中药蒺藜活性成分及质量评价研究
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摘要
根据我们的立题目的，结合药效学的结果，首先确定活性部位，然后对蒺藜的具有活性的水溶性部位进行了系统的化学成分研究，以确定蒺藜的活性物质基础。同时对分离的皂苷和黄酮的单体化合物进性活性研究，确定其心血管活性的前提下，用以作为质量控制的指标成分，使得质量控制方法与活性有机结合，因此我们制定的质量控制方法对蒺藜的质量评价更具确切性和可靠性。具体研究内容如下：
首先对蒺藜水溶性主要成分进行初步研究。根据蒺藜药理方面的文献报道，蒺藜(全草)的水提取物是蒺藜的心脑血管系统主要活性部位(例如心脑舒通胶囊)，因此本文对其具有活性的水溶性成分进行了初步研究。采用比色法和HPLC法分别对蒺藜的总皂苷类成分和总黄酮类成分分别进行含量测定，并对所制定的方法进行了方法学研究。之后，根据所确定的方法进行了含量测定，测定结果表明，蒺藜(全草)的水提取成分中，除了含有总皂苷类成分，还有含量较高的总黄酮类成分，提示蒺藜中总黄酮类成分也可能是蒺藜的活性部位。因此就蒺藜总黄酮在心脑血管系统方面的作用与蒺藜的总皂苷成分做了活性对比实验，药理实验研究结果提示，总黄酮与总皂苷均能显著提高小鼠耐缺氧能力，均能明显延长凝血时间，对离体家兔心脏的冠脉流量明显增加作用。实验结果说明，蒺藜中总黄酮类成分与蒺藜的总皂苷成分一样具有心脑血管系统方面的活性作用。本实验之前并无蒺藜中总黄酮类成分心脑血管系统方面的活性研究报道，蒺藜总黄酮类的活性研究，尚属首次。
由于大孔树脂对于水溶性成分具有良好的富集、纯化作用，因此，对蒺藜的水溶性成分进行有效成分大孔树脂的富集法研究。实验收集了9种常用大孔吸附树脂，对吸附容量进行了对比，确定采用树脂AB-8。又对AB-8树脂富集有效成分的工艺条件进行了考察，采用正交实验以总含量(总皂苷含量+总黄酮含量)为指标进行实验，结果确定富集蒺藜水溶性成分的最佳工艺条件(药液浓度0.5g／mL，流速8／1000BV／min；径高比1：40)。通过本实验得出的提取条件，为蒺藜的进一步研究提供了依据。
其次，化学分离实验主要对50Kg蒺藜，利用前述实验确定的条件，进行大孔树脂富集，对所得提取物(活性部位)进行化学分离。主要采用硅胶柱、葡聚糖凝胶色谱柱等分离手段进行了化学分离，共得到20个单体化合物，通过理化性质及波谱解析鉴定了其中的18个化合物的结构，分别是：terrestris A(TT1)；海柯皂苷元-3-O-β-D-木糖(1-2)-[β-D-葡萄糖-(1-3)]-β-D-葡萄糖(1-4)-β-D-甘露糖(TT2)；海柯皂苷元-3-O-β-D-木糖(1-2)-[β-D-木糖(1-3)]-β-D-葡萄糖(1-4)-[α-鼠李糖(1-2)]-β-D-甘露糖(TT3)；槲皮素-3-O-β-D-葡萄糖苷(TT4)；槲皮素-3-O-龙胆二糖苷(TT5)；山柰酚-3-O-龙胆二糖苷(TT6)；山柰酚-3-O-β-D-葡萄糖苷(TT7)；槲皮素(TT8)；海柯皂苷元(TT9)；25R-spirostan-4-ene-3，12-dione(TT10)；25R-spirostan-3，6，12-trione(TT11)；胡萝卜苷(TT12)；丁二酸(TT13)；β-sitosterol(TT14)；25R-spirostan-2,3-diol-4-ene-12-one(TT15)；7-羰基-△~(5.23)豆甾醇(TT16)；香兰素(TT17)；二十二烷酸(TT18)。其中terrestris A(TT1)是新化
Tribulus terrestris L. are a famous traditional Chinese medicine and has been used for over hundreds years. It can restore the depressed liver for the treatment of fullness in the chest and mastits and also used to dispel the wind and clear the eyes for the treatment of an acute conjunctivitis, headache and vertigo. It has been used for a variety of liver, kidney, dizziness, psoriasis, eczema, premature ejaculation and cardiovascular diseases. The Greeks used the medicinal herb Tribulus terrestris L.as a diuretic and a mood-enhancer. Eastern Indians used it as a diuretic, antiseptic, and anti-inflammatory. The people of Bulgaria used Tribulus terrestris L.as a sex enhancer and to treat infertility.
Tribulus terrestris L.is used in various herbal formulas to treat headaches eye problems such as itching, conjunctivitis and weak vision, pain relief, enhancement of the immune system, improved mood and sense of well-being, cholesterol reduction, and relief of premenstrual (PMS), menopausal symptoms in women, and nervousness, making it a great menopause herb. In women, hormonal imbalances can lead to the typical unpleasant symptoms associated with PMS and menopause. It is also used to treat high blood pressure and rib pain. Recently, eastern European athletes and strength champions have used it as well. Many athletes and bodybuilders have found Tribulus terrestris L. a safe and natural alternative to anabolic steroids.
Tribulus terrestris L.contains three groups of active physiochemicals:
Dioscin, protodioscin, diosgenin and similar, these substances have effect on sexual performance and may treat various sexual disorders, they regulate sexual energy level and strength by increasing the percentage of free testosterone level for men and they effect pregnenolone, progesterone and estrogen. The hormone balancing effects of Bulgarian Tribulus terrestris L.for women makes this herb suitable for premenstrual syndrome and menopausal syndrome. Sterols like betasitosterols or stigma. These substances protect the prostate from swelling and in combination with the steroidal saponins, protect the prostate from cancer. Steroidal saponins currently referred to as steroidal saponins. These steroidal saponins effect the complete immune system. They have been demonstrated to possess anti-bacterial and anti-viral effects. Bulgarian Tribulus terrestris L.may be used internally and externally to treat herpes, and virus infections such as influenza and the common cold.
In this studies, firstly, in order to do more studying on Tribulus terrestris L., we investigated its water-solubility constituents by using UV-spectral method to determine the content of the total saponins and HPLC method to to determine the total flavonoid glycosides. Methods studying results shows that the two methods were convenient, rapid and accurate, and could be used for quality control of quality of Tribulus terrestris L. According these two methods, we took study on Tribulus terrestris L. The 3 samples results show that the total Saponins in Tribulus terrestris L. are 1.55、1.59、1.67, average is 1.60%, and the total flavonoid glycosides are0.97、 1.12、1.23, average 1.11%. The resins are more proper used to exacting water-solubility constituents in herbs these days. We compared nine kinds of different resin. AB-8, performed a good reputation, was being adapted. The optimum condition of using AB-8 were concentration: 0.5g/mL , velocity of flow:8/1000BV/min, ratio of diameter and high: 1: 40, and were development by using orthogonal tests.
Secondly, We investigated Tribulus terrestris L.chemical constituents of the fruits of Tribulus terrestris L by using SiO_2, polyamide, gel, LH-20. Chemical constituents from Tribulus terrestris L. 20compounds were isolated from it and 17 of them were structurally elucidated. With the compounds name: terrestrisA(TTl); hecogenin-3-O-.(3-D-xylopyransyl(1-2)-[β-D-glucopyranosyl
首先对蒺藜水溶性主要成分进行初步研究。根据蒺藜药理方面的文献报道，蒺藜(全草)的水提取物是蒺藜的心脑血管系统主要活性部位(例如心脑舒通胶囊)，因此本文对其具有活性的水溶性成分进行了初步研究。采用比色法和HPLC法分别对蒺藜的总皂苷类成分和总黄酮类成分分别进行含量测定，并对所制定的方法进行了方法学研究。之后，根据所确定的方法进行了含量测定，测定结果表明，蒺藜(全草)的水提取成分中，除了含有总皂苷类成分，还有含量较高的总黄酮类成分，提示蒺藜中总黄酮类成分也可能是蒺藜的活性部位。因此就蒺藜总黄酮在心脑血管系统方面的作用与蒺藜的总皂苷成分做了活性对比实验，药理实验研究结果提示，总黄酮与总皂苷均能显著提高小鼠耐缺氧能力，均能明显延长凝血时间，对离体家兔心脏的冠脉流量明显增加作用。实验结果说明，蒺藜中总黄酮类成分与蒺藜的总皂苷成分一样具有心脑血管系统方面的活性作用。本实验之前并无蒺藜中总黄酮类成分心脑血管系统方面的活性研究报道，蒺藜总黄酮类的活性研究，尚属首次。
由于大孔树脂对于水溶性成分具有良好的富集、纯化作用，因此，对蒺藜的水溶性成分进行有效成分大孔树脂的富集法研究。实验收集了9种常用大孔吸附树脂，对吸附容量进行了对比，确定采用树脂AB-8。又对AB-8树脂富集有效成分的工艺条件进行了考察，采用正交实验以总含量(总皂苷含量+总黄酮含量)为指标进行实验，结果确定富集蒺藜水溶性成分的最佳工艺条件(药液浓度0.5g／mL，流速8／1000BV／min；径高比1：40)。通过本实验得出的提取条件，为蒺藜的进一步研究提供了依据。
其次，化学分离实验主要对50Kg蒺藜，利用前述实验确定的条件，进行大孔树脂富集，对所得提取物(活性部位)进行化学分离。主要采用硅胶柱、葡聚糖凝胶色谱柱等分离手段进行了化学分离，共得到20个单体化合物，通过理化性质及波谱解析鉴定了其中的18个化合物的结构，分别是：terrestris A(TT1)；海柯皂苷元-3-O-β-D-木糖(1-2)-[β-D-葡萄糖-(1-3)]-β-D-葡萄糖(1-4)-β-D-甘露糖(TT2)；海柯皂苷元-3-O-β-D-木糖(1-2)-[β-D-木糖(1-3)]-β-D-葡萄糖(1-4)-[α-鼠李糖(1-2)]-β-D-甘露糖(TT3)；槲皮素-3-O-β-D-葡萄糖苷(TT4)；槲皮素-3-O-龙胆二糖苷(TT5)；山柰酚-3-O-龙胆二糖苷(TT6)；山柰酚-3-O-β-D-葡萄糖苷(TT7)；槲皮素(TT8)；海柯皂苷元(TT9)；25R-spirostan-4-ene-3，12-dione(TT10)；25R-spirostan-3，6，12-trione(TT11)；胡萝卜苷(TT12)；丁二酸(TT13)；β-sitosterol(TT14)；25R-spirostan-2,3-diol-4-ene-12-one(TT15)；7-羰基-△~(5.23)豆甾醇(TT16)；香兰素(TT17)；二十二烷酸(TT18)。其中terrestris A(TT1)是新化
Tribulus terrestris L. are a famous traditional Chinese medicine and has been used for over hundreds years. It can restore the depressed liver for the treatment of fullness in the chest and mastits and also used to dispel the wind and clear the eyes for the treatment of an acute conjunctivitis, headache and vertigo. It has been used for a variety of liver, kidney, dizziness, psoriasis, eczema, premature ejaculation and cardiovascular diseases. The Greeks used the medicinal herb Tribulus terrestris L.as a diuretic and a mood-enhancer. Eastern Indians used it as a diuretic, antiseptic, and anti-inflammatory. The people of Bulgaria used Tribulus terrestris L.as a sex enhancer and to treat infertility.
Tribulus terrestris L.is used in various herbal formulas to treat headaches eye problems such as itching, conjunctivitis and weak vision, pain relief, enhancement of the immune system, improved mood and sense of well-being, cholesterol reduction, and relief of premenstrual (PMS), menopausal symptoms in women, and nervousness, making it a great menopause herb. In women, hormonal imbalances can lead to the typical unpleasant symptoms associated with PMS and menopause. It is also used to treat high blood pressure and rib pain. Recently, eastern European athletes and strength champions have used it as well. Many athletes and bodybuilders have found Tribulus terrestris L. a safe and natural alternative to anabolic steroids.
Tribulus terrestris L.contains three groups of active physiochemicals:
Dioscin, protodioscin, diosgenin and similar, these substances have effect on sexual performance and may treat various sexual disorders, they regulate sexual energy level and strength by increasing the percentage of free testosterone level for men and they effect pregnenolone, progesterone and estrogen. The hormone balancing effects of Bulgarian Tribulus terrestris L.for women makes this herb suitable for premenstrual syndrome and menopausal syndrome. Sterols like betasitosterols or stigma. These substances protect the prostate from swelling and in combination with the steroidal saponins, protect the prostate from cancer. Steroidal saponins currently referred to as steroidal saponins. These steroidal saponins effect the complete immune system. They have been demonstrated to possess anti-bacterial and anti-viral effects. Bulgarian Tribulus terrestris L.may be used internally and externally to treat herpes, and virus infections such as influenza and the common cold.
In this studies, firstly, in order to do more studying on Tribulus terrestris L., we investigated its water-solubility constituents by using UV-spectral method to determine the content of the total saponins and HPLC method to to determine the total flavonoid glycosides. Methods studying results shows that the two methods were convenient, rapid and accurate, and could be used for quality control of quality of Tribulus terrestris L. According these two methods, we took study on Tribulus terrestris L. The 3 samples results show that the total Saponins in Tribulus terrestris L. are 1.55、1.59、1.67, average is 1.60%, and the total flavonoid glycosides are0.97、 1.12、1.23, average 1.11%. The resins are more proper used to exacting water-solubility constituents in herbs these days. We compared nine kinds of different resin. AB-8, performed a good reputation, was being adapted. The optimum condition of using AB-8 were concentration: 0.5g/mL , velocity of flow:8/1000BV/min, ratio of diameter and high: 1: 40, and were development by using orthogonal tests.
Secondly, We investigated Tribulus terrestris L.chemical constituents of the fruits of Tribulus terrestris L by using SiO_2, polyamide, gel, LH-20. Chemical constituents from Tribulus terrestris L. 20compounds were isolated from it and 17 of them were structurally elucidated. With the compounds name: terrestrisA(TTl); hecogenin-3-O-.(3-D-xylopyransyl(1-2)-[β-D-glucopyranosyl
引文
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[1] Bensky D, Gamble A. Chinese Herbal Medicine Materia Medica. Eastland Press, Seattle, 1986, p607-8.
[2] Kapoor LD. CRC Handbook of Ayurvedic Medicinal Plants. CRC Press, Boca Raton, 1990, p 325.
[3] Jayaram S, et al. Tribulus. Indian Drugs 1993; 30(10): 498-500.
[4] Gjulemetowa R, Tomowa M, Simowa M et al. Concerning the determination of furanostanol saponins in the preparation Tribestan(?). Pharmazie 1982; 37(4): 296.
[5] Perepelitsa V, et al. A chemical, study of the steroid glycosides of Tribulus terrestris. Ⅳ. Steroid saponins. Khimya Prirodnykh Soedinenii 1975; 2: 260-1.
[6] Mahato SB, Sahu NP, Ganguly AN et al. Steroidal glycosides of Tribulus terrestris. J Chem Soc Perk Trans 1981; 1(9): 2405-10.
[7] Milanov S, Maleeva E, Taskov M. Tribestan effect on the concentration of some hormones in the serum of healthy volunteers. Med-Biol Inf 1985; 4: 27-9.
[8] Georgiev P, Dimitrov M, Vitanov S. The effect of the preparation "Tribestan" on the plasma concentration of testosterone and spermogenesis of lambs and rams. Vet Sb 1988; 3: 20-2.
[9] Zarkova S. Tribestan: Experimental and Clinical Investigations. Chemical Pharmaceutical Research Institute, Sofia, 1982.
[10] Taskov M. Vitaton. Med-Biol Inf 1988; 1: 3.
[11] Dikova N, Ognyanova V. Pharmacokinetic studies on Tribestan-.Anniversary scientific session 35 Years Chemical Pharmaceutical Research Institute, Sofia, 1983.
[12] Sheitanov M, Khristov T, Taskov M, et al. Vitaton: comparative studies on its effect on incorporation intensity of labeled amino acids in cellular proteins. Med-Biol Inf 1988; 1: 20-3.
[13] Toshkov A, Dimov V, Zarkova S. Tribestan: immunostimulating properties. Med-Biol Inf 1985; 28-31.
[14] Tomova M, Gyulemetova R. Steroidsapogenine: Ⅵ. Furostanol bisglykosid aus Tribulus terrestris L. Planta medica 1978; 34: 188-91.
[15] Protich M, Tsvetkov D, Nalbanski B, et al. Clinical trial of the preparation Tribestan in infertile men. Akush Ginekol 1983; 22(4): 326-9.
[16] Kumanov F, Bozadzhieva E, Andreeva M, et al. Clinical trial of the drug "Tribestan." Savr Med 1982; 4: 211-5.
[17] Viktorov Ⅳ, Kaloyanov AL, Lilov L, et al. Clinical investigation on Tribestan in males with disorders in the sexual function. Med-Biol Inf 1982. Cited in Zarkova S. Tribestan: Experimental and Clinical Investigations. Chemical Pharmaceutical Research Institute, Sofia, 1982.
[18] Zarkova S. Tribestan: Experimental and Clinical Investigations.. Chemical Pharmaceutical Research Institute, Sofia, 1983.
[19] Wang B, Ma L, Liu T. 406 cases of angina pectoris in coronary heart disease treated with saponin of Tribulus terrestris. Chung His I Chieh Ho Tsa Chih 1990; 10(2): 85-7.
[20] Tanev G, Zarkova S. Toxicological studies on Tribestan. Cited in Zarkova S. Tribestan: Experimental and Clinical Investigations. Chemical Pharmaceutical Research Institute, Sofia, 1985.
[21] Gendzhev Z. The carcinogenicity of Tribestan. Tr Nauchnoizsled Khim Farm Inst 1985; 15: 241-50.
[22] Graydon RJ, Hamid H, Zahari P, Gardiner C. Photosensitisation and crystal-associated cholangiohepatopathy in sheep grazing Brachiaria decumbens. Aust Vet J 1991 Jul; 68(7): 234-6.
[1] 张菊红，王志伟．蒺藜全草中总皂苷总含量的测定[J]．中国临床药学杂志．2001，10(4)：244．
[2] 李敏，费曜，李丽霞，方道硕．天冬中总皂苷含量测定方法的探讨[J]．成都中医药大学学报2004，27(4)：46．
[3] 国家药典委员会．中国药典，Ⅰ部[S]．北京化学工业出版社，2005．
[1] 褚书地，瞿伟菁，李穆 蒺藜化学成分及药理作用研究进展 中国野生植物资源．2003，22(4)：4-6
[2] 王淑君，候薇，张天政，毕殿洲HPLC法测定银杏叶缓释胶囊中总黄酮的含量中草药2004，3(2)：163
[3] 国家药典委员会．中国药典，Ⅰ部[S]．北京化学工业出版社，2005．．
[4] 国家中成药标准汇编 中成药地方标准上升国家标准部分 内科心系分册513
[1] 张菊红，王志伟．蒺藜全草中总皂苷总含量的测定[J]．中国临床药学杂．2001，10(4)：244
[2] 王淑君，候薇，张天政HPLC法测定银杏叶缓释胶囊中总黄酮的含量[J1．中草药2004，3(2)：163
[3] 国家药典委员会．中国药典，Ⅰ部[S]．北京化学工业出版社，2005．
[4] 国家中成药标准汇编，中成药地方标准上升国家标准部分，内科心系分册：513
[5] 李敏，费曜，李丽霞，方道硕．天冬中总皂苷含量测定方法的探讨[J]．成都中医药大学学报2004，27(4)：46
[1] 王光忠，胡迪，陈敬炳．正交设计优选黄芪总黄酮和总皂苷的提取工艺[J]．中成药，2004，10(26)：3．
[2] 陈发奎．常用中草药有效成分含量测定[M]．北京：人民卫生出版社，1997，620-621．
[1] 于德泉，杨俊山．分析化学手册，第七分册，北京，化学出版社：393：
[2] Yoshihiyo Mimaki, Toshihiro Kanmoto, Minpei Kuroda. et al. Steroialal saponins from underground part of Hosta Longipes and their inhibitory activity on tumor promoter-enduced phospholipid, Chem. pharm. Bull. 1995. 43(7)1190-1196
[3] phytochemistry, 1990, 29, 3893-3898
[4] phytochemistry, 1996, 41, 1405-1410
[5] 槲皮素-3-O-葡萄糖苷
[6] Nabiel A. M. Saleh, Ahmed A. Ahemed and Mohamed F. Abdalla. Flavonoid glycosids of tribulus pentandrus and T. terrestris. Phytochemist, 1982, 21(8)1995-2000
[7] Petler A, ward R, Lansgray T. Ther Carbon-13 nuclear magnetic resonance spectra of flavonoids and related compounds. J of the chemical society, perkin Trans. 1976. 2475-2483
[8] Yi-Xin Xu, Hai-Sheng Chen, Wen-Yong Liu et al, Two sopogenins from Tribulus terrestris, Phytochemistry 1998. 49(1)199-201.
[9] 王艳、陆蕴如．刺蒺藜的化学成分的研究．西北药学杂志．1990．5(4)14-17
[10] 胡萝卜苷
[11] 丁二酸
[12] 黄金文、蒋山好、谭昌恒等，三个新甾体皂苷元的经构鉴定．有机化学2002．22(11)：917-921
[2] 宣伟东等．HPLC-ELSD法测定刺蒺藜中甾体皂苷TTS-12含量[J]第二军医大学学报，2005，02
[3] 牛伟等RP-HPLC比较蒺藜全草和果实中的主要甾体皂苷元成分[J]中成药2005，04
[4] 柳文媛等藜产地与薯蓣皂苷元含量[J]中药材2001，06
[5] 张菊红等蒺藜全草中总皂苷总含量的测定[J]中国临床药学杂志2001，04
[6] 师勤等反相高效液相色谱法测定蒺藜类3种水解黄酮苷元的含量[J]药物分析杂志1999，02
[7] 吴琼诗蒺藜全草中总黄酮的含量测定[J]广东药学院学报1998，04
[8] 师勤比色法测定蒺藜中多糖的含量[J]中国药科大学学报1997，05
[9] 王学东等蒺藜中氨基酸的含量测定[J]氨基酸和生物资源1994，02
[10] S. P. Bhutani, et al Phytochemisty 1969,8,299
[11] N. A. M,salch et al, Phytochemistry 1982, 21(8), 1995
[12] M. P. Tomova, et al, C. A., 1974, 81: 23083r
[13] E. D, Perplitsa,et al,C,A, 1975,8.3:12865Q
[14] S. B. Mahato,et al,J.Chem. Soc.Perkin TransI, 1981, (9),2405
[15] H. Aehenbach et al, Phytochemistry 1994; 35(6), 1527
[16] Y. Wang, et al, Phytochemistry 1996; 42(5), 1417
[17] Y. Wang, et al, Phytochemistry 1997;45(4),811-817
[18] G. Wu, et al, Phytochemistry 1996; 42(6), 1671
[19] 蔡利锋等蒺藜化学成分的研究[J]药学学报1999，10
[20] Y-XXu, et al, Phytochmistry2000, (66), 545-550
[21] S. B. Mahato, et al, J. Chem. Soc.Perkin TransI, 1981,(9),2405
[22] 2 2Y-X Xu, et al, Phytochmistry1998; 49(1),199-201
[23] 黄金文等蒺藜甾体皂苷元化学成分研究[J]天然产物研究与开发2003，02
[24] 师勤等，Planta Medical998，64，628-631
[25] T-SWu, et al, Phytochemistry 1999, 50, 1411-1415
[26] 任渝江等刺蒺藜果中一种新桂皮酰胺类成分的分离和鉴定[J]药学学报1994，29(3)，204-206
[27] 刘杰等中药刺蒺藜化学成分的研究[J]第二军医大学学报2003，02
[28] 陈海生等蒺藜中一个新的有机酸成分的分离与鉴定[J]第二军医大学学报2004，11
[28] 刘杰等中药刺蒺藜化学成分的研究[J]第二军医大学学报2003，02
[29] 殷翔等蒺藜皂苷水提工艺研究[J]上海医药2004，05
[30] 金京玲等大孔吸附树脂法提取蒺藜总皂苷[J]延边大学医学学报1999，01
[31] 李树珍等蒺藜不同提取工艺的研究[J]中草药1994，09
[32] 苏瑞强等蒺藜粗皂苷提取工艺研究[J]山东医药工业1997，05
[33] 吴家宽等JL-GE对SD大鼠血液流变性的影响[J]中国微循环2002，6(1)：33
[34] 罗兰，殷惠军，史大卓心脑舒通胶囊对小鼠心肌营养性血流影响的研究[J]中国中医药科技2005，12(4)
[35] 姜宗文等．蒺藜果总皂苷对大鼠实验性脑缺血的保护作用[J]中草药2002，33(11)：1020
[36] 吕文伟等．蒺藜果总皂苷对实验性脑缺血作用的研究[J]中国老年学杂志2003，23(2)：54
[37] 王丽岩，祁功才蒺藜果总皂苷抗血栓形成作用[J]中华中西医学杂志，2005，10
[38] 王显刚，殷惠军，郭艳白蒺藜皂苷对高血脂大鼠心肌梗死后血脂及炎性因子表达影响的研究[J]中医药信息2005，22(5)：85
[39] 罗兰，殷惠军，史大卓．心脑舒通胶囊对脂质代谢及高半胱甘氨酸水平影响的研究[J]中国中医药科技，2005，12(4)：237
[40] 郝爱功蒺藜全草皂苷对果蝇寿命的影响．
[41] 张涛等．刺蒺藜对衰老小鼠红细胞SOD活性和MDA含量的影响[J]
[42] 张健等，蒺藜提取物的药理研究进展[J]中医药研究，1998，14(5)：56-5
[43] 钱本余，蒺藜研究概述[J]中成药1990，12(11)：3
[44] Mitra, S. K., Muralidhar, T. S., et al. Experimental assessment of relative efficacy of drugs of herbal origin on sexual performance and hormone levels in alcohol-exposed and normal rats, Chemical Abstracts, 1996, 125: 132506d.
[45] 陈子等．防风与刺蒺藜的药理实验研究[J]中成药，2003，25(9)：737
[46] 李艳莉，钟理，梁丽红6种中药抑制酪氨酸酶活性的实验研究[J]时珍国医国药，2002，03：131
[47] 朱铁君白癜风的病因、诊断和治疗[J]中国全科医学2005，8(6)：105
[48] 邓燕，杨柳，安胜利．不同浓度的中药刺蒺藜对黑素细胞和酪氨酸酶的作用[J]第一军医大学学报，2002，22(11)：1017
[49] 李洪苷，朱文元，夏明玉赤芍等几种上调酪氨酸酶活性中药乙醇提取液对棕黄色豚鼠背部皮肤的增色作用[J]临床皮肤科杂志，2004，30(2)
[50] 李明娟等．蒺藜皂苷的降血糖作用[J]中药材，25(6)：420
[51] 李明娟等蒺藜水煎剂对小鼠糖代谢中糖异生的作用[J]中药材，24(8)：588．
[52] 长华，蒋幼芹，江冰白蒺藜醇苷对混合培养鼠视网膜神经节细胞的作用叶[J]眼视光学杂志2001，3(3)：149
[53] 公惟畏，公衍鹤蒺藜食醋浸泡液热敷治疗跟骨骨刺[J]山东中医杂志2005，1024(10)：597
[54] Sangeeta蒺藜水提取物对雄性成年大鼠尿草酸盐过多的治疗效应Phytothera-py Research[J]1993，7：116-119
[1] Bensky D, Gamble A. Chinese Herbal Medicine Materia Medica. Eastland Press, Seattle, 1986, p607-8.
[2] Kapoor LD. CRC Handbook of Ayurvedic Medicinal Plants. CRC Press, Boca Raton, 1990, p 325.
[3] Jayaram S, et al. Tribulus. Indian Drugs 1993; 30(10): 498-500.
[4] Gjulemetowa R, Tomowa M, Simowa M et al. Concerning the determination of furanostanol saponins in the preparation Tribestan(?). Pharmazie 1982; 37(4): 296.
[5] Perepelitsa V, et al. A chemical, study of the steroid glycosides of Tribulus terrestris. Ⅳ. Steroid saponins. Khimya Prirodnykh Soedinenii 1975; 2: 260-1.
[6] Mahato SB, Sahu NP, Ganguly AN et al. Steroidal glycosides of Tribulus terrestris. J Chem Soc Perk Trans 1981; 1(9): 2405-10.
[7] Milanov S, Maleeva E, Taskov M. Tribestan effect on the concentration of some hormones in the serum of healthy volunteers. Med-Biol Inf 1985; 4: 27-9.
[8] Georgiev P, Dimitrov M, Vitanov S. The effect of the preparation "Tribestan" on the plasma concentration of testosterone and spermogenesis of lambs and rams. Vet Sb 1988; 3: 20-2.
[9] Zarkova S. Tribestan: Experimental and Clinical Investigations. Chemical Pharmaceutical Research Institute, Sofia, 1982.
[10] Taskov M. Vitaton. Med-Biol Inf 1988; 1: 3.
[11] Dikova N, Ognyanova V. Pharmacokinetic studies on Tribestan-.Anniversary scientific session 35 Years Chemical Pharmaceutical Research Institute, Sofia, 1983.
[12] Sheitanov M, Khristov T, Taskov M, et al. Vitaton: comparative studies on its effect on incorporation intensity of labeled amino acids in cellular proteins. Med-Biol Inf 1988; 1: 20-3.
[13] Toshkov A, Dimov V, Zarkova S. Tribestan: immunostimulating properties. Med-Biol Inf 1985; 28-31.
[14] Tomova M, Gyulemetova R. Steroidsapogenine: Ⅵ. Furostanol bisglykosid aus Tribulus terrestris L. Planta medica 1978; 34: 188-91.
[15] Protich M, Tsvetkov D, Nalbanski B, et al. Clinical trial of the preparation Tribestan in infertile men. Akush Ginekol 1983; 22(4): 326-9.
[16] Kumanov F, Bozadzhieva E, Andreeva M, et al. Clinical trial of the drug "Tribestan." Savr Med 1982; 4: 211-5.
[17] Viktorov Ⅳ, Kaloyanov AL, Lilov L, et al. Clinical investigation on Tribestan in males with disorders in the sexual function. Med-Biol Inf 1982. Cited in Zarkova S. Tribestan: Experimental and Clinical Investigations. Chemical Pharmaceutical Research Institute, Sofia, 1982.
[18] Zarkova S. Tribestan: Experimental and Clinical Investigations.. Chemical Pharmaceutical Research Institute, Sofia, 1983.
[19] Wang B, Ma L, Liu T. 406 cases of angina pectoris in coronary heart disease treated with saponin of Tribulus terrestris. Chung His I Chieh Ho Tsa Chih 1990; 10(2): 85-7.
[20] Tanev G, Zarkova S. Toxicological studies on Tribestan. Cited in Zarkova S. Tribestan: Experimental and Clinical Investigations. Chemical Pharmaceutical Research Institute, Sofia, 1985.
[21] Gendzhev Z. The carcinogenicity of Tribestan. Tr Nauchnoizsled Khim Farm Inst 1985; 15: 241-50.
[22] Graydon RJ, Hamid H, Zahari P, Gardiner C. Photosensitisation and crystal-associated cholangiohepatopathy in sheep grazing Brachiaria decumbens. Aust Vet J 1991 Jul; 68(7): 234-6.
[1] 张菊红，王志伟．蒺藜全草中总皂苷总含量的测定[J]．中国临床药学杂志．2001，10(4)：244．
[2] 李敏，费曜，李丽霞，方道硕．天冬中总皂苷含量测定方法的探讨[J]．成都中医药大学学报2004，27(4)：46．
[3] 国家药典委员会．中国药典，Ⅰ部[S]．北京化学工业出版社，2005．
[1] 褚书地，瞿伟菁，李穆 蒺藜化学成分及药理作用研究进展 中国野生植物资源．2003，22(4)：4-6
[2] 王淑君，候薇，张天政，毕殿洲HPLC法测定银杏叶缓释胶囊中总黄酮的含量中草药2004，3(2)：163
[3] 国家药典委员会．中国药典，Ⅰ部[S]．北京化学工业出版社，2005．．
[4] 国家中成药标准汇编 中成药地方标准上升国家标准部分 内科心系分册513
[1] 张菊红，王志伟．蒺藜全草中总皂苷总含量的测定[J]．中国临床药学杂．2001，10(4)：244
[2] 王淑君，候薇，张天政HPLC法测定银杏叶缓释胶囊中总黄酮的含量[J1．中草药2004，3(2)：163
[3] 国家药典委员会．中国药典，Ⅰ部[S]．北京化学工业出版社，2005．
[4] 国家中成药标准汇编，中成药地方标准上升国家标准部分，内科心系分册：513
[5] 李敏，费曜，李丽霞，方道硕．天冬中总皂苷含量测定方法的探讨[J]．成都中医药大学学报2004，27(4)：46
[1] 王光忠，胡迪，陈敬炳．正交设计优选黄芪总黄酮和总皂苷的提取工艺[J]．中成药，2004，10(26)：3．
[2] 陈发奎．常用中草药有效成分含量测定[M]．北京：人民卫生出版社，1997，620-621．
[1] 于德泉，杨俊山．分析化学手册，第七分册，北京，化学出版社：393：
[2] Yoshihiyo Mimaki, Toshihiro Kanmoto, Minpei Kuroda. et al. Steroialal saponins from underground part of Hosta Longipes and their inhibitory activity on tumor promoter-enduced phospholipid, Chem. pharm. Bull. 1995. 43(7)1190-1196
[3] phytochemistry, 1990, 29, 3893-3898
[4] phytochemistry, 1996, 41, 1405-1410
[5] 槲皮素-3-O-葡萄糖苷
[6] Nabiel A. M. Saleh, Ahmed A. Ahemed and Mohamed F. Abdalla. Flavonoid glycosids of tribulus pentandrus and T. terrestris. Phytochemist, 1982, 21(8)1995-2000
[7] Petler A, ward R, Lansgray T. Ther Carbon-13 nuclear magnetic resonance spectra of flavonoids and related compounds. J of the chemical society, perkin Trans. 1976. 2475-2483
[8] Yi-Xin Xu, Hai-Sheng Chen, Wen-Yong Liu et al, Two sopogenins from Tribulus terrestris, Phytochemistry 1998. 49(1)199-201.
[9] 王艳、陆蕴如．刺蒺藜的化学成分的研究．西北药学杂志．1990．5(4)14-17
[10] 胡萝卜苷
[11] 丁二酸
[12] 黄金文、蒋山好、谭昌恒等，三个新甾体皂苷元的经构鉴定．有机化学2002．22(11)：917-921
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