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Fatty acid, amino acid, and mineral composition of four common vetch seeds on Qinghai-Tibetan plateau
Accepted Manuscript Fatty acid, amino acid, and mineral composition of four common vetch seeds on Qinghai-Tibetan plateau Zhuxin Mao, Hua Fu, Zhibiao Nan, Changgui Wan PII: DOI: Reference:
S0308-8146(14)01315-6 http://dx.doi.org/10.1016/j.foodchem.2014.08.090 FOCH 16313
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Food Chemistry
Received Date: Revised Date: Accepted Date:
21 April 2014 14 August 2014 21 August 2014
Please cite this article as: Mao, Z., Fu, H., Nan, Z., Wan, C., Fatty acid, amino acid, and mineral composition of four common vetch seeds on Qinghai-Tibetan plateau, Food Chemistry (2014), doi: http://dx.doi.org/10.1016/ j.foodchem.2014.08.090
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1
Fatty acid, amino acid, and mineral composition of four
2
common vetch seeds on Qinghai-Tibetan plateau
3
Zhuxin Mao a, b, Hua Fub,*, Zhibiao Nanb, Changgui Wanb
4
a
5
PR China.
6
b
7
Technology, Lanzhou University, Lanzhou 730000, PR China.
Xi’ an Botanical Garden of Shaanxi Province, Institute of Botany of Shaanxi Province, Xi’an 710061,
State Key Laboratory of Grassland Agro-ecosystems, College of Pastoral Agriculture Science and
8 9 10
11
*Corresponding author. Tel./ Fax: +86 931 8663778. E-mail address: [email protected] (H. Fu).
12
Fatty acid, amino acid, and mineral composition of four
13
common vetch seeds on Qinghai-Tibetan plateau
14
Zhuxin Mao a, b, Hua Fub,*, Zhibiao Nanb, Changgui Wanb
15
a
16
PR China.
17
b
18
Technology, Lanzhou University, Lanzhou 730000, PR China.
Xi’ an Botanical Garden of Shaanxi Province, Institute of Botany of Shaanxi Province, Xi’an 710061,
State Key Laboratory of Grassland Agro-ecosystems, College of Pastoral Agriculture Science and
19 20
Abstract
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The chemical composition of four common vetch (Vicia sativa L.) seeds was
22
investigated to determine their nutrition value. The result shows that the seeds are low
23
in lipid (1.55-2.74% of dry weight), and high in the unsaturated fatty acid
24
(74.51-77.36% of total fatty acid). The ratio of essential amino acid to non-essential
25
amino acid (0.62-0.69) is even higher than the amount (0.38) recommended by World
26
Health Organization. Besides, the seeds are also found rich in Mg, Mn and Cu, but
27
with a low ratio of Ca to P (0.24-0.73), which may increase the risk of the mineral
28
element toxicity. The results indicate that the four common vetch seeds could be taken
29
as an alternative food source, but the possible toxic effect should be taken into
30
consideration.
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Keywords: common vetch seed, fatty acid, amino acid, mineral element,
32
Qinghai-Tibetan plateau
*
Corresponding author. Tel./ Fax: +86 931 8663778. E-mail address: [email protected] (H. Fu).
33
1. Introduction
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Common vetch (Vicia sativa L.) is a potential source of cheap protein for human
35
beings (Akpinar, Ali Akpinar, & Türkoğlu, 2001), although it has been reported to be
36
toxic (Luse, Kang, Fox, & Nangju, 1975; Ressler, Tatake, Kaizer, & Putnam, 1997).
37
Because of the good adaption to adverse environmental conditions, common vetch has
38
attracted much attention from breeders and producers, and thus has been cultivated as
39
a grain legume for seed production in Asia, Africa and Europe (Francis, Enneking, &
40
El Moneim, 2000). The common vetch seed as a rich source of protein could
41
contribute greatly to the protein supply for human beings and livestock. With a
42
growing recognition on the importance of legume plants to human health, much
43
attention has been paid to the chemical composition of the common vetch seed
44
(Acikgoz, Katkat, Omeroglu, & Okan, 1985; Pastor-Cavada, Juan, Pastor, Alaiz, &
45
Vioque, 2009; Samarah & Ereifej, 2009).
46
Fatty acid is important for human health since it’s the precursor for the biosynthesis
47
of eicosanoids which is considered as an important bio-regulator for many cellular
48
metabolic processes (Gressler et al., 2010; Khotimchenko & Yakovleva, 2005). In the
49
common vetch seed, fatty acids have a high degree of unsaturation (65-75%) (Akpinar
50
et al., 2001). It has been found that the major unsaturated fatty acids in common vetch
51
seeds are oleic, linoleic and α-linolenic acid accounting for over 50% of total fatty
52
acid (Akpinar et al., 2001). The high content of protein (24-32%) enriches the seeds
53
more nutrition (Uzun, Gücer, & Acikgoz, 2011). And protein is composed of different
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amino acids and thus the degree of nutrition is largely determined by their relevant
55
proportion (Gressler et al., 2010). Previous studies have revealed that glutamic
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(19.9%) and aspartic (14.8%) were the most predominant amino acids in common
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vetch seeds (Acikgoz et al., 1985; Karadag & Yavuz, 2010).
58
Mineral deficiency is a widespread problem in many developing countries affecting
59
over 2 billion people, which could even cause infant and child mortalities worldwide
60
(Uzun et al., 2011). Through analyzing the minerals on 388 common vetch accessions,
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Uzun et al. (2011) pointed out that the seeds were rich in calcium (Ca), magnesium
62
(Mg), manganese (Mn), Zinc (Zn) and copper (Cu). However, Acikgoz et al. (1985)
63
found that common vetch seeds were extremely low in Ca, Mg and Mn but rich in
64
phosphorus (P). Thus, the mineral composition seems significantly different among
65
the common vetch varieties.
66
The nutrition differed greatly among common vetch seeds from different regions,
67
which was attributed to the genetic factors and environmental conditions of the
68
production areas (Akpinar et al., 2001). To the best of our knowledge, little
69
information was available on vetches in China as one of the largest countries for the
70
production of common vetch in the word. Some research on common vetch was
71
initiated on Qinghai-Tibetan plateau of China in the late 20th century (Nan, Abd
72
El-Moneim, Larbi, & Nie, 2006). The plateau of a vast area up to 2.5 million km2 is
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the best grazing land in Asia. But the short growing season and harsh environment
74
conditions impose constraints on the production of cropping and livestock (Wu, 2001).
75
In recent years, Chinese breeders have cultivated new common vetch cultivars with
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excellent adaptability to the plateau (Nan et al., 2006). However, the previous
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researches mainly concentrated on the agronomic characteristics and herbage nutrition
78
(Mao, Fu, Nan, Wang, & Wan, 2012), few were devoted to the study in terms of the
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chemical composition in the common vetch seeds. Thus, this study is in the purpose to
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analyze and compare the content of fatty acid, amino acid and mineral element in the
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common vetch seeds obtained from the plateau to determine if they possess favorable
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traits from a biochemical point of view.
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2. Materials and methods
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2.1 Seed sample
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The field experiment was carried out at Lanzhou university research farm, Gansu,
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China (35º06′ N, 102º25′ E, 3050 m above sea level), located on the eastern edge of
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the Qinghai-Tibetan plateau. The average annual precipitation is 424 mm, mean daily
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temperature is 2.8 degrees Celsius, and over 60% of the area is grassland. The soil is
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chernozem, slightly acidic (pH is 6.7), low in P (7.5 mg/kg) and high in K (17.5g/kg)
90
(Mao et al, 2012).
91
Four common vetch cultivars were used in this study: 2505, 2556, 2560 and
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V.333/A. The cultivars were selected from Common Vetch Breeding Program of
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Lanzhou University mainly from the Qinghai-Tibetan plateau. The seeds were grown
94
during the 2011 growing season in a randomized complete block design with four
95
replications, and each replication was a 3.6 m2 plot. Harvesting and threshing were
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performed by hand. After cleaning, the matured seeds (500g) were dried in
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a forced-air oven at 50 degrees Celsius to constant weight, and ground through a 0.5
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mm screen by a cyclone mill. These samples were stored at -20 degrees Celsius in
99
refrigerator before the chemical analysis.
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2.2 Chemical analysis
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2.2.1 Fatty acid analysis
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The lipid was measured by the continuous lipid extraction by a soxhlet apparatus.
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The lipid extracted from the seeds flours were mixed with boron trifluoride-methanol
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reagent (25%) and fatty acids were then converted into the methyl ester derivatives
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(Morrison & Smith, 1964). The fatty acid methyl esters analysis was performed on
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Agilent 6890N gas chromatography instrument coupled with an Agilent MS-5973
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inert XL mass selective detector and an Agilent autosampler 7683-B injector (Little
108
Fall, NY, USA). The capillary column was an HP-FFAP MS with dimension of 30 m
109
length ×0.25mm i.d. ×0.25µm film thickness (Palo Alto, CA, USA). Helium was used
110
as carrier gas at a flow rate of 1.1mL/min. Temperature program began by holding
111
temperature at 60℃ for 2 min, then increased to 200℃ at 15℃ /min, and held at
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200℃ for 1 min before rising to 240℃ at 8℃/min with a final holding temperature
113
at 240℃ for 10min. The injection was performed in a split mode (split ratio 100:1).
114
Detector and injector temperature was 250℃ and 230℃, respectively. MS scan range
115
was (m/z): 40-550 atomic mass units (AMU) under electron impact (EI) ionization
116
(70 eV) and ion source temperature was 230℃ (Mehmood et al., 2008).
117
Fatty acid methyl ester peaks were identified by comparing their retention
118
times and spectra with standard compounds which were purchased from Fluka
119
(Sigma-Aldrich, Inc., St. Louis, MO, U.S.A). Standard methyl esters of palmitic,
120
stearic, oleic and linoleic acids were used for the confirmation of GC-MS library
121
results. The percentage of fatty acid composition was decided by the ratio of
122
individual peak area to all fatty acid areas (Table 1). Each analysis was performed at
123
least in triplicate.
124
2.2.2 Amino acid analysis
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Protein and peptides acid hydrolysis was done according to the method of
126
Spackman et al. (1958). A 10-20 mg powdered sample without lipid was transferred
127
to 10×150 mm borosilicate ampoules previously pyrolysed at 420 degrees Celsius for
128
8 h. For each ampoule, 0.5 mL of HCl 6 M with 0.1% of phenol (m/v) was added.
129
Vacuum was applied before the seal of the ampoules and then they were put into an
130
oven at 110 degrees Celsius for 22 h. The hydrolysed samples were analyzed
131
according to the standard of the European Community (98/64/EG). Amino acid
132
samples were separated by ion exchange chromatography and determined by reaction
133
with ninhydrin with photometric detection at 570 nm (440 nm for proline) using the
134
automatic amino acid analyzer LC-3000 (Eppendorf-Bio-tronik; Germany). Amino
135
acid standard solution (A9781 from Sigma-Aldrich, Germany) (0.06 mg/mL) was
136
injected to calibrate the analyzer and to calculate the amount of amino acid in the
137
samples. In addition, nitrogen (N) was determined by the micro-Kjeldahl technique
138
with the duplicate samples of dried seeds for each genotype. Crude protein content
139
was then calculated by 6.25×N (Mariotti, Tomé, & Mirand, 2008).
140
2.2.3 Mineral analysis
141
Potassium (K), P, Mg, sodium (Na), Ca, Zn, Mn and Cu were analyzed for seed
142
minerals. The seeds were washed with deionised water, dried at 75 degrees Celsius,
143
weighed and ashed at 480 degrees Celsius in a muffle furnace. The ashes were
144
dissolved in 5 mL of 20% (v/v) HCl and diluted to a volume of 100 mL with
145
deionised water. The solution was analyzed for K, Mg, Na, Ca, Zn, Mn, and Cu by a
146
Perkin-Elmer 5000 flame (air-acetylene) atomic absorption spectrometer with
147
hollow-cathode lamp tubes (Norwalk, Connecticut, USA) according to Pinheiro et al.
148
(2010). P was measured in 5 mL of the same solution by the colorimetric
149
molybdenum ammonium vanadate method (Motomizu & Li, 2005), using a Hitachi
150
Perkin-Elmer Model-139 UV-vis spectrophotometer (Tokyo, Japan), at the
151
wavelength of 470 nm.
152
2.3 Statistics analysis
153
The results were expressed as means ± standard deviation (SD). All statistical
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analysis was performed using the SPSS software package, version 13.0 (SPSS Inc.,
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Chicago). The data was analyzed by one way analysis of variance (ANOVA).
156
Differences were considered significant at P < 0.05.
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3. Results and Discussion
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3.1 Seed yield, content of lipid, protein and ash
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In this study, the average yield of common vetch seeds was 1392 kg/ha for 2505,
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853 kg/ha for 2556, 1172 kg/ha for 2560, and 890 kg/ha for V. 333/A (Fig. 1A). The
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seed yield of 2505 and 2560 was significantly higher (P<0.05) than that of 2556 and
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V. 333/A. The seed yield of 2505 and 2560 was higher than that (820-1000 kg/ha) in
163
China (Nan et al., 2006). The seed yield of 2556 and V. 333/A was lower than that
164
(1160-1459 kg/ha) in Turkey (Karadag et al., 2010).
165
The seed crude protein (CP) content varied from 29.23% to 30.57% (Fig. 1B), and
166
no significant difference has been found among the cultivars (P<0.05). The CP
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content was in accordance with that in the previous studies ranged from 25.30% to
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31.38% (Karadag et al., 2010; Uzun et al., 2011). In addition, the CP content of four
169
cultivars was close to that of common vetch lines 845 (28.90%), 1448 (30.77%) and
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384 (31.38%) in Turkey (Karadag et al., 2010), but was lower than that of common
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vetch cultivars (37.03%) in Madrid, Spain (Caballero et al., 2001).
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The lipid content of seeds was 1.55% for 2505, 1.74% for 2556, 2.74% for 2560,
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and 2.28% for V. 333/A; the content in 2560 and V. 333/A was significantly higher
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(P<0.05) than that in 2505 and 2556 (Fig. 1C). The lipid content in the current study
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was higher than other common vetch species (Caballero et al., 2001; Milczak et al.,
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2001), but lower than the varieties reported by Darwish Sayed et al. (1980).
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The ash content of seeds was 2.64 g/kg for 2505, 4.10 g/kg for 2556, 3.20 g/kg for
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2560, and 3.32 g/kg for V. 333/A (Fig. 1D), the content ranked as
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2556>2560=V.333/A>2505 (P<0.05). The result was in accordance with the ash
180
content studied by Samarah and Ereifej (2009).
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3.2 Fatty acid
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The fatty acid composition and content are presented in Table 1. Nine fatty acids
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detected in four cultivars are identified as saturated fatty acids (SFA) and unsaturated
184
fatty acids (UFA). The proportion of SFA accounted for 21.14% to 23.22% of total
185
fatty acids (TFA) in the seeds (Table 1), and no significant difference has been found
186
among the cultivars. The most abundant SFA was palmitic acid (C 16:0) accounting
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for 15.79% to 17.42%. Stearic (C 18:0) and arachidic acids (C 20:0) presented in a
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small amount ranging from 2.78% to 3.21% and 1.17% to 1.51%, respectively.
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Myristic (C 14:0) and pentadecanoic (C 15:0) acids were found in a trace amount,
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ranging from 0.28% to 0.67% and 0.33% to 0.65%, respectively. The result that
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palmitic acid was the only major SFA in four common vetch cultivars was in
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agreement with what was found by Pastor-Cavada et al. (2009). But our result was of
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some difference from what was found by Akpınar et al. (2001) who reported that
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palmitic (8.05%) and stearic acids (7.31%) were both predominant in common vetch
195
seed oil in Turkey. In this study, arachhidic acid was present in a small amount but it
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wasn’t detected by Darwish Sayed et al. (1980) in common vetch. Finally, a big
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difference was found in the content of myristic and pentadecanoic acids up to 3.79%
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and 2.91% in common vetch seed in Turkey (Akpinar et al., 2001), which is much
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higher than that in our study. This may be related with the differences in the
200
ecological and geographical conditions as suggested by Aaes-Jorgensen (1961).
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There are four UFA present in this study (Table 1). The amount of UFA in the
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seeds ranged from 74.51% to 77.36% with no significant differences among the
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cultivars. With regard to the individual UFA, the most abundant UFA was linoleic
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(C18:2 cis-9, 12), ranging from 52.56% to 54.39%. Oleic acid (C18:1 trans-9) and
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α-linolenic acid (C18:3 cis-9, 12, 15) were present in a considerable amount up to
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8.33-11.54% and 9.03-12.12%, respectively. Eicosenoic acid (C20:1 cis-9) has been
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detected in a small amount ranging from 1.28% to 2.27%. In this study, the content of
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oleic acid was 23.70% only half of that in the seeds collected in Turkey (Akpinar et
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al., 2001). However, linoleic acid was more than half of TFA in this study, much
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higher than the content (4.33%) revealed by Akpinar et al. (2001). Most notably, the
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content of linolenic and eicosenoic acids was both higher than that (1.95% and 0.34%,
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respectively) in the seeds from Turkey (Akpinar et al., 2001).
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In addition, the ratio of UFA to SFA at 3.30-3.66 in common vetch seeds in the
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current study was much higher than that from Turkey (Akpinar et al., 2001). The
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result could indicate that the seeds harvested from the high plateau have higher
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proportion of UFA, of which, α-linolenic acid of considerable amount was
217
recognized quite beneficial to heart health (Connor, 1999).
218
3.3 Amino acid
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Eighteen amino acids were detected in the common vetch seeds (Table 2),
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10.64-11.83% of them were essential amino acids (EAA) and rest were non-essential
221
amino acids (NEA). There was a small difference in EAA among the common vetch
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cultivars. Arginine and leucine turned out to be the predominant EAA accounting for
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2.15-2.53% and 1.98-2.23%, respectively. Lysine, isoleucine, phenylalanine and
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threonine were in a small amount at 1.60-1.73%, 1.17-1.33%, 1.18-1.25% and
225
0.97-1.11%, respectively. Histidine, methionine and tryptophane were in a trace
226
amount at 0.66-0.74%, 0.44-0.54% and 0.28-0.33%, respectively. Glutamic and
227
aspartic turned out to be the predominant NEA of 5.24-5.70% and 3.48-4.04%,
228
respectively. Valine, serine, alanine, glycine and proline were in a small amount at
229
1.61-1.73%, 1.33-1.42%, 1.16-1.43%, 1.08-1.25% and 1.13-1.27%, respectively.
230
Tyrosine and cystine were in a trace amount at 0.85-0.93% and 0.45-0.56%,
231
respectively. The type and content of amino acids in this study were somewhat
232
different from the previous reports (Hadjipanayiotou & Economides, 2001).
233
According to Hadjipanayiotou et al. (2001), tryptophane and cystine were not found
234
in common vetch seeds in Cyprus. Glutamic acid was the highest NEA (>5.24%),
235
whereas methionine (0.44%) and alanine (0.44%) were the lowest NEA in this study,
236
which may suggest that there was less NEA in common vetch seeds grown on the
237
Qinghai-Tibetan plateau. In addition, the amino acids content in the present study was
238
close to that in Turkey (Karadag et al., 2010), the highest and lowest EAA was
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1.95-2.26% (leucine) and 0-0.02% (methionine), respectively. While the highest and
240
lowest NEA was 3.65-7.74% (aspartic) and 0.73-0.90% (tyrosine), respectively. In
241
contrast to our result, arginine, tryptophane and cystine were not found in the
242
common vetch seeds in Turkey (Karadag et al., 2010).
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In this study, the amount of amino acids ranged from 28.42% to 30.11% (Table 2).
244
The ratio of EAA to the total amino acid for common vetch species was about 0.4.
245
The ratio of EAA to NEA for all cultivars was 0.62-0.69, which was substantially
246
higher than the adult requirement for EAA/NEA (0.38) recommended by WHO (Joint
247
WHO., 2007).
248
3.4 Mineral element
249
Mineral concentrations of common vetch seeds are reported in Table 3. Among the
250
macronutrient minerals, P, Mg and Ca were 1.63-3.96 g/kg, 2.20-2.66 g/kg and
251
0.94-1.20 g/kg, respectively. The highest content of P (3.96 g/kg) and Mg (2.66 g/kg)
252
was found in 2505 and 2556, respectively and they showed no significant difference
253
among other cultivars. The lowest Ca (0.94 g/kg) was found in 2505, and no
254
significant difference was found with other cultivars. Uzun et al. (2011) found that P
255
content of common vetch seeds in Turkey was 4.96 g/kg, and the corresponding Mg
256
and Ca were 1.53 g/kg and 1.08 g/kg, respectively. Similarly, Moraghan and Grafton
257
(2001) reported that the mean content of P in the common bean seeds in America was
258
4.45-5.26 g/kg, and the corresponding content of Mg was 1.02-1.52 g/kg. Our results
259
show that the seeds harvested from plateau are deficient in P but rich in Mg, which
260
may reflect the difference in the chemical property of the local soils. K and Na were
261
9.69-9.98 g/kg and 1.20-1.28 g/kg, respectively, and no significant difference (P<0.05)
262
was found with other cultivars. The content of K in this study is similar to what has
263
been found by Samarah et al. (2009) although the content of Na was higher in their
264
study.
265
Among the micronutrient minerals, Zn, Mn and Cu were 30.01-48.16 mg/kg,
266
12.74-21.78 mg/kg and 3.37-8.77 mg/kg, respectively. The highest level of
267
micronutrient minerals was all found in 2556, while the lowest level was found in
268
2505 (P<0.05). And there was no significant difference between 2560 and V. 333/A.
269
The level of Zn and Mn in this study was consistent with what has been found by
270
Uzun et al. (2011) that Zn and Mn in common vetch seeds were 31.50 mg/kg and
271
14.50 mg/kg, respectively. The similar result has also been reported by Samarah et al.
272
(2009). However, Cu in our study was lower than the level reported by Uzun et al.
273
(2011) and Samarah et al. (2009) at 11.70 mg/kg and 13.02 mg/kg, respectively.
274
Mineral element in food is important for human health, but excessive intake of the
275
mineral element will result in toxicity (Luse et al., 1975). The potential contribution
276
of 100g of common vetch seeds to the Recommended Dietary Allowances (RDA) (US
277
Department of Agriculture, 2004) on mineral elements is present in Table 4. The
278
content of Mg, Mn and Cu in 100g common vetch seeds could contribute RDA as
279
high as 37.44-97.44% to the adult. K, P and Zn contribute at a medium level
280
(20.62-56.57%), Na and Ca contribute at low level (8.01-12.01%). The result suggests
281
that common vetch seeds could be the dietary resource for the mineral elements, but
282
excessive intake of the seeds may result in toxicity to human beings. In addition, the
283
availability of Ca in the body greatly depends on the ratio of Ca to P, which is
284
recommended optimal between 1.0 and 1.3 (Calvo & Park., 1996). However, the ratio
285
of Ca to P in four common vetch seeds was 0.24-0.73, which may disturb the
286
metabolism of Ca and negatively affect bone health (Kemi et al., 2010).
287
4. Conclusions
288
This study analyzed the composition of fatty acid, amino acid and mineral element
289
in the four common vetch seeds on Qinghai-Tibetan plateau of China. The result
290
shows that the seeds are low in lipid with a ratio of UFA/SFA up to 3.30-3.66, which
291
means that the seeds have accumulated more UFA primarily as linoleic, oleic and
292
α-linolenic acids. In addition, the ratio of EAA to NAA (0.62-0.69) is even higher
293
than the amount (0.38) recommended by WHO (Joint WHO., 2007). Mg, Mn and Cu
294
in 100g common vetch seeds may supply 37.44-97.44% of the RDA for adult, but
295
excessive intake of the seeds may result the mineral element toxicity. In addition, the
296
ratio of Ca to P in the seeds was 0.24-0.73 lower than the level (1.0-1.3)
297
recommended by Calvo and Park (1996). It is concluded that four common vetch
298
seeds can be a potential alternative food or feed on Qinghai-Tibetan plateau, but the
299
possible toxic effect should also be kept in mind.
300 301
Acknowledgments
302
This study was supported by National Basic Research Program of China (No.
303
2014CB138703), Special Fund for Agroscientific Research in the Public Interest (No.
304
201203041), and Shaanxi Province Science Foundation for Youths (No. 2013K-19).
305 306 307 308
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Connor, W. E. (1999). α-Linolenic acid in health and disease. The American Journal of Clinical Nutrition, 69, 827-828.
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Darwish Sayed, M., Afifi, M. S., & Hassan, M. A. (1980). A study of lipid content of the leaves, stems
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Francis, C. M., Enneking, D., & El Moneim, A. A. (2000). When and where will vetches have an
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impact as grain legumes?. In Linking Research and Marketing Opportunities for Pulses in the
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21st Century (pp. 375-384). Springer Netherlands.
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Gressler, V., Yokoya, N. S., Fujii, M. T., Colepicolo, P., Torres, R. P., & Pinto, E. (2010). Lipid, fatty
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acid, protein, amino acid and ash contents in four Brazilian red algae species. Food Chemistry,
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Hadjipanayiotou, M., & Economides, S. (2001). Chemical composition, in situ degradability and amino
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acid composition of protein supplements fed to livestock and poultry in Cyprus. Livestock
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Karadag, Y., & Yavuz, M. (2010). Seed yields and biochemical compounds of grasspea (Lathyrus
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sativus L.) lines grown in semi-arid regions of Turkey. African Journal of Biotechnology, 9,
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8343-8348.
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Kemi, V. E., Kärkkäinen, M. U., Rita, H. J., Laaksonen, M. M., Outila, T. A., & Lamberg-Allardt, C. J.
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(2010). Low calcium: phosphorus ratio in habitual diets affects serum parathyroid hormone
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concentration and calcium metabolism in healthy women with adequate calcium intake.
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British journal of nutrition, 103, 561-568.
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Khotimchenko, S. V., & Yakovleva, I. M. (2005). Lipid composition of the red alga Tichocarpus crinitus exposed to different levels of photon irradiance. Phytochemistry, 66, 73-79.
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Luse, R. L., Kang, B. T., Fox, R. L., & Nangju, D. (1975). Protein quality in grain legumes grown in
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the lowland humid tropics, with special reference to West Africa. In Fertilizer use and protein
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production. XIth Colloquium, International Potash Institute (pp. 193-201). Ronne-Bornholm,
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Denmark.
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Mao, Z. X., Fu, H., Nan, Z. B., Wang, J., & Wan, C. G. (2012). Fatty acid content of common vetch
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(Vicia sativa L.) in different regions of Northwest China. Biochemical Systematics and
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Ecology, 44, 347-351.
350 351 352
Mariotti, F., Tomé, D., & Mirand, P. P. (2008) Converting nitrogen into protein-beyond 6.25 and Jones' factors. Critical Reviews in Food Science and Nutrition. 48:177-184. Mehmood, S., Orhan, I., Ahsan, Z., Aslan, S., & Gulfraz, M. (2008). Fatty acid composition of seed oil
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of different Sorghum bicolor varieties. Food Chemistry, 109, 855-859.
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Milczak, M., Pedzinski, M., Mnichowska, H., Szwed-Urbas, K., & Rybinski, W. (2001). Creative
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breeding of grasspea (Lathyrus sativus L.) in Poland. Lathyrus Lathyrism Newsletter, 2, 85-88.
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Moraghan, J. T., & Grafton, K. (2001). Genetic diversity and mineral composition of common bean
357 358 359 360 361 362 363
seed. Journal of the Science of Food and Agriculture, 81, 404-408. Morrison, W. R., & Smith, L. M. (1964). Preparation of fatty acid methyl esters and dimethylacetals from lipids with boron fluoride-methanol. Journal of lipid research, 5, 600-608. Motomizu, S., & Li, Z. H. (2005). Trace and ultratrace analysis methods for the determination of phosphorus by flow-injection techniques. Talanta, 66, 332-340. Nan, Z. B., Abd El-Moneim, A. M., Larbi, A., & Nie, B. (2006). Productivity of vetches (Vicia spp.) under alpine grassland conditions in China. Tropical Grasslands, 40, 177-182.
364
Pastor-Cavada, E., Juan, R., Pastor, J. E., Alaiz, M., & Vioque, J. (2009). Fatty acid distribution in the
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seed flour of wild Vicia species from Southern Spain. Journal of the American Oil Chemists'
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Society, 86, 977-983.
367
Pinheiro, C., Baeta, J. P., Pereira, A. M., Domingues, H., & Ricardo, C. P. (2010). Diversity of seed
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mineral composition of Phaseolus vulgaris L. germplasm. Journal of Food Composition and
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Analysis, 23, 319-325.
370
Ressler, C., Tatake, J. G., Kaizer, E., & Putnam, D. H. (1997). Neurotoxins in a vetch food: stability to
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cooking and removal of γ-glutamyl-β-cyanoalanine and β-cyanoalanine and acute toxicity
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from common vetch (Vicia sativa L.) legumes. Journal of Agricultural and Food Chemistry,
373
45, 189-194.
374 375 376 377
Samarah, N. H., & Ereifej, K. (2009). Chemical composition and mineral content of common vetch seeds during maturation. Journal of Plant Nutrition, 32, 177-186. Spackman, D. H., Stein, W. H., & Moore, S. (1958). Automatic recording apparatus for use in chromatography of amino acids. Analytical Chemistry, 30, 1190-1206.
378
US Department of Agriculture. (2004). USDA nutrient database for standard reference. Available from:
379
< http://www.ars.usda.gov/Services/docs.htm?docid=8964 > (Accessed 02.08.2004).
380
Uzun, A., Gücer, S., & Acikgoz, E. (2011). Common vetch (Vicia sativa L.) germplasm: Correlations of
381
crude protein and mineral content to seed traits. Plant Foods for Human Nutrition, 66,
382
254-260.
383 384
Wu, T. (2001). The Qinghai-Tibetan plateau: how high do Tibetans live? High Altitude Medicine & Biology, 2, 489-499.
385 386
Fig. 1. Seed yield (A), crude protein (B), lipid (C) and ash contents (D) of four
387
common vetch seeds. Vertical bars represent standard deviations. Different letters
388
indicate significant differences at P < 0.05.
389
Table 1
390
Fatty acid composition of four common vetch seeds (% of TFA; n = 3).
391
Table 2
392
Amino acid composition of four common vetch seeds (% of dry weight, n=3).
393
Table 3
394
Mineral element composition of four common vetch seeds (n=3).
395
Table 4
396
Contribution of 100 g of common vetch seeds to the Recommended Dietary
397
Allowance (RDA) for adults per day.
398
399 400
Fig. 1. Seed yield (A), crude protein (B), lipid (C) and ash contents (D) of four common vetch
401
seeds. Vertical bars represent standard deviations. Different letters indicate significant differences
402
at P < 0.05.
403 404
405
Table 1
406 407
Fatty acid composition of four common vetch seeds (% of TFA; n = 3).
2505
2556
2560
V. 333/A
0.30±0.05b
0.28±0.04b
0.58±0.09a
0.67±0.07a
0.33±0.04b
0.64±0.10a
0.52±0.06a
0.65±0.08a
17.42±0.55a
16.67±0.41a
15.79±0.67a
17.31±0.36a
3.03±0.41a
2.78±0.18a
2.92±0.19a
3.21±0.20a
1.51±0.17a
1.39±0.20a
1.17±0.09a
1.28±0.13a
9.85±0.69b
8.33±0.73b
11.11±1.30a
11.54±1.05a
2.27±0.34a
1.39±0.17c
1.75±0.22b
1.28±0.16c
53.03±5.66a
54.17±4.38a
54.39±3.97a
52.56±5.14a
12.12±1.05a
9.03±0.87c
10.53±1.23b
10.26±0.97b
∑SFA
23.16±2.88
21.82±2.70
21.14±3.14
23.22±2.66
∑UFA
76.74±6.71
74.51±8.36
77.36±6.78
76.64±9.17
3.31±0.5
3.41±0.3
3.66±0.2
3.30±0.4
Saturated fatty acid (SFA) Myristic acid (C14:0) Pentadecanoic acid (C15:0) Palmitic acid (C16:0) Stearic acid (C18:0) Arachidic acid C20:0 Unsaturated fatty acid (UFA) Oleic acid (C18:1 trans-9) Eicosenoic acid (C20:1 cis-9) Linoleic acid (C18:2 cis-9-12) α-Linolenic acid (C18:3 cis-9,12,15)
∑UFA/∑SFA
408 409
a,b,c
Values in a row without a common superscript are significantly different (P<0.05).
410
Table 2
411
Amino acid composition of four common vetch seeds (% of dry weight, n=3). 2505
2556
2560
V. 333/A
Arginine
2.50±0.42a
2.46±0.33a
2.53±0.50a
2.15±0.43b
Leucine
2.07±0.31b
2.23±0.52a
2.22±0.60a
1.98±0.44b
Lysine
1.69±0.63a
1.73±0.11a
1.66±0.37a
1.60±0.32a
Isoleucine
1.33±0.26a
1.27±0.25a
1.30±0.43a
1.17±0.37b
Phenylalanine
1.25±0.20a
1.20±0.45a
1.23±0.19a
1.18±0.28a
Threonine
1.11±0.30a
1.09±0.24b
0.97±0.17b
1.03±0.21b
Histidine
0.66±0.10b
0.73±0.09a
0.74±0.13a
0. 67±0.12b
Methionine
0.44±0.05c
0.51±0.08a
0.54±0.10a
0.48±0.06b
Tryptophane
0.30±0.04a
0.32±0.09a
0.33±0.08a
0. 28±0.05a
Essential amino acid (EAA)
Non-essential amino acid (NEA) Glutamic acid
5.51±1.91a
5.70±0.88a
5.63±1.12a
5.24±0.66b
Aspartic acid
3.72±0.52b
4.04±0.77a
3.48±0.63c
3.57±1.11c
Valine
1.66±0.25a
1.73±0.57a
1.70±0.19a
1.61±0.60a
Serine
1.42±0.33a
1.38±0.26a
1.39±0.41a
1.33±0.77a
Alanine
1.16±0.38b
1.39±0.23a
1.43±0.28a
1.35±0.51a
Glycine
1.22±0.60a
1.25±0.47a
1.19±0.72a
1.08±0.28b
Proline
1.13±0.46b
1.22±0.72a
1.27±0.35a
1.15±0.55b
Tyrosine
0.85±0.27a
0.93±0.41a
0.85±0.34a
0.87±0.37a
Cystine
0.45±0.07b
0.56±0.12a
0.52±0.11a
0.49±0.10a
∑EAA
11.83±1.82
11.77±2.95
11.60±1.73
10.64±2.48
∑NEA
17.21±2.70
18.89±2.26
17.56±2.91
16.79±1.83
0.69±0.0
0.62±0.1
0.66±0.0
0.63±0.0
∑EAA/∑NEA
412 413
a,b,c
Values in a row without a common superscript are significantly different (P<0.05).
414
Table 3
415
Mineral element composition of four common vetch seeds (n=3). 2505
2556
2560
V. 333/A
Macronutrient mineral (g/kg) K
9.87±0.72a
9.98±0.60a
9.79±1.17a
9.69±0.95a
P
3.96±0.51a
1.83±0.11b
1.78±0.19b
1.63±0.26b
Mg
2.20±0.10b
2.66±0.19a
2.27±0.26b
2.33±0.41b
Na
1.28±0.23a
1.26±0.10a
1.20±0.21a
1.22±0.09a
Ca
0.94±0.11b
1.17±0.16a
1.20±0.07a
1.19±0.14a
Micronutrient mineral (mg/kg)
416 417
a,b,c
Zn
30.01±8.21b
48.16±7.34a
44.66±7.57a
46.47±9.01a
Mn
12.74±3.18c
21.78±5.21a
15.13±3.40b
16.89±4.73b
Cu
3.37±0.41c
8.77±2.01a
5.33±1.38b
6.47±1.16b
Values in a row without a common superscript are significantly different (P<0.05).
418
Table 4
419
Contribution of 100 g of common vetch seeds to the Recommended Dietary Allowance (RDA) for
420
adults per day. Element
RDA*, mg
% of RDA supplied by 100g common vetch seeds 2505
2556
2560
V. 333/A
Macronutrient mineral
K
4700
21.01
21.23
20.83
20.62
P
700
56.57
26.14
25.43
23.29
Mg
420
52.38
63.33
54.05
55.48
Na
1500
8.53
8.40
8.01
8.13
Ca
1000
9.40
11.70
12.01
11.90
Zn
11
27.28
43.78
40.60
42.25
Mn
2.3
55.39
94.70
65.78
73.43
Cu
0.9
37.44
97.44
59.22
71.89
Micronutrient mineral
421 422
*
US Department of Agriculture (2004).
423
424 425
Highlights
►Different phytochemicals were evaluated in four cultivars of common vetch seeds.
426
► Fatty acid, amino acid and mineral contents were measured.
427
► High content of linoleic (52.56-54.39%) and α-linolenic acid (9.03-12.12%)
428
was found in seeds.
429
► the ratio of unsaturated fatty acid to saturated fatty acid was 3.30-3.66
430
►the ratio of essential amino acid to non-essential amino acid was bigger than
431 432
0.62.
S0308-8146(14)01315-6 http://dx.doi.org/10.1016/j.foodchem.2014.08.090 FOCH 16313
To appear in:
Food Chemistry
Received Date: Revised Date: Accepted Date:
21 April 2014 14 August 2014 21 August 2014
Please cite this article as: Mao, Z., Fu, H., Nan, Z., Wan, C., Fatty acid, amino acid, and mineral composition of four common vetch seeds on Qinghai-Tibetan plateau, Food Chemistry (2014), doi: http://dx.doi.org/10.1016/ j.foodchem.2014.08.090
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1
Fatty acid, amino acid, and mineral composition of four
2
common vetch seeds on Qinghai-Tibetan plateau
3
Zhuxin Mao a, b, Hua Fub,*, Zhibiao Nanb, Changgui Wanb
4
a
5
PR China.
6
b
7
Technology, Lanzhou University, Lanzhou 730000, PR China.
Xi’ an Botanical Garden of Shaanxi Province, Institute of Botany of Shaanxi Province, Xi’an 710061,
State Key Laboratory of Grassland Agro-ecosystems, College of Pastoral Agriculture Science and
8 9 10
11
*Corresponding author. Tel./ Fax: +86 931 8663778. E-mail address: [email protected] (H. Fu).
12
Fatty acid, amino acid, and mineral composition of four
13
common vetch seeds on Qinghai-Tibetan plateau
14
Zhuxin Mao a, b, Hua Fub,*, Zhibiao Nanb, Changgui Wanb
15
a
16
PR China.
17
b
18
Technology, Lanzhou University, Lanzhou 730000, PR China.
Xi’ an Botanical Garden of Shaanxi Province, Institute of Botany of Shaanxi Province, Xi’an 710061,
State Key Laboratory of Grassland Agro-ecosystems, College of Pastoral Agriculture Science and
19 20
Abstract
21
The chemical composition of four common vetch (Vicia sativa L.) seeds was
22
investigated to determine their nutrition value. The result shows that the seeds are low
23
in lipid (1.55-2.74% of dry weight), and high in the unsaturated fatty acid
24
(74.51-77.36% of total fatty acid). The ratio of essential amino acid to non-essential
25
amino acid (0.62-0.69) is even higher than the amount (0.38) recommended by World
26
Health Organization. Besides, the seeds are also found rich in Mg, Mn and Cu, but
27
with a low ratio of Ca to P (0.24-0.73), which may increase the risk of the mineral
28
element toxicity. The results indicate that the four common vetch seeds could be taken
29
as an alternative food source, but the possible toxic effect should be taken into
30
consideration.
31
Keywords: common vetch seed, fatty acid, amino acid, mineral element,
32
Qinghai-Tibetan plateau
*
Corresponding author. Tel./ Fax: +86 931 8663778. E-mail address: [email protected] (H. Fu).
33
1. Introduction
34
Common vetch (Vicia sativa L.) is a potential source of cheap protein for human
35
beings (Akpinar, Ali Akpinar, & Türkoğlu, 2001), although it has been reported to be
36
toxic (Luse, Kang, Fox, & Nangju, 1975; Ressler, Tatake, Kaizer, & Putnam, 1997).
37
Because of the good adaption to adverse environmental conditions, common vetch has
38
attracted much attention from breeders and producers, and thus has been cultivated as
39
a grain legume for seed production in Asia, Africa and Europe (Francis, Enneking, &
40
El Moneim, 2000). The common vetch seed as a rich source of protein could
41
contribute greatly to the protein supply for human beings and livestock. With a
42
growing recognition on the importance of legume plants to human health, much
43
attention has been paid to the chemical composition of the common vetch seed
44
(Acikgoz, Katkat, Omeroglu, & Okan, 1985; Pastor-Cavada, Juan, Pastor, Alaiz, &
45
Vioque, 2009; Samarah & Ereifej, 2009).
46
Fatty acid is important for human health since it’s the precursor for the biosynthesis
47
of eicosanoids which is considered as an important bio-regulator for many cellular
48
metabolic processes (Gressler et al., 2010; Khotimchenko & Yakovleva, 2005). In the
49
common vetch seed, fatty acids have a high degree of unsaturation (65-75%) (Akpinar
50
et al., 2001). It has been found that the major unsaturated fatty acids in common vetch
51
seeds are oleic, linoleic and α-linolenic acid accounting for over 50% of total fatty
52
acid (Akpinar et al., 2001). The high content of protein (24-32%) enriches the seeds
53
more nutrition (Uzun, Gücer, & Acikgoz, 2011). And protein is composed of different
54
amino acids and thus the degree of nutrition is largely determined by their relevant
55
proportion (Gressler et al., 2010). Previous studies have revealed that glutamic
56
(19.9%) and aspartic (14.8%) were the most predominant amino acids in common
57
vetch seeds (Acikgoz et al., 1985; Karadag & Yavuz, 2010).
58
Mineral deficiency is a widespread problem in many developing countries affecting
59
over 2 billion people, which could even cause infant and child mortalities worldwide
60
(Uzun et al., 2011). Through analyzing the minerals on 388 common vetch accessions,
61
Uzun et al. (2011) pointed out that the seeds were rich in calcium (Ca), magnesium
62
(Mg), manganese (Mn), Zinc (Zn) and copper (Cu). However, Acikgoz et al. (1985)
63
found that common vetch seeds were extremely low in Ca, Mg and Mn but rich in
64
phosphorus (P). Thus, the mineral composition seems significantly different among
65
the common vetch varieties.
66
The nutrition differed greatly among common vetch seeds from different regions,
67
which was attributed to the genetic factors and environmental conditions of the
68
production areas (Akpinar et al., 2001). To the best of our knowledge, little
69
information was available on vetches in China as one of the largest countries for the
70
production of common vetch in the word. Some research on common vetch was
71
initiated on Qinghai-Tibetan plateau of China in the late 20th century (Nan, Abd
72
El-Moneim, Larbi, & Nie, 2006). The plateau of a vast area up to 2.5 million km2 is
73
the best grazing land in Asia. But the short growing season and harsh environment
74
conditions impose constraints on the production of cropping and livestock (Wu, 2001).
75
In recent years, Chinese breeders have cultivated new common vetch cultivars with
76
excellent adaptability to the plateau (Nan et al., 2006). However, the previous
77
researches mainly concentrated on the agronomic characteristics and herbage nutrition
78
(Mao, Fu, Nan, Wang, & Wan, 2012), few were devoted to the study in terms of the
79
chemical composition in the common vetch seeds. Thus, this study is in the purpose to
80
analyze and compare the content of fatty acid, amino acid and mineral element in the
81
common vetch seeds obtained from the plateau to determine if they possess favorable
82
traits from a biochemical point of view.
83
2. Materials and methods
84
2.1 Seed sample
85
The field experiment was carried out at Lanzhou university research farm, Gansu,
86
China (35º06′ N, 102º25′ E, 3050 m above sea level), located on the eastern edge of
87
the Qinghai-Tibetan plateau. The average annual precipitation is 424 mm, mean daily
88
temperature is 2.8 degrees Celsius, and over 60% of the area is grassland. The soil is
89
chernozem, slightly acidic (pH is 6.7), low in P (7.5 mg/kg) and high in K (17.5g/kg)
90
(Mao et al, 2012).
91
Four common vetch cultivars were used in this study: 2505, 2556, 2560 and
92
V.333/A. The cultivars were selected from Common Vetch Breeding Program of
93
Lanzhou University mainly from the Qinghai-Tibetan plateau. The seeds were grown
94
during the 2011 growing season in a randomized complete block design with four
95
replications, and each replication was a 3.6 m2 plot. Harvesting and threshing were
96
performed by hand. After cleaning, the matured seeds (500g) were dried in
97
a forced-air oven at 50 degrees Celsius to constant weight, and ground through a 0.5
98
mm screen by a cyclone mill. These samples were stored at -20 degrees Celsius in
99
refrigerator before the chemical analysis.
100
2.2 Chemical analysis
101
2.2.1 Fatty acid analysis
102
The lipid was measured by the continuous lipid extraction by a soxhlet apparatus.
103
The lipid extracted from the seeds flours were mixed with boron trifluoride-methanol
104
reagent (25%) and fatty acids were then converted into the methyl ester derivatives
105
(Morrison & Smith, 1964). The fatty acid methyl esters analysis was performed on
106
Agilent 6890N gas chromatography instrument coupled with an Agilent MS-5973
107
inert XL mass selective detector and an Agilent autosampler 7683-B injector (Little
108
Fall, NY, USA). The capillary column was an HP-FFAP MS with dimension of 30 m
109
length ×0.25mm i.d. ×0.25µm film thickness (Palo Alto, CA, USA). Helium was used
110
as carrier gas at a flow rate of 1.1mL/min. Temperature program began by holding
111
temperature at 60℃ for 2 min, then increased to 200℃ at 15℃ /min, and held at
112
200℃ for 1 min before rising to 240℃ at 8℃/min with a final holding temperature
113
at 240℃ for 10min. The injection was performed in a split mode (split ratio 100:1).
114
Detector and injector temperature was 250℃ and 230℃, respectively. MS scan range
115
was (m/z): 40-550 atomic mass units (AMU) under electron impact (EI) ionization
116
(70 eV) and ion source temperature was 230℃ (Mehmood et al., 2008).
117
Fatty acid methyl ester peaks were identified by comparing their retention
118
times and spectra with standard compounds which were purchased from Fluka
119
(Sigma-Aldrich, Inc., St. Louis, MO, U.S.A). Standard methyl esters of palmitic,
120
stearic, oleic and linoleic acids were used for the confirmation of GC-MS library
121
results. The percentage of fatty acid composition was decided by the ratio of
122
individual peak area to all fatty acid areas (Table 1). Each analysis was performed at
123
least in triplicate.
124
2.2.2 Amino acid analysis
125
Protein and peptides acid hydrolysis was done according to the method of
126
Spackman et al. (1958). A 10-20 mg powdered sample without lipid was transferred
127
to 10×150 mm borosilicate ampoules previously pyrolysed at 420 degrees Celsius for
128
8 h. For each ampoule, 0.5 mL of HCl 6 M with 0.1% of phenol (m/v) was added.
129
Vacuum was applied before the seal of the ampoules and then they were put into an
130
oven at 110 degrees Celsius for 22 h. The hydrolysed samples were analyzed
131
according to the standard of the European Community (98/64/EG). Amino acid
132
samples were separated by ion exchange chromatography and determined by reaction
133
with ninhydrin with photometric detection at 570 nm (440 nm for proline) using the
134
automatic amino acid analyzer LC-3000 (Eppendorf-Bio-tronik; Germany). Amino
135
acid standard solution (A9781 from Sigma-Aldrich, Germany) (0.06 mg/mL) was
136
injected to calibrate the analyzer and to calculate the amount of amino acid in the
137
samples. In addition, nitrogen (N) was determined by the micro-Kjeldahl technique
138
with the duplicate samples of dried seeds for each genotype. Crude protein content
139
was then calculated by 6.25×N (Mariotti, Tomé, & Mirand, 2008).
140
2.2.3 Mineral analysis
141
Potassium (K), P, Mg, sodium (Na), Ca, Zn, Mn and Cu were analyzed for seed
142
minerals. The seeds were washed with deionised water, dried at 75 degrees Celsius,
143
weighed and ashed at 480 degrees Celsius in a muffle furnace. The ashes were
144
dissolved in 5 mL of 20% (v/v) HCl and diluted to a volume of 100 mL with
145
deionised water. The solution was analyzed for K, Mg, Na, Ca, Zn, Mn, and Cu by a
146
Perkin-Elmer 5000 flame (air-acetylene) atomic absorption spectrometer with
147
hollow-cathode lamp tubes (Norwalk, Connecticut, USA) according to Pinheiro et al.
148
(2010). P was measured in 5 mL of the same solution by the colorimetric
149
molybdenum ammonium vanadate method (Motomizu & Li, 2005), using a Hitachi
150
Perkin-Elmer Model-139 UV-vis spectrophotometer (Tokyo, Japan), at the
151
wavelength of 470 nm.
152
2.3 Statistics analysis
153
The results were expressed as means ± standard deviation (SD). All statistical
154
analysis was performed using the SPSS software package, version 13.0 (SPSS Inc.,
155
Chicago). The data was analyzed by one way analysis of variance (ANOVA).
156
Differences were considered significant at P < 0.05.
157
3. Results and Discussion
158
3.1 Seed yield, content of lipid, protein and ash
159
In this study, the average yield of common vetch seeds was 1392 kg/ha for 2505,
160
853 kg/ha for 2556, 1172 kg/ha for 2560, and 890 kg/ha for V. 333/A (Fig. 1A). The
161
seed yield of 2505 and 2560 was significantly higher (P<0.05) than that of 2556 and
162
V. 333/A. The seed yield of 2505 and 2560 was higher than that (820-1000 kg/ha) in
163
China (Nan et al., 2006). The seed yield of 2556 and V. 333/A was lower than that
164
(1160-1459 kg/ha) in Turkey (Karadag et al., 2010).
165
The seed crude protein (CP) content varied from 29.23% to 30.57% (Fig. 1B), and
166
no significant difference has been found among the cultivars (P<0.05). The CP
167
content was in accordance with that in the previous studies ranged from 25.30% to
168
31.38% (Karadag et al., 2010; Uzun et al., 2011). In addition, the CP content of four
169
cultivars was close to that of common vetch lines 845 (28.90%), 1448 (30.77%) and
170
384 (31.38%) in Turkey (Karadag et al., 2010), but was lower than that of common
171
vetch cultivars (37.03%) in Madrid, Spain (Caballero et al., 2001).
172
The lipid content of seeds was 1.55% for 2505, 1.74% for 2556, 2.74% for 2560,
173
and 2.28% for V. 333/A; the content in 2560 and V. 333/A was significantly higher
174
(P<0.05) than that in 2505 and 2556 (Fig. 1C). The lipid content in the current study
175
was higher than other common vetch species (Caballero et al., 2001; Milczak et al.,
176
2001), but lower than the varieties reported by Darwish Sayed et al. (1980).
177
The ash content of seeds was 2.64 g/kg for 2505, 4.10 g/kg for 2556, 3.20 g/kg for
178
2560, and 3.32 g/kg for V. 333/A (Fig. 1D), the content ranked as
179
2556>2560=V.333/A>2505 (P<0.05). The result was in accordance with the ash
180
content studied by Samarah and Ereifej (2009).
181
3.2 Fatty acid
182
The fatty acid composition and content are presented in Table 1. Nine fatty acids
183
detected in four cultivars are identified as saturated fatty acids (SFA) and unsaturated
184
fatty acids (UFA). The proportion of SFA accounted for 21.14% to 23.22% of total
185
fatty acids (TFA) in the seeds (Table 1), and no significant difference has been found
186
among the cultivars. The most abundant SFA was palmitic acid (C 16:0) accounting
187
for 15.79% to 17.42%. Stearic (C 18:0) and arachidic acids (C 20:0) presented in a
188
small amount ranging from 2.78% to 3.21% and 1.17% to 1.51%, respectively.
189
Myristic (C 14:0) and pentadecanoic (C 15:0) acids were found in a trace amount,
190
ranging from 0.28% to 0.67% and 0.33% to 0.65%, respectively. The result that
191
palmitic acid was the only major SFA in four common vetch cultivars was in
192
agreement with what was found by Pastor-Cavada et al. (2009). But our result was of
193
some difference from what was found by Akpınar et al. (2001) who reported that
194
palmitic (8.05%) and stearic acids (7.31%) were both predominant in common vetch
195
seed oil in Turkey. In this study, arachhidic acid was present in a small amount but it
196
wasn’t detected by Darwish Sayed et al. (1980) in common vetch. Finally, a big
197
difference was found in the content of myristic and pentadecanoic acids up to 3.79%
198
and 2.91% in common vetch seed in Turkey (Akpinar et al., 2001), which is much
199
higher than that in our study. This may be related with the differences in the
200
ecological and geographical conditions as suggested by Aaes-Jorgensen (1961).
201
There are four UFA present in this study (Table 1). The amount of UFA in the
202
seeds ranged from 74.51% to 77.36% with no significant differences among the
203
cultivars. With regard to the individual UFA, the most abundant UFA was linoleic
204
(C18:2 cis-9, 12), ranging from 52.56% to 54.39%. Oleic acid (C18:1 trans-9) and
205
α-linolenic acid (C18:3 cis-9, 12, 15) were present in a considerable amount up to
206
8.33-11.54% and 9.03-12.12%, respectively. Eicosenoic acid (C20:1 cis-9) has been
207
detected in a small amount ranging from 1.28% to 2.27%. In this study, the content of
208
oleic acid was 23.70% only half of that in the seeds collected in Turkey (Akpinar et
209
al., 2001). However, linoleic acid was more than half of TFA in this study, much
210
higher than the content (4.33%) revealed by Akpinar et al. (2001). Most notably, the
211
content of linolenic and eicosenoic acids was both higher than that (1.95% and 0.34%,
212
respectively) in the seeds from Turkey (Akpinar et al., 2001).
213
In addition, the ratio of UFA to SFA at 3.30-3.66 in common vetch seeds in the
214
current study was much higher than that from Turkey (Akpinar et al., 2001). The
215
result could indicate that the seeds harvested from the high plateau have higher
216
proportion of UFA, of which, α-linolenic acid of considerable amount was
217
recognized quite beneficial to heart health (Connor, 1999).
218
3.3 Amino acid
219
Eighteen amino acids were detected in the common vetch seeds (Table 2),
220
10.64-11.83% of them were essential amino acids (EAA) and rest were non-essential
221
amino acids (NEA). There was a small difference in EAA among the common vetch
222
cultivars. Arginine and leucine turned out to be the predominant EAA accounting for
223
2.15-2.53% and 1.98-2.23%, respectively. Lysine, isoleucine, phenylalanine and
224
threonine were in a small amount at 1.60-1.73%, 1.17-1.33%, 1.18-1.25% and
225
0.97-1.11%, respectively. Histidine, methionine and tryptophane were in a trace
226
amount at 0.66-0.74%, 0.44-0.54% and 0.28-0.33%, respectively. Glutamic and
227
aspartic turned out to be the predominant NEA of 5.24-5.70% and 3.48-4.04%,
228
respectively. Valine, serine, alanine, glycine and proline were in a small amount at
229
1.61-1.73%, 1.33-1.42%, 1.16-1.43%, 1.08-1.25% and 1.13-1.27%, respectively.
230
Tyrosine and cystine were in a trace amount at 0.85-0.93% and 0.45-0.56%,
231
respectively. The type and content of amino acids in this study were somewhat
232
different from the previous reports (Hadjipanayiotou & Economides, 2001).
233
According to Hadjipanayiotou et al. (2001), tryptophane and cystine were not found
234
in common vetch seeds in Cyprus. Glutamic acid was the highest NEA (>5.24%),
235
whereas methionine (0.44%) and alanine (0.44%) were the lowest NEA in this study,
236
which may suggest that there was less NEA in common vetch seeds grown on the
237
Qinghai-Tibetan plateau. In addition, the amino acids content in the present study was
238
close to that in Turkey (Karadag et al., 2010), the highest and lowest EAA was
239
1.95-2.26% (leucine) and 0-0.02% (methionine), respectively. While the highest and
240
lowest NEA was 3.65-7.74% (aspartic) and 0.73-0.90% (tyrosine), respectively. In
241
contrast to our result, arginine, tryptophane and cystine were not found in the
242
common vetch seeds in Turkey (Karadag et al., 2010).
243
In this study, the amount of amino acids ranged from 28.42% to 30.11% (Table 2).
244
The ratio of EAA to the total amino acid for common vetch species was about 0.4.
245
The ratio of EAA to NEA for all cultivars was 0.62-0.69, which was substantially
246
higher than the adult requirement for EAA/NEA (0.38) recommended by WHO (Joint
247
WHO., 2007).
248
3.4 Mineral element
249
Mineral concentrations of common vetch seeds are reported in Table 3. Among the
250
macronutrient minerals, P, Mg and Ca were 1.63-3.96 g/kg, 2.20-2.66 g/kg and
251
0.94-1.20 g/kg, respectively. The highest content of P (3.96 g/kg) and Mg (2.66 g/kg)
252
was found in 2505 and 2556, respectively and they showed no significant difference
253
among other cultivars. The lowest Ca (0.94 g/kg) was found in 2505, and no
254
significant difference was found with other cultivars. Uzun et al. (2011) found that P
255
content of common vetch seeds in Turkey was 4.96 g/kg, and the corresponding Mg
256
and Ca were 1.53 g/kg and 1.08 g/kg, respectively. Similarly, Moraghan and Grafton
257
(2001) reported that the mean content of P in the common bean seeds in America was
258
4.45-5.26 g/kg, and the corresponding content of Mg was 1.02-1.52 g/kg. Our results
259
show that the seeds harvested from plateau are deficient in P but rich in Mg, which
260
may reflect the difference in the chemical property of the local soils. K and Na were
261
9.69-9.98 g/kg and 1.20-1.28 g/kg, respectively, and no significant difference (P<0.05)
262
was found with other cultivars. The content of K in this study is similar to what has
263
been found by Samarah et al. (2009) although the content of Na was higher in their
264
study.
265
Among the micronutrient minerals, Zn, Mn and Cu were 30.01-48.16 mg/kg,
266
12.74-21.78 mg/kg and 3.37-8.77 mg/kg, respectively. The highest level of
267
micronutrient minerals was all found in 2556, while the lowest level was found in
268
2505 (P<0.05). And there was no significant difference between 2560 and V. 333/A.
269
The level of Zn and Mn in this study was consistent with what has been found by
270
Uzun et al. (2011) that Zn and Mn in common vetch seeds were 31.50 mg/kg and
271
14.50 mg/kg, respectively. The similar result has also been reported by Samarah et al.
272
(2009). However, Cu in our study was lower than the level reported by Uzun et al.
273
(2011) and Samarah et al. (2009) at 11.70 mg/kg and 13.02 mg/kg, respectively.
274
Mineral element in food is important for human health, but excessive intake of the
275
mineral element will result in toxicity (Luse et al., 1975). The potential contribution
276
of 100g of common vetch seeds to the Recommended Dietary Allowances (RDA) (US
277
Department of Agriculture, 2004) on mineral elements is present in Table 4. The
278
content of Mg, Mn and Cu in 100g common vetch seeds could contribute RDA as
279
high as 37.44-97.44% to the adult. K, P and Zn contribute at a medium level
280
(20.62-56.57%), Na and Ca contribute at low level (8.01-12.01%). The result suggests
281
that common vetch seeds could be the dietary resource for the mineral elements, but
282
excessive intake of the seeds may result in toxicity to human beings. In addition, the
283
availability of Ca in the body greatly depends on the ratio of Ca to P, which is
284
recommended optimal between 1.0 and 1.3 (Calvo & Park., 1996). However, the ratio
285
of Ca to P in four common vetch seeds was 0.24-0.73, which may disturb the
286
metabolism of Ca and negatively affect bone health (Kemi et al., 2010).
287
4. Conclusions
288
This study analyzed the composition of fatty acid, amino acid and mineral element
289
in the four common vetch seeds on Qinghai-Tibetan plateau of China. The result
290
shows that the seeds are low in lipid with a ratio of UFA/SFA up to 3.30-3.66, which
291
means that the seeds have accumulated more UFA primarily as linoleic, oleic and
292
α-linolenic acids. In addition, the ratio of EAA to NAA (0.62-0.69) is even higher
293
than the amount (0.38) recommended by WHO (Joint WHO., 2007). Mg, Mn and Cu
294
in 100g common vetch seeds may supply 37.44-97.44% of the RDA for adult, but
295
excessive intake of the seeds may result the mineral element toxicity. In addition, the
296
ratio of Ca to P in the seeds was 0.24-0.73 lower than the level (1.0-1.3)
297
recommended by Calvo and Park (1996). It is concluded that four common vetch
298
seeds can be a potential alternative food or feed on Qinghai-Tibetan plateau, but the
299
possible toxic effect should also be kept in mind.
300 301
Acknowledgments
302
This study was supported by National Basic Research Program of China (No.
303
2014CB138703), Special Fund for Agroscientific Research in the Public Interest (No.
304
201203041), and Shaanxi Province Science Foundation for Youths (No. 2013K-19).
305 306 307 308
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Caballero, R., Alzueta, C., Ortiz, L. T., Rodríguez, M. L., Barro, C., & Rebolé, A. (2001). Carbohydrate
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Connor, W. E. (1999). α-Linolenic acid in health and disease. The American Journal of Clinical Nutrition, 69, 827-828.
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acid, protein, amino acid and ash contents in four Brazilian red algae species. Food Chemistry,
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Hadjipanayiotou, M., & Economides, S. (2001). Chemical composition, in situ degradability and amino
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(2010). Low calcium: phosphorus ratio in habitual diets affects serum parathyroid hormone
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concentration and calcium metabolism in healthy women with adequate calcium intake.
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British journal of nutrition, 103, 561-568.
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Khotimchenko, S. V., & Yakovleva, I. M. (2005). Lipid composition of the red alga Tichocarpus crinitus exposed to different levels of photon irradiance. Phytochemistry, 66, 73-79.
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Moraghan, J. T., & Grafton, K. (2001). Genetic diversity and mineral composition of common bean
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mineral composition of Phaseolus vulgaris L. germplasm. Journal of Food Composition and
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Samarah, N. H., & Ereifej, K. (2009). Chemical composition and mineral content of common vetch seeds during maturation. Journal of Plant Nutrition, 32, 177-186. Spackman, D. H., Stein, W. H., & Moore, S. (1958). Automatic recording apparatus for use in chromatography of amino acids. Analytical Chemistry, 30, 1190-1206.
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Uzun, A., Gücer, S., & Acikgoz, E. (2011). Common vetch (Vicia sativa L.) germplasm: Correlations of
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Wu, T. (2001). The Qinghai-Tibetan plateau: how high do Tibetans live? High Altitude Medicine & Biology, 2, 489-499.
385 386
Fig. 1. Seed yield (A), crude protein (B), lipid (C) and ash contents (D) of four
387
common vetch seeds. Vertical bars represent standard deviations. Different letters
388
indicate significant differences at P < 0.05.
389
Table 1
390
Fatty acid composition of four common vetch seeds (% of TFA; n = 3).
391
Table 2
392
Amino acid composition of four common vetch seeds (% of dry weight, n=3).
393
Table 3
394
Mineral element composition of four common vetch seeds (n=3).
395
Table 4
396
Contribution of 100 g of common vetch seeds to the Recommended Dietary
397
Allowance (RDA) for adults per day.
398
399 400
Fig. 1. Seed yield (A), crude protein (B), lipid (C) and ash contents (D) of four common vetch
401
seeds. Vertical bars represent standard deviations. Different letters indicate significant differences
402
at P < 0.05.
403 404
405
Table 1
406 407
Fatty acid composition of four common vetch seeds (% of TFA; n = 3).
2505
2556
2560
V. 333/A
0.30±0.05b
0.28±0.04b
0.58±0.09a
0.67±0.07a
0.33±0.04b
0.64±0.10a
0.52±0.06a
0.65±0.08a
17.42±0.55a
16.67±0.41a
15.79±0.67a
17.31±0.36a
3.03±0.41a
2.78±0.18a
2.92±0.19a
3.21±0.20a
1.51±0.17a
1.39±0.20a
1.17±0.09a
1.28±0.13a
9.85±0.69b
8.33±0.73b
11.11±1.30a
11.54±1.05a
2.27±0.34a
1.39±0.17c
1.75±0.22b
1.28±0.16c
53.03±5.66a
54.17±4.38a
54.39±3.97a
52.56±5.14a
12.12±1.05a
9.03±0.87c
10.53±1.23b
10.26±0.97b
∑SFA
23.16±2.88
21.82±2.70
21.14±3.14
23.22±2.66
∑UFA
76.74±6.71
74.51±8.36
77.36±6.78
76.64±9.17
3.31±0.5
3.41±0.3
3.66±0.2
3.30±0.4
Saturated fatty acid (SFA) Myristic acid (C14:0) Pentadecanoic acid (C15:0) Palmitic acid (C16:0) Stearic acid (C18:0) Arachidic acid C20:0 Unsaturated fatty acid (UFA) Oleic acid (C18:1 trans-9) Eicosenoic acid (C20:1 cis-9) Linoleic acid (C18:2 cis-9-12) α-Linolenic acid (C18:3 cis-9,12,15)
∑UFA/∑SFA
408 409
a,b,c
Values in a row without a common superscript are significantly different (P<0.05).
410
Table 2
411
Amino acid composition of four common vetch seeds (% of dry weight, n=3). 2505
2556
2560
V. 333/A
Arginine
2.50±0.42a
2.46±0.33a
2.53±0.50a
2.15±0.43b
Leucine
2.07±0.31b
2.23±0.52a
2.22±0.60a
1.98±0.44b
Lysine
1.69±0.63a
1.73±0.11a
1.66±0.37a
1.60±0.32a
Isoleucine
1.33±0.26a
1.27±0.25a
1.30±0.43a
1.17±0.37b
Phenylalanine
1.25±0.20a
1.20±0.45a
1.23±0.19a
1.18±0.28a
Threonine
1.11±0.30a
1.09±0.24b
0.97±0.17b
1.03±0.21b
Histidine
0.66±0.10b
0.73±0.09a
0.74±0.13a
0. 67±0.12b
Methionine
0.44±0.05c
0.51±0.08a
0.54±0.10a
0.48±0.06b
Tryptophane
0.30±0.04a
0.32±0.09a
0.33±0.08a
0. 28±0.05a
Essential amino acid (EAA)
Non-essential amino acid (NEA) Glutamic acid
5.51±1.91a
5.70±0.88a
5.63±1.12a
5.24±0.66b
Aspartic acid
3.72±0.52b
4.04±0.77a
3.48±0.63c
3.57±1.11c
Valine
1.66±0.25a
1.73±0.57a
1.70±0.19a
1.61±0.60a
Serine
1.42±0.33a
1.38±0.26a
1.39±0.41a
1.33±0.77a
Alanine
1.16±0.38b
1.39±0.23a
1.43±0.28a
1.35±0.51a
Glycine
1.22±0.60a
1.25±0.47a
1.19±0.72a
1.08±0.28b
Proline
1.13±0.46b
1.22±0.72a
1.27±0.35a
1.15±0.55b
Tyrosine
0.85±0.27a
0.93±0.41a
0.85±0.34a
0.87±0.37a
Cystine
0.45±0.07b
0.56±0.12a
0.52±0.11a
0.49±0.10a
∑EAA
11.83±1.82
11.77±2.95
11.60±1.73
10.64±2.48
∑NEA
17.21±2.70
18.89±2.26
17.56±2.91
16.79±1.83
0.69±0.0
0.62±0.1
0.66±0.0
0.63±0.0
∑EAA/∑NEA
412 413
a,b,c
Values in a row without a common superscript are significantly different (P<0.05).
414
Table 3
415
Mineral element composition of four common vetch seeds (n=3). 2505
2556
2560
V. 333/A
Macronutrient mineral (g/kg) K
9.87±0.72a
9.98±0.60a
9.79±1.17a
9.69±0.95a
P
3.96±0.51a
1.83±0.11b
1.78±0.19b
1.63±0.26b
Mg
2.20±0.10b
2.66±0.19a
2.27±0.26b
2.33±0.41b
Na
1.28±0.23a
1.26±0.10a
1.20±0.21a
1.22±0.09a
Ca
0.94±0.11b
1.17±0.16a
1.20±0.07a
1.19±0.14a
Micronutrient mineral (mg/kg)
416 417
a,b,c
Zn
30.01±8.21b
48.16±7.34a
44.66±7.57a
46.47±9.01a
Mn
12.74±3.18c
21.78±5.21a
15.13±3.40b
16.89±4.73b
Cu
3.37±0.41c
8.77±2.01a
5.33±1.38b
6.47±1.16b
Values in a row without a common superscript are significantly different (P<0.05).
418
Table 4
419
Contribution of 100 g of common vetch seeds to the Recommended Dietary Allowance (RDA) for
420
adults per day. Element
RDA*, mg
% of RDA supplied by 100g common vetch seeds 2505
2556
2560
V. 333/A
Macronutrient mineral
K
4700
21.01
21.23
20.83
20.62
P
700
56.57
26.14
25.43
23.29
Mg
420
52.38
63.33
54.05
55.48
Na
1500
8.53
8.40
8.01
8.13
Ca
1000
9.40
11.70
12.01
11.90
Zn
11
27.28
43.78
40.60
42.25
Mn
2.3
55.39
94.70
65.78
73.43
Cu
0.9
37.44
97.44
59.22
71.89
Micronutrient mineral
421 422
*
US Department of Agriculture (2004).
423
424 425
Highlights
►Different phytochemicals were evaluated in four cultivars of common vetch seeds.
426
► Fatty acid, amino acid and mineral contents were measured.
427
► High content of linoleic (52.56-54.39%) and α-linolenic acid (9.03-12.12%)
428
was found in seeds.
429
► the ratio of unsaturated fatty acid to saturated fatty acid was 3.30-3.66
430
►the ratio of essential amino acid to non-essential amino acid was bigger than
431 432
0.62.