Pert Pro MPD,

211 Panalytical, Japan) in accordance with the method of Thumrongchote et al. (2012). The organic rice

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flours were subjected to the aluminum sample holders. The X-ray source was Cu Kα1 radiation (λ =
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213 0.15405 nm) with an operating the voltage of 40 kV and a current of 30 mA. The diffraction data

214 was collected over an angular range from 5-40º (2θ) at a scanning rate of 0.1º/ min with a step size
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215 of 0.05°. The crystallinity was computed by the following equation:

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216 Total crystallin e peak area

Percentage of crystallin ity = × 100
Crystallin e and amorphous peak
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217 2.9. Statistical analysis

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218 An analysis of variance (ANOVA) was performed. The mean comparison was carried out by

219 Duncan’s Multiple Range Tests. The significance of difference was defined at p < 0.05. The
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220 analysis was performed by using an SPSS package (SPSS 17.0 for window, SPSS Inc, Chicago, IL).
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226 3. Results and Discussion

227 3.1. Chemical properties

228 3.1.1. Proximate composition

229 The proximate composition of Thai organic rice flours is presented in Table 1. The moisture content

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230 (5.47-9.87%), ash (0.47-1.57%), crude protein (6.51-7.27%), crude fat (1.13-3.60%), and

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231 carbohydrate (77.06-85.58%) were significantly different (p<0.05) for all samples. These results are

232 in agreement with Oko and Ugwu (2011) who reported values of moisture at 3.67-6.67%, ash at

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233 0.50-2.00%, crude fat at 0.50-3.50%, crude protein at 1.58-6.22%, crude fiber at 1.50-2.00%, and

234 carbohydrate at 76.92-86.03% in five rice varieties (Sipi, Faro 14, Faro 15, Awilo, and Canada) in

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South-Eastern Nigeria. This study found that the moisture (5.47%), ash (0.47%), crude protein
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236 (6.51%), crude fat (1.13%), and crude fiber (0.81%) contents of PRF were lower than other rice

237 flours (p<0.05) while PRF had the highest carbohydrate content (85.58%) (p<0.05). Rice flour
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238 compositions generally rely on rice varieties. Higher contents of ash, crude protein, crude fat, and
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239 crude fiber in colored rice flour (RBF, BJF, and RJF) are present because they have a greater outer
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240 layer. However, higher values of those components contribute to lower carbohydrate content

241 (Wang et al., 2016). Likewise, the proximate composition of rice flour could be affected by the
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242 conditions of their processing (dry and wet-milling). Wet-milling processes tend to provide rice

243 flour with higher carbohydrate content (and lower content of other components). This is due to the
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244 soaking and grinding process where the soluble protein, sugars, and non-starches all bind with lipids
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245 and are removed (Chiang and Yeh, 2002). Lower moisture content in rice flour is supported by

246 higher temperature during drying and storage (Oko and Ugwu, 2011). Consequently, colored rice

247 flours (RBF, BJF, and RJF) tend to provide higher nutrients than white rice flour (PRF) because

248 they contain higher values of ash, crude fat, crude protein, and crude fiber.

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249

250 3.1.2. Amylose content

251 The values of amylose content in the organic rice flours were 18.30-33.80% (Table 1). These results

252 are consistent with Ye et al. (2016) who described an amylose content of 10.40-26.50% in Indica

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253 rice flour. This study found that high a amount of carbohydrate contributed to high amylose content

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254 in rice flour. According to the obtained results, the samples were classified into low (RBF, BJF, and

255 RJF) (≤20%) and high (PRF) (>25%) amylose rice flours (Chiang and Yeh, 2002). Higher amounts

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256 of carbohydrate tend to increase amylose content, yet this also results in low values of other

257 components such as ash, protein, lipid, and fiber (Oko and Ugwu, 2011). Low amylose rice flour

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generally offers dampness, softness, and chewiness to product textures. These qualities can also be
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259 applied in meat products, puddings, and soft cakes (Falade and Christopher, 2015). High amylose
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260 rice can provide firmness and crispness to products due to a three-dimensional network formation

261 (Wang et al., 2016). Hence, it could be employed in food products that need a hard texture including
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262 snacks, noodles, and extruded products.

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263

264 3.2. Physical properties

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265 3.2.1. Color attributes

266 The color parameters (L*, a*, and b*) of the samples were significantly different (p<0.05) (Table 1).
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267 The obvious differences in rice flour colors are influenced by their phenolic compounds, including
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268 anthocyanin (blueness and purpleness), proanthocyanidins (redness), and carotenoids (yellowness)

269 (Anggraini et al., 2015). According to Pereira-Caro et al. (2013), total anthocyanins were found in

270 black (3,474 µg/g) and red rices (4.3 µg/g). Total carotenoids were found in black (482 µg/g), red

271 (22 µg/g), brown (21 µg/g), as well as in white rices (24 µg/g). Consequently, higher values of a*

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272 and b* were found in pigmented rice flours (RBF, BJF, and RJF) (p<0.05) while the highest value

273 of L* was observed in white rice flour (PRF) (p<0.05). Nevertheless, trace levels of carotenoids in

274 PRF resulted in lower yellowness value (p<0.05). The pigments in colored rice flours possess the

275 ability to reduce and prevent the risk of chronic diseases such as hypercholesterolemia,

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276 cardiovascular disease, and type II diabetes (Ye at al., 2016). As a consequence, higher amounts of

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277 pigments in colored rice flour (higher a* and b* values) represent higher health benefits than white

278 rice flour.

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279

280 3.2.2 Particle size distribution

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The average particle sizes of RBF (208.93 µm), RPF (79.43 µm), BJF (158.49 µm), and RJF
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282 (158.49 µm) were observed (Fig. 1). Ahmed et al. (2015) reported the average particle size of Indica

283 rice flour (Basmati) to be 297 µm. The particle size of rice flour could be influenced by its
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284 processing. According to Prasad et al. (2012), dry-milling provides a larger particle size of flour
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285 while wet-milling tends to reduce flour particle size. Soaking and grinding rice grains with water
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286 (wet-milling) contribute to a smaller particle size because rice grain breaks down easily. Moreover,

287 a longer period for the rice grinding process (dry and wet milling) tends to decrease the particle size.
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288 The size affects other properties of the rice flour such as the pasting and functional properties.

289 According to Ye et al. (2016), rice flour with small particle size has high values of viscosity,
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290 swelling power, and water absorption due to the large surface area that associates with water. Thus,
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291 RPF can provide smoothness, fineness, cohesiveness, stickiness, and an oily mouth feel to food

292 products. According to Otegbayo et al. (2013), rice flour with large particle size contributes to high

293 values of bulk density. Hence, food products with hardness, fracturability, and porosity are received

294 from larger particle size rice flours.

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295

296 3.3. Pasting properties

297 RVA profile of the Thai organic rice flours is presented in Fig. 2. The highest peak viscosity

298 (4,067.33 cP) was observed in BJF (p<0.05) (Fig. 2). It indicates a high ability of the starch granule

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299 to bind water via hydrogen bonds (Otegbayo et al., 2013). High peak viscosity is promoted by high

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300 amylopectin content because of high water holding capacity (Ye et al., 2016). Besides, large surface

301 area for binding water by small particle size also increases viscosity in rice flour (Prasad et al.,

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302 2012). Therefore, high amylopectin rice flour requires less energy for cooking. Consequently, the

303 lowest pasting temperature (81.73ºC) was found in BJF (p<0.05). Higher peak viscosity is caused

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by higher breakdown because less heat and shear stress resistance during cooking (Hsu et al., 2015).
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305 Therefore, BJF also presented the highest breakdown (2,077.33 cP) (p<0.05). The highest trough

306 (the minimum viscosity at constant temperature) (2,094.00 cP) and final viscosities (the stability of
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307 cooked paste and ability to form gel after cooling) (3,939.00 cP) were observed in RJF (p<0.05)
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308 (Fig. 2). PRF presented the highest setback (1,551.33 cP) (p<0.05). These results are in agreement
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309 with Falade and Christopher (2015). They reported the values of peak at 2,376-6,080 cP, trough at

310 1,618-4,036 cP, breakdown at 636-3,155 cP, final at 4,086-7,581 cP, setback viscosities at 2,279-
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311 3,980 cP, and pasting temperature at 73.88-86.40ºC for Nigerian rice (Faro 21, 40, 44, 46, 52, and

312 55). According to Ahmed et al. (2015), high setback rice flour is properly applied to stick rice
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313 noodles, fried snacks, and extruded products (providing food structure) while high peak viscosity
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314 rice flour is appropriately employed into high viscosity (e.g. gravy and salad dreesing) and soft

315 textured foods (e.g. puddings and soft cakes).

316 The variations in values of pasting properties are influenced by rice flour compositions (Oko and

317 Ugwu, 2011). According to Wang et al. (2016), amylose is considered as an inhibitor of swelling

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318 because of its tightly packed form. Moreover, a three-dimensional network that occurs by re-

319 association of amylose upon cooling can increase the setback value in rice flour (Jamal et al., 2016).

320 Accordingly, high amylose rice flour tends to provide a low peak, trough, and breakdown

321 viscosities; however it provides high setback, final viscosities, and pasting temperature (Ye et al.,

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322 2016). Not only amylose content, but also other components such as lipid and protein, can affect the

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323 pasting properties in rice flour. Amylose-lipid or amylose-protein complex formations can promote

324 high values of setback and final viscosities, while these complexes reduce the values of peak and

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325 breakdown viscosities (Alcázar-Alay and Meireles, 2015). In contrast, the peak viscosity of rice

326 flour is also supported by high amounts of protein in rice flour due to the higher water holding

327
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ability of its hydrophilic groups (Hsu et al., 2015).
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328

329 3.4. Thermal properties

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330 Thermal properties of the organic rice flours are represented in Table 2. There was no significant
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331 difference in gelatinization enthalpy (∆Hg) for all samples (p>0.05). Lower values of onset (To)
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332 (66.50ºC), peak (Tp) (71.68ºC), and conclusion temperatures (Tc) (76.65ºC) were found in white rice

333 (PRF) (p>0.05). These results are consistent with Ahmed et al. (2015) who reported the
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334 gelatinization temperature of Indica rice flour (Basmati) (66-70ºC). The higher values of DSC

335 parameter (To, Tp, and Tc) are encouraged by large amylopectin branches (crystallinity). According
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336 to Alcázar-Alay and Meireles (2015), high energy is required for disrupting the large crystalline
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337 regions of high amylopectin rice flour. Besides, amylose-lipid and amylose-protein complex

338 formations in pigmented rice flour also can increase the gelatinization temperature due to their rigid

339 structures (Morales-Martínez et al., 2014). In contrast, the gelatinization of flour with high

340 amorphous regions (high amylose) is accomplished easily because of weak hydrogen bonds (Jamal

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341 et al., 2016). The small particle size of rice flour advocates low gelatinization temperature because

342 of large surface areas for binding water molecules (Ye et al., 2016). Moreover, white rice flour has a

343 high ability to absorb water and swell, which also promotes gelatinization (Hsu et al., 2015). Thus,

344 the energy required for the gelatinization of white rice flour (PRF) was lower than RBF, BJF, and

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345 RBF (p>0.05).

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346

347 3.5. Functional properties

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348 3.5.1. WAI and WSI

349 WAI represents the ability of rice flour to associate with water molecules (Shafi et al., 2016). The

350
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WAI values of Thai organic rice flours were 5.44-7.14 g/g (Table 2). The results are in agreement
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351 with Jamal et al. (2016) who described the WAI value for Pakistani rice (Fakhr-e-Malakand and

352 Basmati) (5.38-6.26 g/g). The highest WAI was found in RJF (7.14 g/g) (p<0.05). It is contributed
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353 by a high number of hydrophilic groups within starch molecules, providing softness, smoothness,
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354 and viscosity in food products (Aprianita et al., 2014). High contents of carbohydrates and proteins
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355 in rice flour tend to encourage a strong hydrogen bonding because of polar or charged side chains

356 (Prasad et al., 2012). Besides, increasing water binding capacity is produced from the negative
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357 charges of phosphate groups within amylopectin (Wang et al., 2016). In contrast, high lipid content

358 in rice flour contributes to low WAI by interrupting starch granule hydration with hydrophobic parts
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359 (Alcázar-Alay and Meireles, 2015), Moreover, large particle size tends to reduce the value of WAI
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360 (Otegbayo et al., 2013). Accordingly, the lowest WAI (5.44 g/g) was observed in RBF (p<0.05),

361 which resulted in a high gelatinization temperature in rice flour and a rough texture in its products.

362 In addition, the interruption of water binding in rice flour is caused by the presences of amylose-

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363 lipid/amylose-protein complexes, which reduce polar and charged molecules (Falade and

364 Christopher, 2015).

365 The WSI values of the samples (2.97-7.05%) are shown in Table 2. These results are consistent with

366 Jamal et al. (2016) who described the values of WSI for Pakistani rice (Fakhr-e-Malakand and

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367 Basmati) (1.95-4.94%). The highest WSI (7.05%) was found in BJF (p<0.05). This indicates a high

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368 amount of water soluble components that disperse in aqueous during cooking (Shafi et al., 2016). A

369 higher WSI in rice flour indicates a higher value of adhesiveness and stickiness in food products.

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370 However, it also presents a low ability to preserve food structure (Wang et al., 2016). Normally,

371 junction zone formation by amylose encourages a rigid structure of starch granules, providing low

372
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WSI (Chung et al., 2011). In addition, complexation of starch with proteins or lipids can reduce the
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373 WSI value in rice flour because of a decrease in the soluble parts within starch molecules

374 (Keawpeng and Meenune, 2012). Thereby, the lowest WSI was found (2.97%) in RJF (p<0.05),
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375 indicating a high ability to maintain food structures during the cooking process.
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376
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377 3.5.2. Bulk density

378 Bulk density is used to determine flour expansion and the porosity of products. Moreover, it also
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379 indicates the volume of packaging material (Shafi et al., 2016). The values of bulk density of the

380 samples were 0.57-0.73 g/cm3. The results are in good agreement with Falade and Christopher
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381 (2015) who reported a bulk density of 0.53-0.89 g/cm3 for Nigerian rice (Faro 21, 40, 44, 46, 52,
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382 and 55). The larger particle size of rice flour encourages higher values of bulk density (Ye et al.,

383 2016). Consequently, the highest value of bulk density was found in RBF (p<0.05). In contrast, the

384 high compactness and lower porosity of starch granules reduces the bulk density value (Tharise et

385 al., 2014). High bulk density flour is desirable for reducing paste thickness due to its low viscosity

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386 and for food products with rough textures. Whereas, low bulk density flour is properly served for

387 food with smooth and dense textures (Jamal et al., 2016).

388

389 3.5.3. OAI

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390 There were no significant differences in OAI values (p>0.05) for all samples. The OAI values of the

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391 samples were 1.11-1.34 g/g. This indicates the ability of the starch granule to maintain oil. These

392 results are clearly consistent with Sarangapani et al. (2016) who reported the oil holding capacity of

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393 rice flour (Aiyre) to be 0.88-1.39 g/g. High numbers for hydrophobic groups within starch

394 molecules contribute to high values of OAI (Tharise et al., 2014). Improving mouthfeel, flavor

395
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retention, and palatability in food products are promoted by high OAI. Yet, rancidity occurring in
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396 food products is boosted by higher values of OAI (Falade and Chirstopher, 2015).
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397

398 3.6. Structure

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399 3.6.1. Granule morphology

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400 Polyhedral shapes with smooth surfaces were mainly found in the rice starch granules (Fig. 3).

401 However, there were a few small irregular shapes also observed. The length (1.6-6.2 µm) and the
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402 width (1.0-6.0 µm) of the samples were measured. These results are in agreement with Falade and

403 Christopher (2015) who reported the granule shape (polyhedral), length ranges (1.30-15.00 µm),
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404 and width ranges (1.00-13.00 µm) for Nigerian rice (Faro 21, 40, 44, 46, 52, and 55). According to
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405 Otegbayo et al. (2013), the differences in granule shapes (polyhedral, oval, and round shapes) and

406 sizes are due to the biological origin of the plants. Observations of shape and size are used for

407 identifying starch sources. Different granule shapes and sizes are received from different types of

408 starches. The variations in starch granule size could be influenced by amylose and amylopectin

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409 contents. The presence of large a crystalline region reinforces the sizes of starch granules because of

410 its amylopectin branches (Prasad et al., 2012). The highest length (6.2 µm) and width (6.0 µm) was

411 found in RJF.

412

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413 3.6.2. X-ray diffraction pattern and crystallinity

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414 The X-ray diffraction patterns of the samples were A-type. The main diffraction peaks were 15°,

415 17°, 18°, and 23° (Fig. 4). Normally, an A-type pattern is found in cereal flours including rice, waxy

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416 maize, and wheat (Otegbayo et al., 2013). According to Zeng et al. (2015), an A-type pattern is

417 reinforced by a high degree of crystallinity and tightly packed granules in cereals. Flour made from

418
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root or tuber represents B-type patterns because they contain a large proportion of amylopectin
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419 (Wang et al., 2016). Complexation of amylose with lipid, during the extrusion process, was

420 observed in the V-type pattern. The pattern of crystallinity could be changed during food processing
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421 (Prasad et al., 2012).

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422 The crystallinities of RBF (37.02%), RPF (32.50%), BJF (34.71%), and RJF (35.68%) were
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423 examined and are presented in Table 1. These results are consistent with Chung et al. (2011) who

424 reported about the crystallinity of Indica rice starch (29.9-33.5%). A high crystalline region tends to
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425 increase the gelatinization temperature in rice flour because it interrupts water penetration within

426 starch molecules (Alcázar-Alay and Meireles, 2015). A high percentage of crystallinity is supported
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427 by large cluster branches of amylopectin or tightly packed forms of double helices (Keawpeng and
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428 Meenune, 2012). Moreover, higher amounts of other components such as protein and fat also

429 reinforce the percentage of crystallinity. Rigid structures (complexation of amylose with

430 lipid/protein) are contributed to the large crystalline regions in starch molecules (Morales-Martínez

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431 et al., 2014). For that reason, a higher percentage of crystallinity was found in low amylose rice

432 flours (RBF, BJF, and RJF) (p<0.05).

433

434 4. Conclusion

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435 Differences were observed in the physicochemical and functional properties of Thai organic white

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436 and colored rice flours. The variations are a result of various factors such as the proximate

437 composition, amylose content, and particles size. Higher values of pasting properties, including

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438 peak and breakdown viscosities, were presented in colored rice flours (low amylose flours).

439 However, high values of setback was observed in high amylose white rice flour (PRF). Lower

440
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gelatinization temperature in white rice flour (PRF) could be promoted by a lower amount of other
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441 contents (such as lipids and proteins) and by a smaller particle size. A higher water solubility index

442 was observed in low amylose of BJF. The morphology of starch granule was not affected by their
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443 chemical compositions. Higher crystallinity in pigmented rice flour was caused by higher numbers
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444 of amylopectin branches (in the crystalline regions) and by higher contents of lipids and proteins.
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445 The results obtained from this study provide basic information that is beneficial to the food industry.

446
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447 Acknowledgements

448 The authors thank Urmatt Ltd (Chiang Rai, Thailand) for providing organic rice flour (Phitsanulok)
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449 and Scientific & Technological Instruments Center of Mae Fah Luang university (Chiang Rai,
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450 Thailand) for providing the facilities to carry out this study. This research was financially supported

451 by Mae Fah Luang Univerity and the Thailand Research Fund (TRF) under the Royal Golden

452 Jubilee Ph.D. Program (RGJ) (Grant NO. PHD/0087/2558).

453

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545 Figure & Table Legends

546 Fig 1. Particle size distribution of organic rice flours: Riceberry flour (RBF), Phitsanulok rice flour

547 (PRF), brown Jasmine rice flour (BJF), and red Jasmine rice flour (RJF).

548

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549 Fig 2. Pasting properties of organic rice flours: Riceberry flour (RBF), Phitsanulok rice flour (PRF),

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550 brown Jasmine rice flour (BJF), and red Jasmine rice flour (RJF).

551

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552 Fig 3. Scanning electron micrographs of organic rice starches at 2500× magnification: Riceberry

553 flour (A), Phitsanulok rice flour (B), brown Jasmine rice flour (C), and red Jasmine rice flour (D).

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554
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555 Fig 4. X-ray diffraction pattern of organic rice flours: Riceberry flour (RBF), Phitsanulok rice flour

556 (PRF), brown Jasmine rice flour (BJF), and red Jasmine rice flour (RJF).
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557

558 Table 1. Chemical and physicochemical properties of Thai organic rice flours.
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560 Table 2. Theraml and functional properties of Thai organic rice flours.
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Table 1. Chemical and physicochemical properties of Thai organic rice flours.

Proximate composition and amylose content

Rice flour
Moisture Ash Crude protein Crude fat Crude fiber Total Amylose

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(%) (%) (%) (%) (%) carbohydrate (%) (%)
a b a a a b
RBF 9.60±0.09 1.14±0.03 7.54±0.29 3.60±0.80 1.66±0.04 77.06±1.20 18.30±1.77b

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PRF 5.47±0.05b 0.47±0.09c 6.51±0.15b 1.13±0.23bc 0.81±0.34b 85.58±0.42a 33.80±0.05a

BJF 9.04±1.53a 1.57±0.01a 7.61±0.53a 1.89±0.44b 0.91±0.33b 78.19±1.32b 19.18±0.53b

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RJF 9.87±0.13a 1.44±0.30a 7.27±0.0.22ab 2.05±0.10ab 1.35±0.18ab 78.07±0.11b 20.18±0.53b

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Color Granules size

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Rice flour Appearance Crystallinity (%)
Length range Width range
L* a* b*
(µm) (µm)

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RBF 66.69±0.51d 2.80±0.06b 7.71±0.16b 2.0-5.8 1.2-5.8 37.02±0.57a

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PRF 94.98±0.04a -0.02±0.02d 5.15±0.01c 3.2-6.0 1.0-5.6 32.50±0.91c

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BJF 85.57±0.46b 0.78±0.03c 11.25±0.12a 2.0-6.0 1.6-5.8 34.71±0.88b
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RJF 71.34±0.75c 8.31±0.16a 11.22±0.12a 1.6-6.2 1.2-6.0 35.68±0.95b

All values are means of triplicates ± standard deviation. a-d Means with the same superscript letters with in a column are not significantly different at p <0.05
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level. Riceberry flour (RBF), Phitsanulok rice flour (PRF), brown jasmine rice flour (BJF), and red jasmine rice flour (RJF).
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Table 2. Thermal and functional properties of Thai organic rice flours.

Rice flour sample

Functional properties
RBF PRF BJF RJF

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Onset temperature (°C) 69.57 ± 0.37a 66.50 ± 0.14b 70.15 ± 0.77a 69.54 ± 0.76a

75.74 ± 0.45a 71.68 ± 0.63b 75.90 ± 0.01a 75.81 ± 0.29a

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Peak temperature (°C)

Conclusion temperature (°C) 82.41 ± 0.51a 76.65 ± 0.06c 81.18 ± 0.25b 83.18 ± 0.11a

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Gelatinization enthalpy (J/g) 1.65 ± 0.03a 1.04 ± 0.17a 1.24 ± 0.30a 1.70 ± 0.89a

Water absorption index (g/g) 5.44 ± 0.49bc 7.06 ± 0.12a 6.19 ± 0.28ab 7.14 ± 0.89a

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Water solubility index (%) 5.21 ± 0.71b 3.93 ± 0.62c 7.05 ± 0.52a 2.97 ± 0.18cd

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Oil absorption index (g/g) 1.24 ± 0.06 a 1.34 ± 0.12 a 1.24 ± 0.31 a 1.11 ± 0.02 a

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Bulk density (g/cm3) 0.73 ± 0.01a 0.70 ± 0.04a 0.59 ± 0.04b 0.57 ± 0.03b

All values are means of triplicates ± standard deviation. a-d Means with the same superscript letters with in the same row are not significantly different at p<0.05

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Fig 1. Particle size distribution of organic rice flours: Riceberry flour (RBF), Phitsanulok rice
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flour (PRF), brown Jasmine rice flour (BJF), and red Jasmine rice flour (RJF).
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Pasting properties
Rice
Peak viscosity Trough Breakdown Final viscosity Setback Pasting
flour
(cP) (cP) (cP) (cP) (cP) temperature (ºC)
RBF 1,534.33±11.58d 1,125.00±19.55c 409.33±12.04c 2,338.67±15.83d 804.33±3.79b 87.77±0.03a
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c d c b a
PRF 2,259.67±15.31 1,840.00±10.54 419.67±8.62 3,811.00±17.77 1,551.33±10.53 86.80±0.44b
BJF 4,067.33±8.62a 1,990.00±11.06b 2,077.33±17.50a 3,575.67±14.30c -491.67±18.65d 81.73±0.40d
3,597.33±19.00b 2,094.00±17.06a 1,503.33±14.85b 3,939.67±14.36a 342.33±27.90c 84.08±0.40c
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RJF
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Fig 2. Pasting properties of organic rice flours: Riceberry flour (RBF), Phitsanulok rice flour
(PRF), brown Jasmine rice flour (BJF), and red Jasmine rice flour (RJF).
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A B

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C D

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Fig 3. Scanning electron micrographs of organic rice starches at 2500× magnification: Riceberry
flour (A), Phitsanulok rice flour (B), brown Jasmine rice flour (C), and red Jasmine rice flour
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BJF

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PRF

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RBF

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RJF

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Fig 4. X-ray diffraction pattern of organic rice flours: Riceberry flour (RBF), Phitsanulok rice
flour (PRF), brown Jasmine rice flour (BJF), and red Jasmine rice flour (RJF).
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Highlights

Differences were observed in the physicochemical and functional properties of Thai

organic rice flours

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The variations are a result of various factors such as the proximate composition, amylose

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content, etc.

Low amylose flours gave higher values of pasting properties.

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The morphology of starch granule was not affected by their chemical compositions.

Higher crystallinity in pigmented rice flour was caused by higher numbers of

amylopectin branches.
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