Study On Diterpene Esters And Their Effective Components In Seeds Of Euphorbia Lathyris L.

Leptochloa chinensis is the dry and mature seed of Euphorbia Lathyris L..Commonly used in the treatment of snakebite,acute lymphocytic leukemia,childhood epilepsy and other diseases.The various esters of the main pharmacodynamic substances in Leptochloa chinensis,but so far the content determination,biological activity and the constituents as effective components are rarely studied and reported.In this paper,the chemical constituents,preparation method of the effective components of diterpene esters and their contents determination in Leptochloa chinensis,and the biological activity were researched.Firstly,by using the column chromatography and Preparative HPLC,8 diterpene ester compounds were isolated from Leptochloa chinensis,and their chemical structures were identified by NMR spectroscopy and other methods.They are 5,15-diacetyl-3-phenyl-6,17-epoxylathyrol?compound I?,5,15-diacetyl-3,7-dibenzoylz-hydroxylathyrol?compound II?,5,15-diacetyl-3-benzoyl-lathyro?compound III?,15,17-diacetyl-3-cinnamoyl-17-hydroxy-ioslathyrol?compound IV?,5,15,17-tri-acetyl-3-benzoyl-17-hydroxyiso-lathyrol?compound V?,5,15-diacetyl-3-nicotinoyl-lathyrol?compound VI?,5,15-diacetyl-3-phenylacetyl-lathyrol?compound VII?,and 5,15-diacetyl-3-benzoyl-7-nicotinoyl-7hydroxy-lathyrol?compound VIII?.Secondly,the contents determination of seven kinds of diterpene esters in Leptochloa chinensis by HPLC method was established in this paper.These seven kinds of diterpene esters were compound ? compounds VII.The optimum extraction method and the method for determination of HPLC were established by investigating the kinds of extraction solvents,the extraction method,the dosage of solvent extraction and extraction times.The experimental results showed that in the range of 0.005 ⁰.2 mg/ml has a good linear relationship with the concentration c of I^VII in the range of mg/ml,and the detection limit is in the range of 0.03⁰.80 ng.The RSDs values of the precision,repeatability and stability of compound I^VII are less than 2%,the recovery rate were in the range of 96.5%⁹9.2% with the RSDs within 0.26%⁰.75%.The content determination method established in this experiment has the advantages of high sensitivity,good precision and high accuracy.The content determination results showed that the content of compound I-VII in Leptochloa chinensis were 3.26,2.165,4.745,0.705,1.365,0.425,0.615 and 13.3 mg/g.On the basis of clarifying the composition and content of diterpene esters,in this paper,the first invention is to use mechanical press method to make oil,and then use methanol extraction method to obtain diterpene ester effective components,and then the contents determination method of seven kinds of diterpene esters(compounds ? ^VIIIin the effective components by HPLC was established.The results showed that 3 times times the volume of methanol extracted oil 3 times,the recovery of methanol,drying,then can get diterpene ester effective components.The content of compound I-VII in the effective components was reached to 77.21%.Finally,the inhibitory effect of eight kinds of diterpene ester monomers and the effective components of diterpene ester on Tyrosinase was studied.The results showed that the effective components and seven diterpene esters monomers had inhibitory effect on tyrosinase,and the inhibition rate of compound III was the highest.The results suggest that the di-terpene ester compounds and their effective components may have skin whitening effect.To sum up,the experiment was separated from Leptochloa chinensis to identify 8 diterpene ester compound monomer,the effective components of diterpene esters were separated from Leptochloa chinensis by squeezing and methanol extraction for the first time.HPLC method for the determination of seven kinds of diterpene esters in the effective components of Leptochloa chinensis and in the herb for the first time.For the first time,the inhibitory effect of eight kinds of diterpene esters and their effective components on tyrosinase were studied.The results of this study provide a new scientific basis for the improvement of the quality standard of Leptochloa chinensis and the development and utilization of the esters in Leptochloa chinensis.