Presentation on theme: "Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition."— Presentation transcript:
Antioxidant activity and major isoflavonoid contents of White Kwao Krua by elicitors-induced on greenhouse condition.
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.
Found in northern and north eastern. It is considered to be a rejuvenating drug and has a history in folk medicine. Name White Kwao Krua was definitively identified as Pueraria candollei in 1952 from which an unusual estrogenic. White Kwao Krua (WKK) has a high concentration of phytoestrogen. White Kwao Krua [Pueraria candollei Grah. var. mirifica (Airy Shaw et. Suvatabandhu) Niyomdham].
Applications cosmetic industry breast cream eye gel skin moisturizer. Pharmaceuticals industry Replacement hormones Loss bone metabolism Livestock Animals feed
Antioxidant activity Antioxidant compounds play an important role as a health-protecting factor. Antioxidants reduce the risk for chronic diseases including cancer and heart disease. The main characteristic of an antioxidant is its ability to trap free radicals. Cell metabolism & pollutions superoxide anion radical (O 2 ) hydroxyl radical (OH) peroxyl radical (ROO) Gutteridge (1994)
Major Isoflavonoids Genistein –Phytonutrient –Manopause (hormonal replacement) –Loss bone (bone metabolism) Puerarin –Hypertension –antioxidant Igham et al. (1986)
Elicitors Molecules that stimulate secondary metabolism Elicitors may form inside or outside plant cells, and are distinguished as endogenously or exogenously inducers. Depending on their origin, they are classified as biotic or abiotic Heike and Dietrich (1995)
Elicitors Chitosa n Soybean; 1000 ppm increased seed isoflavone % and Daidzein 150 % Tawaha et al. (2005) Salicyli c acid Lupinus ruteus; 800 µM increased Genistein µ g/g DW. Kneer et al. ( 1999) Copper (II) chlorid e - Pueraria montana; 100 ppm increased Genistein 5 folds - Pueraria lobata; 10 mM for 2 hour increased Genistein 10 folds Takashi et al. (1990) Hakanat suka (1991)
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) Water salicylic acid (SA) CuCl 2 CuCl 2 + SA Chitosan Chitosan + SA Chitosan + CuCl 2 Chitosan + CuCl 2 +SA
Antioxidant determination by DPPH Inhibitory concentration at 50% (IC 50 ) Plot between % radical scavenging VS concentration of each treatment Equation of nonlinear regression
Major isoflavonoids determination by HPLC HPLC condition HPLC column RP-C18 Agilent ® (4.6 mm x 15 cm) Detector Diode array 254 nm Flow rate1 ml / min Volume injection10 µl Presser pump111 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) Standard isoflavonoids: puerarin (95 %) and genistein (99.9 %)
ResultCombinations% Moistur e Diam eter Fresh weight Dry weigh twater SA CuCl CuCl2 + SA Chitosan Chitosan + SA Chitosan + CuCl Chitosan + CuCl2 + SA Effect on root growth
Antioxidant activity by DPPH
Major isoflavonoids by HPLC
The results showed that all combinations had no statistically significant effect on the diameter, fresh weight, dry weight and moisture content of the tuberous roots. it had a statistically significant effect on antioxidant activity and the amount of genistein and puerarin. The combination of chitosan (1000ppm) + copper(II)chloride (300ppm) + salicylic acid (100ppm) gave the highest antioxidant activity (IC50 = 1.99 mg/ml by DPPH). Conclusion & Discussion
The combination of chitosan (1000ppm) + copper(II)chloride (300ppm) gave the highest amount of genistein and puerarin were determined by HPLC are 31.9 g/100g in dry weight and g/g in dry weight, respectively. The elicitors can increase antioxidant activity and major isoflavonoids content in tuberous roots of WKK. The elicitors did not have the bad effects on root growth of WKK. Conclusion & Discussion