![Page 1: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/1.jpg)
Antioxidant activity and major isoflavonoid contents of White Kwao Krua
by elicitors-induced on greenhouse condition.
![Page 2: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/2.jpg)
The experimental purpose
Study on the effect of using the elicitors on induce the antioxidant activity and the major isoflavonoids content in White Kwoa Krua in the greenhouse condition.
![Page 3: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/3.jpg)
• F ound in northern and north eastern.• It is considered to be a rejuvenating d
rug and has a history in folk medicine.• N ame “White Kwao Krua” was definiti
vely identified as Pueraria candollei in 1952 from which an unusual estrogeni
c. White Kwao Krua (WKK) has a
high concentration of
phytoestrogen .
White Kwao Krua [Pueraria candollei Grah. var. mirifica (Airy Shaw et. Suvatabandhu) Niyomdham].
![Page 4: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/4.jpg)
Applications
• cosmetic industry • breast cream• eye gel• skin moisturizer.
• Pharmaceuticals industry • Replacement hormones• Loss bone metabolism• Livestock
• Animals feed
![Page 5: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/5.jpg)
Antioxidant activity • Antioxidant compounds play an imp
- ortant role as a health protecting factor.• A ntioxidants reduce the risk for chronic dise
ases including cancer and heart disease.• The main characteristic of an antioxidant is i
ts ability to trap free radicals.
Cell metabolism & pollutions superoxide ani
on radical (O2
) hydroxyl radic
al (OH) peroxyl radical
(ROO)
Gutteridge (1994)
![Page 6: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/6.jpg)
Major Isoflavonoids
• Genistein– Phytonutrient– Manopause (hormonal replacement)– Loss bone (bone metabolism)
• Puerarin– Hypertension– antioxidant
Igham et al. (1986)
![Page 7: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/7.jpg)
Elicitors Molecules that stimulate secondary
metabolism Elicitors may form inside or outside p
lant cells, and are distinguished as e ndogenously or e
xogenously inducers. D epending on their origin, they are c
lassified as biotic or abiotic Heike and Dietrich (1995)
![Page 8: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/8.jpg)
Elicitors
Chitosan
Soybean; 1000
ppm increased seed isoflavone - 1696
%and Daidzein 15 0%
Tawaha et al. (2005)
Salicylic acid
Lupinus ruteus; 80 0 µM increased
Genistein 1212 µ g/g DW.
Kneer et al.
( 1999)
Copper (II)
chloride
- Pueraria montana; 100 ppm increased Genistein 5 folds- Pueraria lobata; 10 mM for 2 hour increased Genistein 10 folds
Takashi et al. (1990)Hakanatsuka (1991)
![Page 9: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/9.jpg)
Material & Methods
• Plant Planting WKK in greenhouse
at SUT farm Selected 48 plant WKK were
age of 1 year old • Experimental plan
CRD 6 replication with 8 treatments
• Elicitor combination
(treatments)
Watersalicylic acid (SA)
CuCl2 CuCl2 + SA
ChitosanChitosan +
SAChitosan +
CuCl2 Chitosan + CuCl2 +SA
![Page 10: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/10.jpg)
Antioxidant determination by DPPH
Inhibitory concentration at 50% (IC50) Plot between % radical scavenging VS concentration of each treatmentEquation of nonlinear regression
![Page 11: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/11.jpg)
• Major isoflavonoids determination by HPLC
HPLC condition• HPLC column RP-C18 Agilent® (4.6 mm x
15 cm)• Detector Diode array 254 nm • Flow rate 1ml / min• Volume injection 10 µl• Presser pump 111 bar • Mobile phase (A): 0.1% (v/v) acetic acid in
water (B): 0.1% (v/v) acetic acid in
acetonitrile
Time (min) mobile phase (%)(A) (B)
0 90 1035 72 28
Standard isoflavonoids: puerarin (95 %) and genistein (99.9 %)
![Page 12: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/12.jpg)
Result
CombinationCombinationss % Moist
ureDiameter
Freshweigh
t Dry w
eight
waterwater73.23
64.56
589.32
147.33
SASA74.65
62.07
611.93
152.98
CuCl2CuCl272.53
77.53
591.30
147.83
CuCl2 + SA CuCl2 + SA77.20
72.30
561.38
140.35
ChitosanChitosan75.97
68.83
614.32
153.58
Chitosan + S Chitosan + SAA 74.73
66.57
622.53
155.63
Chitosan + C Chitosan + CuCl2uCl2 73.36
75.36
542.44
135.61
Chitosan + C Chitosan + C uCl2 + SA uCl2 + SA 72.53
74.85
602.36
150.59
Effect on root growth
![Page 13: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/13.jpg)
Antioxidant activity by DPPH
3.44
2.362.60
2.312.62
2.43 2.49
1.99
0.070.00
0.50
1.00
1.50
2.00
2.50
3.00
3.50
4.00
Elicitors
IC50
(m
g/m
l)Antioxidant activity by DPPH
![Page 14: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/14.jpg)
Major isoflavonoids by HPLC
![Page 15: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/15.jpg)
min26.5 27 27.5 28 28.5 29 29.5
mAU
0.5
1
1.5
2
2.5
3
3.5
DAD1 A, Sig=254,4 Ref=360,100 (RUAM\GCTRT008.D) 2
6.43
2 27.
008
28.
227
29.
657
![Page 16: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/16.jpg)
Isoflavonoids content
31.27
66.54 67.39
46.98 44.93
72.82
106.30
74.78
2.1
12.9
9.8 11
.3 13.4 16
.3
31.9
8.1
0.00
20.00
40.00
60.00
80.00
100.00
120.00
water SACuCl2
CuCl2 + SA
Chitosan
Chitosan + SA
Chitosan + CuCl2
Chitosan + CuCl2 + SA
Elicitors
unit
puerarin (ug/g DW)
genistein (ug/100 g DW)
![Page 17: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/17.jpg)
• The results showed that all combinations h ad no statistically significant effect on the
diameter, fresh weight, dry weight and mo isture content of the tuberous roots.
• it had a statistically significant effect on a ntioxidant activity and the amount of geni
stein and puerarin.
• The combination of chitosan (1000ppm) + copper(II)chloride (300ppm) + salicylic aci
d (100ppm) gave the highest antioxidant a ctivity (IC50 = 1.99 mg/ml by DPPH).
Conclusion & Discussion
![Page 18: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/18.jpg)
• T 1000he combination of chitosan ( ppm) gggg ggg gg+ () (3 0 0 )
ggggg gg gggg gg ggggggggg ggg gggggggg g ggg 319determined by HPLC are . /1 0 0
1063in dry weight and . g/gindrywei ght , r esp .
• Tgg ggggggggg ggg gggggggg ggggggggggg gggggggg ggg ggggg ggggggggggggg ggggggg gg ggggg ousr oot s of WKK.
• The elicitors ggg ggg gggg ggg ggg ggggggg gg g gggggg gg gggg.
Conclusion & Discussion
![Page 19: Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition](https://reader035.vdocuments.us/reader035/viewer/2022070305/55144ee5550346414e8b500b/html5/thumbnails/19.jpg)
Thank you for your attention.