Nutritional Composition in the Chia Seed and Its Processing Properties on Restructured Ham-Like Products

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Available online at www.sciencedirect.com 65 66 1 ScienceDirect 67 2 68 3 69 4 70 5 71 journal homepage: www.jfda-online.com 6 72 7 73 8 74 9 Original Article 75 10 76 11 Nutritional composition in the chia seed and its 77 12 78 13 processing properties on restructured ham-like 79 14 80 15 81 16 products 82 17 83 18 a,1 b,c,1 a d 84 19 Q6 Yi Ding , Hui-Wen Lin , Yi-Ling Lin , Deng-Jye Yang , 85 * 20 Yu-Shan Yu e, Jr-Wei Chen a,f, Sheng-Yao Wang g, Yi-Chen Chen a, 86 21 87 a 22 Department of Animal Science and Technology, National Taiwan University, Taipei, Taiwan 88 23 b Department of Optometry, Asia University, Taichung, Taiwan 89 24 c Department of Medical Research, China Medical University Hospital, China Medical University, Taichung, Taiwan 90 25 d Department of Nutrition, China Medical University, Taichung, Taiwan 91 26 e Health Bureau of Taichung City Government, Taichung, Taiwan 92 27 f Poultry Industry Section, Animal Industry Department, Council of Agriculture, Executive Yuan, Taipei, Taiwan 93 28 94 g Experimental Farm, College of Bioresources and Agriculture, National Taiwan University, Taipei, Taiwan 29 95 30 96 31 article info abstract 97 32 98 33 99 34 Article history: Low-fat meat products always have harder texture, lower juiciness, and worse flavor. Due 100 35 Received 11 September 2016 to their higher water-holding, water absorption, and organic molecule absorption, chia 101 36 Received in revised form seeds (CHIA) have been applied in powders, nutrition bars, breads, and cookies. Hence, the 102 37 19 December 2016 objectives of this study were to: (1) analyze the nutritional compositions in CHIA; and (2) 103 38 Accepted 27 December 2016 look for the possible application of CHIA on restructured ham-like products. CHIA has high 104 39 Available online xxx amounts of a-linolenic acid, crude polysaccharides, and also contains essential amino 105 40 acids, minerals, and polyphenols. Regarding processing properties of CHIA, a combination 106 41 Keywords: of CHIA and carrageenan (CA) increased (p < 0.05) production yield of restructured ham-like 107 42 108 chia seed products. A scanning electron microscope observation indicated that CHIA and CA addition 43 109 nutritional profile can assist an emulsification in this ham-like product. Addition of 0.5% CA and 1.0% CHIA in 44 110 45 physicochemical changes this ham-like product showed the similar overall acceptance as products with added fat. 111 < 46 restructured ham-like product Following storage at 4 C, higher (p 0.05) purge and centrifugation losses, as well as 112 47 scanning electron microscope hardness of this ham-like product can be improved by adding CHIA and CA. CHIA addition 113 48 also resulted in lower (p < 0.05) lipid and protein oxidation, especially a 1.0% addition. In 114 49 summary, due to both nutritional addition and improvements on physicochemical and 115 50 sensorial properties of restructured ham-like products, CHIA seeds have great potential on 116 51 the development of healthy and good-quality meat products. 117 52 Copyright © 2017, Food and Drug Administration, Taiwan. Published by Elsevier Taiwan 118 53 119 LLC. This is an open access article under the CC BY-NC-ND license (http:// 54 120 creativecommons.org/licenses/by-nc-nd/4.0/). 55 121 56 122 57 123 58 124 59 125 60 * Corresponding author. Department of Animal Science and Technology, National Taiwan University, Taipei 106, Taiwan. Q1 126 61 E-mail address: [email protected] (Y.-C. Chen). 127 62 1 These two authors contributed equally as co-first authors. 128 63 http://dx.doi.org/10.1016/j.jfda.2016.12.012 129 64 1021-9498/Copyright © 2017, Food and Drug Administration, Taiwan. Published by Elsevier Taiwan LLC. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Please cite this article in press as: Ding Y, et al., Nutritional composition in the chia seed and its processing properties on restructured ham-like products, Journal of Food and Drug Analysis (2017), http://dx.doi.org/10.1016/j.jfda.2016.12.012 JFDA457_proof ■ 13 February 2017 ■ 2/11

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1 column from Supelco Inc. (Bellefonte, PA, USA). For the amino 66 2 1. Introduction acid profile of CHIA, 1 g of CHIA powder was hydrolyzed in 67 3 68 2 mL methane sulfonic acid solution (4N) for 24 hours. Amino 4 69 Generally, the fat content makes meat products creamy and acids were quantified using the Hitachi L-8900 High Speed 5 70 delicious. However, saturated fatty acid intakes from meat Amino acid Analyzer (Hitachi High-Technologies Co., Tokyo, 6 products increase the occurrence of cardiovascular diseases. 71 7 Japan). The data were described as mg amino acid per 100-g 72 Hence, the development of low-fat meat products has received 8 CHIA. Regarding the mineral profile, the CHIA powder was 73 global attention. In the USA, the claim of low-fat is the fat con- 9 ashed at 550 C for 6 hours. Two-mL nitric acid (70%) was 74 tent <6% in a small serving size food (<50 g) [1]. Ham is generally 10 added. Acidized samples were diluted in double distilled H2O 75 11 recognized as a low-fat meat product in Europe and America. and then filtered. Filtrate was diluted to 50 mL volumetric 76 Although the low-fat meat products are regarded as healthier 12 bottle by double distilled H2O. The mineral profile of CHIA was 77 13 than other meat products, they still have some textural, analyzed by Inductively Coupled Plasma-Optical Emission 78 14 sensorial, and lower yield problems. In order to conquer these Spectrometer (ELEMENT 2*ICP-MS; Thermo Fisher Scientific 79 15 80 problems, fat replacers are candidates to improve the texture Inc., Waltham, MA, USA). 16 and sensorial properties of low-fat meat products. 81 17 Dietary fiber has hydration properties, as well as particle 82 18 2.2.2. Crude polysaccharide and phytochemical contents, and 83 size, density, and surface characteristics; hence, it has been 19 polyphenolic profile of CHIA 84 applied well in baked goods [2] or meat products [3]. Chia 20 The crude polysaccharide content in CHIA was assayed ac- 85 (Salvia hispanica) seed (CHIA) contains rich dietary fibers [4]. 21 cording to the procedure of a previous report [9]. Regarding 86 22 Due to the gum-like property of CHIA, their fiber-rich fraction phytochemical analyses, 50 g CHIA powder was mixed with 87 23 had 56.4 g/100 g dietary fiber, where 53.45 g/100 g is insoluble 1 L of n-hexane in a Waring Laboratory Blender (The Lab 88 24 dietary fiber and the remainder is soluble [5]. In comparison Depot, Inc., Dawsonville, GA, USA) for 3 minutes, and extrac- 89 25 with other fiber sources (soybean, wheat, maze, wheat hulls), ted for 24 hours in the dark. After filtration, the defatted reside 90 26 91 the fiber-rich fraction in CHIA has higher water holding, ab- was dried under nitrogen gas, and then extracted with 1 L of 27 92 sorption, and organic-molecule absorption with high emulsi- 80% ethanol for 24 hours in the dark. After filtration, the sol- 28 93 fying activity (53.26 mL/100 mL) and emulsion stability vent was removed under vacuum at 40C, followed by lyoph- 29 (94.84 mL/100 mL) [5]. Therefore, it has been well applied in 94 30 ilization in a freeze-dryer (Vastech Scientific Co., Ltd., Taipei, 95 desserts or cookies [4,6]. 31 Taiwan) to obtain the phenolic extract. Total phenolic acid, 96 Although CHIA is a potential ingredient in health and diet 32 flavonoid, and condensed tannin contents in CHIA were 97 33 food products on its physicochemical properties [5,7],itis determined according to the methods of a previous report [10]. 98 34 seldom used in meat products to our knowledge. Hence, the The polyphenolic profile in CHIA powders was analyzed by 99 35 purposes of this study were: (1) to understand the nutritional using high-performance liquid chromatography (Shimadzu 100 36 compositions in CHIA; (2) to investigate the effects of CHIA on SCL-10A system controller module; Shimadzu, Kyoto, Japan) is 101 37 processing properties of restructured ham-like products; and composed of a Shimadzu SCL-10AT pump system, a Shimadzu 102 38 103 (3) to look for the effects of CHIA on the physicochemical SPD-10A UV-vis detector, and a 20 mL loop (Rheodyne Inc., 39 104 changes of restructured ham-like products after storage. Cotati, CA, USA). A Inspire C column (250 mm 4.6 mm, 40 18 105 m 41 5 m; Dikma Technologies Inc., Lake Forest, CA, USA) and a 106 42 gradient solvent system consisting of 2% glacial acetic acid 107 43 2. Methods (solvent A) and acetonitrile (solvent B; conditions: A/B ¼ 2/98 108 44 (v/v) from 0 minutes to 25 minutes, A/B ¼ 4/96 (v/v) from 109 45 2.1. Materials 25 minutes to 40 minutes; A/B ¼ 10/90 (v/v) from 40 minutes to 110 46 50 minutes, A/B ¼ 15/85 (v/v) from 50 minutes to 60 minutes, 111 47 CHIA (The Chia Co., Ltd., Port Melbourne, Victoria, Australia) A/B ¼ 20/80 (v/v) from 60 minutes to 115 minutes, A/B ¼ 22/78 112 48 were purchased from a local market and ground to powder. (v/v) from 115 minutes to 135 minutes, and A/B ¼ 25/75 (v/v) 113 49 114 Carrageenan (CA) was purchased from Gemfont Co., Ltd. from 135 minutes to 150 minutes; flow rate ¼ 0.8 mL/min) 50 115 (Taipei, Taiwan). Pork leg meat and back fat were purchased were used for separation of components whose UV spectra 51 116 from Shang Lee Food Co., Ltd. (Nantou County, Taiwan), were recorded at 280 nm. Those phenolic acid and flavonoid 52 117 packaged with polyethylene bags under 20C. All other 53 compounds in CHIA were identified via high-performance 118 54 chemicals used in this study were purchased from Sigma- liquid chromatography based on UV absorbance and reten- 119 e 55 Aldrich Co., LLC (St Louis, MO, USA). tion time compared with the standards for phenolic acid 120 56 compounds (Sigma-Aldrich Co., LLC), and quantified using 121 57 2.2. Nutritional compositions in CHIA standard curves of authentic compounds. 122 58 123 59 2.2.1. Fatty acid, amino acid, and mineral profiles of CHIA 2.3. Preparation of restructured ham-like products 124 60 Lipid in CHIA powders was extracted by chloroform and 125 61 126 methanol (2:1, v/v). based on the previous report [8], the fatty Three individual batches in each formula ham-like product 62 127 acid profile was analyzed by using a gas chromatograph were made in this study and each time seven different 63 128 64 (Model 6890N; Agilent, Santa Clara, CA, USA) and a flame products were manufactured on the same day. The same 129 65 ionization detector fitted with a highly polar stationary phase portion of pork leg meat from the meat packer (Shang Lee 130 SP-2560 (100 m length, 0.25 mm inside diameter, 0.20 mm film) Food Co., Ltd, Nantou County, Taiwan) was minced (~0.2 cm

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1 length width height) by a high-shear emulsifying ma- magnification was 1000 [8]. In the last set of product 66 2 chine (Model#: 334, Talleres Cato, S. A., Barcelona, Spain) at manufactures (7 different products), the sensory evalua- 67 3 68 <10 C and then mixed with sodium chloride (Taiyen Biotech tion, which contained preference test (odor, color, texture, 4 69 Co., Ltd., Tainan City, Taiwan) and polyphosphate (Gemfont and flavor), sensorial test (juiciness), and overall accep- 5 70 Co., Taipei, Taiwan). Pork back fat was added only in high-fat tance, was performed 1 week after restructured ham-like 6 71 7 batch (fat added groups). The minced meat was blended with products were manufactured and stored at 4 C. Forty pan- 72 e 8 iced water and additives containing sodium nitrite/nitrate elists (age, 20 40 years; 20 women and 20 men) were 73 9 (Palatinata Cure PM; Gemfont Co., Taipei, Taiwan), sugar recruited from staff, faculties, and students in the National 74 10 (Taiwan Sugar Co., Tainan City, Taiwan), ascorbic acid/so- Taiwan University, Taipei, Taiwan and pretrained for this 75 11 dium ascorbate (TARI Colpur 40S; Fibrisol Service Australia, panel assessment. The evaluation was done using a five- 76 12 Heatherton, Victoria, Australia), five-spice powder, and pep- point scale (5 ¼ very good and 1 ¼ very bad). The restruc- 77 13 per powder, while CHIA and CA were added in groups except tured ham-like products stored at 4C were prepared in hot 78 14 high-fat and control batches. The blended meat mixtures water (100C) for 10 minutes. After the nylon casings of 79 15 80 were assigned into seven treatment formula (Table 1). Mix- products were peeled off and then cut into sizes of 16 81 tures were stuffed into vacuum-packs (110 mm diameter length ¼ 3.0 cm, width ¼ 3.0 cm, and thickness ¼ 0.5 cm. 17 82 18 nylon casings; Ten Geniuses Enterprise Co., Ltd, Taipei, The restructured ham-like products (2 slices per treatment) 83 19 Taiwan). The restructured ham-like products were cooked in were prepared for tasting. The restructured ham-like 84 20 a water bath (85 C) until the core temperature reached 75 C, products of each treatment were distributed on a white 85 21 and then cooled in an ice water bath (4 C) for 20 minutes. plate for evaluation, and water was provided for cleaning 86 22 After cooling, casings were removed, and restructured ham- the palate. All sensory evaluations were conducted at room 87 23 like products were vacuum-packed in high-density poly- temperature (25C). 88 24 ethylene bags (Taipei Pack Industries Co., Taipei, Taiwan), 89 25 and then stored at 4C for further analyses. 2.5. Physicochemical changes of restructured ham-like 90 26 products after storage 91 27 92 2.4. Production yields, proximate compositions, 28 93 microstructure, and sensory properties of restructured ham- 2.5.1. Texture profile analysis and color measurement of 29 94 30 like products restructured ham-like products 95 31 Texture profile analysis indices of restructured ham-like 96 32 2.4.1. Production yield and proximate composition of products were determined using a texture analyzer (Model 97 33 restructured ham-like products TA.XTplus Texture Analyzer; Stable Micro Systems, God- 98 34 After stuffing the batters into the casings (initial weight) and alming, UK) with a P/50 probe (50 mm diameter cylinder 99 35 cooking at 85C for 40 minutes (processed weight), production aluminum; Stable Micro Systems). The texture profile anal- 100 36 yield (%) of restructured ham-like products was calculated as ysis values were calculated by graphing a curve using force 101 37 processed weight (g)/initial weight (g) 100%. Moisture, ash, and time plots. The units for hardness, adhesiveness, 102 38 103 crude fat, and crude protein contents of restructured ham-like springiness, and cohesiveness of products are N, N s, 39 104 products were determined initially in duplicate for each dimensionless, and dimensionless, respectively. Color mea- 40 105 41 sample [11]. surements were taken in the section of restructured ham-like 106 42 products immediately after opening the package. The 107 43 2.4.2. Scanning electron microscope and sensory evaluation of following color coordinates were determined: lightness (L*), 108 44 restructured ham-like products redness (a*), and yellowness (b*). CIE-L*, a*, and b* values were 109 45 The microstructure of restructured ham-like products was measured by a color checker (Model NR-11A, Nippon Den- 110 46 analyzed by using scanning electron microscope, and the shoku Co., Japan). 111 47 112 48 113 49 114 e 50 Table 1 Ingredient levels (g) for restructured ham-like products with different levels of chia seed (CHIA) and carrageenan 115 51 (CA). 116 52 HF CON CONþ0.5CHIA CONþ1.0CHIA CONþ0.5CA CONþ0.5CAþ0.5CHIA CONþ0.5CAþ1.0CHIA 117 53 118 Minced pork leg meat (g) 1700 1700 1690 1680 1690 1680 1670 54 119 Pork back fat (g) 100 0 0 0 0 0 0 55 Water (g) 200 300 300 300 300 300 300 120 56 Chia seed (g) 0 0 10 20 0 10 20 121 57 Carrageenan (g) 0 0 0 0 10 10 10 122 58 Sodium chloride (g) 30 30 30 30 30 30 30 123 59 Polyphosphate (g) 4 4 4 4 4 4 4 124 60 Sodium nitrite (g) 0.3 0.3 0.3 0.3 0.3 0.3 0.3 125 61 Sugar (g) 40 40 40 40 40 40 40 126 62 Five spices powder (g) 0.2 0.2 0.2 0.2 0.2 0.2 0.2 127 63 Pepper powder (g) 6 6 6 6 6 6 6 128 64 Vitamin C (g) 1 1 1 1 1 1 1 129 65 CON ¼ control (without addition of fat); HF ¼ high fat (addition of 5% pork back fat). 130

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1 2.5.2. Purge loss and centrifugation loss of restructured ham- restructured ham-like products (n ¼ 3). Tested parameters in 66 2 like products each batch per product were carried out with at least three 67 3 68 Water holdingcapacities of restructuredham-like products were analyses. The experiment was conducted by using a 4 69 determined as purge and centrifugation losses according to a completely randomized design with subsampling. Due to the 5 70 only one session of sensory evaluation performed, the previous method with a slight modification [12]. Purge loss was 71 6 7 measured in 2- and 4-week storage intervals at 4 C and calcu- completely randomized design was used, and the sample size 72 8 lated as a percentage of the weight of each sample at each stor- on sensory evaluation was 40. Data were analyzed using 73 9 age period compared to its initial weight. Centrifugation loss of analysis of variance. The significant differences were deter- 74 10 restructured ham-like products was measured immediately mined at 0.05 probability level, and differences between 75 11 after manufacturing (1000 g,1hour,4C), and in 2- and 4-week treatments were tested using the least significant difference 76 12 storage intervals at 4C. The centrifugation loss (%) was calcu- test. All statistical analyses of data were performed using SAS 77 13 lated as the difference in weights before and after centrifugation. 9.0 (SAS Institute Inc., Cary, NC, USA). 78 14 79 15 80 2.5.3. Measurements of lipid and protein oxidation of 16 81 restructured ham-like products 17 3. Results and discussion 82 18 Regarding measurements of lipid and protein oxidation, sam- 83 19 ples (~50 g) of ham-like products from different groups stored at 3.1. Nutritional-composition profiles in CHIA 84 20 4 C were picked up in each storage period (0 weeks, 2 weeks, 85 21 and 4 weeks). The lipid oxidation of ham-like products was Regarding the fatty acid profile in CHIA (Table 2), a-linolenic 86 22 measured by previous method [8]. Protein oxidation was eval- acid (ALA) was in the highest amount, followed by linoleic acid 87 23 88 uated by a sulfhydryl content assay as described by Jia et al [13]. (LA), oleic acid, and stearic acid. Meanwhile, the amount of 24 89 unsaturated fatty acids was almost eight times than that of 25 90 u u 26 2.6. Statistical analysis saturated fatty acids where the ratio of -3 fatty acids and -6 91 27 fatty acids was 2.65. Leucine was the highest content in 92 28 All analysis parameters, except sensory evaluation, were essential amino acids of CHIA. Besides, glutamic acid, argi- 93 29 conducted on three independent batches of CHIA or nine, and aspartic acid made up more than 60% nonessential 94 30 95 31 96 32 Table 2 e Fatty acid and amino acid profiles in chia seeds. Q5 97 33 98 Content Adult DIRs Content FAO/WHO/UNU(1985) 34 99 (male, female)a adults (mg/kg BW/day)b 35 100 36 Fatty acid (g/100g oil) Amino acid (mg/100 g) 101 37 Myristic acid (C 14:0) 0.04 ± 0.01 Threonine 795.33 ± 6.58 7 102 38 Pentadecanoic acid (C 15:0) 0.03 ± 0.00 Valine 940.67 ± 19.84 10 103 39 Palmitic acid (C 16:0) 6.95 ± 0.26 Methionine 467.93 ± 8.82 13c 104 40 Palmitoleic acid (C 16:1) 0.07 ± 0.01 Isoleucine 775.25 ± 20.89 10 105 41 Margaric acid (C 17:0) 0.05 ± 0.01 Leucine 1514.48 ± 37.86 14 106 42 Stearic acid (C 18:0) 4.33 ± 0.34 Phenylalanine 1021.05 ± 39.21 14d 107 43 Oleic acid (C 18:1) 9.17 ± 0.08 Lysine 1183.91 ± 72.11 12 108 44 Linoleic acid (C 18:2 (u-6)) 21.51 ± 0.35 (12, 17) g/day Histidine 663.35 ± 42.85 8e12 109 45 a-Linolenic acid (C 18:3 (u-3)) 56.98 ± 0.77 (1.1, 1.6) g/day Arginine 2380.73 ± 176.40 110 46 Arachidic acid (C 20:0) 0.30 ± 0.01 Aspartic acid 2068.83 ± 88.28 111 ± ± 47 Gadoleic acid (C 20:1) 0.23 0.03 Serine 1197.13 30.57 112 ± ± 48 Eicosadienoic acid (C 20:2) 0.04 0.00 Glutamic acid 3761.49 57.80 113 ± ± 49 cis-11,14,17-Eicosatrienoic 0.04 0.00 Glycine 1182.02 29.07 114 u 50 acid (C 20:3 ( -3)) 115 Behenic acid (C 22:0) 0.09 ± 0.00 Alanine 1163.09 ± 35.18 51 116 Tricosanoic acid (C 23:0) 0.02 ± 0.00 Cysteine 380.35 ± 38.44 52 117 Lignoceric acid (C 24:0) 0.08 ± 0.01 Tyrosine 893.33 ± 45.47 53 118 Saturated fatty acid 11.90 ± 0.42 Proline 528.59 ± 14.70 54 119 Unsaturated fatty acid 88.11 ± 0.47 Essential 7361.97 ± 55.76 55 amino acids 120 56 u-3 fatty acid 57.02 ± 0.77 Nonessential 13555.57 ± 37.29 121 57 amino acids 122 58 u-6 fatty acid 21.51 ± 0.35 123 59 u-3:u-6 2.65 ± 0.08 124 60 125 Data are mean ± standard error of the mean (n ¼ 3). 61 126 DIR ¼ daily reference intake. 62 127 a Values are based on people aged 19e50 years from USDA (2015). 63 b Values are based on people older than 12 years from FAO/WHO/UNU Expert Consultation (1985). 128 64 c Methionine þ cysteine. 129 65 d Phenylalanine þ tyrosine. 130

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1 amino acids. The major minerals of CHIA were Mg, Ca, and K; not as good as animal proteins, CHIA has complete essential 66 2 Fe, Zn, Mn, Co, and Se were also found (Table 3). Total crude amino acids. In a comparison of daily recommended values in 67 3 68 polysaccharide content reached 30.81 g per 100 g CHIA. The essential amino acids from FAO/WHO/UNU Expert Consulta- 4 69 flavonoid content occupied 80.85% in the polyphenols of CHIA tion (1985) [16], CHIA should be a good amino-acid supple- 5 70 (Table 3) where both rutin and hesperidin (Figure 1) are major mentation. According to the adult daily reference intakes in 6 71 7 components. minerals from USDA (2013) [14], CHIA can be also a suggestive 72 8 According to the dietary reference intakes for LA and ALA supplementation for the adults. CHIA improved the water 73 9 per day, suggested by USDA (2015) [14], CHIA is a good choice holding capacity and emulsifying ability in cookies, bread, and 74 10 for a daily supplementation. It was also reported that CHIA other desserts due to their gum-like characteristic of poly- 75 11 can decrease serum triglyceride and increase high-density li- saccharides mainly consisting of crude fiber and carbohydrate 76 12 poprotein contents in rats [15]. This benefit has been attrib- [4,6,17], but published reports relevant to their application on 77 13 uted to ALA contents in CHIA. The high ALA content (56.98 g/ meat products seem lacking. Moreover, the application of 78 14 100 g oil) and a good ratio of u-3 and u-6 fatty acids (2.65) in plant polyphenols on prolonging the shelf life of meat prod- 79 15 80 CHIA should be good for the cardiovascular system in ucts has been studied [12,18e21]. The application of rutin or 16 81 humans. Although the biological values of plant proteins are hesperidin, major flavonoid compounds in CHIA, on food 17 82 18 83 19 84 20 Table 3 e Mineral and polyphenolic profiles in chia seeds. 85 21 86 Content Adult DIRs Content 22 87 (male, female)a 23 88 24 Mineral Total crude polysaccharides (g/100g) 30.81 ± 1.44 89 ± 25 Potassium (K; mg/100g) 13,477.61 56.27 (4.7, 4.7) g/d 90 ± ± 26 Magnesium (Mg; mg/100g) 4963.81 31.80 (420, 320) mg/d Total polyphenols (mg GAE/100g extract) 239.02 7.06 91 ± ± 27 Calcium (Ca; mg/100g) 4221.89 232.44 (1.0, 1.0) g/d Total flavonoids (mg CE/100g extract) 193.24 5.39 92 ± ± 28 Sodium (Na; mg/100g) 11.55 0.87 (2.3, 2.3) g/d Condensed tannins (mg CE/100g extract) 31.15 0.66 93 Iron (Fe; mg/100g) 131.12 ± 14.60 (45, 45) mg/d Rutin (mg/100g extract) 98.56 ± 3.62 29 94 Zinc (Zn; mg/100g) 88.69 ± 5.24 (40, 40) mg p-Anisic acid (mg/100g extract) 8.32 ± 0.36 30 95 Manganese (Mn; mg/100g) 71.01 ± 6.35 (2.3, 1.8) mg/d Hesperidin (mg/100g extract) 56.79 ± 2.53 31 96 Copper (Cu; mg/100g) 26.67 ± 3.47 (10,000, 10,000) mg/d 32 97 Cobalt (Co; mg/100g) 1.73 ± 0.07 NA 33 Nickel (Ni; mg/100g) 4.09 ± 0.30 (1.0, 1.0) mg/d 98 34 Selenium (Se; mg/100g) 45.33 ± 1.67 (55, 55) mg/d 99 35 100 ± ¼ 36 Data are given as mean standard error of the mean (n 3). 101 ¼ ¼ 37 DIR daily reference intake; NA not available. 102 a Values are based on people aged 19e70 years from USDA (2015). 38 103 39 104 40 105 41 106 42 107 43 108 44 109 45 110 46 111 47 112 48 113 49 114 50 115 51 116 52 117 53 118 54 119 55 120 56 121 57 122 58 123 59 124 60 Figure 1 e (A) High-performance liquid chromatograms of flavonoids and phenolic acids in chia seed and standards. (B) The 125 61 126 structures of rutin and hesperidin. Peaks: (1) gallic acid; (2) gentisic acid; (3) p-hydroxybezoic acid; (4) catechin; (5) 62 127 chlorogenic acid; (6) vanillic acid; (7) caffeic acid; (8) syringic acid; (9) epicatechin; (10) p-coumaric acid; (11) ferulic acid; (12) 63 128 64 sinapic acid; (13) rutin; (14) p-anisic acid; (15) quercitrin; (16) myricetin; (17) naringin; (18) hesperidin; (19) rosmarinic acid; 129 65 (20) diosmin; (21) neohesperidin; (22) morin; (23) daidzein; (24) eriodictyol; (25) glycitein; (26) quercetin; (27) luteolin; (28) 130 narigenin; (29) genistein; (30) apigenin; (31) kaempferol; (32) hesperetin; (33) isorhamnetin. Q7

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1 products has been characterized as oxidative stability of oil in possibly due to the scanning electron microscopy dehydration 66 2 water emulsions [22], a native stabilizer for protein emulsions process. Therefore, it corresponds to the better emulsifying 67 3 68 [23], and improvements of meat quality (i.e., lipid/protein status and production yields in ham-like products added with 4 69 oxidation), water holding capacity for supplementing lambs both CHIA and CA compared to others without fat addition 5 70 [24] and broilers [25]. Hence, it is speculative that a CHIA (Figure 2A and 2B). Pork back fat (5.0%) in recipe of restruc- 6 71 7 addition not only adds nutritional value but also improves tured ham-like products got the best overall acceptance 72 8 processing properties of low-fat meat products. because of its better emulsifying properties, water holding 73 9 capacity, mouth feel, and aroma. Due to smaller fat globules 74 10 3.2. Application of CHIA on manufacture of restricted (Figure 2A) and higher consumers' acceptance (Figure 2B), an 75 11 ham-like products extra addition of combination of 0.5% CA and 1.0% CHIA is 76 12 recommended to the recipes of restructured ham-like prod- 77 13 3.2.1. Effects of CHIA on processing properties and sensorial ucts without the fat addition. 78 14 evaluation of restructured ham-like products 79 15 80 HF, CONþ0.5CAþ0.5CHIA, and CONþ0.5CAþ1.0CHIA products 3.2.2. Physicochemical properties of CHIA on restructured 16 81 had higher (p < 0.05) production yields than other products; ham-like products during refrigerator storage (4 C) 17 82 < < 18 meanwhile, the HF group had the lowest (p 0.05) moisture HF products had the highest (p 0.05) hardness when prod- 83 < 19 and the highest (p 0.05) fat contents among all groups (Table ucts were tested immediately after manufacturing, as well as 84 20 4). Generally, CONþ1.0CHIA and CONþ0.5CAþ1.0CHIA prod- cohesiveness during 4 weeks of storage (Table 4). Generally, 85 21 ucts had higher crude protein and ash contents, respectively, CHIA, CA, or a combination of CHIA and CA resulted in softer 86 22 among different-recipe products. Via the microstructural texture in restructured ham-like product without fat addition 87 23 observation, HF product had the smallest fat globules uni- after storage. The patterns of adhesiveness and springiness of 88 24 formly, and CON product had the largest ones among groups all products after storage did not seem to change greatly, 89 25 (Figure 2A). The fat globule sizes in products with CHIA, CA, or whereas the CONþ0.5CAþ1.0CHIA product always kept the 90 26 91 CHIA þ CA were smaller than that of CON products. Larger lowest (p < 0.05) springiness. In color measurements, HF 27 92 areas of rock-like cracks were also observed in products had the highest (p < 0.05) L* value among groups 28 93 CONþ0.5CAþ0.5CHIA and CONþ0.5CAþ1.0CHIA groups than during the storage (Figure 3A). Regarding the color parameters 29 94 þ < 30 others (Figure 2A). Overall, the HF group demonstrated the of products, the CON 0.5CA product had the lowest (p 0.05) 95 < < 31 best (p 0.05) odor, color, texture, flavor, and overall accep- a* value but the highest (p 0.05) b* value after storage. The 96 32 tance among groups while the CON group had the least purge losses (%) of products were not (p > 0.05) different after 97 33 (p < 0.05) scores in all parameters except color (Figure 2B). In a 2 weeks of storage, but after 4 weeks of storage the HF product 98 34 comparison of all sensorial parameters among products, had the lowest purge (Figure 3B). Although no (p > 0.05) dif- 99 35 CONþ0.5CAþ1.0CHIA product showed a similar acceptance to ferences on purge loss were detected among products without 100 36 HF product, and even better juiciness than HF ones. fat addition, there was a tendency towards lower purge loss in 101 37 According to the recipes of restructured ham-like products products added with CHIA or CA. Regarding centrifugation 102 38 103 (Table 1), the HF group includes additions of 5% pork back fat loss, CON products showed higher losses at any measurable 39 104 and 10% water compared to other groups with no additions of time of storage. The HF, CONþ1.0CHIA, CONþ0.5CAþ0.5CHIA, 40 105 þ þ < 41 pork fat and addition of 15% water. Therefore, it is reasonable and CON 0.5CA 1.0CHIA products showed lower (p 0.05) 106 42 that HF products had the highest fat content but the lowest centrifugation loss after the 4 weeks of storage compared to 107 43 moisture content. In addition, the higher crude protein and CON product. Figure 3C illustrates that the HF group had the 108 44 ash contents in an extra 1.0% CHIA addition in the recipe may highest (p < 0.05) TBARS value and the lowest thiol group 109 45 result from the amino acid and mineral contents in CHIA, content during the storage period. CONþ0.5CHIA, 110 46 accordingly. Emulsion stability and yield are similar concepts CONþ1.0CHIA, CONþ0.5CAþ0.5CHIA, and 111 47 that indicate the abilities of holding water and oil when CONþ0.5CAþ1.0CHIA products showed lower (p < 0.05) TBARS 112 48 products being cooked [26]. A combination of 0.7% CA and values after storage. Especially, CONþ1.0CHIA and 113 49 114 pectin show the good yield on low-fat beef frankfurters [27]. CONþ0.5CAþ1.0CHIA products showed the lowest (p < 0.05) 50 115 They also reported that high-fat meat products own lower TBARS values among products during the storage. Generally, 51 116 exudates and higher production yields because of lower or the changes of thiol groups in the seven product groups during 52 117 53 even no water addition. Similar observations were also the refrigerator storage were in contrast with those of TBARS 118 54 demonstrated in our results (Figure 1). Overall, a combination values in the products. 119 55 of CHIA and CA in the recipe of restructured ham-like prod- The ratio of protein, fat, and water is the major factor 120 56 ucts showed the better water binding capacities. A better transforming the textural properties of meat products. If part 121 57 emulsifying ability is considered in that smaller fat globules of protein or fat is substituted by water in meat products, the 122 58 disperse in the products homogeneously [28]. The inconsis- texture of products becomes overly tender. Hence, the hardest 123 59 tent particle sizes of fat globule make worse mouth feel in texture of HF products possibly results from less water addi- 124 60 meat and even dairy products [28]. The restructured ham-like tion. An addition of CA increased water holding capacity in 125 61 126 products are emulsified products, and the fat globules do in- low-fat frankfurters, thus improving textural properties [29]. 62 127 fluence overall acceptance of products significantly. Accord- Upon storage, the lower hardness of restructured ham-like 63 128 64 ing to the results, CHIA and CA improved the emulsifying products added with CHIA, CA, or combination of CHIA and 129 65 effect of ham-like products. The rock-like cracks in ham-like CA than that without them is possible due to CHIA and CA with 130 products depict the vacancy of water retention in products a higher water holding capacity, which retains the water in the

Please cite this article in press as: Ding Y, et al., Nutritional composition in the chia seed and its processing properties on restructured ham-like products, Journal of Food and Drug Analysis (2017), http://dx.doi.org/10.1016/j.jfda.2016.12.012 65 64 63 62 61 60 59 58 57 56 55 54 53 52 51 50 49 48 47 46 45 44 43 42 41 40 39 38 37 36 35 34 33 32 31 30 29 28 27 26 25 24 23 22 21 20 19 18 17 16 15 14 13 12 11 10 9 8 7 6 5 4 3 2 1 laect hsatcei rs s igY ta. urtoa opsto nteci edadispoesn rpriso restructured on properties processing its and seed chia http://dx.doi.org/10.1016/j.jfda.2016.12.012 the (2017), in Analysis composition Drug Nutritional and al., Food et of Y, Journal Ding as: products, press ham-like in article this cite Please

Table 4 e Effects of different levels of chia seeds (CHIA) and carrageenan (CA) on production yield and proximate composition of restructured ham-like products, and changes of textural proﬁles of restructured ham-like products during the storage period. HF CON CONþ0.5CHIA CONþ1.0CHIA CONþ0.5CA CONþ0.5CAþ0.5CHIA CONþ0.5CAþ1.0CHIA ora ffo n rgaayi x 21)1 (2017) xxx analysis drug and food of journal Production yield (%) 99.22 ± 0.04a 96.57 ± 0.38bc 97.05 ± 0.53b 95.43 ± 1.55c 96.87 ± 0.17b 99.64 ± 0.21a 98.94 ± 0.47a Proximate compositions Moisture (%) 69.05 ± 0.32c 73.79 ± 0.94a 73.46 ± 0.90a 71.79 ± 0.77ab 73.14 ± 0.62a 73.37 ± 0.43a 70.35 ± 1.42bc Crude fat (%) 10.42 ± 0.29a 5.55 ± 0.33c 6.13 ± 0.33c 6.82 ± 0.33bc 6.41 ± 0.47c 6.29 ± 0.10c 8.31 ± 1.19b Crude protein (%) 15.86 ± 0.03ab 16.21 ± 0.82ab 15.90 ± 0.57ab 16.99 ± 0.32a 15.79 ± 0.28ab 15.44 ± 0.37b 16.27 ± 0.36ab Ash (%) 2.13 ± 0.03b 2.12 ± 0.06b 2.16 ± 0.08ab 2.21 ± 0.03ab 2.12 ± 0.07b 2.21 ± 0.04ab 2.29 ± 0.05a Storage period Textural properties 0 week Hardness (N) 18.17 ± 0.92a 16.38 ± 0.57ab 14.62 ± 1.52b 16.47 ± 0.68ab 14.89 ± 0.93b 15.38 ± 0.39b 14.79 ± 0.57b Adhesiveness (Nxs) 0.05 ± 0.01a 0.06 ± 0.00a 0.05 ± 0.01a 0.04 ± 0.01a 0.04 ± 0.01a 0.04 ± 0.01a 0.04 ± 0.00a Springiness 0.93 ± 0.01a 0.92 ± 0.01a 0.91 ± 0.01ab 0.88 ± 0.00bc 0.86 ± 0.01c 0.89 ± 0.00bc 0.83 ± 0.01d Cohesiveness 0.70 ± 0.01a 0.70 ± 0.00a 0.70 ± 0.03a 0.64 ± 0.01b 0.59 ± 0.01b 0.69 ± 0.01a 0.62 ± 0.02b 2 week Hardness (N) 16.92 ± 1.25a 18.05 ± 1.98a 15.35 ± 0.45a 17.07 ± 0.59a 15.47 ± 0.68a 15.02 ± 1.05a 15.22 ± 1.57a Adhesiveness (Nxs) 0.05 ± 0.02ab 0.08 ± 0.01b 0.03 ± 0.01a 0.04 ± 0.01a 0.05 ± 0.01ab 0.05 ± 0.00a 0.04 ± 0.01a Springiness 0.86 ± 0.07a 0.92 ± 0.00a 0.91 ± 0.00a 0.90 ± 0.00a 0.87 ± 0.01a 0.89 ± 0.01a 0.86 ± 0.02a Cohesiveness 0.75 ± 0.02a 0.70 ± 0.01bc 0.72 ± 0.01ab 0.65 ± 0.00d 0.61 ± 0.02e 0.69 ± 0.01c 0.64 ± 0.00de 4 week Hardness (N) 19.03 ± 0.89a 17.75 ± 0.72ab 16.26 ± 0.22bcd 17.14 ± 0.67bc 15.09 ± 0.58d 15.68 ± 0.55cd 14.70 ± 0.48d Adhesiveness (Nxs) 0.06 ± 0.02b 0.03 ± 0.00a 0.05 ± 0.01ab 0.06 ± 0.01b 0.05 ± 0.01ab 0.04 ± 0.00ab 0.06 ± 0.00ab Springiness 0.91 ± 0.00ab 0.91 ± 0.00ab 0.93 ± 0.01a 0.90 ± 0.02ab 0.84 ± 0.02ab 0.89 ± 0.01bc 0.87 ± 0.01cd JFDA457_proof ± a ± bc ± ab ± ab ± cd ± cd ± d e Cohesiveness 0.69 0.01 0.64 0.02 0.68 0.02 0.67 0.01 0.60 0.01 0.63 0.01 0.59 0.00 11

Data are given as mean ± standard error of the mean (n ¼ 3). e a d Mean values without the same letters in each testing parameter are signiﬁcantly different by using least signiﬁcant difference (p < 0.05). CON ¼ control (without addition of fat); HF ¼ high fat (addition of 5% pork back fat). ■ 3Fbur 2017 February 13 ■ 7/11 7 100 120 123 122 129 128 126 125 104 106 124 105 109 108 130 127 103 102 107 121 101 112 110 116 115 114 113 119 118 117 111 66 68 69 75 90 89 88 87 86 85 84 83 82 81 80 79 78 99 98 96 95 94 93 92 67 70 77 73 72 97 71 91 76 74 JFDA457_proof ■ 13 February 2017 ■ 8/11

8 journal of food and drug analysis xxx (2017) 1e11

1 66 2 67 3 68 4 69 5 70 6 71 7 72 8 73 9 74 10 75 11 76 12 77 13 78 14 79 15 80 16 81 17 82 18 83 19 84 20 85 21 86 22 87 23 88 24 89 25 90 26 91 27 92 28 93 29 94 30 95 31 96 32 97 33 98 34 99 35 100 36 101 37 102 38 103 39 104 40 105 41 106 42 107 43 108 44 109 45 110 46 111 47 112 48 113 49 114 50 115 51 116 52 117 53 118 54 119 55 120 56 121 57 122 58 123 59 124 60 125 61 126 62 Figure 2 e Effects of different levels of chia seed (CHIA) and carrageenan (CA) on (A) changes in scanning electron 127 63 128 micrographs (magnification, 1000£) and (B) sensorial attributes of restructured ham-like products. Data are given as 64 e 129 mean ± standard error of the mean (n ¼ 40 for sensory evaluation). a c Mean values without the same letters in each testing 65 130 parameter are significantly different by using least significant difference (p < 0.05). CON ¼ control (without addition of fat); HF ¼ high fat (addition of 5% pork back fat). Please cite this article in press as: Ding Y, et al., Nutritional composition in the chia seed and its processing properties on restructured ham-like products, Journal of Food and Drug Analysis (2017), http://dx.doi.org/10.1016/j.jfda.2016.12.012 JFDA457_proof ■ 13 February 2017 ■ 9/11

journal of food and drug analysis xxx (2017) 1e11 9

1 66 2 67 3 68 4 69 5 70 6 71 7 72 8 73 9 74 10 75 11 76 12 77 13 78 14 79 15 80 16 81 17 82 18 83 19 84 20 85 21 86 22 87 23 88 24 89 25 90 26 91 27 92 28 93 29 94 30 95 31 96 32 97 33 98 34 99 35 100 36 101 37 102 38 103 39 104 40 105 41 106 42 107 e 43 Figure 3 Effect of different levels of chia seed (CHIA) and carrageenan (CA) on (A) color parameters (L*, a*, and b* value), (B) 108 44 water holding capacities [purge and centrifugation (%)], and (C) lipid/protein oxidation levels (TBARS value and thiol-group 109 45 contents) of restructured ham-like products during the storage period. Data are given as mean ± standard error of the mean 110 e 46 (n ¼ 3). a c Mean values without the same letters on data bars in the same storage period are significantly different by using 111 47 least significant difference (p < 0.05). CON ¼ control (without addition of fat); HF ¼ high fat (addition of 5% pork back fat); 112 48 a* ¼ greenness (-a*) to redness (a*), b* ¼ blueness (-b*) to yellowness (b*); L* ¼ lightness. 113 49 114 50 115 51 116 products (Figure 3B). Color plays an important role in both the storage. The recipe added with 1.0% CHIA (CONþ1.0CHIA and 52 117 ' þ þ 53 quality and consumer s preference of meat products. Gener- CON 0.5CA 1.0CHIA products) performed the lower centri- 118 54 ally, the formations of metmyoglobin and lipid oxidation make fugation loss during the storage period (Figure 3B). Oxidation 119 55 meat discolor. The group with pork back fat added increased causes rancidity and deterioration in meat products. Many 120 56 the lightness (L* value) of ham-like products (Figure 3A) which studies have been available for a retarded effect of plant ex- 121 57 assumed that white pork fat brightens the products. The high tracts on lipid oxidation in meat products, such: as tea cate- 122 58 ratio of pork fat makes the pork patties lighter and also has the chins in red meat, poultry, and fish patties [18]; rosemary and 123 59 better color preference to panelists [30]. Moreover, ham-like lemon balm extracts in pork patties or packaged beef [19]; 124 60 products with CA addition made the yellowness (b* value) grape seed flour in frankfurters [13]; and ethanolic grape-seed 125 61 126 lower to the value as similar as ham-like products in this extract in dry cured sausage (chorizo) [20]. It was reported that 62 127 experiment (Figure 3A). Purge and centrifugation losses are rutin or hesperidin has a good effect against lipid oxidation 63 128 e 64 common indicators for water holding capacity of meat prod- [23 25]. Hence, the lower TBARS values in restructured ham- 129 65 ucts [12]. According to the results, the CON group had the like products with CHIA partially results from their poly- 130 higher purge and centrifugation losses upon the refrigerator phenols, especially rutin and hesperidin (Table 3 and Figures 1

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