| Artemisinic acid has important signifcance for its important medical value that it isthe premise in synthesis of artemisinin and artemisinin B. In this paper, the contentchanges of Artemisinic acid were studied in the Artemisia annua plant as theexperimental material growning different stages in Wuling Mountains. Artemisinic acidpreparation technology was optimized. And the biological stability of Artemisinic acidwas analyzed. The results would provide the experimental and theoretical reference forthe development and utilization of Artemisinic acid in Wuling Mountains. The workincludes the following sections:1. Analysis of Artemisinic acid content of Artemisia annua at different growthstagesContent of Artemisinic acid was analyzed using HPLC and TLC methods in A.annua at different growth stages around Wuling Mountains including Jishou Hunan,Zhangjiajie Hunan, Shataping Hunan, Xiushan Chongqing, Tongren Guizhou, GuilinGuangxi. The results showed the content of Artemisinic acid around Wuling Mountainsfrom May to September in2011,2012and2013years. The Artemisinin acid content isclosely related to its growing season of plant. And the average contents of Artemisinicacid in June were highest amount mouths. April and May were minimum（0.13mg/g~0.21mg/g）, then contents increased gradually from June to July（0.27mg/g~0.23mg/g）,but reduced from August to Septembe（r0.22mg/g~0.15mg/g）. Artemisinic acidcontent was the highest in the mature growth period (from June to July) of plant, growthperiod (from August to September) followed by, and the minium content was at younggrowth period (from April to May), So the best harvest season was the mature growthperiod of A. annua around Wuling Mountains which benefiting to the utilization ofArtemisinic acid. Artemisinic acid content also had some certain relevance with thealtitude, which Artemisinic acid content growing low altitude was higher than that athigh altitude.2. Preparation process optimization of Artemisinic acidUsing Artemisia anncua as preparation material of Artemisinic acid in WulingMountains, we optimized the process of operation through the extraction of Artemisinicacid preparation technology, crude separation, fine separation and crystallization.Results showed that:(1) the Artemisinic acid extraction rate with methanol,0.12%(w/w),was highest amount six kinds of solvent. The effect of solvent extraction timesand temperature on Artemisinic acid extraction yield showed that it was by three timesprocess that Artemisinic acid could be extracted completely,while under the conditionof room temperature that the highest extraction yield.(2) The crude separation oforganic solvent, acid, alkali, organic solvent had higher Artemisinic acid yield and better effect on removing impurity. Artemisinic acid of4.21mg was collected,with81.0%(w/w) Artemisinic acid yield and85.0%(w/w) the total rate of removingimpurity. The fine points process was optimized. And the elution effect of differentmobile phase to Artemisinic acid and the separation laboratory of adsorbent dosage toArtemisinic acid showed that petroleum ether, ethyl acetate=90:10(v/v)as mobilephase. Artemisinic acid yield was maximum,6.90mg, adsorbent silica gel: sample=15:1(w/w). Artemisinic acid separation effect was best. Eight kinds of solvent grainswere relatively stable and the results showed that petroleum ether precipitation crystalcolour and lustre was transparent. Further, under the condition of-70oC,-20oC,crystalline grain color and shape were better than that at4oC.3. Study on preparation and biologycal stability of Artemisinic acidAs the important medicinal value, the preparation technology and the biologicalstability of Artemisinic acid were explored from Artemisia annua.The preparationtechnology named Solvent method-Adsorptive separation and the biological stability ofArtemisinic acid were analyzed using the methods of Thin Layer Chromatography(TLC) and High Performance Liquid Chromatography (HPLC). Crystal of Artemisinicacid was successfully obtained by the Solvent method-Adsorptive separationpreparation technology, and the yield of product reached to60percents. Preparation ofArtemisinic acid purity of96%, the maximum UV absorption wavelength was220nm,melting point is130oC~132oC, mass spectrometry for M+234.The biological stabilityof Artemisinic acid was very stable saving under the conditions of4oC, roomtemperature,60oC and natural lighting for30d. Furthermore, Artemisinic acid wasstably preserved under the conditions of in plant vivo or in vitro for a year, the contentof Artemisinic acid was the same. The structure of Artemisinic acid was easilydestructed by UV light, and the content undetected after lighting for6h. The product ofArtemisinic acid could be successfully obtained by the Solvent method-Adsorptiveseparation preparation technology. The biological stability of Artemisinic acid could bemaintained under the conditions of the certain temperature, natural lighting and in vivoor vitro. However, the structure of Artemisinic acid was easily destroyed by UV light. |
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