陕西地方习用药材落新妇的质量标准及岩白菜素的提取工艺研究
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摘要
落新妇(Astilbe chinensis (Maxim.) Franch.et Sav.)系虎耳草科(Saxifragaceae)落新妇属(Astilbe)多年生直立草本植物，根茎供药用。现代大量的药理研究和临床应用证明其具有抗肿瘤、镇痛、活血祛瘀、抗炎利尿、止咳祛痰、退热、抗菌、抗内毒素的生理活性。落新妇有效化学成分岩白菜素具有止咳、抗炎、护肝、抗溃疡、提高免疫等作用，目前已经被开发成各种剂型药物，应用于临床，其治疗支气管炎有独到之处。落新妇产于我国多省，其在陕西省的自然资源量较大，秦岭—大巴山地区是落新妇的主产区之一。
本论文主要对陕西地方习用药材落新妇的质量标准及其有效化学成分岩白菜素的提取工艺进行研究，为该药的资源开发利用和质量标准研究提供科学依据。主要结果如下：
1、通过常规化学成分预试验研究，结果表明，落新妇根茎中含有黄酮类、鞣质、氨基酸类、三萜类、酚性成分、香豆素类成分，不含有蒽醌类成分。
2、本试验探究了回流提取法与超声波辅助提取法提取落新妇中岩白菜素的最佳工艺条件，通过单因素试验和正交试验，确定最佳工艺为超声波辅助提取，提取条件是：以45%乙醇为提取溶剂，提取时间为10min，料液比1∶15，提取温度为60℃，提取功率为50W，提取次数为2次。在此最佳工艺条件下，岩白菜素得率为2.68%。各因素对岩白菜素得率的影响顺序为：乙醇体积分数＞提取时间＞料液比＞提取温度。
3、落新妇根茎横切面显微特征是：皮层组织内分布有维管束。无限外韧型维管束多而小，呈环状排列，髓射线较宽，具有明显发达的髓部。药材粉末棕黄色，淀粉粒较多，呈椭圆型或杆状，草酸钙簇晶较大且明显，螺纹或梯纹导管，并有大量黄棕色物质分布。
4、落新妇药材水分含量不宜超过16.5%，总灰分不宜高于11.0%，酸不溶性灰分不宜高于1.0%。水溶性浸出物测定中，热浸法测得的含量高于冷浸法，因此，水溶性浸出物测定应宜采用热浸法，且含量不低于11.5%。醇溶性浸出物测定中，热浸法测得的含量远高于冷浸法，且60%乙醇为最佳溶媒，应选择热浸法以60%乙醇作为溶剂进行醇溶性浸出物测定，含量不应低于29.0%。
5、建立高效液相色谱（HPLC）法来测定岩白菜素含量，岩白菜素进样量在0.004～0.06mg·mL~(-1)范围内，275nm波长处有良好的线性关系（R~2=0.9997）。岩白菜素含量平均值为4.17%，最低1.99%，最高值为7.60%。平均加样回收率为99.86%，RSD为0.98%(n=5)。该方法操作简单、准确、重现性好，可用于落新妇药材中岩白菜素的含量测定。为了保证落新妇药材质量，拟定：岩白菜素含量不得低于2.0%。
6、本实验中有关落新妇质量研究的内容和各项指标，可以作为鉴定落新妇和制定落新妇药材质量标准的科学参考依据。
Astilbe chinensis (Maxim.) Franch.et Sav. is a genus of perennial vertical herb, whichbelongs to the family Astilbe of Saxifragaceae, and the rhizome can be used for medicine. Itsmain efficacy are antitumor, analgesia, promoting blood circulation, anti-inflammatory anddiuretic, relieving cough and reducing sputum, defervesce, antibiosis and antiendotoxin whichwere demonstrated by a lot of pharmacological researches and clinical applicationes.Bergenin, known as the effective chemical composition has been developed into a variety ofdosage form drugs due to its functions, such as relieving cough, anti-inflammatory, protectingliver, anti-ulcer, strengthening immune system and so on. It is an exclusive drug treating thebronchitis. China has an abundant resource of Astilbe chinensis (Maxim.) Franch.et Sav., themost abundant natural resource is located in Shaanxi Province, and Qinling-Daba mountain isproduction area.
This study was mainly to evaluate the quality standards and the extraction of thebergenin which in order to provide the scientific basis of development, utilization and qualitystandards. The main results showed as follows:
1. According to the method of physical and chemical identification for determining thechemical constituents, the results showed that the rhizome of Astilbe chinensis (Maxim.)Franch.et Sav. contain flavonoids, tannins, amino acids, triterpenes, phenol-ic constituents,coumarins and no anthraquinones.
2. To compare the ultrasound extraction and continuous reflux extraction process ofbergenin from Astilbe chinensis(Maxim.)Franch.et Sav. by single factor experiments andorthogonal test, then determined the ultrasound extraction was better. The best optimalconditions were as follows, ethanol concentration45%, extraction time10min, solid-liquidratio1:15, extraction temperature60℃, extraction intensity50W, extraction times2. Thehigh extraction ratio of bergenin is2.68%. The sequence of all factors are ethanolconcentration> extraction time> solid-liquid ratio> extraction temperature.
3. The microstructure characteristics of rhizome transverse section were as follows:vascular bundles can be found in cortical, the type of infinite external tough vascular are many but small which showed annular arrangement, the pith rays are wide and the pith aresignificantly developed. There are many ellipse or rod starch grains in powder, calciumoxalate cluster crystals are large and obvious, thread and ladder stria catheter, and a largenumber of yellow-brown substance distribution.
4. The moisture in Chinese Astilbe Rhizome was less than16.5%, with the total ashescontent within11.5%, acid insoluble ash content was that lower than1.0%, the content ofwater extract with hot soak was higher than11.0%, that of60%ethanol extract with hot soakwas higher no less than29.0%.
5. According to the method of the high performance liquid chromatography (HPLC) fordetermining the bergenin. It was good linear range of bergenin in the range from0.004～0.06mg·mL~(-1)and275nm (R~2=0.9997). The average recovery was99.86%and the RSD was0.98%.This method is simple, accurate and reproducible which could be used for the bergenindetermination of Astilbe chinensis (Maxim.) Franch.et Sav..In order to ensure the quality,bergenin content was no less than2.0%.
6. The identification from quality standards research of Astilbe chinensis (Maxim.)Franch.et Sav. can be used as the basis for identification, which could provide a reference forsetting up the quality standards research.
本论文主要对陕西地方习用药材落新妇的质量标准及其有效化学成分岩白菜素的提取工艺进行研究，为该药的资源开发利用和质量标准研究提供科学依据。主要结果如下：
1、通过常规化学成分预试验研究，结果表明，落新妇根茎中含有黄酮类、鞣质、氨基酸类、三萜类、酚性成分、香豆素类成分，不含有蒽醌类成分。
2、本试验探究了回流提取法与超声波辅助提取法提取落新妇中岩白菜素的最佳工艺条件，通过单因素试验和正交试验，确定最佳工艺为超声波辅助提取，提取条件是：以45%乙醇为提取溶剂，提取时间为10min，料液比1∶15，提取温度为60℃，提取功率为50W，提取次数为2次。在此最佳工艺条件下，岩白菜素得率为2.68%。各因素对岩白菜素得率的影响顺序为：乙醇体积分数＞提取时间＞料液比＞提取温度。
3、落新妇根茎横切面显微特征是：皮层组织内分布有维管束。无限外韧型维管束多而小，呈环状排列，髓射线较宽，具有明显发达的髓部。药材粉末棕黄色，淀粉粒较多，呈椭圆型或杆状，草酸钙簇晶较大且明显，螺纹或梯纹导管，并有大量黄棕色物质分布。
4、落新妇药材水分含量不宜超过16.5%，总灰分不宜高于11.0%，酸不溶性灰分不宜高于1.0%。水溶性浸出物测定中，热浸法测得的含量高于冷浸法，因此，水溶性浸出物测定应宜采用热浸法，且含量不低于11.5%。醇溶性浸出物测定中，热浸法测得的含量远高于冷浸法，且60%乙醇为最佳溶媒，应选择热浸法以60%乙醇作为溶剂进行醇溶性浸出物测定，含量不应低于29.0%。
5、建立高效液相色谱（HPLC）法来测定岩白菜素含量，岩白菜素进样量在0.004～0.06mg·mL~(-1)范围内，275nm波长处有良好的线性关系（R~2=0.9997）。岩白菜素含量平均值为4.17%，最低1.99%，最高值为7.60%。平均加样回收率为99.86%，RSD为0.98%(n=5)。该方法操作简单、准确、重现性好，可用于落新妇药材中岩白菜素的含量测定。为了保证落新妇药材质量，拟定：岩白菜素含量不得低于2.0%。
6、本实验中有关落新妇质量研究的内容和各项指标，可以作为鉴定落新妇和制定落新妇药材质量标准的科学参考依据。
Astilbe chinensis (Maxim.) Franch.et Sav. is a genus of perennial vertical herb, whichbelongs to the family Astilbe of Saxifragaceae, and the rhizome can be used for medicine. Itsmain efficacy are antitumor, analgesia, promoting blood circulation, anti-inflammatory anddiuretic, relieving cough and reducing sputum, defervesce, antibiosis and antiendotoxin whichwere demonstrated by a lot of pharmacological researches and clinical applicationes.Bergenin, known as the effective chemical composition has been developed into a variety ofdosage form drugs due to its functions, such as relieving cough, anti-inflammatory, protectingliver, anti-ulcer, strengthening immune system and so on. It is an exclusive drug treating thebronchitis. China has an abundant resource of Astilbe chinensis (Maxim.) Franch.et Sav., themost abundant natural resource is located in Shaanxi Province, and Qinling-Daba mountain isproduction area.
This study was mainly to evaluate the quality standards and the extraction of thebergenin which in order to provide the scientific basis of development, utilization and qualitystandards. The main results showed as follows:
1. According to the method of physical and chemical identification for determining thechemical constituents, the results showed that the rhizome of Astilbe chinensis (Maxim.)Franch.et Sav. contain flavonoids, tannins, amino acids, triterpenes, phenol-ic constituents,coumarins and no anthraquinones.
2. To compare the ultrasound extraction and continuous reflux extraction process ofbergenin from Astilbe chinensis(Maxim.)Franch.et Sav. by single factor experiments andorthogonal test, then determined the ultrasound extraction was better. The best optimalconditions were as follows, ethanol concentration45%, extraction time10min, solid-liquidratio1:15, extraction temperature60℃, extraction intensity50W, extraction times2. Thehigh extraction ratio of bergenin is2.68%. The sequence of all factors are ethanolconcentration> extraction time> solid-liquid ratio> extraction temperature.
3. The microstructure characteristics of rhizome transverse section were as follows:vascular bundles can be found in cortical, the type of infinite external tough vascular are many but small which showed annular arrangement, the pith rays are wide and the pith aresignificantly developed. There are many ellipse or rod starch grains in powder, calciumoxalate cluster crystals are large and obvious, thread and ladder stria catheter, and a largenumber of yellow-brown substance distribution.
4. The moisture in Chinese Astilbe Rhizome was less than16.5%, with the total ashescontent within11.5%, acid insoluble ash content was that lower than1.0%, the content ofwater extract with hot soak was higher than11.0%, that of60%ethanol extract with hot soakwas higher no less than29.0%.
5. According to the method of the high performance liquid chromatography (HPLC) fordetermining the bergenin. It was good linear range of bergenin in the range from0.004～0.06mg·mL~(-1)and275nm (R~2=0.9997). The average recovery was99.86%and the RSD was0.98%.This method is simple, accurate and reproducible which could be used for the bergenindetermination of Astilbe chinensis (Maxim.) Franch.et Sav..In order to ensure the quality,bergenin content was no less than2.0%.
6. The identification from quality standards research of Astilbe chinensis (Maxim.)Franch.et Sav. can be used as the basis for identification, which could provide a reference forsetting up the quality standards research.
引文
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