Determination Of The Main Active Components In Ampelopsis Grossdentatae Stems And The Fingerprint Study

Objective: To establish a method for the determination of the main active components and analyze the content of dihydromyricetin,rutin and myricetin in Ampelopsis grossdentatae stems from different areas of Guangxi.To study the influence of different harvest time on the quality of Ampelopsis grossdentatae stems medicinal materials,and determine the content of dihydromyricetin and myricetin in the stems for different time,and determine the best harvest time for Ampelopsis grossdentatae stems medicinal materials.Determination of Ampelopsis grossdentatae stems and leaves HPLC fingerprint from different areas of Guangxi,to establish the HPLC fingerprint mutual model of the Ampelopsis grossdentatae stems and leaves,and show the stems and leaves main differences in chemical composition,and provide experimental basis for further the herb the quality control.Methods:?1?HPLC conditions: The column: Agilent Poroshell EC-C18?4.6mmx250 mm,4.0?m?;the mobile phase: acetonitrile-0.1%H3P O4,gradient elution?0⁴0min,acetonitrile12%?20%,0.1%H₃PO₄88%?80%;40⁵0min,acetonitrile20%?24%;0.1%H₃PO₄80%?76%?.The flow rate:1ml · min-1,the column temperature: 25?,the detection wavelengths: 0²3min,330nm;23⁵0min,375nm;Sample quantity: 5?L.?2?HPLC fingerprint conditions: The column:Agilent Poroshell EC-C18?4.6mm × 250 mm,4.0?m?;the mobile ph ase: acetonitrile-0.1%H₃PO₄,gradient elution?0²0min,acetonitrile10%?15%,0.1%H₃PO₄90%?85%;20⁴0min,acetonitrile15%?19%,0.1%H₃PO₄85%?81%;40⁶6min,acetonitrile19%?32%,0.1%H₃PO₄81%?68%?;The flow rate:1ml · min-1,the column temperat ure: 25?,the detection wavelengths: 365nm;Sample quantity: 5?L?Results:?1?The linear ranges were 0.8005⁹.606?g?r=0.9999?for dihydromyricetin,0.2016 2.419?g?r=0.9998?for rutin and 0.100 ¹.202?g?r=0.9997?for myricetin,and the average recovery were 98.1%,101% and 99.9%,RSD<3%.The stability,repeatability and precision of the method were all better in all cases?RSD<3%?.The content of dihydromyricetin,rutin and myricetin in Ampelopsis grossdentatae stems was quite different,The content of dihydromyricetin was slightly higher than that of rutin,and the content of myricetin was the lowest.The determined results showed that the content of dihydromyricetin,rutin and myricetin in Ampelopsis grossdentatae stems from Ling Chuan of Gui Lin was higher,and the content of dihydromyricetin and myricetin from Shuang Jiang of Wu Ming and Hu Zhu Cun of Jin Xiu was lower,and the content of rutin from Si Ding Zhen of Liu Zhou was lower.?2?The experiment used Ampelopsis grossdentatae stems as the research object,which planted in Nanning,Guangxi,and compared content ofdihydromyricetin and myricetin in Ampelopsis grossdentatae stems in different time,the content was relatively high in May,June,July and August,and the others were low;the content of dihydromyricetin and myricetin was the highest in July,The best harvest time for stems medicinal materials was determined in July.?3?This experiment determined the fingerprint of Ampelopsis grossdentatae stems for the 10 batches of Guangxi,established the HPLC fingerprint mutual model of Ampelopsis grossdentatae stems in different areas of Guangxi,calibrated10 common peaks,the results were not less than 0.89.This experiment measured HPLC fingerprint of Ampelopsis grossdentatae leaves from 15 different regions of Guangxi,established the HPLC fingerprint mutual model of Ampelopsis grossdentatae leaves,calibrated 10 common peaks,and the similarity results were not less than 0.9,between 0.938 and 0.99.Conclusion: The content of dihydromyricetin,rutin and myricetin in the Ampelopsis grossdentatae stems from different regions of Guangxi was quite different.The established method can be separated from the co-existing components in the sample,and the separation effect was good,the results were accurate and repeatable.In the same Ampelopsis grossdentatae stems medicinal materials and different harvest time content of dihydromyricetin and myricetin had significant differences,the trend of them broadly changed in line in one year,and from the overall,the content of dihydromyricetin in the stems was significantly higher thanmyricetin,determined the optimal harvest time for Ampelopsis grossdentatae stems in July.This experiment determined the HPLC fingerprint of Ampelopsis grossdentatae stems and leaves respectively,established the HPLC fingerprint mutual model of Ampelopsis grossdentatae stems and leaves,the method was of high precision,stability and repeatability,and can provide reference for quality evaluation of Ampelopsis grossdentatae medicinal materials.