| In this paper,the antifungal activities of different bamboo leaves extract against Gibberella sanbinetti were tested using bioassay methods;a HPLC methed was optimizeded for isoorientin and kresoxim-methyl in bamboo leaves extract,10% bamboo leaves extracts water emulsions and 10.7% bamboo leaves extract·kresoxim-methyl complex aqueous emulsion.The accuracy of the method was evaluated.The content of isoorientin in different bamboo leaves extract was detected.And the decomposition rate of isoorientin and kresoxim-methyl was detected after the thermal storage and cold storage of 10% bamboo leaves extracts water emulsions and 10.7% bamboo leaves extract·kresoxim-methyl complex aqueous emulsion to evaluate the stability of pesticide preparation.And toxicological assessment of bamboo leaves extract was investigaed to make preliminary safety evaluation.The main results were following: 1.The antifungal activities of bamboo leaves extract from different sources.The antifungal activities of bamboo leaves extract from different sources against Gibberella sanbinetti was examined.Among different locations,bamboo of huoshan,ningguo and shitai shows the best antifungal activities against Gibberella sanbinetti.The inhibition rates were 92.31%,83.98% and 92.31% respectively;the inhibition rates of bamboo in hunan Shaodong,fujian jianou,guizhou chishui,qianshan and fanchang were 75.69%,72.66%,68.13%,65.52% and 62.36%;Among different collection times,the inhibition rates bamboo in July 2014 was all 100% after 48 h and 72 h,the inhibition rates of bamboo in May and November was all 100% after 48 h,86.05%,78.72% and 89.98% after 72 h respectively;the inhibition rates of bamboo in June,September and January 2015 were 88.10%,83.98%,77.06% and 88.10% after 48 h respectively;among different ages,the antifungal activities of old bamboo leaves is better than new bamboo leaves,the inhibition rates of old bamboo in June and July were 89.61%,100%,annual 76.62%,100% respectively.2.Analysis of isoorientin in bamboo leaves extract by HPLC.An HPLC method was optimized and validated for the determination of isoorientin in bamboo leaves extract.Extraction was carried out with 60% methanol.The separation was performed using a C18 column(4.6×250mm,5?m)at 30?.The solvent system consisted of a mixture of water with 0.5%(v/v)glacial acetic acid and acetonitrile(86/14,v/v)at a flow rate of 1.0 mL/min.The detection wavelength was set at 270 nm.The optimized HPLC methed proved to be linear in the concentration range tested(0.5~100?g/mL),accurate(93.1~102.8%),The proposed method was validated to be simple and reliable.3.The content of isoorientin in bamboo leaves extract from different sources.By HPLC method to compare content of isoorientin in bamboo leaves extract from different sources.Among different locations,the content isoorientin of bamboo leaves extracts in ningguo,huoshan shitai and fujian jianou were 0.41%,0.36%,0.35% and 0.36% respectively,the content isoorientin of bamboo leaves extracts in hunn shaodong,fanchang,guizhou chishui and qianshan were 0.16%,0.16%,0.15% and 0.16% respectively;among different collection times,the content isoorientin of bamboo leaves extracts in July 2014 was 0.72%,and in march,may,September and November were respectively to 0.52%,0.42%,0.38%,0.36% and 0.24%,the content isoorientin of bamboo leaves extracts in January were 0.06%;among different ages,the content isoorientin of old bamboo leaves extracts(0.24% and 0.72%)were higher than new bamboo leaves(0.06% and 0.46%).4.Analysis of kresoxim-methyl in bamboo leaves extract water emulsions by HPLC.An HPLC method was optimized and validated for the determination of isoorientin and kresoxim-methyl in bamboo leaves extract water emulsions.Samples was carried out with 60% methanol.The separation was performed using a C18 column(4.6×250mm,5?m)at 30?.Isoorientin: The solvent system consisted of a mixture of water with 0.5%(v/v)glacial acetic acid and acetonitrile(86/14,v/v)at a flow rate of 1.0 mL/min.The detection wavelength was set at 270 nm.The optimized HPLC methed proved to be linear in the concentration range tested(0.5~100?g/mL),accurate(92.9 ~ 102.4%).Kresoxim-methyl: The solvent system consisted of a mixture of water with 0.5%(v/v)glacial acetic acid and acetonitrile(20/80,v/v)at a flow rate of 1.0 mL/min.The detection wavelength was set at 234 nm.The optimized HPLC methed proved to be linear in the concentration range tested(0.05~20?g/mL),accurate(95.2~98.3%).The proposed method was validated to be simple and reliable.5.The stability of 10% bamboo leaves extracts water emulsions and 10.7% bamboo leaves extract·kresoxim-methyl complex aqueous emulsion.By HPLC method to compare content of isoorientin and kresoxim-methyl in 10% bamboo leaves extracts water emulsions and 10.7% bamboo leaves extract·kresoxim-methyl complex aqueous emulsion.After cold storage,the decomposition rate of isoorientin in 10% bamboo leaves extracts water emulsions was 0.91%,the decomposition rate of isoorientin and kresoxim-methyl in 10.7% bamboo leaves extract·kresoxim-methyl complex aqueous emulsion were 1.03% and 1.04% respectively;after thermal storage,the decomposition rate of isoorientin in 10% bamboo leaves extracts water emulsions was 2.22%,the decomposition rate of isoorientin and kresoxim-methyl in 10.7% bamboo leaves extract·kresoxim-methyl complex aqueous emulsion were 2.61% and 2.46% respectively.According to the product stability evaluation standard,effective physical decomposition rate were within the scope of water emulsion heat storage performance,preparation stability is good.6.Preliminary safety evaluation of bamboo leaves extract.Toxicological assessment of bamboo leaves extract was investigaed to make preliminary safety evaluation.The research results show that the bamboo leaf extract of rat acute oral and dermal half lethal dose(LD50)are greater than 5000 mg/kg,the 95% confidence interval were greater than 5000 mg/kg,according to the classification standard,the bamboo leaf extract belongs to micro toxicity material.Bamboo leaf extract of rabbit acute skin irritation of integral index is 0,the average index is 0,without excitant.Bamboo leaf extract of rabbit acute eye irritation when undeveloped integral index is 11.0,the average was 4.5(less than 5 after 4 d),7 d returned to normal,mild to moderate irritation;4 s acute eye irritation when flushing integral index was 6.5,with an average of 1.5(less than 5)after 48 h,72 h returned to normal,mild stimulant;30 s acute eye irritation when flushing integral index was 7.5,with an average of 3.0(less than 5)after 48 h,72 h returned to normal,mild irritation. |
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