Studies On Chemical Constituents And Quality Evaluation Of Citri Reticulatae Semen And Atractylodes Japonica Koidz.ez Kitam

In this paper,a variety of modern chromatographic separation techniques were used to study the chemical composition of the Citri Reticulatae Semen and Atractylodes japonica Koidz.ex Kitam Studies on quality evaluation of them and some compounds were screened for activity in vitro.I.Studies on the Chemical Composition of Citri Reticulatae SemenA total of 21 compounds were identified from the Citri Reticulatae Semen by various chromatographic separation techniques.Including 15 limonoids;compounds?J1-J15?,2 flavonoids?J16-J17?;3 phenolic acid compounds?J18,J19,J21?;One fatty acid compound?J20?.Among them,the compounds J1-J4 and J21 are new compounds.They are 21,23-dihydro-21-hydroxy-23-oxoichangin?J1??21,23-dihydro-23-ethoxy-21-oxolimonexic acid?J2??Tetrahydro-20,22-dihydroxy-21,23-diethoxy-nomilin?J3??21,23-dihydro-23-ethoxy-21-oxoobacunone?J4?,Tinosporaic acid C?J21?.J8-J14and J16-J20 in known compounds were first separated in the orange core.The effects of15 limonoids Compounds on the proliferation inhibition of Hep G2 hepatoma cells were determined by CCK-8 method.The results showed that compounds J2,J6,J10,J12,J1,J3,J7,and J9 all produced different levels of cytotoxic activity.Combining the structural characteristics of these compounds,we speculate that the cytotoxic activity of the Ring-A,D-seco-limonoids that forms a 5-membered cyclolactone in A ring cracking plan stronger effects than that does not form a lactone and ring A cracking to form a7-membered cyclolactone.2.Studies on the quality evaluation of Citri Reticulatae SemenIn the current Pharmacopoeia standard,there is no qualitative and quantitative method about the Citri Reticulatae Semen.In view of this,based on the study of chemical constituents,the qualitative and quantitative methods of Citri Reticulatae Semen were studied in this paper.The TLC qualitative identification method of Citri Reticulatae Semen was established with silica gel as the stationary phase and petroleum ether-ethyl acetate-85%formic acid?1:2:0.1?as the spreading agent.At the same time,the HPLC method was used to establish a QAMS of five kinds of limonoids in the Citri Reticulatae Semen?obacunone,Limonin,nomilin,deacetyl nomilin,ichangin?.And,limonin was used as the internal reference substance with the ralative correlation factors of the limonoids.The contents of these five limonoids were calculated by the RCF,thus,to make the QAMS possible.At the same time,the content of these five components was determined by external standard method,and the difference between the calculated value and the measured value was compared by SPSS software,verified the rationality and feasibility of QAMS.3.Studies on the Chemical Composition of Atractylodes japonica Koidz.ez Kitam.A total of 42 compounds were isolated and identified by various chromatographic separation techniques.Including 9 Sesquiterpenes?G1-G9?,10 polyacetylenes?G11-G20?,2 benzoquinones?G21,G22?,5 glycosides,and 16 other compounds?G24-G42?.Among them,the compounds G11,G12,and G13 are new compounds,which are(?4E,6E,12E?-4,6,12-tetradecatrien-3-hydroxy-8,10-diynyl Ester,?2E?-?9CI??G11?,?4E,6E,12E?-4,6,12-tetradecatrien-14-hydroxy-8,10-diynyl Ester,?2E?-?9CI??G12?,?8E,15E?-heptadeca-8,15-dien-11,13-diynoic acid?G13?.4.Studies on the quality evaluation of Atractylodes japonica Koidz.ez KitamAtractylodes japonica Koidz.ez Kitam is the base plant of Atractylodes macrocephala,and Atractylodes macrocephala is an inverted species in the current Pharmacopoeia standard.The standard of this species has been formulated of our research group.Based on the previous research,the quantitative method has been revised and the content determination method of selina-4?14?,7?11?-dien-8-one by HPLC has been established.The chromatographic conditions:column:Preverse C18?250 mm?4.6 mm,5?m?;Mobile phase:acetonitrile-water?70:30?;Detect wavelength 254 nm;Flow rate 1.0 m L.min^-1;Column temperature 25?;Sampling quantity 10?L.The number of theoretical plates should not be less than 4000.The selina-4?14?,7?11?-dien-8-one is Y=12.833 X-25.982,R=0.99984,and the linear range is 8.43?530?g.The precision,stability,repeatability and sample recovery of the method were all good.The content of selina-4?14?,7?11?-dien-8-one in 20 batches of Atractylodes japonica Koidz.ez Kitam was determined.According to the dry product calculation,the content of selina-4?14?,7?11?-dien-8-one was not less than 0.2%.