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International Conference on , Marine and Environmental Sciences (PMES-2015) Jan. 1-2, 2015 Kuala Lumpur () Study of Glutinous and Non-Glutinous ( Sativa) Varieties on Their Antioxidant Compounds

Widiastuti Setyaningsih, Nikmatul Hidayah, Irfan Estiono Saputro, Miguel Palma Lovillo, and Carmelo García Barroso

The phenolics compounds described in rice grains are Abstract—Cultivated rice () is a global cereal crop phenolic acids [5] and its aldehydes [6]. The most common in . It serves as staple food, thus has a major forms of phenolic acids in rice are derived from contribution to the calorie intake. Furthermore, a number of hydroxybenzoic (C6−C1; an aromatic ring linked to a carbon antioxidant substances have been identified in rice including phenolic atom) and hydroxycinnamic (C6−C3; an aromatic ring linked compounds and melatonin. The concentration and composition of to a three-carbon chain) acids. Along with these two major these antioxidant compounds were studied on glutinous and non- groups, aldehyde analogues such as protocatechuicaldehyde, glutinous grains for both pigmented and non-pigmented rice varieties. Additionally, several medium-amylose rice grains of three p-hydroxybenzaldehyde [7] and vanillin [6], were also found Indonesian varieties (IR64, umbul-umbul and pandan wangi) were in rice grains. also evaluated. Finding indicates that the composition of phenolic As well as phenolic compounds, the presence of melatonin compounds are noticeably different between glutinous and non- [N-acetyl-3-(2-aminoethyl)-5-methoxyindole] in rice grain has grains. The level of both melatonin and total phenolics been identified in a wide range of varieties [8]. However, both in non-glutinous rice was higher than its glutinous variety. Hence, the level and composition of these compounds in different higher amylose content exhibits relatively higher amount of varieties of rice grains may diverge in phenolics and melatonin antioxidant compounds.  concentrations. This discrepancy reveals that one would also expect changes due to the differences on their type of Keywords—Rice, amylose, glutinous, antioxidants that have been distinguished as glutinous (waxy) and non- glutinous variety. I.INTRODUCTION Glutinous differs from the regular (non-glutinous) rice ICE (Oryza sativa) is a pivotal cereal crop as subsists mainly in having low (<5%) or almost no amylose in its starch Rcalories for Southeast Asian livelihoods. This grain is but basically high in amylopectin [9]. Amylose is essentially remarkably essential for the nutrition status of a large long linear chains composed of (1→4)-linked α-D- number of the population within this region. In addition to glucopyranosyl units with a few (1→6)-α-linkages branches nutrition purposes, rice contains secondary metabolites that whilst amylopectin has a higher molecular weight and much are possessing several antioxidant compounds namely shorter chains of (1→4)-linked α-D-glucose units that are melatonin [1] and phenolic compounds [2]. These compounds highly branched through additional (1→6)-α-linkages (Fig. 1). attracted a growing attention in recent years due to their health In general, the amylose content of rice starch varies from 0- benefits, as their consumption has been linked to lowering the 2% in waxy (glutinous), 20-25% in normal or medium and up risk of diseases related with oxidative stress i.e. inhibitory to 30% in high-amylose rice grains. effects on carcinogenesis and mutagenesis [3], [4]. Recent It has been reported higher production of antioxidant research has led to an increase in the production of antioxidant compounds from high-amylose genotypes versus the waxy compounds in transgenic rice seeds and antioxidant-rich rice (glutinous) ones for corn (Zea mays L.) grains [10], further have been successfully generated. Functional rice work will be necessary to study the level and composition of products have also become more widely appreciated in the antioxidants compounds in other foods, including rice. Hence, current market. the objective of the study described here was to assess the levels of antioxidant compounds, including melatonin and phenolics on some rice varieties taking into account their Widiastuti Setyaningsih, is with Department of Analytical Chemistry, Faculty of Sciences, University of Cadiz, Puerto Real, Spain. Department of amylose content. Food and Agricultural Product Technology, Gadjah Mada University, Yogyakarta, . Email: [email protected], [email protected]. Nikmatul Hidayah, is with Indonesian Center of Agricultural Postharvest Research and Development, Bogor, Indonesia. Irfan Estiono Saputro, Miguel Palma Lovillo, and Carmelo García Barroso, are with Department of Analytical Chemistry, Faculty of Science, University of Cadiz, Puerto Real, Spain. Email: [email protected], [email protected], [email protected], [email protected]

http://dx.doi.org/10.15242/IICBE.C0115068 27 International Conference on Plant, Marine and Environmental Sciences (PMES-2015) Jan. 1-2, 2015 Kuala Lumpur (Malaysia)

mL capacity). A cellulose filter was inserted at the outlet end of the extraction cell. An accurately weighed sample of rice powder (2.5 g) and washed sea sand were loaded into the extraction cell. The Pressurized Liquid Extraction (PLE) condition for phenolic extractions was 60% (v/v) EtOAc in MeOH, static time of 10 min in 3 cycles at 190 ºC under a pressure of 200atm, flushing 100% and purge for 60 s; while for melatonin extraction was 70% (v/v) EtOAc in MeOH, static time of 5 min in 2 cycles at 200 ºC under a pressure of 200atm, flushing 50% and purge for 60 s. The resulting extract was dried under vacuum on a rotary evaporator. The residue was reconstituted with methanol and adjusted to a final volume of 5 mL. The liquid

Fig. 1 Molecular representation of amylose and amylopectin. was then passed through a 0.45 µm nylon filter prior to injection on the HPLC-FD system. II. MATERIALS AND METHODS E. Determination of Phenolic Compounds High performance liquid chromatography (HPLC) A. Materials and Chemicals analyses were carried out on Dionex HPLC system comprised HPLC-grade methanol (MeOH), acetic acid, and ethyl a Dionex P680 HPLC Pump, Dionex ASI 100 Automated acetate (EtOAc) were purchased from Merck (Darmstadt, Sample Injector, Dionex PDA-100 Photodiode Array Germany). Melatonin (MEL) standard, M-5250, was obtained Detector, Dionex UCI-50 Universal Chromatography from Sigma-Aldrich (St. Louis, MO, USA). Water was Interface, and Dionex TCC-100 Thermostatic Column purified with a Milli-Q purification system (Millipore, Compartment. Separations were performed on a reversed Billerica, MA, USA). Phenolic compound standards of the phase RP 18 Lichrospher Column (LiChroCART 250 × 4 highest available purity were used. Protocatechuic acid (PRO), (5μm) Merck KgaA). Gradient elution was carried out at a protocatechuic aldehyde (PRA), vanillic acid (VAA), p- flow rate of 1.0 mL min–1. A PDA-100 Photodiode Array hydroxybenzoic acid (p-HBA), vanillin (VAN), p- Detector was used for UV-Vis and the 3D mode was set at hydroxybenzaldehyde (p-HB), ferulic acid (FER) and collection rate of 1.0 Hz, 3D wavelength scan range 250-600 sinapic acid (SIN) were obtained from Fluka (Buchs, nm, 3D bunch width 1 nm and band width 50 nm. The column Switzerland). Guaiacol (GUA), p-Coumaric acid (p-COU), compartment thermostat was set at 25oC. Injection volume caffeic acid (CAF), chlorogenic acid (CHL), 5- was set to 25 μL. hydroxymethyl-2-furaldehyde (HMF), and 5-methylfurfural A gradient elution program was used with two mobile (MF) were obtained from Sigma–Aldrich (St. Louis, MO, phases: A (2% acetic acid and 5% MeOH in water) and B (2% USA). Ellagic acid (ELL) and iso-vanillic acid (IVA) were acetic acid and 88% MeOH in water). The mobile phases were purchased from Sarsynthese (Merignac, France). filtered through a 0.45 µm membrane filter (Millipore) and B. Rice Samples were degassed for 15 min prior to use. The gradient applied was as follows: (time, solvent B): 0 min, 0%; 10 min, 25%; 25 Rice samples including non-pigmented and black min, 40%; 30 min, 50%; 35 min, 50%. The identification of pigmented of both glutinous and non-glutinous rice grains phenolic acids in the samples was achieved by spiking and by were obtained from an Asian market in Spain. Three varieties comparison of retention times and maximum UV-Vis of Indonesian medium-amylose rice produced using absorptions with those of standards. conventional farming systems were obtained randomly from various smallholder rice mills in Central area (umbul- F. Determination of Melatonin umbul, pandan wangi and IR64). Chromatographic separations were carried out on an C. Sample Preparation Alliance HPLC 2695 system, controlled by an Empower Pro 2002 data station (Waters, Milford, MA) and coupled with a A rice sample (20 g) was placed in a plastic cylinder and fluorescence detector (Waters 474 Fluorescence Detector). rice grains were milled with an Ultraturrax homogenizer The column separation and mobile phase preparation were (IKA® T25 Digital, Germany) for 10 min prior to extraction. similar with phenolics determination. The applied gradient The milling process was stopped every 1 min to avoid was as follows: (time, solvent B): 0 min, 0%; 5 min, 35%; 12 excessive heating of the sample. The fine grain was then min, 40%; 15 min, 40%; 20 min, 45%; 25 min, 50%. The homogenized by stirring and then stored in a closed labelled established conditions for fluorescence detector were as bottle. follows: excitation wavelength was set at 290 nm, emission D. Extraction Technique wavelength was set at 330 nm, sensitivity was set at gain Phenolic compounds and melatonin extractions were 1000, attenuation was fixed at 16, and injection volume was performed in a Dionex ASE 200 extractor (Dionex, set to 10 µL. Sunnyvale, CA, USA) equipped with incorporating stainless steel extraction cells (11 mL volume) and collection vials (60

http://dx.doi.org/10.15242/IICBE.C0115068 28 International Conference on Plant, Marine and Environmental Sciences (PMES-2015) Jan. 1-2, 2015 Kuala Lumpur (Malaysia)

G. Determination of Amylose Content The precision of the method was evaluated by performing Amylose content of the samples was determined by repeatability (intra-day) and intermediate precision (extra- Iodometric method based on official methods of analysis of day). Repeatability was assessed by ten independent analyses AOAC. In summary, 100 mg of sample were weighed and of the same samples on the same day while intermediate mixed with 1 mL 95% ethanol and 9 mL of 1N NaOH. The precision was determined by five independent analyses on samples were diluted and the iodine solution was added. After three consecutive days. Precision was expressed as coefficient 10 min incubation at room temperature, the absorbance at 625 of variance (CV) of retention time and peak signal. The CV nm was analysed with a spectrophotometer and the amylose values for both repeatability and intermediate precision were content was calculated based on the standard curve. The less than 6% showing that the methods have adequate samples were analysed in triplicate. precisions. The analytical properties for the determination of phenolic H. Performance of the Method compounds and melatonin are listed in Table I. The analytical procedures for the chromatographic methods I. Data Analysis for melatonin and phenolic compounds were carried out according to the recommendations of ICH Guideline Q2 (R1) The experimental results in single- and two-factor and suggestions in ISO 17025. Linearity, range, precision, experiments were analysed using Gnumeric. The Analysis of detection and quantification limits of the method were Variance (ANOVA) and Least Significant Difference (LSD) established. Linearity was evaluated in order to express the test were used to determine the significant differences (p < ability of the method to obtain test results that are directly 0.05) between the means. proportional to the concentration of melatonin in two different ranges. Appropriate dilution from a stock solution of III. RESULT AND DISCUSSION melatonin was carried out to give concentrations ranging from 0.75 to 15 µg L–1 and 15 to 750 µg L–1. A series of dilution A. Phenolics in Glutinous and Non-glutinous Rice from stock solution of phenolic compounds was carried out to In order to evaluate the phenolic levels and compositions on give concentrations from 0.15 to 12 mg L-1. Gnumeric 1.12.17 glutinous and non-glutinous rice, two clusters of rice varieties was used to generate the regression analysis. Calibration i.e. non-pigmented and black pigmented rice grains were curves were obtained from these regression analyses and the extracted and analysed in duplicate (Table II). Result shows melatonin and phenolic compounds in the extracts were that the content of phenolic compounds in rice samples with quantified. The standard deviation (σ) obtained for the higher amylose content (non-glutinous) were higher than in response and the slope (a) from the regression were then used the glutinous samples for both clusters. The highest to calculate the limit of detection (LOD) and limit of concentration of phenolics presented in pigmented-rice was quantification (LOQ). 211.94±18.65 µg g-1 owned by black non-glutinous rice then followed by its glutinous type (209.00±18.39 µg g-1).

TABLE I. ANALYTICAL CHARACTERISTICS FOR THE DETERMINATION OF MELATONIN AND PHENOLIC COMPOUNDS Intra-day, CV (%) Inter-day, CV (%), Concentration Compounds Linear equation R2 LOD LOQ n = 10 n = 3 × 5 range RT Peak RT Peak * * * PRO 0.15 – 12 y = 1.32x − 0.05 0.9997 0.23 0.77 0.00 0.33 0.26 2.33 * * * PRA 0.15 – 12 y = 5.52x + 0.03 0.9999 0.17 0.56 0.00 0.33 0.21 2.22 * * * p-HBA 0.15 – 12 y = 7.43x − 0.23 0.9997 0.22 0.74 0.00 0.38 0.21 2.22 * * * p-HB 0.15 – 12 y = 3.44x − 0.07 0.9998 0.23 0.75 0.00 0.49 0.18 1.85 * * * VAA 0.15 – 12 y = 1.34x + 0.06 0.9996 0.26 0.87 0.08 0.32 0.10 0.45 * * * IVA 0.15 – 12 y = 1.37x + 0.08 0.9994 0.32 1.07 0.07 0.24 0.06 0.23 * * * VAN 0.15 – 12 y = 1.85x − 0.02 0.9998 0.20 0.67 0.00 0.48 0.22 1.83 * * * GUA 0.15 – 12 y = 0.35x + 0.03 0.9993 0.37 1.24 0.00 1.43 0.24 0.71 * * * p-COU 0.15 – 12 y = 7.52x + 0.16 0.9998 0.18 0.61 0.04 0.36 0.16 2.19 * * * FER 0.15 – 12 y = 4.96x − 0.27 0.9996 0.27 0.90 0.04 0.65 0.14 2.10 * * * CAF 0.15 – 12 y = 3.74x – 1.05 0.9963 0.84 2.80 0.05 0.40 0.06 0.63 * * * CHL 0.15 – 12 y = 10.56x – 3.85 0.9916 1.27 4.23 0.06 5.99 0.11 3.33 * * * SIN 0.15 – 12 y = 4.72x – 0.29 0.9996 0.27 0.90 0.04 0.45 0.14 1.65 * * * HMF 0.15 – 12 y = 24.87x – 1.95 0.9998 0.18 0.61 0.16 0.17 0.21 0.60 * * * MF 0.15 – 12 y = 0.32x + 0.28 0.9954 0.94 3.12 0.06 0.42 0.48 1.36 * * * ELL 0.15 – 10 y = 1.35x + 0.12 0.9827 1.59 4.83 0.05 3.32 0.15 5.49 ** ** ** Melatonin 0.75 to 15 y = 580.79x – 1806.8 0.9957 1.15 3.84 0.99 0.93 1.23 2.05 15 to 750** y = 615.11x − 961.13 0.9994 * Range in the unit of mg/Kg-1 ** Range in the unit of µg/Kg-1

http://dx.doi.org/10.15242/IICBE.C0115068 29 International Conference on Plant, Marine and Environmental Sciences (PMES-2015) Jan. 1-2, 2015 Kuala Lumpur (Malaysia)

The difference of phenolics content between black C. Phenolics and Melatonin in Medium-Amylose Rice glutinous and black non-glutinous rice was not as impressive Three rice varieties from conventional farming were used as if compared to the non-pigmented rice. Thus, in this in this study. In addition to being considered as , particular case, phenolics concentration in rice appears to be together with umbul-umbul, pandan wangi represents the local strongly related to the pigment of rice in their . Both rice varieties of Indonesia. IR64 is a rice variety that was black pigmented glutinous and non-glutinous rice grains were developed in a major advance in rice production as it provided produced without a bran removal process called polishing. higher yield potential, greater yield stability and more Hence, as the phenolic compounds are mainly associated with efficient management practices. Amylose content in these the pericarp in the grain, the milling process to produce tested rice varieties was determined (Fig. 2). Subsequently, in polished grain reduces the level of these compounds in the order to study the effect of amylose content on the level of grain. antioxidant compounds including phenolic and melatonin, a In contrary, the levels of phenolics in non-pigmented single-factor ANOVA has been used (p.0.05). ANOVA (polished) grains were prominently remarkable for different revealed that the amylose content has significant effect on the amylose content: 35.58±3.13 µg g-1 for non-glutinous rice -1 level of antioxidant compounds for both phenolics and whilst 16.15±1.42 µg g for glutinous rice. The level of melatonin. It is, therefore, Least Significant Differences phenolics in non-glutinous was roughly two folds higher than (LSDs) for these factors were also estimated (Fig. 2). its glutinous type. This finding agrees the result revealed by The levels of both phenolics and melatonin for the three Chi et al. [11] who conducted a research related to phenolics tested varieties were directly corresponded to the amount of analysis of Korean rice grain varieties. These researchers amylose content wherein the higher the amylose content, the confirmed that polished non-glutinous rice (Ilpumbye) extract higher the amount of antioxidant compounds. This finding is has a higher level of phenolics than the glutinous type consistent with that reported by Li et al.[10], who found that (Hwasunchalbyo). plant samples with high amylose contents exhibit better B. Melatonin in Glutinous and Non-glutinous Rice antioxidant activity than their glutinous genotypes. Despite significant research effort, the specific antioxidant enhanced The level of melatonin in rice with different amount of by amylose has not been investigated further and phenolics as amylose content was studied on the same sample used in well as melatonin are considered as a potential molecule that previous experiment of phenolics. However, unlike the result has a high antioxidant activity [13]. for phenolics, the difference in melatonin contents between black glutinous (182.04±2.79 µg Kg–1) and non-glutinous rice IV. CONCLUSION (73.81±1.13µg Kg–1) was notably more significant. The level of melatonin in black non-glutinous rice was more than Antioxidant compounds in glutinous and non-glutinous rice double than in its glutinous type. were studied. The results suggest a positive correlation This variation is reasonable when taking into consideration between the amylose content and the level of antioxidant the specific type of carbohydrates in rice support the synthesis compounds i.e. melatonin and phenolic compounds. However, of the neurotransmitter serotonin, which is later transformed particularly in pigmented variety, rice bran contributes greater into melatonin [12]. Thus, the presence of lower amounts of effect on the level of phenolics in rice grain than the amylose starch may not enhance the synthesis of melatonin. content.

TABLE II. PHENOLIC COMPOUNDS IN TESTED SAMPLES

MEL Phenolic compounds (mg Kg-1)* Cluster Rice Sample Picture (µg (Amylose Level) -1 Kg ) PRO PRA p-HBA p-HB VAA IVA VAN GUA p-COU FER CAF CHL SIN ELL HMF MF SUM

Black 182.04 74.58 7.16± 2.81± 0.49± 59.56± 1.04± 2.79± 45.00± 1.38± 5.71± 2.53± 0.85± 0.48± 7.40± 0.16± 211.94 Non-glutinous ND (Medium Amylose ±2.79 ±1.97 0.20 0.12 0.01 6.94 0.16 0.07 2.67 0.04 0.34 0.13 0.05 0.01 0.49 0.01 ±18.65 Pigmented Content) Rice Black glutinous 73.81± 91.47 6.77± 3.34± 1.97± 34.50± 1.74± 1.96± 29.65± 4.74± 5.87± 1.55± 0.92± 24.35± 0.16± 209.00 ND ND (Low Amylose 1.13 ±2.42 0.19 0.15 0.05 4.02 0.28 0.05 1.76 0.14 0.35 0.08 0.06 1.62 0.01 ±18.39 Content)

White 1.65± 1.13± 3.64± 3.47± 1.29± 10.68± 0.98± 2.16± 0.96± 0.83± 0.17± 6.56± 0.17± 1.88± 35.58± Non-glutinous - ND ND (Medium Amylose 0.04 0.03 0.16 0.10 0.03 0.63 0.03 0.13 0.05 0.05 0.00 0.44 0.01 0.08 3.131 Non- Content) pigmented Rice White 0.81± 0.70± 1.63± 4.10± 0.28± 0.97± 3.28± 0.31± 1.02± 0.84± 0.76± 1.29± 0.17± 16.15± Glutinous - ND ND ND (Low Amylose 0.00 0.02 0.07 0.11 0.03 0.02 0.19 0.01 0.06 0.04 0.05 0.09 0.01 1.422 Content) * ND: Not Detected (< LOD)

http://dx.doi.org/10.15242/IICBE.C0115068 30 International Conference on Plant, Marine and Environmental Sciences (PMES-2015) Jan. 1-2, 2015 Kuala Lumpur (Malaysia)

[11] H.-Y. Chi, C.-H. Lee, K.-H. Kim, S.-L. Kim, I.-M. Chung, "Analysis of phenolic compounds and antioxidant activity with H4IIE cells of three different rice grain varieties," Eur. Food Res. Technol., vol. 225, pp. 887–893. 2007. http://dx.doi.org/10.1007/s00217-006-0498-3 [12] USA Rice® Federation, "Rice Nutrition Information," http://www.usarice.com/index.php?option=com_content&view=article &id=1727&Itemid=660 (accessed September 23, 2014). [13] I. Gulcin, M.E. Buyukokuroglu, M. Oktay, I. Kufrevioglu, "On the in vitro antioxidative properties of melatonin," J. Pineal Res.,vol. 33,pp. 167–171. 2002. http://dx.doi.org/10.1034/j.1600-079X.2002.20920.x

Widiastuti Setyaningsih was born in Klaten, Indonesia on July 21, 1984. She graduated cum laude from Gadjah Mada University (UGM) with a Bachelor of Food Technology. She received her M.Sc. from Gdansk University of Technology (GUT) Poland, University of Barcelona (UB) and University of Cadiz (UCA), Spain under the frame of European Master in Quality in Analytical Laboratories (EMQAL). She is a LECTURER at the Department of Food and Agricultural Product Technology, Faculty of Agricultural Technology, UGM, Indonesia. Presently, she is pursuing her doctoral studies in field of food analytical chemistry at the Department of Analytical Chemistry, Faculty of Sciences, UCA, Spain. She held an undergraduate scholarship from UGM for the period of 2004–2005, an Erasmus Mundus Scholarship from European Union Commission (Education, Fig. 2 Melatonin (1) and phenolics (2) levels in medium-amylose rice Audiovisual and Culture Executive Agency-EACEA) during the period 2010– 2011, a MAEC-AECID (Agencia Española de Cooperación Internacional para varieties. LSD: bars followed by the same letter are indicated as not el Desarrollo) scholarship from the Ministry of Spain for 2011–2012, and a significantly different (p = 0.05) conclusion Ph.D. studentship from the CIMB Foundation from 2012 to date. Her research is focused on analytical method development and validation for bio-active Acknowledgment compounds in foods. W. S. thanks the CIMB Foundation for a Ph.D. studentship Nikmatul Hidayah was born in Magelang, Indonesia in February 1982. through the CIMB Regional Scholarships She graduated from Gadjah Mada University (UGM) with a bachelor of Food Technology. At present, she is a RESEARCHER at Indonesian Center of .REFERENCES Agricultural Postharvest Research & Development, Bogor, Indonesia. Her research is focused on postharvest handling and processing of agricultural product such as enzymatic processing of rice and modification of [1] W. Setyaningsih, M. Palma, C.G. Barroso, "A new microwave-assisted based on local agricultural product in Indonesia. extraction method for melatonin determination in rice grains," J. Cereal

Sci., vol. 56, pp. 340–346. 2012. Irfan Estiono Saputro was born in Jakarta, Indonesia on 25 April 1984. http://dx.doi.org/10.1016/j.jcs.2012.02.012 His desire to pursue a career in science was reinforced after opting for Food [2] S. Tian, K. Nakamura, H. Kayahara, "Analysis of Phenolic Compounds Quality Assurance as his major at Bogor Agricultural University (IPB), in , , and ," J. Agric. Indonesia. In order to advance his knowledge in food analytical chemistry, he Food Chem., vol. 52, pp. 4808–4813. 2004. is currently pursuing a European Master in Quality in Analytical Laboratories http://dx.doi.org/10.1021/jf049446f (EMQAL) at the University of Barcelona (Spain). [3] S. Yafang, Z. Gan, B. Jinsong, "Total phenolic content and antioxidant

capacity of rice grains with extremely small size," African J. Agric. Professor Miguel Palma Lovillo was born in Olvera, Spain on January Res., vol. 6, pp. 2289–2293. 2011. 14, 1968. He obtained his degree in Chemistry at the University of Cadiz [4] R. Sompong, S. Siebenhandl-Ehn, G. Linsberger-Martin, E. Berghofer, (UCA) and later obtained his Ph.D. in Science at the same University in 1995. "Physicochemical and antioxidative properties of red and He is a FULL PROFESSOR of Analytical Chemistry at the Faculty of varieties from , and Sri Lanka," Food Chem., vol. 124. Sciences, UCA, Spain. He was visiting scientist at the Department of Pharma- pp. 132–140. 2011. http://dx.doi.org/10.1016/j.foodchem.2010.05.115 ceutical Sciences at the Southern School of Pharmacy in Mercer University, [5] M.A. Al-Farsi, C.Y. Lee, "Optimization of phenolics and dietary fibre Atlanta, GA, USA and also at the Department of Chemistry, at Virginia Tech, extraction from date seeds," Food Chem. vol. 108, pp. 977–985. 2008. Blacksburg, VA, USA. http://dx.doi.org/10.1016/j.foodchem.2007.12.009 Professor Palma received the Premio Extraordinario de Doctorado (the [6] Y. Qiu, Q. Liu, T. Beta, "Antioxidant properties of commercial wild Highest Distinction at PhD level) at the Faculty of Sciences (UCA) and the rice and analysis of soluble and insoluble phenolic acids," Food Chem., Premio Extraordinario de Licenciatura “Octavio Comes” (Distinction at vol. 121, pp. 140–147. 2010. Bachelor degree). His research interests include food analytical methods, food http://dx.doi.org/10.1016/j.foodchem.2009.12.021 science, functional foods and bioactive compounds in foods. [7] W. Setyaningsih, I.E. Saputro, M. Palma, C.G. Barroso, Optimisation

and validation of the microwave-assisted extraction of phenolic Professor Carmelo García Barroso was born in Arcos, Spain on compounds from rice grains, Food Chem., vol. 169, pp. 141–149. December 9, 1955. He obtained his degree in Chemistry at the University of 2015. http://dx.doi.org/10.1016/j.foodchem.2014.07.128 Cadiz (UCA) and then his Ph.D. in Sciences at the same University in 1985. [8] W. Setyaningsih, N. Hidayah, I.E. Saputro, M.P. Lovillo, C.G. He is a FULL PROFESSOR of Analytical Chemistry at Faculty of Barroso, "Melatonin Profile during Rice (Oryza Sativa) Production," J. Sciences, UCA, Spain. His research is focus on food and wine science and Adv. Agric. Technol., vol. 1, pp. 60–64. 2014. food analytical methods. [9] H.-J. Chung, Q. Liu, L. Lee, D. Wei, "Relationship between the Professor García co-founded the Spanish Society for Wine Research structure, physicochemical properties and in vitro digestibility of rice (GIENOL) and was elected President of this Society in 1999. He is with different amylose contents," Food Hydrocoll., vol. 25, pp. responsible for the postgraduate program in Agrifood Sciences, a common 968–975. 2011. http://dx.doi.org/10.1016/j.foodhyd.2010.09.011 teaching program between UCA and the University of Cordoba (Spain) since [10] W. Li, C.V. Wei, P.J. White, T. Beta, "High-amylose corn exhibits 2006. In 2001 he became Chairman of the Andalusian Center for Wine better antioxidant activity than typical and waxy genotypes.," J. Agric. Research, a research center at UCA sponsored by the local Andalusian Food Chem., vol. 55, pp. 291–8. 2007. Government. http://dx.doi.org/10.1021/jf0622432

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