| Uncaria rhynchophylla was used as the test materials.The response surface methodology?RSM?was used to optimize the extraction conditions of alkaloids,and the measurement conditions of alkaloids were screened by high performance liquid chromatography?HPLC?.By studying the differences in the main medicinal components of U.rhynchophylla which was grown in different parts of Guizhou,and using different degrees of shading treatment on U.rhynchophylla,and the main medicinal components of U.rhynchophylla with different growing years were compared to determine the suitable cultivated conditions and areas in Guizhou.Determine the appropriate light intensity for the accumulation of major medicinal components and the optimal harvest period for different growing years of U.rhynchophylla.The main results are as follows:1.By RSM,the optimal extraction conditions of rhynchophylline and isorhynchophylline in the leaves of U.rhynchophylla were 54.95?of drying temperature,60 min of ultrasound time,73.35%of methanol,and the extraction value of rhynchophylline was 133.342?g·g-1and of isorhynchophylline was 429.746?g·g-1.In order to facilitate the practice,the conditions were modified as follows:drying temperature was 55?,ultrasound time was 60min,the volume of methanol was 73%.From the verification test,the average yield of rhynchophylline was 130.8?g·g-1 and of isorhynchophylline was 425.6?g·g-1,which was close to the model predicted value.This proved that RSM can be used to optimize the extraction conditions of rhynchophylline and isorhynchophylline in the leaves of U.rhynchophylla.In the conditions of Agilent ZORBAX SB-C18?4.6 mm×250 mm,5?m?,mobile phase:methanol-0.1%ammonia?74:26,V/V?,flow rate:1.0 mL·min-1,column temperature:30°C,detection wavelength:246 nm,the U.rhynchophylla were measured and found that the recoveries of various substances of corynoxeine,isorhynchophylline,isocorynoxeine and rhynchophylline were 98.71%,100.59%,98.90%and 105.76%,RSD were all below 3%,indicating this method was suitable for the determination of the contents of corynoxeine,isorhynchophylline,isocorynoxeine and rhynchophylline in U.rhynchophylla.2.The content of alkaloids in U.rhynchophylla was affected by different environmental factors.For mature U.rhynchophylla,appropriate shading helped the accumulation of secondary metabolites in plants.The stronger shading degree,the higher content of alkaloids accumulated in various parts of U.rhynchophylla.However,long-term shading gradually reduced the content of alkaloids.The shading treatment also affected the accumulation of alkaloids in seedlings,and was more serious impact on the accumulation of alkaloids in the leaves.However,shading was too strong to form hooks.The higher altitude,the lower temperature.The higher content of alkaloids in U.rhynchophylla accumulating at altitudes from 800 to 1100 m?Danzhai and Changshun?than those planted at altitudes from 1100 to1300 m?Guiyang and Pingba?.Therefore,the areas below 1100 m above sea level are suitable for planting U.rhynchophylla.The U.rhynchophylla in Danzhai and Changshun are planted in forest lands with good soil fertility and high contents of total nitrogen,alkali-hydrolyzable N and available P in the soil.These have a certain role in accelerating the accumulation of alkaloids.When it comes to cultivated fileds,fertile soil should be chosen.In summary,altitude,light intensity,and soil fertility play an important role to accumulate of alkaloids in U.rhynchophylla.U.rhynchophylla is suitable for planting in forest land below 1100 m above sea level with fertile soil and a certain shade?medium shading?.Therefore,Danzhai and Changshun are a good choice for planting.3.Different ages of U.rhynchophylla had different harvest periods.The suitable harvest period for all parts of the annual U.rhynchophylla is in August.All parts of biennial is in December.All parts of triennial are in August,September and November.Quadrennial is in August.Quadrennial except for the leaves,the rest is suitable for the harvest period in August,but leaves can be harvested from August to January,which basically meet the requirements of the pharmacopoeia for harvesting in autumn and winter.As the age of U.rhynchophylla increase,the content of alkaloids gradually decreased. |
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