steroids.ppt

单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,steroid,Section 1 introduction,categories and structure characteristics of natural steroids,Name,A/B,B/C,C/D,Substituent on C,17,Phytosterol,Cis,trans,Trans,Trans,liphatic hydrocarbon with 810 carbon,Bile acid,Cis,Trans,Trans,amyl acid,C,21,sterol,Trans,Trans,Cis,C,2,H,5,Moulting hormones,Cis,Trans,Trans,liphatic hydrocarbon with 810 carbon,Cardiac glycosides,Cis,trans,Trans,Cis,unsaturated lactone,bufogenin,Cis,trans,Trans,Trans,six-membered unsaturated lactone,Steroid saponins,Cis,trans,Trans,Trans,loop-coil containing oxygen,Steroid alkaloids,Section 2 cardiac glycosides,steroid glycoside,physiological activity to heart.,No cardiac glycosides from animal.Toad venom also show physiological activity,but it doesnt belong to glycosides,belong to ester of liphatic acids.,Cino bufogenin(蟾酥）,structure and classification,1.,the structure of aglycone,five-membered unsaturated lacone,(,-lactone),six-membered unsaturated lacone(,-lactone),structure characteristics:,(1),A/B:cis or trans,but most are cis;B/C:trans;C/D:most cis,(2)C,3,-OH,C,14,-OH;,C,3,-OH,few are-OH(called epi-),C,3,-O-saccharide;,C,14,-OH,dewater with C,15,-H.,(3)C,10,CH,3,CHO,CH,2,OH,COOH;C,13,-CH,3,(4)C,11,C,12,C,19,C=O,Digitoxigenin,（洋地黄毒苷元）,3,14-dihydroxy-5-card-20(22)-enolide,Gitoxigenin,（羟基洋地黄毒苷元）,digoxigenin diginatigenin,（异羟基洋地黄毒苷元）（双羟基洋地黄毒苷元）,scilliglaucosidin,（绿海葱苷元）,3,14-dihydroxy-19-oxo-scilla-4,20,22 trienolide,bufotalin,（蟾毒素）,2.,the structure of saccharide,2.1-OH saccharide,D-quinovose D-antiarose,(,D-,鸡纳糖）（,D-,弩箭子糖）,6-deoxy saccharide,2.2,-deoxy saccharide,D-digitoxose D-cymarose,（,D-,洋地黄毒糖）（,D-,加拿大麻糖）,3.,the connecting type of aglycone and saccharide,Type aglycone-(2,6-deoxysaccharide),x,-(D-glucose),y,Type aglycone-(6-deoxysaccharide),x,-(D-glucose),y,Type aglycone-(D-glucose),y,purpurea glycoside A （紫花洋地黄毒苷）,thevetin A （黄夹苷A）,Scilliglaucoside （绿海葱苷）,the relationships between structure and activity,1.,steroid part:,C/D:cis,，,C,14,-OH.,if A/B cis,the C,3,-OH must be-OH,if A/B trans,C,3,-OH C,3,-OH.,2.unsaturated latone,C,17,-unsaturated lactone.,If the lactone is saturated,the activity decrease.,3.,substituent,C,10,-CH,3,change to C,10,-CHO or CH,2,OH,the activity increase,C,10,-CH,3,change to C,10,-C0OH or H,the activity decrease,4.saccharide part,甲型：,trisaccharides cardiac glycoside bisaccharides cardiac glycoside cardiac aglycone,乙型:,cardiac aglycone monosaccharides cardiac glycoside bisaccharides cardiac glycoside,Properties,1.Description,colorless crystal or amorphous powder;optical activity;,C,17,-,：,bitter,；,C,17,-,：,not bitter,no physiological activity;,stimulation to mucosa.,2.Solubility,cardiac glycoside can dissolve in H,2,O,alcohol,acetone,；,slightly dissolve in EtOAc,CHCl,3,containing alcohol,cant dissolve in Et,2,O,benzene,p.e.,The solubility is related to the number,kind of saccharide and the number,site of hydroxy in aglycone.,The more the number of hydroxy,the stronger the hydrophilicity.,e.g.,ouabain is a monosaccharide glycoside with 8 hydroxys,the solubility in water is 1:75,difficultly dissolve in CHCl,3,.,digitoxin is a trisaccharide glycoside with 5 hydroxys,the solubility in water is 1:100000,easily dissolve in CHCl,3,(1:40).,+3D-digitoxose,3.dehydration,4.hydrolysis,4.1 acid hydrolysis,(1)niddle acid hydrolysis,(2)strong acid hydrolysis,(3)Mannich and siewert methods,(1)niddle acid hydrolysis,0.020.05mol/L HCl or H,2,SO,4,Type cardiac glycoside,hydrolysis on-deoxysugar,0.02-0.05mol/L,aglycone+2 D-digitoxose+D-digitoxose-D-glucose,(2)strong acid hydrolysis,3%5%HCl,pressure and heat,type,type,cause dehydration,+3D-digitoxose,(3)Mannich and siewert methods,1HCl/acetone,most type,4.2 enzyme hydrolysis,the plant containing cardiac glycoside only have the enzyme which can,hydrolyze glucose,havnt the enzyme which can hydrolyzed-deoxysugar.,Snail enzyme almost can hydrolyze all glycosidic linkage.,4.3 base enzyme,can hydrolyze the glycosidic linkage,(1)hydrolysis on acyl,（酰基）,NaHCO,3,KHCO,3,Ca(OH),2,Ba(OH),2,These four bases only can hydrolyze acyl,cant hydrolyze lactone ring.,(2)hydrolysis on lactone ring,NaOH and KOH can hydrolyze both acyl and lactone ring.,.color reaction,1 reaction act on steroid,Liebermann-Burchard reaction;,Salkowski reaction;,Tschugaev reaction;,Rosen-Heimer reaction;,Kahlenberg reaction,2 reaction act on,-unsaturated lactone ring,can distinguish,甲型,and,乙型,2.1 Legal reaction,1-2mg sample/2-3drops pyridine+1 drop 3%nitrosyl sodium ferricyanide+1 drop 2mol/L NaOH deep red fading,2.2 Raymond reaction,1mg/50%EtOH+0.1ml m-dinitrobenzene+20%NaOHviolet red,2.3 Kedde reaction,sample/MeOH+3,5-nitro benzoic acid reagent red or violet red,2.4 Baljet reaction,sample/MeOH+basic picric acid reagent orange or orange red,3.reactions act on-deoxysaccharide,3.1 Keller-Kiliani(K-K)reaction,1mg/5ml glacial acetic acid+20%FeCl,3,+conc.H,2,SO,4,free-deoxysaccharide or-deoxysaccharide connecting with aglycone:the acetic acid layer show blue.,3.2 Xanthydrol reaction,sample/Xanthydrol reagents,heated on water bath for 3 minutes,free-deoxysaccharide or associated-deoxysaccharide,show red.,3.3-dimethylaminobenzal dehyde,sample/EtOH spotted on filter paper,spraying-dimethylaminobenzal dehyde,heated on 90 for 30 seconds,free-deoxysaccharide or associated-deoxysaccharide,show grayish-red.,3.4 periodate-nitroaniline,sample/EtOH spotted on filter paper or thin plate,spraying sodium perodate reagents,10 minutes later,spraying-nitroaniline reagents,show deep yellow spot on grayish yellow background,then spraying 5%NaOH/MeOH,the spot show green,.,Extraction and isolation of cardiac glycoside,1.Extraction,70%80%EtOH.,2.Isolation,Extraction,（萃取）,with solvents and chromatography,1.UV,217-220nm(lg4.20-4.24)295-300nm(lg3.93),270nm 330nm,isolated carbonyl 290-300nm(lg1.8),C=O 18001700cm-1 two strong peaks,1783 cm-1 unnormal peak,1756 cm-1 normal peak,the intensity of unnormal peak decrease or the peak disappear with the increasment of solvents polarity.,2,、,IR,1740 cm-1 unnormal peak,1718 cm-1 normal peak,3,、,1,H-NMR,C,21,上的两个,H,：,H,d,；,H d,4.55.0,C,22,br.,5.66.0,(1)H on lactone,H-21 s 7.20,H-22 d 7.80,H-23 d 6.30,（）,CH,3,and,CH,3,s,，,1.0,（）,m,，,.,，,shift to low field after glycosylation,1-H,dd,1-H,d,-去氧糖的2H比-羟基糖的处于高场,（）H on saccharides,4.,13,C-NMR,primary carbon伯碳 1224,secondary carbon仲碳 2041,tertiary carbon叔碳 3557,quarternary carbon季碳 2743,R-C-OH醇碳 6591,olefinic carbon烯碳 119-172,carbon on carbonyl group羰基碳,177220,Section 3 steroid saponin,Introduction,steroidal saponin=spirostane+saccharide,structure and classification,1.structure,2.1,.spirostanol 2.2 isospirostanol,(,螺甾烷醇型,)(,异螺甾烷醇型,),、,classification,2.3 furostanol 2.4 pseudo-spirostanol,(,呋甾烷醇型,),(,变形螺甾烷醇型,),properties,1.description,steroidal saponin:colorless or white amorphous powder.,aglycone:crystal.,High melting point,the melting point of aglycone increase with the increasement of hydroxy.,Both steroidal saponins and their aglycones have optical activity,levo-rotatory.,、,solubility,H,2,O,MeOH,EtOH,CHCl,3,Et,2,O,Steroidal saponin,aglycone,、,precipitation reaction,Steroidal saponin/EtOH,sterol,（,cholesterin,）,ppt,reflux with Et,2,O,Et,2,O,ppt,（,cholesterin,）（,steroidal saponin,）,除胆甾醇外，皂苷可与其它含有,-OH,的甾醇形成沉淀。,4,、,color reaction,Liebermann,Burchard reaction Rosen-Heimer reaction,Steroidal saponins last show green show color heated to 60,Triterpenoid saponins last show red show color heated to 100,extraction and isolation of steroidal saponin,1.extraction,Steroidal saponin,are often extracted by MeOH or EtOH,the solvents are recovered and then the extracts can be purified by:,formed ppt in acetone or Et,2,O,extracted with n-BuOH saturated with H,2,O,purified by macroreticular resin,Steroidal aglycone:,first extract the steroidal saponin,then hydrolyze the saponin,extract the aglycone by benzene,CHCl,3,etc.organic solvents.,First hydroylze the drug,then extract the hydrolysates by organic solvents.,2.Isolation,ppt by solvents(Et,2,O,acetone);,ppt by cholesterin;,Girard reagents to extract the steroidal aglycone with carbonyl;,Column chromatography:,Silica column chromatography(often eluted by CHCl3-MeOH H2O system);,Macroreticular resin column chromatography;,Sephdex LH-20 column chromatography;,DCCC;,