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Amaranth Seeds and Products – the Source of Bioactive Compounds

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Amaranth Seeds and Products – the Source of Bioactive Compounds Pol. J. Food Nutr. Sci., 2014, Vol. 64, No. 3, pp. 165-170 DOI: 10.2478/v10222-012-0095-z http://journal.pan.olsztyn.pl Original article Section: Food Chemistry Amaranth Seeds and Products – The Source of Bioactive Compounds Dorota Ogrodowska1, Ryszard Zadernowski1, Sylwester Czaplicki1*, Dorota Derewiaka2, Beata Wronowska1 1Chair of Food Plant Chemistry and Processing, Faculty of Food Science, University of Warmia and Mazury, Plac Cieszyński 1, 10–726 Olsztyn, Poland 2Division of Food Quality Evaluation, Department of Biotechnology, Microbiology and Food Evaluation, Faculty of Food Sciences, Warsaw University of Life Sciences, ul. Nowoursynowska 159, 02–787 Warsaw, Poland Key words: amaranth, expanded seeds, fl akes, tocopherols, sterols, squalene In recent years, new products obtained from amaranth seeds have entered the food market including expanded “popping” seeds and fl akes. Lipids and biologically-active substances dissolved in these products are susceptible to changes. Additionally, due to the fact that fat quality has high dietary importance, there is a need to conduct detailed quality and quantity studies on the lipid composition of Amaranthus cruentus. For the samples under analysis, protein, fat, starch and ash content were determined. Fatty acids and sterols were analysed by gas chromatogra- phy. The analysis of tocopherols and squalene content was carried out with the application of high-performance liquid chromatography coupled with photodiode array and fl uorescence detectors (HPLC-DAD-FLD). Protein, fat and starch content did not change during seed processing. However in the case of tocopherols, the total tocopherol content was 10.6 mg/100 g for seeds, while in “popping” and in fl akes it was reduced by approximately 35%. The squalene content ranged from 469.96 mg/100 g for seeds to 358.9 mg/100 g for fl akes. No signifi cant differences were observed in the fatty acid profi le of seeds and products, but differences were observed in the sterol content. INTRODUCTION amaranth seeds instead of amaranth fl our might be used to improve the nutritional value of bread [Bodroža-Solarov “Pseudocereals” is a term which has not been properly et al., 2008]. Amaranth is rich in minerals including: magne- defi ned in the literature to date. This term covers plants such sium, calcium, potassium and phosphorus, iron and sodium as amaranth, quinoa, tartary buckwheat, buckwheat and mil- [Shukla et al., 2006; Gajewska et al., 2002]. Moreover, ama- let. Unlike plants that are recognised as true cereals, which ranth seeds and their products are a rich source of bioactive are monocotyledonous plants, these are dicots. Owing to substances showing antioxidant properties [Worobiej et al., their chemical composition, seeds of these plants, just like 2009; Klim czak et al., 2002]. The non-saponifi able substanc- cereal grains, are used for human nutrition [Ceglińska & es of the grain lipid fraction include: squalene, tocopherols, Cacak-Pietrzak, 1999]. sterols and others [Gamel et al., 2007; Gorinstein et al., 2007; Amaranthus cruentus, a member of the Amaranthaceae León-Camacho et al., 2001; Sun et al., 1997]. family, is an unconventional plant that has gained popularity, The scope of this study was to examine the content of se- both among consumers and farmers. The use of its seeds has lected nutrients in the seeds, popped seeds and fl akes of am- also increased; it is now used as a source of lipids and a mate- aranth. Moreover, due to the importance of the fat quality rial for the production of fl our, fl akes, popped seeds, several on the products’ dietary value and its infl uence on shelf life, sorts of bread [Januszewska-Jóźwiak & Synowiecki, 2008] a detailed examination of the lipid content was carried out and confectionery [Sindhuja et al., 2005]. The amaranth- to determine the fatty acid profi le and the content of lipid- -based products are a valuable source of nutrients and miner- -soluble bioactive substances. als. In addition, the amino acid profi le of amaranth makes it an attractive source of protein. If consumed with other MATERIALS AND METHODS cereals, it is a balanced source of proteins; therefore, ama- ranth fl our is often used in mixtures with maize or wheat Materials [Alvarez-Jubete et al., 2010; Escudero et al., 2004]. It has Amaranth (Amaranthus cruentus) seeds and commercially also been reported that the addition of 10–20% expanded available products (expanded seeds - popping and fl akes) were obtained from “Szarłat” company (Łomża, Poland). Seeds, popping and fl akes were produced from the same lot * Corresponding Author: Tel./Fax: +48 89 523 34 66; E-mail: [email protected] (S. Czaplicki) of material. © Copyright by Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences © 2014 Author(s). This is an open access article licensed under the Creative Commons Attribution-NonCommercial-NoDerivs License (http://creativecommons.org/licenses/by-nc-nd/3.0/). 166 Amaranth Seeds and Products – The Source of Bioactive Compounds Proximate chemical composition the same manufacturer. Separations were performed on a Li- Assessment of the nutritional value of amaranth seeds Chrospher Si60 column (250 mm 4 mm 5 m, Merck). included determination of starch content by the polarimet- Next, 20 μL of sample was injected into the chromatographic ric method in a hydrochloric acid solution, in accordance column. A solution of 0.7% iso-propyl alcohol in hexane was to the procedure of the AOAC, following the guidelines pro- used as a mobile phase at a fl ow rate of 1 mL/min. Excitation vided by Bagdach, detailed in Rutkowska [1981]. Dry matter, and emission wavelengths were set at 296 nm and 330 nm, protein and ash contents were determined following AACC respectively. Peaks were identifi ed on the basis of retention methods 44–15, 46–10 and 08–03 [AACC, 1994], respectively. times determined for -, -, - and -tocopherol standards Protein content was calculated on the basis of total nitrogen (Calbiochem, UK) analysed separately, and their content was using a 6.25 multiplication factor. Total fat was determined calculated using external calibration curves prepared for all by the method described by Folch et al. [1957]. analysed tocopherols. Analysis of the lipid fraction of amaranth seeds and ama- ranth products obtained as total lipids using the method de- Determination of phytosterols scribed by Folch et al. [1957] involved characterisation of fat- Sterol content in oils was determined by gas chroma- ty acids, squalene, tocopherols and sterols. tography coupled with mass spectrometry (GC-MS) ac- cording to the method described by Vlahakis & Hazebroek Determination of fatty acids [2000]. The unsaponifi ables were extracted with diethyl Fatty acid methyl esters were prepared according to ether, and the extract was subsequently evaporated with ro- the method described by Zadernowski & Sosulski [1978]. tary evaporator type R210 (Büchi Labortechnik AG, Post- The methyl esters were dried under a nitrogen stream, purifi ed fach, Switzerland) under nitrogen. Dry residues were re- by hexane extraction and analysed by gas liquid chromatog- dissolved in 1.5 mL of n-hexane, and 0.2 mL (0.4 mg/mL) raphy (GLC) on a Fisons 8000 gas chromatograph equipped of 5-cholestane internal standard solution was used. The ex- with a fl ame ionization detector (FID) (Carlo Erba, Italy). tract was transferred into a vial and evaporated with a ni- Separations were performed on a J&W Scientifi c DB-225 trogen stream. The residues were re-dissolved in 100 μL capillary column (30 m 0.25 mm 0.15 μm) using helium of pyridine and 100 μL of BSTFA (N,O-bis (trimethylsilyl) as a carrier gas. The GC oven was operated isothermally at trifl uoroacetamide) with 1% TMCS (trimethylchlorosilane) 190oC. Peaks were identifi ed on the basis of retention times and left in the dark for 24 h to complete the derivatization re- determined for fatty acids standards mixture (Supelco® action. This was followed by the addition of 1 mL of hexane, 37 Component FAME Mix) (Milwaukee, WI, USA). and 1 μL of obtained mixture was used for GC-MS analysis. Analyses were conducted on a GCMS-QP2010S gas Determination of squalene chromatograph coupled with mass spectrometer manufac- The squalene content was determined with high perfor- tured by Shimadzu Corporation (Japan). A DB-5ms (30 m mance liquid chromatography (HPLC), according to meth- 0.25 mm 0.25 μm fi lm thickness) Agilent J&W Scientifi c od described by Czaplicki et al. [2009] with modifi cations. (Perlan Technologies, Poland) capillary column was used Briefl y, 0.01 g of oil (±0.0001 g) was transferred into a 10 mL for separation of phytosterols with helium as a carrier gas measuring fl ask, and fi lled up with n-hexane. Next, 20 mL at a fl ow rate of 0.9 mL/min. The injector temperature was of diluted oil sample was injected into the chromatograph- 230°C, and the column temperature was programmed as ic system. Analyses were conducted on a 1200 series high follows: 50ºC for 2 min, a subsequent increase to 230ºC at performance liquid chromatograph, manufactured by Agi- the rate of 15ºC/min, to 310ºC at the rate of 3ºC/min, 10 min lent Technologies (Palo Alto, CA, USA) and equipped with hold. The interface temperature of GC-MS was 240°C. Tem- a photodiode detector (PDA). Chromatographic separations perature of the ion source was 220°C and the electron energy were conducted on a LiChrospher RP-18 column (250 mm was 70 eV. The total ion current (TIC) mode was used for 4.6 mm 5 m, Merck, Germany), at 30oC. The mobile phase quantifi cation (100–600 m/z range). contained acetonitrile, isopropyl alcohol and hexane at a fl ow rate of 1 mL/min. An analytical wavelength was set at 218 nm.
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