蟾酥质量分析及其炮制前后指纹图谱研究
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摘要
目的:
通过对山东临沂、河南、河北等产地的蟾酥有效成分的提取方法、含量、炮制工艺、指纹图谱、炮制前后指纹图谱变化方面进行研究,建立华蟾酥毒基和脂蟾毒配基的最佳提取方法和含量测定方法、蟾酥最佳酒炮制工艺、蟾酥TLC指纹图谱与HPLC指纹图谱,并比较蟾酥不同炮制样品之间及炮制前后指纹图谱的差别,为蟾酥的炮制规范化、质量控制标准化和进一步的应用开发等提供科学依据。
方法:
采用HPLC法,以华蟾酥毒基和脂蟾毒配基的提取率为评价指标,分别进行不同溶剂加热回流提取和超声提取比较考察、不同浓度乙醇单因素考察,以及L_9(3~4)正交试验,筛选蟾酥最佳提取工艺,并对蟾酥中的华蟾酥毒基和脂蟾毒配基进行含量测定。采用HPLC法,以华蟾酥毒基和脂蟾毒配基的含量为评价指标,选择市售白酒、牛奶、滑石粉作蟾酥炮制辅料单因素考察,并通过L_9(3~4)正交试验进一步考察酒炮制工艺,筛选蟾酥药材最佳炮制工艺。采用TLC法对12批蟾酥药材的薄层图谱进行分析。采用HPLC法进行梯度洗脱,对12批蟾酥药材的指纹图谱进行分析。采用TLC和HPLC指纹图谱法比较蟾酥不同方法及炮制前后指纹图谱的变化。
结果:
1.蟾酥有效成分最佳提取方法:加入50倍量80%乙醇70℃提取两次,每次90 min。
2.建立了华蟾酥毒基和脂蟾毒配基的含量测定方法,并测定了12批蟾酥药材,华蟾酥毒基和脂蟾毒配基的含量范围分别是0.937%～4.893%和0.198%～6.692%。
3.蟾酥经三种辅料炮制后有效成分含量均有所下降,其中酒炮制后成化变化较小,选择对酒炮制作进一步研究。优选出蟾酥酒炮制最佳工艺是:加入2倍量55%乙醇(白酒),在60℃下炮制12小时,干燥,粉碎。蟾酥经酒炮制后,有效成分的总含量比原药材降低约20%。
4.蟾酥TLC指纹图谱以V环己烷:V二氯甲烷:V丙酮(2:1:1)为展开剂,二次展开结果最佳,可分离出12个紫外荧光斑点和9个可见光斑点,斑点分离度好。不同批次产地的蟾酥样品的斑点数目相同,斑点颜色和大小因产地不同而各有差异。
5.测定了12批蟾酥的HPLC指纹图谱,从中标定了26个共有峰,并划分为2个区域,第Ⅰ区(RT=0～20 min)由9个共有峰组成,不同产地蟾酥在此区间峰貌相似;第Ⅱ区(RT=20～40 min)由17个共有峰组成,是体现蟾酥产地差异的主要特征区域。对12批蟾酥进行相似度评价均大于0.9,整体相似度较好,初步建立了蟾酥药材的共有峰模式。
6.不同炮制品的TLC指纹图谱薄层行为与蟾酥药材相似。同一批次蟾酥酒炮制和牛奶炮制样品的斑点数目与炮制前基本相同,但斑点颜色较淡;滑石粉炮制样品斑点比该产地生品、酒炮制品和牛奶炮制品的斑点颜色淡,且部分斑点有明显变化。同法对多个批次蟾酥进行对比,结果相似。同一批次蟾酥经不同酒度炮制后,样品斑点数目与生品基本相同,炮制品的斑点颜色较浅,炮制品之间的斑点颜色没有明显差异。
7.不同炮制品的HPLC指纹图谱整体面貌与蟾酥药材相似,各共有峰基本存在,保留时间与相对保留时间基本一致,但共有峰强度差别较大。同一批次蟾酥经牛奶和滑石粉炮制后,共有峰相对峰面积变化较大,酒炮制后的共有峰相对峰面积变化较小。同法考察了多个批次蟾酥,结果相似。同一批次蟾酥经不同酒度炮制后,除6号峰的相对峰面积经炮制后有所增加外,其他共有峰的相对峰面积均呈降低的趋势。
结论:
1.新建的提取方法对蟾酥华蟾酥毒基和脂蟾毒配基的提取率高于2005版《中国药典》的方法,能够充分提取蟾酥中的有效成分。
2.新建的蟾酥有效成分含量测定方法优于2005版《中国药典》的测定方法。
3.规范了蟾酥的酒炮制工艺参数,为蟾酥炮制进一步研究提供实验依据。
4.蟾酥TLC指纹图谱能分离药材中多个成分,可用于蟾酥药材鉴别及质量评价。
5.蟾酥HPLC指纹图谱能够对蟾酥的总体成分进行系统分析,提供了丰富的信息,可用于评价蟾酥药材品质及对质量的全面控制。
6.蟾酥TLC指纹图谱和HPLC指纹图谱可用于评价炮制前后蟾酥的成分变化,能从整体上对蟾酥炮制前后各成分的变化情况进行分析。蟾酥炮制后各成分基本呈下降趋势,这可能是蟾酥酒炮制后降毒的原因。
7.初步建立了蟾酥炮制品质量标准。若能得到进一步验证,可作为行业中蟾酥炮制质量控制的参考基础和依据。
Objective:
Study on the component extraction technology, contents, processing technology, fingerprints of Venenum Bufonis which come from Shan Dong, He Nan, He Bei and so on. And research the changes of their fingerprint of the crude and processed. To build up the best method to extract and a method to determine the Venenum Bufonis contents Cinobufagin and resibufogenin. To establish the best wine processing method, the TLC fingerprint method and the HPLC fingerprint method of Venenum Bufonis. Compare with various processed samples and the crude to find out the differences by fingerprint methods, in order to offer scientific basis to standardize the processing technology, control the quality and for further develop and apply of Venenum Bufonis.
Method:
Measure the total extraction rate of Cinobufagin and Resibufogenin by HPLC to investigate the best extraction technology through selecting different solvents by heating reflux or ultrasonic extraction, designing single factor exploration of different ethanol concentration, and applying the L_9(3~4) of orthogonal test.
Measure the content of Cinobufagin and Resibufogenin by HPLC.
Study the wine, milk and talcum powder as the processing excipient in single factor experiment. Select the wine as the suitable excipient. Then apply the L_9(3~4) of orthogonal test and determine the total content of Cinobufagin and Resibufogenin by HPLC to build up the optimal processing technology.
To analysis 12 groups Venenum Bufonis by TLC method.
To measure 12 groups Venenum Bufonis by HPLC method and gradient elution, and analysis the fingerprint by the computer similarity evaluation system.
Compare with various processed samples and the crude by TLC & HPLC fingerprint methods to find out the differences between before and after processed.
Result:
1. The optimal extraction technology of Cinobufagin and Resibufogenin of Venenum Bufonis is: add 50 times 80% ethanol and heating reflux twice, 90min per time.
2. A new method for measuring the quality of Cinobufagin and Resibufogenin is established. 12 groups of Venenum Bufonis quantitative determination results show that, the contents circumscription of Cinobufagin is 0.937% to 6.520%. And the contents circumscription of Resibufogenin is 0.198% to 7.550%.
3. The contents of Venenum Bufonis are both decline after wine, milk and talcum power processing. Compared with the milk processing and talcum power processing, the change of Venenum Bufonis component is more stable by wine processed. Choose wine as the processing excipient for further study. The results of the orthogonal test, the signal factor experiment of different ethanol concentration and the verification test show that the optimal processing technology is: add 2 times 55% ethanol (wine), heating processing in 60℃for 12 hours, aired and comminuted. Compare to the crude Venenum Bufonis, the quality of the effective component are decrease about 20% after wine processing.
4. TLC fingerprint developing solvent composition is cyclohexane, dichloromethane and acetone. Proportion is 2:1:1. Developed two times and the spots were clear. There are 12 spots can be saw under the 365nm ultraviolet light and 9 spots can be seen under the visible light. Separation degree of the spots is great. Different origins of Venenum Bufonis samples have the same spots number. But the color and the size of the spots have difference according to different origin.
5. HPLC fingerprints of 12 groups Venenum Bufonis consisted of 26 common characteristic peaks. It separate into two areas. One area is consisted of 9 common peaks. Its retention time range is 0-20 min. Different Venenum Bufonis origins have the similar peak appearance in this area. Another area is consisted of 17 common peaks. Its retention time range is 20-40 min. It is the mostly important characteristic area. The similarity degree of 12 groups samples both over 0.9, which has a great integer similarity degree.
6. The TLC fingerprints of different processing samples have similar TLC action with the crude samples. The wine processing samples and the milk processing samples of the same group Venenum Bufonis both have the same spots numbers with the crude ones from different groups. But the color is weaker than the crude one. The spots of talcum powder processing samples are weakest, and some have taken great changed. It has the same phomonena in different groups. After different concentration ethanol processing of the same group, the samples have the same spot numbers with the crude ones, and the spot color is weaker than the crude ones. Each wine processing sample does not have evidence differences.
7. The HPLC fingerprints of different processing samples have similar action with the crude samples. Both have the common peaks and the retain time and the relative retain time are identically with the crude ones. But there is great discrimination in the intension of the common peaks. The relative peak areas of the common peaks are great changed after the milk processing and talcum powder processing while which have less changed after the wine processing samples of the same origin. It has the same phomonena in different groups. After different concentration ethanol processing to the same origin group Venenum Bufonis, both the relative retain areas of common peaks are decreased except the No.6 peak.
Conclusion:
1. Compared to the extraction method of Venenum Bufonis contents in《Chinese Pharmacopoeia》, the new established extraction technology of this study has a higher extracting rate. This method can extract the Venenum Bufonis components more sufficient.
2. Compared to the determination method of Venenum Bufonis contents in《Chinese Pharmacopoeia》, the new established determination method of this study is better.
3. It makes the processing parameters into specific and identify. It offers basis for further study of the Venenum Bufonis processing.
4. The TLC fingerprint can separate several components of the Venenum Bufonis. It can be used to identify Venenum Bufonis and evaluate its quality.
5. The HPLC fingerprint can systematic analysis the hold components of Venenum Bufonis. It can be used to evaluate the Venenum Bufonis quality and offer abundant information to control its quality.
6. Both the TLC and HPLC fingerprint methods can be used to evaluate the change between the processing and the crude of Venenum Bufonis. The results show that the content of most components are delined after processed. It is the likely reason that the toxicity become lower after wine processing.
7. Preliminary establishment of quality standards of Venenum Bufonis processing.If it could be further comfirm, it would be offer a reference basis to the quality control of Venenum Bufonis processing in the vocation.
通过对山东临沂、河南、河北等产地的蟾酥有效成分的提取方法、含量、炮制工艺、指纹图谱、炮制前后指纹图谱变化方面进行研究,建立华蟾酥毒基和脂蟾毒配基的最佳提取方法和含量测定方法、蟾酥最佳酒炮制工艺、蟾酥TLC指纹图谱与HPLC指纹图谱,并比较蟾酥不同炮制样品之间及炮制前后指纹图谱的差别,为蟾酥的炮制规范化、质量控制标准化和进一步的应用开发等提供科学依据。
方法:
采用HPLC法,以华蟾酥毒基和脂蟾毒配基的提取率为评价指标,分别进行不同溶剂加热回流提取和超声提取比较考察、不同浓度乙醇单因素考察,以及L_9(3~4)正交试验,筛选蟾酥最佳提取工艺,并对蟾酥中的华蟾酥毒基和脂蟾毒配基进行含量测定。采用HPLC法,以华蟾酥毒基和脂蟾毒配基的含量为评价指标,选择市售白酒、牛奶、滑石粉作蟾酥炮制辅料单因素考察,并通过L_9(3~4)正交试验进一步考察酒炮制工艺,筛选蟾酥药材最佳炮制工艺。采用TLC法对12批蟾酥药材的薄层图谱进行分析。采用HPLC法进行梯度洗脱,对12批蟾酥药材的指纹图谱进行分析。采用TLC和HPLC指纹图谱法比较蟾酥不同方法及炮制前后指纹图谱的变化。
结果:
1.蟾酥有效成分最佳提取方法:加入50倍量80%乙醇70℃提取两次,每次90 min。
2.建立了华蟾酥毒基和脂蟾毒配基的含量测定方法,并测定了12批蟾酥药材,华蟾酥毒基和脂蟾毒配基的含量范围分别是0.937%～4.893%和0.198%～6.692%。
3.蟾酥经三种辅料炮制后有效成分含量均有所下降,其中酒炮制后成化变化较小,选择对酒炮制作进一步研究。优选出蟾酥酒炮制最佳工艺是:加入2倍量55%乙醇(白酒),在60℃下炮制12小时,干燥,粉碎。蟾酥经酒炮制后,有效成分的总含量比原药材降低约20%。
4.蟾酥TLC指纹图谱以V环己烷:V二氯甲烷:V丙酮(2:1:1)为展开剂,二次展开结果最佳,可分离出12个紫外荧光斑点和9个可见光斑点,斑点分离度好。不同批次产地的蟾酥样品的斑点数目相同,斑点颜色和大小因产地不同而各有差异。
5.测定了12批蟾酥的HPLC指纹图谱,从中标定了26个共有峰,并划分为2个区域,第Ⅰ区(RT=0～20 min)由9个共有峰组成,不同产地蟾酥在此区间峰貌相似;第Ⅱ区(RT=20～40 min)由17个共有峰组成,是体现蟾酥产地差异的主要特征区域。对12批蟾酥进行相似度评价均大于0.9,整体相似度较好,初步建立了蟾酥药材的共有峰模式。
6.不同炮制品的TLC指纹图谱薄层行为与蟾酥药材相似。同一批次蟾酥酒炮制和牛奶炮制样品的斑点数目与炮制前基本相同,但斑点颜色较淡;滑石粉炮制样品斑点比该产地生品、酒炮制品和牛奶炮制品的斑点颜色淡,且部分斑点有明显变化。同法对多个批次蟾酥进行对比,结果相似。同一批次蟾酥经不同酒度炮制后,样品斑点数目与生品基本相同,炮制品的斑点颜色较浅,炮制品之间的斑点颜色没有明显差异。
7.不同炮制品的HPLC指纹图谱整体面貌与蟾酥药材相似,各共有峰基本存在,保留时间与相对保留时间基本一致,但共有峰强度差别较大。同一批次蟾酥经牛奶和滑石粉炮制后,共有峰相对峰面积变化较大,酒炮制后的共有峰相对峰面积变化较小。同法考察了多个批次蟾酥,结果相似。同一批次蟾酥经不同酒度炮制后,除6号峰的相对峰面积经炮制后有所增加外,其他共有峰的相对峰面积均呈降低的趋势。
结论:
1.新建的提取方法对蟾酥华蟾酥毒基和脂蟾毒配基的提取率高于2005版《中国药典》的方法,能够充分提取蟾酥中的有效成分。
2.新建的蟾酥有效成分含量测定方法优于2005版《中国药典》的测定方法。
3.规范了蟾酥的酒炮制工艺参数,为蟾酥炮制进一步研究提供实验依据。
4.蟾酥TLC指纹图谱能分离药材中多个成分,可用于蟾酥药材鉴别及质量评价。
5.蟾酥HPLC指纹图谱能够对蟾酥的总体成分进行系统分析,提供了丰富的信息,可用于评价蟾酥药材品质及对质量的全面控制。
6.蟾酥TLC指纹图谱和HPLC指纹图谱可用于评价炮制前后蟾酥的成分变化,能从整体上对蟾酥炮制前后各成分的变化情况进行分析。蟾酥炮制后各成分基本呈下降趋势,这可能是蟾酥酒炮制后降毒的原因。
7.初步建立了蟾酥炮制品质量标准。若能得到进一步验证,可作为行业中蟾酥炮制质量控制的参考基础和依据。
Objective:
Study on the component extraction technology, contents, processing technology, fingerprints of Venenum Bufonis which come from Shan Dong, He Nan, He Bei and so on. And research the changes of their fingerprint of the crude and processed. To build up the best method to extract and a method to determine the Venenum Bufonis contents Cinobufagin and resibufogenin. To establish the best wine processing method, the TLC fingerprint method and the HPLC fingerprint method of Venenum Bufonis. Compare with various processed samples and the crude to find out the differences by fingerprint methods, in order to offer scientific basis to standardize the processing technology, control the quality and for further develop and apply of Venenum Bufonis.
Method:
Measure the total extraction rate of Cinobufagin and Resibufogenin by HPLC to investigate the best extraction technology through selecting different solvents by heating reflux or ultrasonic extraction, designing single factor exploration of different ethanol concentration, and applying the L_9(3~4) of orthogonal test.
Measure the content of Cinobufagin and Resibufogenin by HPLC.
Study the wine, milk and talcum powder as the processing excipient in single factor experiment. Select the wine as the suitable excipient. Then apply the L_9(3~4) of orthogonal test and determine the total content of Cinobufagin and Resibufogenin by HPLC to build up the optimal processing technology.
To analysis 12 groups Venenum Bufonis by TLC method.
To measure 12 groups Venenum Bufonis by HPLC method and gradient elution, and analysis the fingerprint by the computer similarity evaluation system.
Compare with various processed samples and the crude by TLC & HPLC fingerprint methods to find out the differences between before and after processed.
Result:
1. The optimal extraction technology of Cinobufagin and Resibufogenin of Venenum Bufonis is: add 50 times 80% ethanol and heating reflux twice, 90min per time.
2. A new method for measuring the quality of Cinobufagin and Resibufogenin is established. 12 groups of Venenum Bufonis quantitative determination results show that, the contents circumscription of Cinobufagin is 0.937% to 6.520%. And the contents circumscription of Resibufogenin is 0.198% to 7.550%.
3. The contents of Venenum Bufonis are both decline after wine, milk and talcum power processing. Compared with the milk processing and talcum power processing, the change of Venenum Bufonis component is more stable by wine processed. Choose wine as the processing excipient for further study. The results of the orthogonal test, the signal factor experiment of different ethanol concentration and the verification test show that the optimal processing technology is: add 2 times 55% ethanol (wine), heating processing in 60℃for 12 hours, aired and comminuted. Compare to the crude Venenum Bufonis, the quality of the effective component are decrease about 20% after wine processing.
4. TLC fingerprint developing solvent composition is cyclohexane, dichloromethane and acetone. Proportion is 2:1:1. Developed two times and the spots were clear. There are 12 spots can be saw under the 365nm ultraviolet light and 9 spots can be seen under the visible light. Separation degree of the spots is great. Different origins of Venenum Bufonis samples have the same spots number. But the color and the size of the spots have difference according to different origin.
5. HPLC fingerprints of 12 groups Venenum Bufonis consisted of 26 common characteristic peaks. It separate into two areas. One area is consisted of 9 common peaks. Its retention time range is 0-20 min. Different Venenum Bufonis origins have the similar peak appearance in this area. Another area is consisted of 17 common peaks. Its retention time range is 20-40 min. It is the mostly important characteristic area. The similarity degree of 12 groups samples both over 0.9, which has a great integer similarity degree.
6. The TLC fingerprints of different processing samples have similar TLC action with the crude samples. The wine processing samples and the milk processing samples of the same group Venenum Bufonis both have the same spots numbers with the crude ones from different groups. But the color is weaker than the crude one. The spots of talcum powder processing samples are weakest, and some have taken great changed. It has the same phomonena in different groups. After different concentration ethanol processing of the same group, the samples have the same spot numbers with the crude ones, and the spot color is weaker than the crude ones. Each wine processing sample does not have evidence differences.
7. The HPLC fingerprints of different processing samples have similar action with the crude samples. Both have the common peaks and the retain time and the relative retain time are identically with the crude ones. But there is great discrimination in the intension of the common peaks. The relative peak areas of the common peaks are great changed after the milk processing and talcum powder processing while which have less changed after the wine processing samples of the same origin. It has the same phomonena in different groups. After different concentration ethanol processing to the same origin group Venenum Bufonis, both the relative retain areas of common peaks are decreased except the No.6 peak.
Conclusion:
1. Compared to the extraction method of Venenum Bufonis contents in《Chinese Pharmacopoeia》, the new established extraction technology of this study has a higher extracting rate. This method can extract the Venenum Bufonis components more sufficient.
2. Compared to the determination method of Venenum Bufonis contents in《Chinese Pharmacopoeia》, the new established determination method of this study is better.
3. It makes the processing parameters into specific and identify. It offers basis for further study of the Venenum Bufonis processing.
4. The TLC fingerprint can separate several components of the Venenum Bufonis. It can be used to identify Venenum Bufonis and evaluate its quality.
5. The HPLC fingerprint can systematic analysis the hold components of Venenum Bufonis. It can be used to evaluate the Venenum Bufonis quality and offer abundant information to control its quality.
6. Both the TLC and HPLC fingerprint methods can be used to evaluate the change between the processing and the crude of Venenum Bufonis. The results show that the content of most components are delined after processed. It is the likely reason that the toxicity become lower after wine processing.
7. Preliminary establishment of quality standards of Venenum Bufonis processing.If it could be further comfirm, it would be offer a reference basis to the quality control of Venenum Bufonis processing in the vocation.
引文
1. (五代)吴越,尚志钧辑释.日华子本草/蜀本草[M].合肥:安徽科学技术出版社,2005,07:473.
2. 梁·陶弘景编,尚志钧,尚元胜辑校.本草经集注(辑校本)[M].北京:人民卫生出版社,1994,02:440.
3. 叶定江,原思通.中药炮制学辞典[M].上海:上海科学技术出版社.2005:549.
4. 唐·陈藏器撰,尚志钧辑释.《本草拾遗》辑释[M].合肥:安徽科学技术出版社,1998,02:642.
5. 明·李时珍.本草纲目[M].北京:中国中医药出版社.1998,02:986-988.
6. 国家药典委员会.中国药典(一部) [S] 北京:化工出版社,1963,02:316-317.
7. 国家药典委员会.中国药典(一部) [S] 北京:化工出版社,1977,02:668-669.
8. 国家药典委员会.中国药典(一部) [S] 北京:化工出版社,1985,02:344.
9. 国家药典委员会.中国药典(一部) [S] 北京:化工出版社,1990,02:346.
10. 国家药典委员会.中国药典(一部) [S] 北京:化工出版社,1995,02:340-341.
11. 国家药典委员会.中国药典(一部) [S] 北京:化工出版社,2000,02:316-317.
12. 国家药典委员会.中国药典(一部) [S] 北京:化工出版社,2005,02:25-266.
13. 吕侠卿.中药炮制大全[M].湖南科学技术出版社,2003,3:134-135.
14. 中华人民共和国卫生部药政管理局,中国药品生物制品检定所主编.中药材手册[M].北京:人民卫生出版社,1959:710-712.
15. 袁敬.蟾酥的采制技术[J].时珍国医国药,2001,12(8):675.
16. 高本刚.蟾酥的采集与加工[J].中国农村科技,1999,11:41.
17. 丁欣芳,陈圣茂.蟾酥的采集与加工[J].农村发展论丛(实用版),1999.15(8):40.
18. 邓厚群.蟾酥的采集与加工[J].农技服务,2005,04,9-3.
19. 彭洪光.蟾酥采集与加工[J].农村新技术,1996,12:18-19.
20. 安铁生.蟾酥采集加工中应注意的问题[J].中药材, 1994,17(4):51.
21. 谢麟.蟾酥的采集与加工[J].中国农村科技,2003,9:44
22. 明·陈嘉谟.本草蒙荃[M].北京:人民卫生出版社,1988:419-420.
23. 明·李时珍.本草纲目[M].北京:人名卫生出版社影印,1988:卷四十二 虫之四.
24. 钟少伟.蟾酥的采集与加工[J].农技服务,1997,9:37-38.
25. 郭津伍.蟾酥的采集与加工[J].农家参谋,1996,10:31.
26. 清·赵学敏.本草纲目拾遗[M].北京:人民卫生出版社,1963:440-441.
27. 明·龚延贤.寿世保元(校点本)[M].天津:天津科学技术出版社,2000:663-665.
28. 清·许克昌,毕法.外科证治全集[M].北京:人民卫生出版社,1987,4:113.
29. 明·张景岳.景岳全书[M].上海:上海科学技术出版社,1959:四十四卷.
30. 杨祖山,王宁,蟾蜍的药用和炮制[J].基层中药杂志,1995,9(4):11-12
31. 清·黄宫绣.本草求真[M].上海:上海科学技术出版社,1959:258-259.
32. 付兰,何树芸,潘德敏.蟾酥不同炮制品蟾毒内酯的含量测定[J].中药材,1990; 13(2):25.
33. 葛莉,张兆瑞,马莹,等.蟾酥炮制前后的质量比较[J].中成药,1993,15(2):16.
34. 王志巍,王力,李玉山.蟾酥的炮制方法讨探[J].中医药学报,1994,1:43-44.
35. 许保军,夏华玲,赵新杰,等.炮制对蟾酥药材中蟾毒内酯的影响[J].中国中药杂志,1998,23(12):722-723.
36. 肖崇厚.中药化学[M].上海:上海科学技术出版社,1987:170.
37. 金向群.蟾蜍化学与药理作用的研究进展[J].中草药,1996,27(4):246-249.
38. 周学敏,蟾酥成分的分析方法[J].中草药,1992,23(9):490-494.
39. 杨素娟,等.中医药信息1992,9(5):12.
40. Bick RT, Poindexter BT, Sweney RR, etal. Effects of Chan, a traditional Chinese medicine,on the calcium transients of isolated cardiomyocytes:Cariotoxicity due to more than Na,K-ATP ase blocking [J] Life Sci,2002,72(6):699- 709.
41. 张屏,崔征,刘雅妹等.蟾酥中吲哚烷胺类生物碱的分离与结构鉴定[J].沈阳药科大学学报,2006,23(4):216-219.
42. 欧明,林励,李衍文.简明中药成分手册[M].中国医药科技出版社:2003,395-396.
43. 李素琴,Wilhelm SCHONER,符云峰.从蟾酥中纯化一种内源性钠泵抑制因子[J].中国生物化学与分子生物学报,2002,18(2):250- 253.
44. 王学勇.蟾酥中含有免疫调节剂[J].国际中医中药杂志,2006,28(3):150.
45. 程国华.蟾酥质量研究及其药理临床应用进展[J].中草药,2001,32(2):184-186.
46. 朱天忠.蟾酥的药理研究和临床应用现状[J].浙江中医杂志,1992, 22(12):565.
47. 王锋,张薇.蟾酥的药理研究及其临床应用概况[J].国医论坛,2003,18(5):50-52.
48. 齐刚,张莉,何文形,等. Bufalin的急性毒性研究[J].武警医学院学报,2003,13(1):10-12.
49. 程国华.蟾酥质量研究及其药理临床应用进展[J].中草药,2001,32(2):184-186.
50. Low SK, Gwee MC,Yuen R,et al.Stonustoxin: effects on neuromuscular function in vitro and in vivo[J].Toxicorr,1994,32(5):573.
51. 梁克军.蟾酥的化学与药理作用[J].医药工业,1985,(1):29.
52. Low SK, Gwee MC,Yuen R,et al.Stonustoxin: effects on neuromuscular function in vitro and in vivo[J].Toxicorr,1994,32(5):573.
53. Iiasu A, Gomes A, Dasnupta SC, et al.The effect of arenobufanin in potassium currents in nastric antral circular myocytes of rabits [J].Indian J Exp Biol,1990,28(5):451.
54. Yei CC, Deng JF. Toad or toad cake intoxication in Taiwan:report of four cases[J].Torrnos Med Assoc,1993,92Sup (3):135.
55. 李邓.蟾酥的采集与加工(一)[J].中草药,1985,(6):44.
56. 禹永春,陆书广,王雷,等.四乙基胺抑制蟾酥的强心效应.延边大学医学学报,2001,24(3):164-168.
57. Bick RJ,Poindexter BJ,Sweney RR,et al. Effect of Chan Su, a traditional Chinese medicine, on the calcium transients of isolated cardiomyocytes:Cardiotoxicity due to more than Na,K-ATPaseblocking[J].Life Sci,2002,72(6):699-709.
58. 久保道德.六神丸中蟾酥、牛黄、麝香对于血栓诱发的缺血性心脏障碍的作用[J].生药学杂志, 1984,38(1):59.
59. 李安国.蟾酥提取物的药理研究[J].湖南中医学院学报, 1986,6(4):52-53.
60. 谢景田,N Belic, J McCullounh,等.脂蛤毒配基对人和犬心肌细胞膜反应性与后电位的影响[J].中国药理学报,1985,6(4):254- 257.
61. 韩永晶,崔荣芬,靳珠华.蟾蜍灵对豚鼠心房的影响[J].中国药理学与毒理学杂志,1991,4(1): 71-72.
62. 钮亚珍,薛平贵.蟾酥的药理作用研究进展[J].张家口医学院学报,2002,19(3):58-59.
63. 韩水晶,靳珠华,侯庆昌,等.蟾毒灵促进离体豚鼠输精管释放去甲肾上腺素[J].中国药理学报, 1986,7(4):314-316.
64. 郝继亭.蟾酥现代药理研究进展[J].中国医药报,2004,9,23.
65. 韩水晶,张力,崔荣芬.华蟾蜍毒素对离体豚鼠输精管的作用[J].药学学报,1992,27(4):252- 255.
66. 姜延良,严达常.我国抗癌中草药研究现状和展望[J].中西医结合杂志1986,6:698一700.
67. 高瑞霖, 邵义祥.蟾酥提取物对荷瘤小鼠肿瘤细胞形态学实验研究[J].交通医学,2006,120 (4):376-378.
68. Ji ZN, Ye WC, Rui J et al.Patensin-induced apoptosis accrmpanied by decreased bcl-2 expression and telrmerase activity in HL-60 cells[J].Asian Nat Prod Res 2004,6(2):107-114.
69. Huang ST, Yang RC, Chen MY, et al. Phyllanthus urinaria induces the Fas recepto/ligand expression and ceramide-mediated apoptosis in HL-60 cells[J].Life Sci,2004,75(3):339-35.
70. 刘云鹏,曲秀娟,王萍萍,等.蟾酥灵诱导K562细胞分化和凋亡过程中WT1表达的下调[J].中华血液学杂志,2002,23(7):356-359.
71. Yeh JY, Huang WJ, Kan SF,et al. Effect of bufalin and cinobufagin on the proliferationof androgen dependent and independent prostate cancer cells [J].Prostate,2003,54 (2):112-124.
72. Chow J, Johnson M, Wells A, et al. Effect of the traditional Chinese medicines Chan Su Liu Shen-Wan,Dan Shen and Asian ginseng on serum digoxin measurement by Tinaquant Roche (and Syncbron LX systcrn Beckman)digoxin immunoassays,[J].Clin Lab Anal 2003,17(1):22-27.
73. 李墨林,舒晓宏,刘用析,等.蟾酥对小鼠膀胱癌的抗癌作用研究[J].中医药学刊,2004,22(4): 651-665.
74. 徐瑞成,陈小义,陈莉,等.蟾蛛灵诱导人白血病HL60细胞凋亡的初步研究[J].中国中药杂志, 2001,26(1):59-61.
75. 高晓东,陈烨,丁润生.蟾酥灵对HL-60/ADR作用的研究[J].交通医学,2002,6(5):506-507.
76. Rem R Rer/abl does not prevent apoptotic death induced by human natural killer or lymohokine-activated killer cells[J].Blood,1996,7:1113.
77. 戴锡孟,徐芳,郭义.六神丸诱学自血病细胞凋亡及调控凋亡相关基因的实验研究[J].辽宁中医杂志,2003,30:431-433.
78. 卢香兰,许崇安,王萍,等.蟾蜍毒素粗提物对自血病K562细胞的杀伤作用[J].中国医科人学学报,2003,32(1):19.
79. 刘云鹏,曲秀娟,王萍萍,等.蟾蜍灵诱学K562细胞分化和凋亡过程中WT1表达的下调[J].中华血液学杂志,2002,23(7):356.
80. 贾彩云,呼文亮,徐瑞成,等.蟾蜍灵对K562细胞生长抑制及凋亡诱学作用[J].武警医学院学报, 2003,12(4):265.
81. Zhang L, Nakava K, Yoshida T,et al. Induction of hufalin of ferentiation of human leukemia cells HL-60, U937,and ML1 toward maerophage/monocyte-like cells and its potent synergistic effcet on the differenlion of human lcukemia cells in combination with other inducers[J].Cancer Res,1992,52(17):4634.
82. Masuda Y, Ksawazoc N,Nakajo S,et al. Bufalin inducesis apoptosis and influences the expression of apoptosi-related genes in human leukemia cells[J]. Leuk Res, 1995,19 (8):549.
83. Watabc M,Kawazoc N,Nasuda Y,et al.Bca-2 protein inhibits bufalin-induced apoptosis through inhibition of mitogen-acaivated protein Kinasc a caivation in human leukemia U937 cells[J].Cance,Res,1997,57(15):3097.
84. 赵建斌,崔勤,张雪,等.华蟾毒精抗癌作用的体外研究[J].第四军医大学学报,2001,22(16): 1504-1507.
85. Guschtscha LH, Bartier WA. Ross CEA, et al. Characteristic of cancel cell death after exposure to cptotoxic drugs in vitro. Br J Cancen,1996,73:54.
86. 尹军强,沈靖南.蟾酥水提取物抗骨肉瘤作用的体外研究[J].中国骨肿瘤骨病,2004,3 (5): 303-305.
87. 邢增涛,卫功庆,张爱武,等.蟾酥的研究与应用[J].经济动物学报,1998,2(2):56-60.
88. 药学杂志(日),1968,88(11):1367.
89. 张薇,潘明辉,张豁中,等.花背蟾蜍耳后腺分泌物的药效学研究[J].沈阳药学院学报,1992,9 (1):71.
90. 孙关林,王振义,陈淑容,等.蟾酥制剂对免疫功能作用的初步研究[J].中西医结合杂志,1984, 4(5):297- 299.
91. 刘嘉湘,许德凤,范忠泽.蟾酥4膏缓解癌性疼痛的临床疗效观察[J].中医杂志,1993,34(5): 281- 282.
92. 陈庆强.冰膏外敷治疗癌痛45例[J].新中医,1997,29(6):48-52.
93. 李雪岩,王滨,彭如心,金香兰.蟾酥对蟾蜍坐骨神经干复合动作电位的影响[J].中国中医药科技,1996(6):365-366.
94. 徐叔云,卞如濂,陈修主编.药理实验方法学[M].第2版,北京:人民卫生版社,1994:968-969.
95. 杨华中,刘英姿,张镭. 蟾酥和维拉帕米镇痛协同作用的实验研究. 湖南师范人学学报(医学版), 2004,1(1):50-52.
96. 张薇,刘玉兰,徐从石,等.蟾酥的镇痛活性成分[J].沈阳药科大学学报,1998,15(4):268-271.
97. 薛开先.新医药学杂志1975:(1):39.
98. 李雪岩,王滨,彭如心,金香兰.蟾酥对蟾蜍坐骨神径干复合动作电位的影响[J].中国中医药科技1995,6(6):365.
99. 江苏新医学院.中药大词典(下册)[M].上海:上海人民出版社,1977:7-34.
100. 《中华本草》编委会.中华本草(第9册)[M].上海:上海科学技术出版社,1999:365.
101. [日]筱原达雄.淫羊着配合蟾酥制剂的抗疲劳作用.和汉医药学杂志,1996,13(3):260- 267.
102. 孙非,张淑芹,刘志屹,等.蟾毒内醋抗柯萨奇病毒、腺病毒和流感病毒作用的研究[J].中国药理学通报,2004,20(4):478-479.
103. 曲莉,孙非,张淑芹,等.蟾毒内酯抗流感病毒的实验研究[J].长春中医学院学报,2004,20 (1):37-38.
104. 杜平,朱关福,刘湘云.现代临床病毒学[M].北京:人民军医出版社,1991,233-285.
105. 杨秀伟,刑增涛,帐景荣.华蟾毒精和轻基华蟾毒精的人肠内细菌代谢研究[J].北京人学学报(医学版),2001,33(3):199-204.
106. 王浴生,等.中药药理与应用[M].北京:学苑出版社,1992.311.
107. Chan WY, Ng TB, Yeung HW.Examination for toxicity of a Chinese drug, the load glandular secretory product Chan Su, in pregnant mice and embryos [J]. Bilo Neonate, 1995,67(S):376-380.
108. 张瑜瑶.蟾酥水溶性总成分的实验研究[J].南京中医学院学报,1988,(2):33
109. 李树林.毒物的毒理与毒物分析[M].北京:人民卫生出版社,1987,5:596-597.
110. 钱伟平.蟾酥对实验动物循环系统毒副作用的研究[J]. 浙江中医学院学报,2001,25(6): 46-47.
111. G.哈贝尔梅尔.有毒动物和毒物毒[M].北京:科学技术出版社.1981,6:48-49.
112. Yeh J Y, Huang W J, Kan S F, et al. Effect of bufalin and cinobufagin on the proliferation of androgen dependent and independent prostate cancer cells [J]. Prostate,2003,54(2):112-124.
113. Brubacher J R, Ravikumar P R, Bania T, et al. Treatment of toad venom poisoning with digoxin-specific Fab fragments [J].Chest,1996,110(5):1139-1141.
114. 赵强,孟凡静,刘安西.蟾酥的研究进展[J].中草药,2004,35(IO):4-7.
115. 韩莉.中药蟾酥临床应用简介[J].实用医药杂志,2003,20(11):854
116. 齐刚,张莉,何文形,等. Bufalin的急性毒性研究[J].武警医学院学报,2003,13(1):10-12.
117. 陈端艺,林巧艺,高淑琼,等.服六神丸中毒一例[J].中国基层医药,2001,8(2):106.
118. 陈强,高翠萍.服六神丸出现过敏反应一例[J].中成药,2002,27(10):798.
119. 张萍,文家福.新生儿六神丸中毒一例[J].中华急诊医学杂志,2001,10(3):172.
120. 张凡景,马东成,赵霞.女童口服六神丸引起雷诺现象一例[J].中国中药杂志,1999,24(9): 570.
121. 龚岳亭.生物活性肤[M].上海:上海科学技术出版社.1985.10,161-162.
122. 梁秀兰,许廷生,黄新.蟾酥及其成方制剂的合理应用[J].中国药物与临床,2004,4(3):217.
123. 谭道玉,王绍琼,李昌平,等.纷妥拉明顶防华蟾素所致局部静脉反应的临床观察[J].中国中西医结合杂志,2003:23(12):940.
124. 杨勇,奉建芳,芦洁,等. 蟾酥固体脂质纳米粒冻干针剂体内药代动力学研究[J].中成药. 2006,28(6):787-790.
125. 范健,黄慰魁,武福乐,等.华蟾毒精微球制剂肝动脉栓塞的实验研究[J].癌症,1995,14(6): 434.
126. 韩洪刚,刘云海,谢委.蟾酥β-环糊精包合物的研究现状[J]. 医药导报,2005,24(11): 1039-1041.
127. 张梅,阳长明.冠心微丸中脂蟾毒配基的含量测定[J].湖南中医杂志,2001,17(2):59-60.
128. 唐青华,劳三申,高明珠.等.金蟾定痛微粒丸下艺的研究[J].中成药.1997,19(3):3-5.
129. 虞金宝,程怡,金克宁,等蟾酥透皮吸收研究[J].中国中药杂志,1995,20(9):539.
130. 朱盛山,吕志平,刘强.氮酮对蟾酥中蟾毒内酯类成分透皮吸收的影响[J].中成药. 2003, 25(9):694-685.
131. 杨莉娅,丁训诚.巴布剂及橡胶剂蟾酥膏对家兔完整皮肤刺激试验的比较[J].中国药业,2000, 9(5):11.
132. 王隶书,程东岩,刘永宏.华蟾毒精气雾剂的研制[J].中成药,1998,20(4):1-2.
133. 周学敏,程培元,许美娟,等.HPLC法分离测定蟾酥脂质体中三种蟾毒内酯成分[J].中国药科大学学报1993.24(2):116.
134. 李勤,谢艳华,孙纪元,等.中药安迪粉针剂对离体肿瘤细胞的细胞毒作用[J].西安交通大学学报(医学版),2003,24(8):390-392.
135. 王彦泓, 胡俊敏.中药消毒栓杀菌和杀精子性能的试验观察[J].河北医药,2001,23(5): 346-348.
136. 孟庆增,袁青春,姚元庚,等.蟾酥复方消毒剂性能的实验观察[J].中国消毒学杂志, 1998,15 (1):26-29.
137. 孔凤利.中药蟾酥的鉴别[J].浙江中医学院学报,2004,28(6):78
138. 陈美珍.中药蟾酥的鉴别[J].中草药,2002,33(3):272.
139. 赖新华.蟾酥的真伪鉴别法[J].基层中药杂志,2002,16(2):34.
140. 伊惠贤.用多次层析法鉴别中药成方制剂中的蟾酥[J].中草药,1997,28(5):277-278.
141. 祝君.牛黄消炎片的薄层色谱鉴别[J].中国药业,2001,10(2):19-20.
142. 邵陆.康肤灵配鉴别试验的研究[J].山东医药工业,1996,15(2).
143. 石俊英.哈蟆油、鸡内金等动物类中药及其混淆品的电泳鉴别研究[J].山东中医杂志,1994, 13(2):73-74.
144. 霍建中.微波消解ICP-AES法测定澳洲蟾酥中的常量及微量元素[J].天津中医学院学报,2003, 22(1):19-20.
145. 于萍.优劣蟾酥的毛细管气相色谱鉴别方[J].分析测试技术与仪器,1998,4(4):245-250.
146. 工伟,乔文礼,塔娜.薄层扫描法测定梅花点舌丸中华蟾蜍毒基含量[J].国现代应用药学杂志, 2001,18(2):129-131.
147. 王莉华.双波长薄层扫描法测定风油精中华蟾毒精的含量[J].中国药科大学学报,1995,26 (5):279-281.
148. 钟镜金.薄层--分光光度法测定制剂中脂蟾毒配基的含量[J].中药材,1995,18(6):311-312.
149. 段亚丽.紫外分光光度法控制酥黄注射液中蟾酥质量[J].广西农业科学,2005,36(2):158-159.
150. 杨锡,罗兴平.本底校正分光光度法测定蟾酥及其制剂中蟾毒内脂的含量[J].药物分析杂志, 1994,14(4):19-21.
151. 蟾酥的研究(1):用比色计测定商品蟋酥的化学成分差异〔日〕/川原一仁…//Natural Medines,1999,53(5).242 – 248.
152. 何丽霞.成方制剂中蟾酥的质量控制[J].中国兽药杂志,2003,37(4):28-30.
153. 王京辉.高效液相色谱法测定蟾酥中有效成分的含量[J].中国中药杂志,1998,23(11): 651-653.
154. 乔延江,吴刚,王玺,等.中药蟾酥质量的人工神经网络化学模式识别研究[J].分析化学 1995, 23(6)630-634.
155. 乔延江,王玺,毕开顺,等.人工神经网络在中药蟾酥化学模式识别特征提取中的应用[J].药学学报,1995;30(9):698-701.
156. 王玺,马丽娟,潘卫东.等.蟾酥薄层色谱数据的化学模式识别[J].沈阳药科大学学报,1995, 12(4):50-254.
157. 葛勤,康钦树.蟾酥的化学及制剂质量研究近况[J].中成药,1998,20(1):38-40.
158. 王永刚,吴忠,魏凤环,等.中药指纹图谱研究的现状与未来[J].中药材,2003,26(11): 820-825.
159. 国家药品监督管理局.中药注射液指纹图谱研究的技术要求(暂行)[J].中成药,2000,22(10): 671.
160. 江苏新医学院[M].中药大词典(下册).上海:上海人民出版社,1977,27-34.
161. 杨基森主编.中国药酒学[M].贵州:贵州科技出版社,2002,8.
2. 梁·陶弘景编,尚志钧,尚元胜辑校.本草经集注(辑校本)[M].北京:人民卫生出版社,1994,02:440.
3. 叶定江,原思通.中药炮制学辞典[M].上海:上海科学技术出版社.2005:549.
4. 唐·陈藏器撰,尚志钧辑释.《本草拾遗》辑释[M].合肥:安徽科学技术出版社,1998,02:642.
5. 明·李时珍.本草纲目[M].北京:中国中医药出版社.1998,02:986-988.
6. 国家药典委员会.中国药典(一部) [S] 北京:化工出版社,1963,02:316-317.
7. 国家药典委员会.中国药典(一部) [S] 北京:化工出版社,1977,02:668-669.
8. 国家药典委员会.中国药典(一部) [S] 北京:化工出版社,1985,02:344.
9. 国家药典委员会.中国药典(一部) [S] 北京:化工出版社,1990,02:346.
10. 国家药典委员会.中国药典(一部) [S] 北京:化工出版社,1995,02:340-341.
11. 国家药典委员会.中国药典(一部) [S] 北京:化工出版社,2000,02:316-317.
12. 国家药典委员会.中国药典(一部) [S] 北京:化工出版社,2005,02:25-266.
13. 吕侠卿.中药炮制大全[M].湖南科学技术出版社,2003,3:134-135.
14. 中华人民共和国卫生部药政管理局,中国药品生物制品检定所主编.中药材手册[M].北京:人民卫生出版社,1959:710-712.
15. 袁敬.蟾酥的采制技术[J].时珍国医国药,2001,12(8):675.
16. 高本刚.蟾酥的采集与加工[J].中国农村科技,1999,11:41.
17. 丁欣芳,陈圣茂.蟾酥的采集与加工[J].农村发展论丛(实用版),1999.15(8):40.
18. 邓厚群.蟾酥的采集与加工[J].农技服务,2005,04,9-3.
19. 彭洪光.蟾酥采集与加工[J].农村新技术,1996,12:18-19.
20. 安铁生.蟾酥采集加工中应注意的问题[J].中药材, 1994,17(4):51.
21. 谢麟.蟾酥的采集与加工[J].中国农村科技,2003,9:44
22. 明·陈嘉谟.本草蒙荃[M].北京:人民卫生出版社,1988:419-420.
23. 明·李时珍.本草纲目[M].北京:人名卫生出版社影印,1988:卷四十二 虫之四.
24. 钟少伟.蟾酥的采集与加工[J].农技服务,1997,9:37-38.
25. 郭津伍.蟾酥的采集与加工[J].农家参谋,1996,10:31.
26. 清·赵学敏.本草纲目拾遗[M].北京:人民卫生出版社,1963:440-441.
27. 明·龚延贤.寿世保元(校点本)[M].天津:天津科学技术出版社,2000:663-665.
28. 清·许克昌,毕法.外科证治全集[M].北京:人民卫生出版社,1987,4:113.
29. 明·张景岳.景岳全书[M].上海:上海科学技术出版社,1959:四十四卷.
30. 杨祖山,王宁,蟾蜍的药用和炮制[J].基层中药杂志,1995,9(4):11-12
31. 清·黄宫绣.本草求真[M].上海:上海科学技术出版社,1959:258-259.
32. 付兰,何树芸,潘德敏.蟾酥不同炮制品蟾毒内酯的含量测定[J].中药材,1990; 13(2):25.
33. 葛莉,张兆瑞,马莹,等.蟾酥炮制前后的质量比较[J].中成药,1993,15(2):16.
34. 王志巍,王力,李玉山.蟾酥的炮制方法讨探[J].中医药学报,1994,1:43-44.
35. 许保军,夏华玲,赵新杰,等.炮制对蟾酥药材中蟾毒内酯的影响[J].中国中药杂志,1998,23(12):722-723.
36. 肖崇厚.中药化学[M].上海:上海科学技术出版社,1987:170.
37. 金向群.蟾蜍化学与药理作用的研究进展[J].中草药,1996,27(4):246-249.
38. 周学敏,蟾酥成分的分析方法[J].中草药,1992,23(9):490-494.
39. 杨素娟,等.中医药信息1992,9(5):12.
40. Bick RT, Poindexter BT, Sweney RR, etal. Effects of Chan, a traditional Chinese medicine,on the calcium transients of isolated cardiomyocytes:Cariotoxicity due to more than Na,K-ATP ase blocking [J] Life Sci,2002,72(6):699- 709.
41. 张屏,崔征,刘雅妹等.蟾酥中吲哚烷胺类生物碱的分离与结构鉴定[J].沈阳药科大学学报,2006,23(4):216-219.
42. 欧明,林励,李衍文.简明中药成分手册[M].中国医药科技出版社:2003,395-396.
43. 李素琴,Wilhelm SCHONER,符云峰.从蟾酥中纯化一种内源性钠泵抑制因子[J].中国生物化学与分子生物学报,2002,18(2):250- 253.
44. 王学勇.蟾酥中含有免疫调节剂[J].国际中医中药杂志,2006,28(3):150.
45. 程国华.蟾酥质量研究及其药理临床应用进展[J].中草药,2001,32(2):184-186.
46. 朱天忠.蟾酥的药理研究和临床应用现状[J].浙江中医杂志,1992, 22(12):565.
47. 王锋,张薇.蟾酥的药理研究及其临床应用概况[J].国医论坛,2003,18(5):50-52.
48. 齐刚,张莉,何文形,等. Bufalin的急性毒性研究[J].武警医学院学报,2003,13(1):10-12.
49. 程国华.蟾酥质量研究及其药理临床应用进展[J].中草药,2001,32(2):184-186.
50. Low SK, Gwee MC,Yuen R,et al.Stonustoxin: effects on neuromuscular function in vitro and in vivo[J].Toxicorr,1994,32(5):573.
51. 梁克军.蟾酥的化学与药理作用[J].医药工业,1985,(1):29.
52. Low SK, Gwee MC,Yuen R,et al.Stonustoxin: effects on neuromuscular function in vitro and in vivo[J].Toxicorr,1994,32(5):573.
53. Iiasu A, Gomes A, Dasnupta SC, et al.The effect of arenobufanin in potassium currents in nastric antral circular myocytes of rabits [J].Indian J Exp Biol,1990,28(5):451.
54. Yei CC, Deng JF. Toad or toad cake intoxication in Taiwan:report of four cases[J].Torrnos Med Assoc,1993,92Sup (3):135.
55. 李邓.蟾酥的采集与加工(一)[J].中草药,1985,(6):44.
56. 禹永春,陆书广,王雷,等.四乙基胺抑制蟾酥的强心效应.延边大学医学学报,2001,24(3):164-168.
57. Bick RJ,Poindexter BJ,Sweney RR,et al. Effect of Chan Su, a traditional Chinese medicine, on the calcium transients of isolated cardiomyocytes:Cardiotoxicity due to more than Na,K-ATPaseblocking[J].Life Sci,2002,72(6):699-709.
58. 久保道德.六神丸中蟾酥、牛黄、麝香对于血栓诱发的缺血性心脏障碍的作用[J].生药学杂志, 1984,38(1):59.
59. 李安国.蟾酥提取物的药理研究[J].湖南中医学院学报, 1986,6(4):52-53.
60. 谢景田,N Belic, J McCullounh,等.脂蛤毒配基对人和犬心肌细胞膜反应性与后电位的影响[J].中国药理学报,1985,6(4):254- 257.
61. 韩永晶,崔荣芬,靳珠华.蟾蜍灵对豚鼠心房的影响[J].中国药理学与毒理学杂志,1991,4(1): 71-72.
62. 钮亚珍,薛平贵.蟾酥的药理作用研究进展[J].张家口医学院学报,2002,19(3):58-59.
63. 韩水晶,靳珠华,侯庆昌,等.蟾毒灵促进离体豚鼠输精管释放去甲肾上腺素[J].中国药理学报, 1986,7(4):314-316.
64. 郝继亭.蟾酥现代药理研究进展[J].中国医药报,2004,9,23.
65. 韩水晶,张力,崔荣芬.华蟾蜍毒素对离体豚鼠输精管的作用[J].药学学报,1992,27(4):252- 255.
66. 姜延良,严达常.我国抗癌中草药研究现状和展望[J].中西医结合杂志1986,6:698一700.
67. 高瑞霖, 邵义祥.蟾酥提取物对荷瘤小鼠肿瘤细胞形态学实验研究[J].交通医学,2006,120 (4):376-378.
68. Ji ZN, Ye WC, Rui J et al.Patensin-induced apoptosis accrmpanied by decreased bcl-2 expression and telrmerase activity in HL-60 cells[J].Asian Nat Prod Res 2004,6(2):107-114.
69. Huang ST, Yang RC, Chen MY, et al. Phyllanthus urinaria induces the Fas recepto/ligand expression and ceramide-mediated apoptosis in HL-60 cells[J].Life Sci,2004,75(3):339-35.
70. 刘云鹏,曲秀娟,王萍萍,等.蟾酥灵诱导K562细胞分化和凋亡过程中WT1表达的下调[J].中华血液学杂志,2002,23(7):356-359.
71. Yeh JY, Huang WJ, Kan SF,et al. Effect of bufalin and cinobufagin on the proliferationof androgen dependent and independent prostate cancer cells [J].Prostate,2003,54 (2):112-124.
72. Chow J, Johnson M, Wells A, et al. Effect of the traditional Chinese medicines Chan Su Liu Shen-Wan,Dan Shen and Asian ginseng on serum digoxin measurement by Tinaquant Roche (and Syncbron LX systcrn Beckman)digoxin immunoassays,[J].Clin Lab Anal 2003,17(1):22-27.
73. 李墨林,舒晓宏,刘用析,等.蟾酥对小鼠膀胱癌的抗癌作用研究[J].中医药学刊,2004,22(4): 651-665.
74. 徐瑞成,陈小义,陈莉,等.蟾蛛灵诱导人白血病HL60细胞凋亡的初步研究[J].中国中药杂志, 2001,26(1):59-61.
75. 高晓东,陈烨,丁润生.蟾酥灵对HL-60/ADR作用的研究[J].交通医学,2002,6(5):506-507.
76. Rem R Rer/abl does not prevent apoptotic death induced by human natural killer or lymohokine-activated killer cells[J].Blood,1996,7:1113.
77. 戴锡孟,徐芳,郭义.六神丸诱学自血病细胞凋亡及调控凋亡相关基因的实验研究[J].辽宁中医杂志,2003,30:431-433.
78. 卢香兰,许崇安,王萍,等.蟾蜍毒素粗提物对自血病K562细胞的杀伤作用[J].中国医科人学学报,2003,32(1):19.
79. 刘云鹏,曲秀娟,王萍萍,等.蟾蜍灵诱学K562细胞分化和凋亡过程中WT1表达的下调[J].中华血液学杂志,2002,23(7):356.
80. 贾彩云,呼文亮,徐瑞成,等.蟾蜍灵对K562细胞生长抑制及凋亡诱学作用[J].武警医学院学报, 2003,12(4):265.
81. Zhang L, Nakava K, Yoshida T,et al. Induction of hufalin of ferentiation of human leukemia cells HL-60, U937,and ML1 toward maerophage/monocyte-like cells and its potent synergistic effcet on the differenlion of human lcukemia cells in combination with other inducers[J].Cancer Res,1992,52(17):4634.
82. Masuda Y, Ksawazoc N,Nakajo S,et al. Bufalin inducesis apoptosis and influences the expression of apoptosi-related genes in human leukemia cells[J]. Leuk Res, 1995,19 (8):549.
83. Watabc M,Kawazoc N,Nasuda Y,et al.Bca-2 protein inhibits bufalin-induced apoptosis through inhibition of mitogen-acaivated protein Kinasc a caivation in human leukemia U937 cells[J].Cance,Res,1997,57(15):3097.
84. 赵建斌,崔勤,张雪,等.华蟾毒精抗癌作用的体外研究[J].第四军医大学学报,2001,22(16): 1504-1507.
85. Guschtscha LH, Bartier WA. Ross CEA, et al. Characteristic of cancel cell death after exposure to cptotoxic drugs in vitro. Br J Cancen,1996,73:54.
86. 尹军强,沈靖南.蟾酥水提取物抗骨肉瘤作用的体外研究[J].中国骨肿瘤骨病,2004,3 (5): 303-305.
87. 邢增涛,卫功庆,张爱武,等.蟾酥的研究与应用[J].经济动物学报,1998,2(2):56-60.
88. 药学杂志(日),1968,88(11):1367.
89. 张薇,潘明辉,张豁中,等.花背蟾蜍耳后腺分泌物的药效学研究[J].沈阳药学院学报,1992,9 (1):71.
90. 孙关林,王振义,陈淑容,等.蟾酥制剂对免疫功能作用的初步研究[J].中西医结合杂志,1984, 4(5):297- 299.
91. 刘嘉湘,许德凤,范忠泽.蟾酥4膏缓解癌性疼痛的临床疗效观察[J].中医杂志,1993,34(5): 281- 282.
92. 陈庆强.冰膏外敷治疗癌痛45例[J].新中医,1997,29(6):48-52.
93. 李雪岩,王滨,彭如心,金香兰.蟾酥对蟾蜍坐骨神经干复合动作电位的影响[J].中国中医药科技,1996(6):365-366.
94. 徐叔云,卞如濂,陈修主编.药理实验方法学[M].第2版,北京:人民卫生版社,1994:968-969.
95. 杨华中,刘英姿,张镭. 蟾酥和维拉帕米镇痛协同作用的实验研究. 湖南师范人学学报(医学版), 2004,1(1):50-52.
96. 张薇,刘玉兰,徐从石,等.蟾酥的镇痛活性成分[J].沈阳药科大学学报,1998,15(4):268-271.
97. 薛开先.新医药学杂志1975:(1):39.
98. 李雪岩,王滨,彭如心,金香兰.蟾酥对蟾蜍坐骨神径干复合动作电位的影响[J].中国中医药科技1995,6(6):365.
99. 江苏新医学院.中药大词典(下册)[M].上海:上海人民出版社,1977:7-34.
100. 《中华本草》编委会.中华本草(第9册)[M].上海:上海科学技术出版社,1999:365.
101. [日]筱原达雄.淫羊着配合蟾酥制剂的抗疲劳作用.和汉医药学杂志,1996,13(3):260- 267.
102. 孙非,张淑芹,刘志屹,等.蟾毒内醋抗柯萨奇病毒、腺病毒和流感病毒作用的研究[J].中国药理学通报,2004,20(4):478-479.
103. 曲莉,孙非,张淑芹,等.蟾毒内酯抗流感病毒的实验研究[J].长春中医学院学报,2004,20 (1):37-38.
104. 杜平,朱关福,刘湘云.现代临床病毒学[M].北京:人民军医出版社,1991,233-285.
105. 杨秀伟,刑增涛,帐景荣.华蟾毒精和轻基华蟾毒精的人肠内细菌代谢研究[J].北京人学学报(医学版),2001,33(3):199-204.
106. 王浴生,等.中药药理与应用[M].北京:学苑出版社,1992.311.
107. Chan WY, Ng TB, Yeung HW.Examination for toxicity of a Chinese drug, the load glandular secretory product Chan Su, in pregnant mice and embryos [J]. Bilo Neonate, 1995,67(S):376-380.
108. 张瑜瑶.蟾酥水溶性总成分的实验研究[J].南京中医学院学报,1988,(2):33
109. 李树林.毒物的毒理与毒物分析[M].北京:人民卫生出版社,1987,5:596-597.
110. 钱伟平.蟾酥对实验动物循环系统毒副作用的研究[J]. 浙江中医学院学报,2001,25(6): 46-47.
111. G.哈贝尔梅尔.有毒动物和毒物毒[M].北京:科学技术出版社.1981,6:48-49.
112. Yeh J Y, Huang W J, Kan S F, et al. Effect of bufalin and cinobufagin on the proliferation of androgen dependent and independent prostate cancer cells [J]. Prostate,2003,54(2):112-124.
113. Brubacher J R, Ravikumar P R, Bania T, et al. Treatment of toad venom poisoning with digoxin-specific Fab fragments [J].Chest,1996,110(5):1139-1141.
114. 赵强,孟凡静,刘安西.蟾酥的研究进展[J].中草药,2004,35(IO):4-7.
115. 韩莉.中药蟾酥临床应用简介[J].实用医药杂志,2003,20(11):854
116. 齐刚,张莉,何文形,等. Bufalin的急性毒性研究[J].武警医学院学报,2003,13(1):10-12.
117. 陈端艺,林巧艺,高淑琼,等.服六神丸中毒一例[J].中国基层医药,2001,8(2):106.
118. 陈强,高翠萍.服六神丸出现过敏反应一例[J].中成药,2002,27(10):798.
119. 张萍,文家福.新生儿六神丸中毒一例[J].中华急诊医学杂志,2001,10(3):172.
120. 张凡景,马东成,赵霞.女童口服六神丸引起雷诺现象一例[J].中国中药杂志,1999,24(9): 570.
121. 龚岳亭.生物活性肤[M].上海:上海科学技术出版社.1985.10,161-162.
122. 梁秀兰,许廷生,黄新.蟾酥及其成方制剂的合理应用[J].中国药物与临床,2004,4(3):217.
123. 谭道玉,王绍琼,李昌平,等.纷妥拉明顶防华蟾素所致局部静脉反应的临床观察[J].中国中西医结合杂志,2003:23(12):940.
124. 杨勇,奉建芳,芦洁,等. 蟾酥固体脂质纳米粒冻干针剂体内药代动力学研究[J].中成药. 2006,28(6):787-790.
125. 范健,黄慰魁,武福乐,等.华蟾毒精微球制剂肝动脉栓塞的实验研究[J].癌症,1995,14(6): 434.
126. 韩洪刚,刘云海,谢委.蟾酥β-环糊精包合物的研究现状[J]. 医药导报,2005,24(11): 1039-1041.
127. 张梅,阳长明.冠心微丸中脂蟾毒配基的含量测定[J].湖南中医杂志,2001,17(2):59-60.
128. 唐青华,劳三申,高明珠.等.金蟾定痛微粒丸下艺的研究[J].中成药.1997,19(3):3-5.
129. 虞金宝,程怡,金克宁,等蟾酥透皮吸收研究[J].中国中药杂志,1995,20(9):539.
130. 朱盛山,吕志平,刘强.氮酮对蟾酥中蟾毒内酯类成分透皮吸收的影响[J].中成药. 2003, 25(9):694-685.
131. 杨莉娅,丁训诚.巴布剂及橡胶剂蟾酥膏对家兔完整皮肤刺激试验的比较[J].中国药业,2000, 9(5):11.
132. 王隶书,程东岩,刘永宏.华蟾毒精气雾剂的研制[J].中成药,1998,20(4):1-2.
133. 周学敏,程培元,许美娟,等.HPLC法分离测定蟾酥脂质体中三种蟾毒内酯成分[J].中国药科大学学报1993.24(2):116.
134. 李勤,谢艳华,孙纪元,等.中药安迪粉针剂对离体肿瘤细胞的细胞毒作用[J].西安交通大学学报(医学版),2003,24(8):390-392.
135. 王彦泓, 胡俊敏.中药消毒栓杀菌和杀精子性能的试验观察[J].河北医药,2001,23(5): 346-348.
136. 孟庆增,袁青春,姚元庚,等.蟾酥复方消毒剂性能的实验观察[J].中国消毒学杂志, 1998,15 (1):26-29.
137. 孔凤利.中药蟾酥的鉴别[J].浙江中医学院学报,2004,28(6):78
138. 陈美珍.中药蟾酥的鉴别[J].中草药,2002,33(3):272.
139. 赖新华.蟾酥的真伪鉴别法[J].基层中药杂志,2002,16(2):34.
140. 伊惠贤.用多次层析法鉴别中药成方制剂中的蟾酥[J].中草药,1997,28(5):277-278.
141. 祝君.牛黄消炎片的薄层色谱鉴别[J].中国药业,2001,10(2):19-20.
142. 邵陆.康肤灵配鉴别试验的研究[J].山东医药工业,1996,15(2).
143. 石俊英.哈蟆油、鸡内金等动物类中药及其混淆品的电泳鉴别研究[J].山东中医杂志,1994, 13(2):73-74.
144. 霍建中.微波消解ICP-AES法测定澳洲蟾酥中的常量及微量元素[J].天津中医学院学报,2003, 22(1):19-20.
145. 于萍.优劣蟾酥的毛细管气相色谱鉴别方[J].分析测试技术与仪器,1998,4(4):245-250.
146. 工伟,乔文礼,塔娜.薄层扫描法测定梅花点舌丸中华蟾蜍毒基含量[J].国现代应用药学杂志, 2001,18(2):129-131.
147. 王莉华.双波长薄层扫描法测定风油精中华蟾毒精的含量[J].中国药科大学学报,1995,26 (5):279-281.
148. 钟镜金.薄层--分光光度法测定制剂中脂蟾毒配基的含量[J].中药材,1995,18(6):311-312.
149. 段亚丽.紫外分光光度法控制酥黄注射液中蟾酥质量[J].广西农业科学,2005,36(2):158-159.
150. 杨锡,罗兴平.本底校正分光光度法测定蟾酥及其制剂中蟾毒内脂的含量[J].药物分析杂志, 1994,14(4):19-21.
151. 蟾酥的研究(1):用比色计测定商品蟋酥的化学成分差异〔日〕/川原一仁…//Natural Medines,1999,53(5).242 – 248.
152. 何丽霞.成方制剂中蟾酥的质量控制[J].中国兽药杂志,2003,37(4):28-30.
153. 王京辉.高效液相色谱法测定蟾酥中有效成分的含量[J].中国中药杂志,1998,23(11): 651-653.
154. 乔延江,吴刚,王玺,等.中药蟾酥质量的人工神经网络化学模式识别研究[J].分析化学 1995, 23(6)630-634.
155. 乔延江,王玺,毕开顺,等.人工神经网络在中药蟾酥化学模式识别特征提取中的应用[J].药学学报,1995;30(9):698-701.
156. 王玺,马丽娟,潘卫东.等.蟾酥薄层色谱数据的化学模式识别[J].沈阳药科大学学报,1995, 12(4):50-254.
157. 葛勤,康钦树.蟾酥的化学及制剂质量研究近况[J].中成药,1998,20(1):38-40.
158. 王永刚,吴忠,魏凤环,等.中药指纹图谱研究的现状与未来[J].中药材,2003,26(11): 820-825.
159. 国家药品监督管理局.中药注射液指纹图谱研究的技术要求(暂行)[J].中成药,2000,22(10): 671.
160. 江苏新医学院[M].中药大词典(下册).上海:上海人民出版社,1977,27-34.
161. 杨基森主编.中国药酒学[M].贵州:贵州科技出版社,2002,8.