鎖陽化學(xué)成分的初步研究

鎖陽化學(xué)成分的初步研究

鎖陽是鎖定科的植物，具有除花序過程中干燥的肉質(zhì)莖，具有益腎陽、益血、潤腸排便的功效。由于大量采挖,資源瀕臨枯竭,本實(shí)驗(yàn)對(duì)鎖陽花序中黃酮類化學(xué)成分進(jìn)行初步研究,以更加合理地利用鎖陽資源。1柱層析硅膠、聚酰胺薄膜X4型顯微熔點(diǎn)測(cè)定儀(北京第三光學(xué)儀器廠),Unity-Inova600超導(dǎo)核磁共振譜儀(美國Varian公司),柱層析硅膠(100～200目,青島海洋化工廠),薄層層析硅膠G(青島海洋化工廠),柱層析聚酰胺(30～60目,浙江省臺(tái)州市路橋四甲生化塑料廠),聚酰胺薄膜(薄層層析,浙江省臺(tái)州市路橋四甲生化塑料廠)。鎖陽藥材采自新疆和碩地區(qū),其品種由新疆醫(yī)科大學(xué)天藥/生藥教研室鑒定。2方法和結(jié)果2.1萃取物的制備干燥的鎖陽花序切片1kg,用95%乙醇回流提取3次(3h、3h、1h),回收乙醇后得浸膏120g。聚酰胺30～60目碾磨至60～100目,處理待用。將提取浸膏120g上聚酰胺柱(60～100目),氯仿∶甲醇(8∶2～0∶1)梯度洗脫,共得到100個(gè)流份(Fr.J1-100);Fr.J11-85以水混懸,用乙酸乙酯萃取,得乙酸乙酯萃取物;將乙酯萃取物經(jīng)硅膠柱層析,石油醚∶乙酸乙酯(1∶0～0∶1)、氯仿∶甲醇(1∶0～0∶1)梯度洗脫,得到化合物Ⅰ、化合物Ⅱ和含有化合物Ⅲ的混合物以及化合物Ⅳ;將含有化合物Ⅲ的混合物2g上聚酰胺柱(60～100目),水∶丙酮(1∶0～0∶1)梯度洗脫,得到化合物Ⅲ。2.2結(jié)構(gòu)評(píng)價(jià)2.2.1c-nmr5.2-2,4h,2,4h,2,4h,2,4h,2,4h,2,4h,2,4h,2,4h,3,5,10.4h,4h,3,5,4.3,4.3,4.3,4.4h,3,4.3,4.3,4.4h,3,4.3,4.4h,3,4.3,3,4.3,3.3,3,4.3,3,4.4,3,3,3,3,3,3,4.4,3,3,3,3,3,3,3,3,4.3,3,3,3,3,3,3,4.3,3,4.3,3,3,3,4.3,3.3,4.3,4.3,4.3,4.3,3,3,3,3,4.3,3,3.3,4.3,3,3,3,4.3,3,3,3.3,3,4.3,3,3,3,3,3,3,3,3,3,3,3.3,3.3,3.3,3.3,3.3,3.3,4.3,4.3,4.3,3.3,3.3,3.3,3.3,3.3,3.3,3.3,3.3,3.3,3.3,3.3,3.3,3.3,3.3,3.3,3.3,3.3,3.3,3.3,3.35,3.3,3黃色粉末(甲醇),mp176～178℃,鹽酸-鎂粉反應(yīng)陽性,Molish反應(yīng)陽性。1H-NMR(600MHz,CD3OD),δH:1.02(3H,d,J=6.6Hz,-CH3),4.42(1H,d,J=7.2Hz,H-1?),3.17～3.53(16H,m,糖質(zhì)子峰),5.01(1H,d,J=7.8Hz,H-1″),6.11(1H,d,J=2.4Hz,H-6),6.30(1H,d,J=1.8Hz,H-8),6.78(1H,d,J=8.4Hz,H-5′),7.54(1H,d,J=2.4Hz,H-6′),7.57(1H,d,J=2.4Hz,H-2′)。13C-NMR(150MHz,CD3OD),δC:178.1(C-4),164.6(C-7),161.6(C-5),157.9(C-2),157.1(C-9),148.4(C-4′),144.4(C-3′),134.2(C-3),122.1(C-6′),121.8(C-1′),116.3(C-5′),114.6(C-2′),104.1(C-10),103.2(C-1″),100.9(C-1?),98.5(C-6),93.4(C-8),76.8(C-3″),75.8(C-5″),74.3(C-2″),72.5(C-4″),70.8(C-5?),70.7(C-3?),69.9(C-4?),68.2(C-6″),67.1(C-2?),16.4(C-6?);結(jié)合上述圖譜數(shù)據(jù)并查閱文獻(xiàn),確定該化合物為蕓香苷(Rutin),結(jié)構(gòu)式如下:2.2.2糖酸水質(zhì)分布淡黃色粉末(甲醇),mp193～195℃。鹽酸-鎂粉反應(yīng)陽性,Molish反應(yīng)陽性。1H-NMR(600MHz,CD3OD),δH:7.73(1H,d,J=2.4Hz,H-2′),7.62(1H,dd,J=2.4,8.1Hz,H-2′),6.89(1H,d,J=8.4Hz,H-5′),6.42(1H,d,J=1.5Hz,H-8),6.23(1H,d,J=1.5Hz,H-6),5.29(1H,d,J=7.2Hz,H-1″),3.33～3.75(5H,m,糖質(zhì)子峰)。氫譜中的6.23d,6.42d峰為5,7-二羥基黃酮類中H6,H8的特征峰,說明可能是5,7-二羥基黃酮類化合物。與蕓香苷相比,異槲皮苷的氫譜中少了鼠李糖在δ1.02處-CH3的二重峰和端基質(zhì)子在δH4.42處的峰。結(jié)合上述圖譜并查閱文獻(xiàn),最后鑒定其結(jié)構(gòu)式為槲皮素-3-O-β-D-葡萄糖,故確定該化合物為異槲皮苷(isoquerecitrin),結(jié)構(gòu)式如下:2.2.3周延甲51h,d,d,nmr,c白色針狀結(jié)晶(水),mp254～256℃。TLC展開,紫外(254nm)下顯紫黑色,噴FeCl3顯色為深藍(lán)色,提示結(jié)構(gòu)中有酚羥基;鹽酸鎂粉反應(yīng)呈陰性。1H-NMR(600MHz,CD3OD),δH:6.86(1H,d,J=1.8Hz,H-2′),6.78(1H,d,J=8.4Hz,H-5′),6.75(1H,dd,J=8.1,2.4Hz,H-6′),5.95(1H,d,J=2.4Hz,H-8),5.88(1H,d,J=2.4Hz,H-6),4.58(1H,d,J=7.2Hz,H-2),3.99(1H,m,H-3),2.87(1H,dd,J=16.2,5.4Hz,H-4a),2.53(1H,dd,J=16.2,7.8Hz,H-4b)。13C-NMR(150MHz,CD3OD),δC:156.40(C-5),156.15(C-7),155.47(C-9),144.79(C-3′,4′),130.73(C-1′),119.09(C-6′),115.11(C-2′),114.35(C-5′),99.34(C-10),94.80(C-6),93.85(C-8),81.35(C-2),67.39(C-3),27.30(C-4)。結(jié)合上述圖譜并查閱文獻(xiàn),確定該化合物為(+)-兒茶素[(+)-catechin],結(jié)構(gòu)式如下:2.2.4c-nmr5.2值,c黃色油狀物(乙酸乙酯)。TLC展開,紫外燈(254nm)下顯紫黑色,噴FeCl3顯色為深藍(lán)色,提示結(jié)構(gòu)中有酚羥基;鹽酸鎂粉反應(yīng)呈陰性。1H-NMR(600MHz,CD3OD),δH:2.894(1H,dd,J=2.4,17.1Hz,H-4b),3.031(1H,dd,J=4.8,17.4Hz,H-4a),5.051(1H,brs,H-2),5.560(1H,brd,J=2.4Hz,H-3),6.006(1H,d,J=2.4Hz,H-8),6.010(1H,d,J=1.8Hz,H-6),6.737(1H,d,J=8.4Hz,H-5′),6.843(1H,dd,J=2.4,8.1Hz,H-6′),6.974(1H,d,J=1.8Hz,H-2′),6.993(1H,s,H-2″,6″)。13C-NMR(150MHz,CD3OD),δC:25.44(C-4),68.57(C-3),77.18(C-2),94.49(C-6),95.13(C-8),98.00(C-10),113.68(C-3′),114.60(C-5′),118.00(C-6′),130.04(C-1′),144.48(C-4′),144.50(C-5′),155.78(C-7),155.83(C-5),156.39(C-9);108.80(C-2″,6″),120.05(C-1″),138.35(C-4″),144.86(C-3″,5″),166.19(C=O)。結(jié)合上述圖譜并查閱文獻(xiàn),確定該化合物為(-)-表兒茶素-3-O-沒食子酸酯[(-)-epicatechin-3-O-gallate],結(jié)構(gòu)式如下:3黃酮類化合物+-表兒茶素[+-catechen,和[--表兒茶素]分離,未被完全分離的植物中,總黃酮類化合物2-5-o-gaha-3-o-garate,2,4-3-o-garate,5,5,5,5,5-o,5,5,5,5,5,5,5,5,5,5,5,5,5-o,5,5-o,5,5-o,5,5,5-o,5,5,5-o,5,5,5-o,5,5-o,5-o,5,5,5-o,5,5-o,5-o,5-o,5-o,5-o,5-o,5-o,5-o,5-garate,5-3-o,5-garate,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,5,本實(shí)驗(yàn)從新疆鎖陽花序95%的乙醇浸膏中分離鑒定得到4種黃酮類化合物,它們分別為:蕓香苷(rutin,I)、異槲皮苷(isoquer