Studies On Chemical Constituents From Seeds Of Euphorbia Lathyris Linn

"Qianjinzi"is the dry and mature seed of Euphorbia Lathyris L.. It mainly grows in Henan and Zhejiang provinces in China,and is used to remedy hydropsy, amenorrhea , coprostasis, scabies ,wart and snakebites. Recent research indicate it can be used to cure leucocythemia,esophagus cancer,skin cancer and very effective.As"Qianjinzi"serves as a very useful Chinese medicinal plant, it is know recorded by Chinese pharmacopoeia. However, up to now, there is no report concerning about the content determination of diterpene esters in the quality standard of Semen Euphorbiae but for the dedermination of esculetin, esculin, Euphorbetin were reported. .In the experiment,"Qianjinzi"was extracted by heat reflux and then was isolated with Silica gel, sephadex gel, ODS column chromatography, preparative HPLC and recrystallization. All the compounds were identified respectively by the spectroscopic methods including IR, MS, and NMR as 6 , 20-epoxylathyrol- 5,15-diacetate-3-phenylacetate(I),lathyrol-5,15-diacetate-3-benzoate(II),7-hydroxyla- thyrl-5,15-diacetate-3,7-dibenzoate(III), 17-hydroxyiso-lathyrol 5,15,17-triacetate-3- benzoate (IV),esculetin(V),β-sitosterol(VI)。Moreover, the method has been established for content determination of compounds I,II,III in seeds of Euphorbia Lathyris L. in this paper. Extraction and SeparationSeeds of Euphorbia Lathyris L. (2kg) were extracted by heat reflux in ethyl acetate(15L) for three times (2h, 1.5h, 1h, respectively). The extraction was subjected to silica gel column and ODS column eluted repeatedly to afford compounds I(2.1g),II(2.0g),III(1.6g), IV(30mg),V(30mg)and VI(100mg).Compounds I～VI were identified respectively by IR, MS and NMR as 6,20-epoxylathyrol-5, 15-diacetate-3-phenylacetate,lathyrol-5,15-diacetate-3-benzoate,7-hydroxylathyrol -5,15-diacetate-3,7-dibenzoate,17-hydroxyiso-lathyrol-5,15,17-triacetate-3-benzoate, esculetin,β-sitosterol. Determination of diterpene ester in seeds of Euphorbia Lathyris L.In this paper, we present a more sensitive quantificational method by HPLC for determining the main diterpene ester (Ⅰ～Ⅲ) in seeds of Euphorbia Lathyris L.. Separation was carried out on Agilent 1100 HPLC instrument with a Prevail SB-C18 column (150 mm×4.6 mm,5μm). The mobile phase was methanol-water(75∶25) . The flow rate was 1.0 mg·mL^-1 at 25℃,and the detector was set at 280 nm. ResultsⅰThe linearity was in the range of 0.10-4.0μg forⅠ,Ⅱ,Ⅲ, and the correlation coefficients were 0.9998,0.9999 and 0.9996 respectively. Y=4.859×103X+13.40 r=0.9998 Y=3.815×103X+10.79 r=0.9999 Y=7.655×103X+14.30 r=0.9996ⅱWe injected the solution ofⅠ,ⅡandⅢinto HPLC continuously six times and the results of precision tests were RSD<2%.ⅲWe measured the contents ofⅠ,Ⅱ,Ⅲ, seeds of Euphorbia Lathyris L.. and the results were RSD<2% respectively.ⅳS/N was 3 after the concentration ofⅠ,Ⅱ,Ⅲdiluted to 2.0×10-5, 2.0×10-5, 4.0×10-5 mg·mL^-1, so the detectability were 0.4,0.4,0.8ng respectively.ⅴThe stability ofⅠ,Ⅱ,Ⅲwere good in ten hours because the results were RSD<2% .ⅵThe average recoveries (n=6) forⅠ,Ⅱ,Ⅲare 97.9% (RSD=1.5%),100.9%(RSD =1.5%),99.6%(RSD =1.2%)(n=6) respectively.ⅶThe content ofⅠ,Ⅱ,Ⅲin seeds of Euphorbia Lathyris L. purchased from jilin pharmacy were determined by the above method, the results showed that their contents were different. The content ofⅢis lower thanⅠ,Ⅱ. The average contents ofⅠ,Ⅱ,Ⅲwere 0.38%,0.40%and 0.20% respectively.The results of this paper can offer a new index on content determination for establishment of quality standards and provide science foundation of the development and utilization of seeds of Euphorbia Lathyris L.