| Artemisinin is derived from A.annua.As the current drug of choice for the treatment of malaria[1],artemisinin solves the problem of resistance of quinoline to Plasmodium falciparum,recognized the first-line antimalarial drug.In recent years,the global artemisinin dosage is about 180 tons[2],and it is mainly extracted from the leaves of A.annua..The artemisinin content in A.annua,directly affects its extraction cost,which is the key to restrict the development of artemisinin downstream industry.At present,there are many studies on the content of artemisinin,artemisinic acid,artemisinin B and dihydroartemisinic acid,but no ultra-high performance liquid chromatography-tandem quadrupole mass spectrometry(UPLC-ESI-QQQ-MS)has been used to have the comprehensive detection of A.annua,in different habitats,different developmental stages and different plant organs.In this paper,UPLC-ESI-QQQ-MS was used to study the content characteristics of artemisinin,artemisinin B,artemisinic acid and dihydroartemisinic acid in different habitats,different developmental stages and different plant organs in order to obtain the above.The changes of the content of four artemisinin compounds in the place of origin,time and plant organs provide reference and data to support for the related research of large-scale breeding,high-efficiency harvesting and biosynthetic pathway analysis of A.annua..In this experiment,UPLC-ESI-QQQ-MS multiple reaction monitoring MRM and SIM models were used to determine the content of artemisinin,artemisinin B,artemisinic acid and dihydroartemisinic acid.Quantitative methods were verified by linear,precision(RSD:2.07%,2.85%,3.09%,3.27%),repeatability(RSD:5.45%,5.40%,6.73%,5.69%)stability(RSD:7.85%,6.86%),2.84%,2.68%)recovery(RSD:1.83%,2.82%,1.65%,2.80%)etc.Samples from different habitats,different developmental stages,and different plant organs were analyzed by one-way analysis of variance,correlation analysis,and cluster analysis.In this experiment,9 wild provenances and 3 artificial breeding sources were planted in Beijing Huairou experimental base on March 10,2018,and samples of A.annua,from different sources were harvested on July 30;Samples of different developmental stages were collected from June 28 to September 23;samples of different plant organs were collected on October 12,19,and November 27.The results show:Among the 12 different samples of A.annua.,the artemisinin content of the artificially selected provenance was significantly higher than that of the wild provenance,both above 1%,and the highest YQ7 content reached 1.55%.The content of artemisinin in the field of A.annua.was less than 1%.The content of artemisinin from north to south was on the rise,the content of artemisinin B was below 0.2%,and the content of artemisinin B in the south is very low,most of which are below 0.1%.Artemisinic acid content was not significantly different among 12 samples from different sources.The artemisinic acid contents of Jilin,Beijing,Henan,Yunnan,YQ2 and YQ1 were all above 0.1%,and the rest were below 0.1%.Dihydroartemisinic acid is significantly different in germplasm from different sources,and dihydroartemisinic acid is also higher in germplasm with high artemisinin content.The correlation between the artemisinin compounds and the ecological factors was found to be significantly negatively correlated with the latitude(P<0.01),and significantly negatively correlated with the longitude(P<0.05).The absolute values of the correlation coefficients were respectively.It was 0.908 and 0.714,and there was a significant positive correlation between artemisinin and dihydroartemisinic acid(P<0.01).The absolute value of the correlation coefficient was 0.927.In the nine wild areas tested in this experiment,the content of artemisinin and dihydroartemisinic acid from north to south increased roughly.In the results of the determination of artemisinin compounds in different developmental stages of A.annua,at six sources,there is a significant difference of the artemisinin content from Hubei,Hainan,YQ2,YQ1,YQ7 at different times,except for Jilin.Artemisinin content is positively correlated with time.The content of artemisinin reached the highest in September,and the highest content of A.annua.from Jilin,Hubei,Hainan,YQ2,YQ1 and YQ7 was 0.09%,1.02%,2.08%,2.08%,2.07%,2.94%.In addition to Jilin,the A.annua,had the highest dihydroartemisinic acid content of 0.02%after August,and the other five provenances of dihydroartemisinic acid reached the highest level at the end of June.The highest content of dihydroartemisinic acid in Hubei,Hainan,YQ2,YQ1 and YQ7 were 0.42%,0.48%,0.77%,0.71%and 0.75%,and then decreased to varying degrees.Artemisinin B and artemisinic acid showed no obvious change trend.In different plant organs of A.annua.,the content of artemisinin in leaves was generally higher than that in flowers.The content of artemisinin in leaves and flowers of Jilin was 0.02%,the stem was 0.01%,and the roots and seeds were almost undetectable.The content of artemisinin in the leaves of A.annua.in Guangxi,Hainan,YQ2,YQ1 and YQ7 was higher than that of other plant organs,which were 0.97%,1.20%,1.78%,1.93%and 2.42%,respectively.Secondly,the content of artemisinin in flowers was higher,0.11%,0.26%,0.30%,0.24%,0.21%.A small amount of artemisinin was also present in the stem,and the artemisinin content was 0.03%,0.04%,0.03%,0.02%,and 0.04%.Artemisinin,artemisinin B,artemisinic acid and dihydroartemisinic acid in roots and seeds were almost undetectable,and the proportion of artemisinin B and artemisinic acid in the leaves and flowers of A.annua.from Jilin was higher.Among the other five sources,artemisinin and dihydroartemisinic acid account for a large proportion.In addition,the content of dihydroartemisinic acid in the stems of six species of A.annua,was much higher than that of the other three compounds.The contents of dihydroartemisinic acid in the stems of Jilin,Guangxi,Hainan,YQ2,YQ1 and YQ7 were 0.06%,0.50%,0.47%,0.73%,0.18%,0.96%.The content of four artemisinin compounds in different sources,different developmental stages and different plant organs was obtained.Based on the basic data theory,it is expected to provide reference and data support for the research on the breeding,harvesting and artemisinin biosynthesis pathways of A.annua. |
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