锁阳化学成分的初步研究

锁阳化学成分的初步研究

锁阳是锁定科的植物，具有除花序过程中干燥的肉质茎，具有益肾阳、益血、润肠排便的功效。由于大量采挖,资源濒临枯竭,本实验对锁阳花序中黄酮类化学成分进行初步研究,以更加合理地利用锁阳资源。1柱层析硅胶、聚酰胺薄膜X4型显微熔点测定仪(北京第三光学仪器厂),Unity-Inova600超导核磁共振谱仪(美国Varian公司),柱层析硅胶(100～200目,青岛海洋化工厂),薄层层析硅胶G(青岛海洋化工厂),柱层析聚酰胺(30～60目,浙江省台州市路桥四甲生化塑料厂),聚酰胺薄膜(薄层层析,浙江省台州市路桥四甲生化塑料厂)。锁阳药材采自新疆和硕地区,其品种由新疆医科大学天药/生药教研室鉴定。2方法和结果2.1萃取物的制备干燥的锁阳花序切片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,处理待用。将提取浸膏120g上聚酰胺柱(60～100目),氯仿∶甲醇(8∶2～0∶1)梯度洗脱,共得到100个流份(Fr.J1-100);Fr.J11-85以水混悬,用乙酸乙酯萃取,得乙酸乙酯萃取物;将乙酯萃取物经硅胶柱层析,石油醚∶乙酸乙酯(1∶0～0∶1)、氯仿∶甲醇(1∶0～0∶1)梯度洗脱,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;将含有化合物Ⅲ的混合物2g上聚酰胺柱(60～100目),水∶丙酮(1∶0～0∶1)梯度洗脱,得到化合物Ⅲ。2.2结构评价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黄色粉末(甲醇),mp176～178℃,盐酸-镁粉反应阳性,Molish反应阳性。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,糖质子峰),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‴);结合上述图谱数据并查阅文献,确定该化合物为芸香苷(Rutin),结构式如下:2.2.2糖酸水质分布淡黄色粉末(甲醇),mp193～195℃。盐酸-镁粉反应阳性,Molish反应阳性。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,糖质子峰)。氢谱中的6.23d,6.42d峰为5,7-二羟基黄酮类中H6,H8的特征峰,说明可能是5,7-二羟基黄酮类化合物。与芸香苷相比,异槲皮苷的氢谱中少了鼠李糖在δ1.02处-CH3的二重峰和端基质子在δH4.42处的峰。结合上述图谱并查阅文献,最后鉴定其结构式为槲皮素-3-O-β-D-葡萄糖,故确定该化合物为异槲皮苷(isoquerecitrin),结构式如下:2.2.3周延甲51h,d,d,nmr,c白色针状结晶(水),mp254～256℃。TLC展开,紫外(254nm)下显紫黑色,喷FeCl3显色为深蓝色,提示结构中有酚羟基;盐酸镁粉反应呈阴性。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)。结合上述图谱并查阅文献,确定该化合物为(+)-儿茶素[(+)-catechin],结构式如下:2.2.4c-nmr5.2值,c黄色油状物(乙酸乙酯)。TLC展开,紫外灯(254nm)下显紫黑色,喷FeCl3显色为深蓝色,提示结构中有酚羟基;盐酸镁粉反应呈阴性。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)。结合上述图谱并查阅文献,确定该化合物为(-)-表儿茶素-3-O-没食子酸酯[(-)-epicatechin-3-O-gallate],结构式如下:3黄酮类化合物+-表儿茶素[+-catechen,和[--表儿茶素]分离,未被完全分离的植物中,总黄酮类化合物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,本实验从新疆锁阳花序95%的乙醇浸膏中分离鉴定得到4种黄酮类化合物,它们分别为:芸香苷(rutin,I)、异槲皮苷(isoquer