【作者】 郑莹；

【导师】 金永日；

【作者基本信息】 吉林大学 ， 药物化学， 2004， 硕士

【摘要】 三七[Panax Notoginseng(Burk.)EH.chen]又名参三七、田七、三七、金不换，为五加科人参属多年生草本植物，是一种用途比较广泛的名贵中药材，主要分布于我国西南部。药理研究表明，三七具有抗氧化、抗衰老、抗炎、护肝利胆、止血、抗冠心病、抗心律失常、扩张血管和降压等作用。至今为止，三七及三七茎叶化学成分的研究主要集中在皂苷类成分上，而关于对治疗心脑血管疾病有明显疗效的黄酮类成分的分离鉴定、含量测定、药理活性研究却鲜见报道。 本实验从三七茎叶中提取、分离到了七个黄酮类单体化合物，根据其理化性质和光谱分析鉴定出了其中六种的结构，分别是山柰酚-3-O-β-D-半乳糖(2→1)葡萄糖苷(SA)、槲皮素-3-O-B-D-半乳糖(2→1)葡萄糖苷(SB)、山柰酚-3-O-β-D-半乳糖苷(SC)、山柰酚(SG)、槲皮素(SH)、山柰酚-7-O-α-L-鼠李糖苷(SI)。除山柰酚与槲皮素之外，其余五种化合物皆为首次从本植物中分离得到。本实验还建立了山柰酚-3-O-β-D-半乳糖(2→1)葡萄糖苷与槲皮素-3-O-β-D-半乳糖(2→1)葡萄糖苷这两种主要黄酮成分的HPLC含量测定方法，并且对三七叶中所含的这两种成分进行了含量测定。 一．黄酮类单体化合物的提取、分离、鉴定 1．实验材料与仪器 ABI公司(美国应用生物系统公司)Q-STAR质谱仪，BRUKER AV600型质谱仪(600HZ)，美国尼高立公司AVATAR 330型红外光谱仪，Unico UV-2102 PCS型紫外可见全波长扫描仪，Koffiler显微熔点测定仪(未校正)，硅胶(柱层析，200-300目)青岛海洋化工厂吉林大学硕士学位论文出品，ODS OUYA一RP18(40一60um)北京金欧亚科技发展公司出品，AB一8大孔吸附树脂(0 .3一1.25mm)南开大学化工厂出品，其余试剂皆为分析纯。三七茎叶采于云南省文山县，经吉林大学药学院生药教研室王广树鉴定为三七【Panax Nofoginseng(B urk.少F.日.Chen】的茎叶。2.提取分离 将干燥的三七茎叶ZKg粉碎后加水煎煮提取三次(用水量分别为3OL、24L、20L;煎煮提取时间分别为2h，，.sh，，h)，合并三次提取液通过大孔吸附树脂柱吸附，水洗，95%的乙醇解吸。浸膏用硅胶柱以氯仿一甲醇为洗脱剂梯度洗脱得到化合物SA(4 29)、sB(3 .19)、sG(195mg)、sH(22mg)、51(35mg)。以oDS柱以甲醇水梯度洗脱得到SC(68mg)、SD(gmg)。3.结构鉴定 通过其理化性质、紫外光谱、红外光谱、ESI一MS、，HNMR、，℃NMR鉴定并与文献对照确定SA为山奈酚一3一O一p一O一半乳糖一(2-+1)葡萄糖普，SB为檄皮素一3一O一p一O一半乳糖一(2一1)葡萄糖普、SC为山奈酚一3一O一p一O一半乳糖普、SG为山奈酚、S日为榭皮素、Sl为山奈酚一7一O一a一L一鼠李糖普。通过50理化性质、紫外光谱、ESI一MS、IHNMR、13CNMR鉴定SD为山奈酚的C3一OH上连有两分子糖的化合物，而且糖的某个O日被乙酞化，其结构正在进一步鉴定当中。二.三七茎叶中SA、SB的含量测定2.，仪器及试药仪器设备:Agilent 1100 Series高效液相色谱仪，ZORBAX Extend一C18 4.6x250mm ODS，spm色谱柱试药:三七叶采自云南省文山县，甲醇为色谱纯，乙酸为分析纯.吉林大学硕士学位论文2.2方法与结果2.2.1色谱条件:流动相为甲醇:水(35:65)，含0.1%的乙酸;流速 1.2m}/min;检测波长268nm;柱温25oC。22.2标准曲线的绘制 在0.5一20阳范围内SA、SB进样量与峰面积均呈现良好的线性关系。 SA标准曲线为:y=1312.5x+92.5r=0.9999; SB标准曲线为:y=1157.6x一75.7r=0.9999。22.3重现性实验 SA及S日的6次进样的峰面积RSO值SA为0.26%，S日为0.44%，表明方法重现性良好.2.2.4精密度实验 同一样品五次平行测定，SA含量的RSO值为0.41%，SB含量的RSO值为0.43%，表明方法精密度良好。2.2.5稳定性实验 同一样品溶液每2小时进样一次，计算其峰面积RSO值SA为0.42%，SB为0.54%，表明样品水溶液在0一12小时内稳定性良好，。2.2.6回收率实验 SA平均回收率为97.5%，RSO为0.78%;SB平均回收率为10，.8%，RSO为1.，0%，表明方法回收率良好。2.2.7三七叶黄酮类成分SA、SB含量测定 利用水煎煮，大孔吸附树脂吸附提取，测得三七叶中SA含量为0.17%，SB含量为0.30%;利用甲醇回流提取测得三七叶中SA含量为0.加%，S日含量为0.38%，水提取率为甲醇提取率的80%，更多还原

【Abstract】 Panax Notoginseng(Burk.) F.H.chen, a well-known medicinal plant , which is widely used for treatment of cardiovascular diseases, inflammation, different body pains, trauma , bleeding caused by internal and external injury. It was been also used as a tonic and haemostatic agent. Due to the demand for its important medicinal use in China, it was largely cultivated in Yunnan Province.The saponins in this plant have been reported more before, but the flavonoids reported fewer. For the sake of the better use of this herb medicine, this paper describes the isolation, structural determinations and the analysis of the flavonids from the dried stems and leaves of Panax Notoginseng. Extraction and isolationDried stems and leaves (2kg)were extracted with water three times(30L ,24L,20L;2h,1.5h, lh).The extracted solution was absorbed by macroporous resin, then eluented with 95%EtOH. The extraction was subjected to silica gel column chromatography using methanol-chloroform gradient aseluent to afford SA (4.2g) ,SB (3.1g) ,SG (195mg) ,SH ( 22mg )and SI( 35mg ). The extraction was subjected to ODS column chromatography using methanol-water gradient as eluent to afford SC (68mg) and SD (9mg) .Compounds SA, SB, SC, SQ SH and SI was identified as kaempferol 3 - O - ( 2" - p - D - glucopyranosyl) - |3 - D- galactopyranoside, quercetin 3 - O - ( 2" - (3 - D -glucopyranosyl) - |3 - D - galactopyranoside, kaempferol 3- O - P - D - galactoside, kaempferol, quercetin, kaempferol 7-O-a-L- rhamnoside respectively by UV, IR, MS,JHNMR and 13CNMR.Compound SD is not identified now and hoped to be identified in the future .But for SH and SG ,all the compounds were isolated from Panax Notoginseng for the first time.Analysis offlavonids in the leaves of Panax Notoginseng In this paper we present a more sensitive quantificationmethod by HPLC for analyzing the main flavonoids (SA andSB) in the leaves of Panax Notoginseng. HPLC was performed on a ZORBAX C18 column (250vmmx4.6 mm,5 JJL m )at 25 癈 with the rate of 1.2 ml/mm , using 35% methanol and 65% water/acetic acid mixture (100/1 v/v) as mobile phase.The flavonol compounds SA and SB were extracted with different solvents: methanol and water .Two extraction methods have been compared in this paper. The results show that the content of SA and SB extracted with methanol was 0.20% and 0.38% respectively compared to the results 0.17% and 0.30% extracted with water.更多还原