l o g o a comparative assessment of total phenolic content, ferric reducing-anti-oxidative power,...
TRANSCRIPT
L o g o
A comparative assessment of total phenolic content, ferric reducing-anti-oxidative power, free radical scavenging activity, vitamin C and isoflavones content in soybean with varying seed coat colour
汇报人：胡冬梅指导老师：薛照辉副教授
L o g o
2011-10-23学术报告
Introduction
Materials and methods
Results and discussion
Conclusions
1
2
3
4
Contents
L o g o
2011-10-23学术报告
1. Introduction
vitamin C
isoflavones
tocopherols
lecithin
saponins
soy peptides
soybean
L o g o
2011-10-23学术报告
1. Introduction
green
yellow
black
Furuta et al. (2003) soybean-boiled extracts
and boiled soybean
homogenate of all three types
of soybean
Xu et al. (2007) 2 yellow and 1
black
Xu and Chang (2008a, 2008b) 28 yellow and
2black
FRAPFRSATPC
L o g o
2011-10-23学术报告
Diagram
vitamin C is astrong scavenger of free radicals
the number of black soybeangenotypes analysed were too few
soybean with green seed coat were not included
.
6 genotypes from each type of soybeanFRAPFRSATPCforms of isoflavonesvitamin C
L o g o
2011-10-23
2. Materials and methods
学术报告
the seeds were dried inoven at 70 ℃ till the weight
became constant.Moisture free seeds
were secured in screw-capped vials
L o g o
2011-10-23学术报告
2.1. Extraction of antioxidants from the seeds
ground into flourOven dried seeds pass through 100-mesh sieve
extracted with 15 ml aqueous acetone at 25 ℃
in the dark overnight
centrifuged at3000r for 10 min
residues were re-extracted with 5 ml 70% acetone
Both the extracts were combined and stored
at 4 ℃ in dark for analyses
L o g o
2011-10-23学术报告
2.2 Determination of TPC
0.05 ml 70% acetone soy extract
0.5 ml F–C reagent and 0.5 ml 20% Na2CO3
final volume was 5 ml with distilled water
incubation for 30 min at room temperature
700nm distilled water gallic acid equivalents
L o g o
2011-10-23学术报告
2.3 Ferric reducing-antioxidant power assay
absorbance was recorded at 593 nm.0.1 ml of soybean flour
extract + 3ml FRAP reagent
incubated for 15 min at 37 ℃
300 mM acetate buffer 10 mM TPTZ in 40 mM HCl
and 20 mM FeCl3· 6H2O in the ratio 10:1:1
L o g o
2011-10-23
2.4. DPPH free radical-scavenging activity0.1 ml of the extraction solvent
adding DPPH solution
incubation10 min at room temperature
Decrease in absorbance at 517 nm
(ethanolic as blank)学术报告
L o g o
2011-10-23学术报告
where Acontrol is absorbance of the control Asample is the absor-bance of the sample after.
L o g o
2011-10-23
2.5 HPLC determination of isoflavones
2.5.1. Sample preparation
学术报告
drie in an oven at 70
℃
ground andpass
through 100-mesh
sieve
Extract with 80% ethanol (5
ml) and concentrated
HCl(1 ml) for 2h in a boiling water
bath
centrifuge at 10,000
rpmfor 10 min
suspension was
colleted
Further test
L o g o
2011-10-23
2.5.2. HPLC conditions20ulC-18 silica columncolumn oven was maintained at 40℃flow rate:0.8 ml/min for 25 minsolvent A (10% ACN) ( 乙腈 )solvent B (38% ACN)The solvent system(% solvent
A/solvent B): 0 min (0/100), 5 min(10/90), 20 min (0/100), and 25
min (0/100).
260 nm学术报告
L o g o
2011-10-23
2.5.2.1. Vitamin C
Vitamin C was quantified by using standard 2,6dichlorophenol-indophenol dye method
学术报告
还原型抗坏血酸能还原染料 2.6- 二氯酚靛酚，该染料在酸性中呈红色，被还原后红色消失。还原型抗坏血酸还原 2.6- 二氯酚靛酚后，本身被氧化成脱氢抗坏血酸。在没有杂质干扰时，一定量的样品提取
液还原标准 2.6- 二氯酚靛酚的量与样品中所含维生素 C 的量成正比。
L o g o
2011-10-23
3. Results and discussion
学术报告
1.06-1.54
0.96-2.89
0.81-5.89
1.10-3.11
4.02-6.44
2.87-6.15
2.55
5.14
4.56
1.38
1.61
2.82
16.42-44.05
21.08-32.55
59.71-83.47
31.19
26.37
75.10
L o g o
2011-10-23学术报告
1:1.1:1.5
1.5:1:2.2
1:1.4 ：1.9
282.5
358.4
250.5
478.8
220.9
321.9
309.6
453.4
475.4
L o g o
2011-10-23学术报告
类胡萝卜素， 叶黄素
L o g o
2011-10-23学术报告
L o g o
2011-10-23学术报告
4. Conclusions
black seed exhibited higher FRSA than yellow and green soybean.
yellow soybean exhibited lower FRAP value than black and green soybean.
antioxidant constituents other than isoflavones contribute to the higher FRSA in black soybean.
L o g o
2011-10-23学术报告
4. Conclusions
•every black genotype does not necessarily contain higher level of TPC than yellow or green soybean.
•some black genotypes despite having low TPC than green soybean showed higher levels of FRSA.
?
L o g o