杭白菊根茎叶有效成分的提取分离及其应用研究
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摘要
杭白菊是我国著名的药用和饮用菊，对其花的有效成分和药理作用
已进行了较多研究，结果表明其中存在黄酮、多糖、萜类等化合物并具
有广泛的生理活性，而对杭白菊根、茎、叶的相关研究尚属空白。浙江
桐乡是“中国杭白菊之乡”，每年有大量杭白菊根茎叶得不到有效利用
而废弃。鉴于次生代谢产物在植株中分布的广泛性，本文特对杭白菊根
茎叶的有效成分及其生物活性进行相关研究。结果如下：
1.对六种桐乡产菊花的花序（4种杭白菊、1种黄菊和1种野菊）
进行热回流提取，并测定其总酚和总黄酮含量及清除自由基活性。结果
表明不同品种间含量存在较大差异，异种大白菊的总酚和总黄酮含量
（以菊花干基计）最高，分别为9.76％和4.46％；最低的是黄山种野菊，
分别为3.21％和1.99％。不同菊花均显示了良好的抗自由基活性，清~·OH
和O~·_2~-的IC_(50)在0.54～1.98mg/ml和24.2～54.5μg/ml之间。其中清
O~·_2~-活性最强是大洋菊，最弱是软梗小洋菊；而清~·OH能力最强为早小
洋菊，最弱为异种大白菊。
2.通过L_9（3~4）表设计的正交试验得出杭白菊主要品种——小洋菊
叶中总黄酮提取的优化工艺参数为：乙醇浓度60％，料液比1:20，提取
时间1h。在此基础上，进行了小洋菊花及根茎叶有效成分提取的中试
规模试验，分别得到了四种提取物。全样分析显示，各提取物中常规成
分含量不存在显著差异，其重金属、农药残留、微生物指标等均符合食
用安全要求。
3.进一步研究表明，小洋菊花序及根茎叶的水提物中均存在黄酮、
酚酸、萜类及多糖等生物活性成分，其中总酚含量较高，且花序中高于
其它部位。分别用两种生物化学和化学发光体系来测定杭白菊提取物的
清除自由基活性。结果表明，植株不同部位的提取物均具有一定的清除
自由基活性，其清~·OH和O~·_2~-的IC_(50)分别在126～206μg/ml和8.25～
15.38μg/ml之间，并呈一定的量效关系。
4．安全毒理学评价结果表明，根茎叶提取物的大鼠30天喂养试验
无明显毒性反应，大体解剖未见异常病理改变，对大鼠体重、进食
量、食物利用率、血象、血生化及脏器无损害性影响，其无作用剂量
为 5刀叭g体重；对大、小鼠经口 LDS。，雌雄均大于 20 e体重，属无
毒类；对小鼠骨髓嗜多染红细胞微核试验、小鼠精子畸形试验以及
Anies试验检测结果均为阴性。
5．通过生物测定分析，进行了小洋菊叶浸提物对桃蚜杀虫活性的
时间－剂量．死亡率模型的研究。结果表明，其苯、甲醇、乙醚浸提物
对该模型具有良好的适合性。其中，甲醇提取物对桃蚜杀虫活性最
强，并具有速效性，但对小菜蛾幼虫仅有微效的毒杀作用。
6．根茎叶提取物作为鳖饲料添加剂的应用研究初步表明，在饲料
中添加 0．2％的提取物对稚鳖饲料适口性无影响，可使稚鳖成活率提高
2．3％，但对其生长无明显影响。
综上所述，杭白菊根茎叶提取物是一种纯天然、具有一定生物活
性、对人体安全无毒的新型植物制剂。通过进一步的深入研究，有望
将其应用于食品、保健食品‘、饲料添加剂、生物杀虫剂等领域，对提
高杭白菊产业的综合效益将大有稗益。
The flower of Hangbaiju is a kind of famous chrysanthemum
traditionally used as Chinese medicine and tea drink, of which effective
components and pharmacological effects have been studied by many
researchers. Those results showed that the flower contains many bio-active
compounds such as flavonoids, terpenoids and polysaccharids. However,
there is still a vacancy in the related research about its root, stem and leaf up
to now. Tongxiang city is 搕he kingdom of hangbayu?in China, in which a
mass of root, stem and leaf is abandoned annually due to a lack of effective
utilization. This paper was located on conducting the bio-active components
in the root, stem and leaf of Hangbaiju. Some results are as follows:
1) Six species of chrysanthemum flowers were collected from Tongxiang
city, microwave-dried and extracted individually by thermal reflux method
with 70% ethanol-water solution. The contents of total phenols (TP) and total
flavonoids (TF) of above extracts were measured by spectrophotometry.
Obvious difference was found in these chrysanthemums: Yizhongdabayu has
the highest contents at 9.76% TP and 4.46 %TF (in dry base of flower), and
Yeju is the lowest one at 3.21% TP and 1.99 %TF, respectively. Using the
Chemiluminescence method to evaluate the scavenging capacities of six
extracts on active oxygen free radical, their IC50 ranges are located in 0.54挆
1.98 mg/mi to 慜H and 24.2---54.5~gIml to ~2, respectively. Among those
chrysanthemums, Dayangju has the strongest scavenging activity to 02, and
Ruangengxiaoyang/u is the weakest one; similarly, Zaoxiaoyangju has the
strongest scavenging activity to 慜H and Yizhongdabaqu is the weakest one.
2) The extracting technology of total flavonoids from the leaves of
Xiaoyangju was optimized by means of a L~(34) orthogonal test. Ethanol
concentration 60%, solid/liquid ratio 1:20 and extracting time lhour were
regarded as optimized parameters. A series of large-scale extracting tests
were carried out using the flower, root, stem and leaf of Xiaoyangju as raw
material and 4 extracts were given out then. Chemical Analysis showed that
there is not significant difference of macro- and micro- nutrients among those
extracts. And other indexes such as heavy metal, pesticide residue and
microorganism are also according to hygienic demand.
3) Further studies revealed that all of the water-distilled extracts of
flowers and root-stem-leaf of Xiaoyangju contained?abundant flavonoids,
terpenoids and polysaccharide et al. Especially, the content of TP was much
high and the TP in flower was higher than in other part. The scavenging
capacity of these extracts on free radicals was also measured by two bio-
chemiluminescence systems.Their ranges of IC50 against 0H and 02 were
in 126---206~tgIml and 8.25?15.38j.tglml, respectively. And the scavenging
capacity displayed a concentration-dependent relationship.
4) The results of safety evaluation of root-stem-leaf extract conducted by
a rat 30-days-feeding test showed that the root-stem-leaf extract of
Xiaoyang/u did not present any significant toxic effect. Anatomising
examination did not show any abnormal pathologic changes. At the same
time, there were no harmful effects on body weight, appetite, feed efficiency,
viscera, and blood morphologic and biochemical indexes. So the no-effect
dosage was regarded as 5.Og/kg body weight. Acute toxic test showed that
the LI)50 both for rat and mice ,as well as male and female, was all above
2OgIkg body weights which meant that t
已进行了较多研究，结果表明其中存在黄酮、多糖、萜类等化合物并具
有广泛的生理活性，而对杭白菊根、茎、叶的相关研究尚属空白。浙江
桐乡是“中国杭白菊之乡”，每年有大量杭白菊根茎叶得不到有效利用
而废弃。鉴于次生代谢产物在植株中分布的广泛性，本文特对杭白菊根
茎叶的有效成分及其生物活性进行相关研究。结果如下：
1.对六种桐乡产菊花的花序（4种杭白菊、1种黄菊和1种野菊）
进行热回流提取，并测定其总酚和总黄酮含量及清除自由基活性。结果
表明不同品种间含量存在较大差异，异种大白菊的总酚和总黄酮含量
（以菊花干基计）最高，分别为9.76％和4.46％；最低的是黄山种野菊，
分别为3.21％和1.99％。不同菊花均显示了良好的抗自由基活性，清~·OH
和O~·_2~-的IC_(50)在0.54～1.98mg/ml和24.2～54.5μg/ml之间。其中清
O~·_2~-活性最强是大洋菊，最弱是软梗小洋菊；而清~·OH能力最强为早小
洋菊，最弱为异种大白菊。
2.通过L_9（3~4）表设计的正交试验得出杭白菊主要品种——小洋菊
叶中总黄酮提取的优化工艺参数为：乙醇浓度60％，料液比1:20，提取
时间1h。在此基础上，进行了小洋菊花及根茎叶有效成分提取的中试
规模试验，分别得到了四种提取物。全样分析显示，各提取物中常规成
分含量不存在显著差异，其重金属、农药残留、微生物指标等均符合食
用安全要求。
3.进一步研究表明，小洋菊花序及根茎叶的水提物中均存在黄酮、
酚酸、萜类及多糖等生物活性成分，其中总酚含量较高，且花序中高于
其它部位。分别用两种生物化学和化学发光体系来测定杭白菊提取物的
清除自由基活性。结果表明，植株不同部位的提取物均具有一定的清除
自由基活性，其清~·OH和O~·_2~-的IC_(50)分别在126～206μg/ml和8.25～
15.38μg/ml之间，并呈一定的量效关系。
4．安全毒理学评价结果表明，根茎叶提取物的大鼠30天喂养试验
无明显毒性反应，大体解剖未见异常病理改变，对大鼠体重、进食
量、食物利用率、血象、血生化及脏器无损害性影响，其无作用剂量
为 5刀叭g体重；对大、小鼠经口 LDS。，雌雄均大于 20 e体重，属无
毒类；对小鼠骨髓嗜多染红细胞微核试验、小鼠精子畸形试验以及
Anies试验检测结果均为阴性。
5．通过生物测定分析，进行了小洋菊叶浸提物对桃蚜杀虫活性的
时间－剂量．死亡率模型的研究。结果表明，其苯、甲醇、乙醚浸提物
对该模型具有良好的适合性。其中，甲醇提取物对桃蚜杀虫活性最
强，并具有速效性，但对小菜蛾幼虫仅有微效的毒杀作用。
6．根茎叶提取物作为鳖饲料添加剂的应用研究初步表明，在饲料
中添加 0．2％的提取物对稚鳖饲料适口性无影响，可使稚鳖成活率提高
2．3％，但对其生长无明显影响。
综上所述，杭白菊根茎叶提取物是一种纯天然、具有一定生物活
性、对人体安全无毒的新型植物制剂。通过进一步的深入研究，有望
将其应用于食品、保健食品‘、饲料添加剂、生物杀虫剂等领域，对提
高杭白菊产业的综合效益将大有稗益。
The flower of Hangbaiju is a kind of famous chrysanthemum
traditionally used as Chinese medicine and tea drink, of which effective
components and pharmacological effects have been studied by many
researchers. Those results showed that the flower contains many bio-active
compounds such as flavonoids, terpenoids and polysaccharids. However,
there is still a vacancy in the related research about its root, stem and leaf up
to now. Tongxiang city is 搕he kingdom of hangbayu?in China, in which a
mass of root, stem and leaf is abandoned annually due to a lack of effective
utilization. This paper was located on conducting the bio-active components
in the root, stem and leaf of Hangbaiju. Some results are as follows:
1) Six species of chrysanthemum flowers were collected from Tongxiang
city, microwave-dried and extracted individually by thermal reflux method
with 70% ethanol-water solution. The contents of total phenols (TP) and total
flavonoids (TF) of above extracts were measured by spectrophotometry.
Obvious difference was found in these chrysanthemums: Yizhongdabayu has
the highest contents at 9.76% TP and 4.46 %TF (in dry base of flower), and
Yeju is the lowest one at 3.21% TP and 1.99 %TF, respectively. Using the
Chemiluminescence method to evaluate the scavenging capacities of six
extracts on active oxygen free radical, their IC50 ranges are located in 0.54挆
1.98 mg/mi to 慜H and 24.2---54.5~gIml to ~2, respectively. Among those
chrysanthemums, Dayangju has the strongest scavenging activity to 02, and
Ruangengxiaoyang/u is the weakest one; similarly, Zaoxiaoyangju has the
strongest scavenging activity to 慜H and Yizhongdabaqu is the weakest one.
2) The extracting technology of total flavonoids from the leaves of
Xiaoyangju was optimized by means of a L~(34) orthogonal test. Ethanol
concentration 60%, solid/liquid ratio 1:20 and extracting time lhour were
regarded as optimized parameters. A series of large-scale extracting tests
were carried out using the flower, root, stem and leaf of Xiaoyangju as raw
material and 4 extracts were given out then. Chemical Analysis showed that
there is not significant difference of macro- and micro- nutrients among those
extracts. And other indexes such as heavy metal, pesticide residue and
microorganism are also according to hygienic demand.
3) Further studies revealed that all of the water-distilled extracts of
flowers and root-stem-leaf of Xiaoyangju contained?abundant flavonoids,
terpenoids and polysaccharide et al. Especially, the content of TP was much
high and the TP in flower was higher than in other part. The scavenging
capacity of these extracts on free radicals was also measured by two bio-
chemiluminescence systems.Their ranges of IC50 against 0H and 02 were
in 126---206~tgIml and 8.25?15.38j.tglml, respectively. And the scavenging
capacity displayed a concentration-dependent relationship.
4) The results of safety evaluation of root-stem-leaf extract conducted by
a rat 30-days-feeding test showed that the root-stem-leaf extract of
Xiaoyang/u did not present any significant toxic effect. Anatomising
examination did not show any abnormal pathologic changes. At the same
time, there were no harmful effects on body weight, appetite, feed efficiency,
viscera, and blood morphologic and biochemical indexes. So the no-effect
dosage was regarded as 5.Og/kg body weight. Acute toxic test showed that
the LI)50 both for rat and mice ,as well as male and female, was all above
2OgIkg body weights which meant that t
引文
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