常山碱的提取、分离及其药效学研究
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摘要
鸡球虫病是一种重要的寄生虫病,分布广,感染率高,常发生在15～50日龄的雏鸡,若不能有效控制,其现场发病率可达50 %～70 %,死亡率可达20 %～30 %,严重爆发时其死亡率可高达80 %。据统计,全球养鸡业每年仅球虫病造成经济损失就达20亿英镑。目前对该病的防治仍以化学合成药及抗生素为主,但由于长期应用,易产生抗药性,且在畜产品中因药物残留而影响人类的健康。鉴于此,人们将目光转向非抗生素防治研究,如中草药。
迄今为止,关于常山中单体的分离报道较为深入,而常山中生物碱提取工艺和分离方法的报道较少,并且在常山的药理作用的研究中,关于常山中生物碱对畜禽生产常见的球虫病的研究未见报道。因此,本文由此出发,对常山中生物碱开发为新型兽药的几个关键的步骤进行了一些研究,包括生物碱的提取、分离及其抗球虫活性的研究,以期为常山的工业化生产和进一步开发利用提供理论依据。主要结果如下:
1常山碱含量测定方法的确定。利用常山碱在碱水条件下水解,产生邻氨基苯甲酸并将其与N-1-萘基-乙二胺盐酸盐溶液显色的原理,用紫外分光光度计比色法测定得出,常山碱提取物的总碱含量为0.327 %。
2常山总生物碱提取工艺的研究。运用正交实验,分别以浸膏重量和常山碱含量作为评价指标,考察了超声波提取工艺的效果,运用SPSS 14.0软件对结果进行分析,得到优化的提取工艺如下:其提取条件为提取温度50℃,料液比1:5,提取时间1 h。
3为了评价常山碱的安全性,对其进行了小白鼠的急性毒性试验。预试验初步确定用药剂量,正式试验取60只昆明种小白鼠,灌胃给药后,连续观察7天,记录小白鼠的急性毒性反应,并通过改良寇氏法计算常山碱对小白鼠的半数致死量(LD50)及LD50的95 %可信限。结果表明,给药组部分小白鼠出现中毒反应并死亡,剖检主要脏器未见病理改变,测得LD50为18.16 g/kg体重,LD50的95 %可信限为15.35 g/kg～21.49 g/kg体重。实验结果初步提示,常山碱的毒性很低,临床用药安全可靠。
4在常山总生物碱抗鸡球虫病和预防鸡球虫试验中,用艾美耳球虫接种雏鸡人工引发鸡球虫病。通过不同剂量常山总生物碱防治鸡球虫病作比较,确定鸡球虫病最佳临床治疗和预防剂量,其中以20 mg/kg给药可以达到良好的治疗鸡球虫病效果,其抗球虫指数(ACI为166.31)。
Avian coccidiosis is an important disease that widely spreads with a high incidence in chicken farms and is often found in chicks at 15 to 50 days old. If not effectively controlled, the prevalence of infection will be high up to 50 %～70 % with a mortality of 20 %～30 % or 80 % in severe cases. Statistical analysis indicated that the annual cost caused by the disease is estimated at￡200 million in the world. Current control of coccidosis mainly depends on synthetic chemicals or antibiotics. The durable use of the chemical drugs in the poultry industry easily leads to the resistant strains as well as the drug residues in the chickens and their products which are harmful to humans. Therefore, more and more research has shifted to non-antibiotic approaches such as medicinal herbs.
So far, study on monomer abstraction from Dichroa Febrifuga deeply, discovered that until now the alkaloid extraction process and the abstraction method study report are few about Dichroa Febrifuga. It is really micro studying on the common avian coccidiosis of livestock and fowl among the numerous pharmacologicalactions of alkaloid extraction from Dichroa Febrifuga. According to the above, this article study on each committed step for new veterinary medicine, including the alkaloid extraction, abstraction and the research of anticoccdidal activity in chicken, to provide some basic theory for a large-scale industry. The main results were indicated as follows:
First, the content of febrifugine.The present work presents a colorimetric method based on the principle that o-aminobenzoic acid is produced by the alkaline hydrolysis of the quinazolone structure of the febrifugine molecule, which can be determined colorimetrically by coupling with N-1-naphthyl ethylene diamine after diazotization. The content of febrifugine determinated by UV spectrophotometer and is 0.327 %.
Second, research on the technical of alkaloid extraction from Dichroa Febrifuga. Using the orthogonal experimental designed method, taken the evaluating target by the extracting weight and the content of febrifugine, analyzed by software SPSS 14.0, inspected extraction process of the ultrasonic wave method . The condition is the extraction temperature 50℃, the ratio of dosage liquor 1:5, extraction time 1 h.
Third, in order to evaluate the security of dichrone, the acute toxicity of dichrone was tested in mice. After preliminary experiment to determine the dosage, 60 mice were selected and divided into 6 groups. After giving medicine lavage and observing for 7 days, the acute toxic reaction was recorded and the LD50 value and 95 % confidence limit of LD50 were determined by the modified Karber’s method. The results showed that some mice appeared toxic reaction and death. No pathological change was found in the main organs by necropsy of dead mice. The LD50 was 18.16 g/kg and its 95% confidence limit was 15.35～21.49 g/kg. The results demonstrated that the dichrone has lower toxicity and safety for clinical application.
Forth, in the experiments of prevention and treatment of chicken coccidiosis, we made chicken coccidiosis with Eimeria coccidia. We uesed different doses of the febrifugine to prevent and treatchicken coccidiosis, so we confirmed the best dose of clinical treatment and prevention of febrifugine. It is good therapeutic effect to treat chicken coccidian with 20 mg/kg of the febrifugine, and its anticoccidial index (ACI 166.31).
迄今为止,关于常山中单体的分离报道较为深入,而常山中生物碱提取工艺和分离方法的报道较少,并且在常山的药理作用的研究中,关于常山中生物碱对畜禽生产常见的球虫病的研究未见报道。因此,本文由此出发,对常山中生物碱开发为新型兽药的几个关键的步骤进行了一些研究,包括生物碱的提取、分离及其抗球虫活性的研究,以期为常山的工业化生产和进一步开发利用提供理论依据。主要结果如下:
1常山碱含量测定方法的确定。利用常山碱在碱水条件下水解,产生邻氨基苯甲酸并将其与N-1-萘基-乙二胺盐酸盐溶液显色的原理,用紫外分光光度计比色法测定得出,常山碱提取物的总碱含量为0.327 %。
2常山总生物碱提取工艺的研究。运用正交实验,分别以浸膏重量和常山碱含量作为评价指标,考察了超声波提取工艺的效果,运用SPSS 14.0软件对结果进行分析,得到优化的提取工艺如下:其提取条件为提取温度50℃,料液比1:5,提取时间1 h。
3为了评价常山碱的安全性,对其进行了小白鼠的急性毒性试验。预试验初步确定用药剂量,正式试验取60只昆明种小白鼠,灌胃给药后,连续观察7天,记录小白鼠的急性毒性反应,并通过改良寇氏法计算常山碱对小白鼠的半数致死量(LD50)及LD50的95 %可信限。结果表明,给药组部分小白鼠出现中毒反应并死亡,剖检主要脏器未见病理改变,测得LD50为18.16 g/kg体重,LD50的95 %可信限为15.35 g/kg～21.49 g/kg体重。实验结果初步提示,常山碱的毒性很低,临床用药安全可靠。
4在常山总生物碱抗鸡球虫病和预防鸡球虫试验中,用艾美耳球虫接种雏鸡人工引发鸡球虫病。通过不同剂量常山总生物碱防治鸡球虫病作比较,确定鸡球虫病最佳临床治疗和预防剂量,其中以20 mg/kg给药可以达到良好的治疗鸡球虫病效果,其抗球虫指数(ACI为166.31)。
Avian coccidiosis is an important disease that widely spreads with a high incidence in chicken farms and is often found in chicks at 15 to 50 days old. If not effectively controlled, the prevalence of infection will be high up to 50 %～70 % with a mortality of 20 %～30 % or 80 % in severe cases. Statistical analysis indicated that the annual cost caused by the disease is estimated at￡200 million in the world. Current control of coccidosis mainly depends on synthetic chemicals or antibiotics. The durable use of the chemical drugs in the poultry industry easily leads to the resistant strains as well as the drug residues in the chickens and their products which are harmful to humans. Therefore, more and more research has shifted to non-antibiotic approaches such as medicinal herbs.
So far, study on monomer abstraction from Dichroa Febrifuga deeply, discovered that until now the alkaloid extraction process and the abstraction method study report are few about Dichroa Febrifuga. It is really micro studying on the common avian coccidiosis of livestock and fowl among the numerous pharmacologicalactions of alkaloid extraction from Dichroa Febrifuga. According to the above, this article study on each committed step for new veterinary medicine, including the alkaloid extraction, abstraction and the research of anticoccdidal activity in chicken, to provide some basic theory for a large-scale industry. The main results were indicated as follows:
First, the content of febrifugine.The present work presents a colorimetric method based on the principle that o-aminobenzoic acid is produced by the alkaline hydrolysis of the quinazolone structure of the febrifugine molecule, which can be determined colorimetrically by coupling with N-1-naphthyl ethylene diamine after diazotization. The content of febrifugine determinated by UV spectrophotometer and is 0.327 %.
Second, research on the technical of alkaloid extraction from Dichroa Febrifuga. Using the orthogonal experimental designed method, taken the evaluating target by the extracting weight and the content of febrifugine, analyzed by software SPSS 14.0, inspected extraction process of the ultrasonic wave method . The condition is the extraction temperature 50℃, the ratio of dosage liquor 1:5, extraction time 1 h.
Third, in order to evaluate the security of dichrone, the acute toxicity of dichrone was tested in mice. After preliminary experiment to determine the dosage, 60 mice were selected and divided into 6 groups. After giving medicine lavage and observing for 7 days, the acute toxic reaction was recorded and the LD50 value and 95 % confidence limit of LD50 were determined by the modified Karber’s method. The results showed that some mice appeared toxic reaction and death. No pathological change was found in the main organs by necropsy of dead mice. The LD50 was 18.16 g/kg and its 95% confidence limit was 15.35～21.49 g/kg. The results demonstrated that the dichrone has lower toxicity and safety for clinical application.
Forth, in the experiments of prevention and treatment of chicken coccidiosis, we made chicken coccidiosis with Eimeria coccidia. We uesed different doses of the febrifugine to prevent and treatchicken coccidiosis, so we confirmed the best dose of clinical treatment and prevention of febrifugine. It is good therapeutic effect to treat chicken coccidian with 20 mg/kg of the febrifugine, and its anticoccidial index (ACI 166.31).
引文
[1]李国清.兽医寄生虫学[M].广州:广东高等教育出版社,1999:228.
[2]孔繁瑶.家畜寄生虫学[M].北京:中国农业大学出版社,1997:328-334.
[3]汪明.兽医寄生虫学(第3版)[M].北京:中国农业出版社,2003:284-288.
[4] Augustine PC, Danforth HD. Influence of betaine and salinomycin on intestinal absorption of methionine and glu-cose and on the ultrastructure of intestinal cells and para-site developmental stages in chickens infected with Eimeria acervulina[J]. Avian Dis, 1999, 43(1): 89-97.
[5]夏丽英.现代中药毒理学[M].天津:天津科技翻译出版公司,2005:712-713.
[6]张雅,李春,雷国莲.常山化学成分研究[J].中国实验方剂学杂志,2010,16(5):40-42.
[7]南京中医药大学.中药大辞典下册(第2版)[M].上海:上海科学技术出版社,2006: 2942-2944.
[8]宋世荣,李良荣,高学军,等.常山粉对鸡球虫病的防治效果[J].中国兽医科技,2000,30(5):37-38.
[9]颜正华.中药学(第2版)[M].北京:人民卫生出版社,2006:1100-1101.
[10] Deng Yonghong, Xu Rensheng, Ye Yang. A New Quinazolone Alkaloid from Leaves of Dichroa febrifuga [J]. Journal of Chinese Pharmaceutical Sciences, 2000, 9(3): 116.
[11]江苏新医学院.中药大辞典(下册)[M].上海:上海人民出版社,1977:1201.
[12]孙红祥,吴军.中药常山炮制历史沿革[J].山东中医学院学报,1993,17(6):53.
[13]叶定江,丁安伟,蔡宝昌,等.常山炮制方法的研究[J].中成药研究,1981,(2):19-21.
[14]叶定江,赵蕴馥.常山饮片中常山碱含量差异初步研究[J].中成药研究,1985,(7):22- 23.
[15]陈国佩,刘华钢,覃丽秋.常山炮制的实验研究[J].中药材,1998,21(l):18-20.
[16]阴健.中药现代研究与临床应用[M].北京:学苑出版社,1993:244-248.
[17]张昌绍.现代的中药研究[M].北京:中国科学图书仪器公司,1953:139.
[18]福建医学院药物研究室.八仙花的抗鸡瘧作用及其初步药理观察[J].福建医学院学报, 1959,(l):93.
[19]王进英.甜菜抗疟药之效能[J].医药学,1950,3(l0):255.
[20]唐汝愚.常山碱乙与槟榔碱的抗疟试验[J].上海中医药杂志,1958,(2):37.
[21]赵灿熙.常山提取物对氯喹敏感株和抗氯喹株鼠疟原虫的效应观察[J].同济医科大学报,1986,15(2):129.
[22]赵灿熙.宿主和耐氯喹株疟原虫在常山治疗中的酶学变化[J].同济医科大学学报,1987 ,16(5):334.
[23]张覃沐,吕富华.几种抗疟药(常山碱乙,氯胍和环氯胍)及黄芩素等的抗阿米巴作用[J].武汉医学院学报,1958,(l):11.
[24]李广勋.中药药理毒理与临床[M].天津:天津科技翻译出版公司,1992:135-136.
[25]张昌绍,黄琪章.常山碱的药理[J].生理学报,1956,20(l):30.
[26]郭晓庄.有毒中草药大辞典[M].天津:天津科技翻译出版公司,1992:186.
[27]江文德,张昌绍,杨藻宸.常山碱催吐作用的研究-Ⅰ.常山碱对鸽催吐作用的机制[J].上海第一医学院学报,1957,(3):253-258.
[28]江文德.常山碱对狗催吐作用的机制[J].生理学报,1961,(3-4):180.
[29]王本祥.现代中药药理学[M].天津:天津科技出版社,1997:764-765.
[30]吴葆杰.中草药药理学[M].北京:人民卫生出版社,1983:278.
[31]徐小洲.“常山合剂”治疗小儿上呼吸道感染63例疗效观察[J].新医药学杂志,1979, (5):33.
[32]夏斌.梅核参常滚痰汤治梅核气60例[J].陕西中医,1958,(l):11.
[33]丁书文,焦华琛,尹柱汉.青蒿、常山对大鼠急性心肌缺血所致心律失常的影响[J].山东中医杂志,2003,22(12):742-743.
[34]丁书文,焦华琛,解砚英,等.青蒿常山对冠脉结扎所诱发犬急性心肌缺血所致心律失常的保护作用[J].中华中医药学刊,2008,26(8):1613-1614.
[35]徐景藩.常山治疗蓝氏贾第虫病的体会[J].江苏中医中医,1962,(l0):17.
[36]孔荣顺.常山治寒热效佳[J].甘肃中医学院学报,1962,(l0):17.
[37]晋爱兰,陈秀真,张供领,等.中药常山及其组方与西药对鸡球虫病的疗效对比试验[J].中国动物检疫,2009,26(8):56-57.
[38]郭永和.中草药体外抗阴道毛滴虫试验及临床应用研究[J].济宁医学院学报,2000,23 (l):58.
[39]褚建军.高速逆流色谱分离中药中的生物碱[D].浙江工业大学硕士学位文,2004,5:1.
[40]玄玉实.高效制备色谱法制备菱角壳中生物碱的研究[D].东北师范大学硕士学位论文, 2005,5:1.
[41]肖崇厚.中药化学[M].上海:上海科学技术出版社,1987:71.
[42]陈晓青,蒋新宇,刘佳佳.中草药成分分离分析技术与方法化学[M].北京:化工工业出版社,2006:1-3.
[43]肖崇厚,杨松松,洪筱坤.中药化学[M].上海:上海科学技术出版社,1997:84-85.
[44]刘成梅,游海.天然产物有效成分的分离与应用[M].北京:化学工业出版社,2003:148- 150.
[45]张德华,黄仁术,左露,等.生物碱的提取方法研究进展[J].中国野生植物资源,2010,29 (5):15-20.
[46]周贤春,何春霞,苏力坦·阿巴白克力.生物碱的研究进展[J].生物技术通讯,2006,17(3):476-479.
[47]李倩霞,赵青,蒋林,等.酶法提取岩黄连总生物碱的研究[J].现代中药研究与实践, 2009,23(4):47-49.
[48]何际婵,董志超,王建荣,等.纤维素酶在提取长春花碱中的应用[J].时珍国医国药, 2010,21(3):544-545.
[49]魏金莹,朱宏吉,等.酶法提取贝母中总生物碱的工艺研究[J].中草药,2007,38(9): 1344-1346.
[50]邹建国,刘飞,徐小龙,等.微波辅助提取马钱子中总生物碱工艺[J].食品科学,2011,31(18):116-119.
[51]赵华,李会鹏,廖克俭.微波辅助提取汉防己总生物碱及其中汉防己碱含量的测定[J].化学与生物工程,2011,28(3):91-94.
[52]莫肖云,黄丽秀,田芳年,等.超声波提取两面针中的总碱[J].中国野生植物源,2009, 28(5):58-59.
[53]田明,朴胜华,栗德林.超临界萃取法提取草乌总生物碱有效成分的研究[J].中医药信息,2004,21(5):66.
[54]赵宋亮,陶春元,谢宝华.超临界CO2萃取菊三七生物碱的工艺研究[J].中药材,2008,31 (11):1749-1751.
[55]张良,袁瑜,李玉锋.CO2超临界萃取川贝母游离生物碱工艺研究[J].西华大学学报(自然科学版),2008,27(1):39-41.
[56]刘军红,廖国玲.生物碱提取、分离和纯化的研究进展[J].时珍国医国药,2007,18(5) :1230-1231.
[57]张兆旺,孙秀梅,尉小慧,等.川乌2种方法提取液的成分比较[J].中成药,1999,21(1):5 .
[58]卢艳花.中药有效成分提取分离技术[M].北京,化工工业出版社,2007:136.
[59]李霞,熊远福,文祝友,等.大孔吸附树脂纯化石蒜中石蒜碱[J].应用化学,2009,26(3): 325-328.
[60]王震,常金花,刘翠哲.离子交换树脂法提取野罂粟总生物碱的工艺优化[J].承德医学院学报,2010,27(1):14-15.
[61]霍建新.膜分离法的应用[J].安徽农学通报林产化学与工业,2007,13(14):41-42.
[62]陈立娜,都述虎,马坤芳,等.分子印记技术在葛根素分离中的应用及溶剂对聚合物识别能力的影响[J].林产化学与工业,2008,28(3):18-22.
[63] Xu S L, Wang J W, Xu S M,et al. Purif ication of octacosano by agitated short- path distillation[J]. Chinese Journal of Chemical Engineering, 2003, 11(4): 480-482.
[64]郑来丽,田平芳,李永生,等.分光光度法测定附桂骨痛复方中总生物碱的含量[J].北京化工大学学报(自然科学版),2009,36(4):80-83.
[65]何蓉,马惠芬,杨卫,等.高效液相色谱法测定大黄藤中生物碱的研究[J].安徽农业科学,2009,37(21):9980-9981.
[66]罗忠勇,曾建国,黄敬,等.RP-HPLC法测定小果博落回中7种异喹啉类生物碱[J].中草药,2010,41(7):1188-1190.
[67]许燕娟,白长敏,钟科军,等.气相色谱/质谱分析烟草中的主要生物碱[J].分析化学, 2006,34(3):382-384.
[68]张素梅,李耀维.交流示波极谱滴定法测定三黄片中盐酸小檗碱的含量[J].中国中药杂志,1997,22(4):224-226.
[69]薛燕,陈世智.乌头炮制品中酯型生物碱含量的红外光谱定量分析研究[J].中医药刊, 2005,23(6):987-988.
[70]庞志功,汪宝琪,王翔,等.用荧光猝灭法测定苦参碱和氧化苦参碱[J].药物分析杂志, 1998,18(6):378-380.
[71]金阳.薄层荧光扫描法测定黄柏胶囊中小蘗碱的含量[J].沈阳药科大学学报,2009,7: 113.
[72]姜秀娟,孟庆艳,余明星,等.双波长薄层扫描法测定刺山柑不同部位盐酸水苏碱的含量[J].光谱实验室,2010,27(5):1959-1963.
[73]庞志功,汪宝琪,范春.薄层-胶束荧光法测定北豆根中的两种生物碱[J].分析化学, 1995,23(5):539-542.
[74]蔡梅超,周洪雷,王真,等.高效毛细管电泳法测定黄柏干皮和枝皮中盐酸小檗碱的含量[J].西北药学杂志,2010,25(4):270-271.
[75]许雪琴,黄李彦,陈国南.化学发光法测定枳壳中生物碱含量[J].光谱实验室,2008,25 (5):955-958.
[76]鹿尘,曾华金,杨冉,等.流动注射化学发光法测定药品中青藤碱[J].现代预防医学, 2007,37(15):2883-2884.
[77]陈焕文,郑健,王伟萍,等.电喷雾解吸电离质谱快速测定吴茱萸中生物碱[J].分析化学,2009,37(2):237-241.
[78] El-Abasy M., M. Motobu, et al. Protective effects of sugar cane extracts (SCE) on Eimeria tenella infection in chickens[J]. J Vet Med Sci, 2003, 65(8): 65-71.
[79] Guo F C, K wakkel R P, Williams B A.et, al. Effects of mushroom and herb polysaccharides on cellular and humoral immune responses of Eimeria tenella infected chickens[J]. Poultry Science, 2004, 83(7): 1124-1132.
[80] Allen PC, Lydon J, Danforth HD. Effects of components of Artemisia annua on coccidian infections in chickens[J].Poultry Science, 1997, 76: 1156-1163.
[81]陈蒲丹,刘娟.青篙及其提取物对鸡柔嫩艾美耳球虫的抑制作用[J].中兽医医药杂志, 2008,2:24-26.
[82]马廷法,孟祥民,宋宪波,等.青篙素分散体在畜禽球虫病防治中的应用[J].当代畜牧, 2004,(8):32.
[83] Giannenas 1. Efect of dietary supplementation with oregano essential oil on performance broilers after experimental infection with Eimeria tenella[J]. Arch Tierernahr, 2003, 57(2): 99-106.
[84]刘国华,陈犬国,曾荣,等.牛至油对肉鸡球虫的防治效果研究[J].畜禽业,2006(204): 8-10.
[85]韩春来,郑明学,范瑞文.甜菜碱在防治鸡球虫病中的作用[J].中国家禽,2001,23(8): 32-33.
[86]张伟武,谢正,金明升,等.应用甜菜碱及球虫疫苗防治肉鸡球虫病的效果试验[J].浙江畜牧兽医,2006,5:30-31.
[87]李宝春,俞浩.白头翁水提液预防鸡球虫病作用的研究[J].安徽科技学院学报,2006,20(3):1-4.
[88]李禄先,区嘉伟,谢淑敏.常山碱的比色测定法[J].化学学报,1965,31(6):482-485.
[89]徐叔云,卞如濂,陈修.药理实验方法学(第3版)[M].北京:人民卫生出版社,2002:227-231.
[90]沈建忠.动物毒理学[M].北京:中国农业出版社,2002:83-97.
[91] Morehouse N F, Baron D R. Coccidiosis evaluation of cocidiostats by mortality, weight gains and fecal score[J]. Exp Parasital, 1970, (28): 5-29
[92] Johnson J, Reid W M. Anticoccidia drugs:lesin scoring techniques in battery and floor-pan experiments[J]. Exp Parasitol, 1970, (28): 30-36.
[93]索勋,李国清.鸡球虫病学[M].北京:中国农业大学出版社,1998:158-167.
[94]角田清,陈谊,明如镜译.鸡球虫病[M].上海:上海科学技术文献出版社,1986,101-103.
[95] Merk sharp and Dohome Laboratory, Anticoccidial Index,1976.
[96]刘其容.常山防治雏鸡球虫病[[J].中兽医医药杂志,1985,(1):17.
[97]孙浴东.常山酮对火鸡球虫病的效果观察[[J].家禽辑要,1989,(3):33.
[2]孔繁瑶.家畜寄生虫学[M].北京:中国农业大学出版社,1997:328-334.
[3]汪明.兽医寄生虫学(第3版)[M].北京:中国农业出版社,2003:284-288.
[4] Augustine PC, Danforth HD. Influence of betaine and salinomycin on intestinal absorption of methionine and glu-cose and on the ultrastructure of intestinal cells and para-site developmental stages in chickens infected with Eimeria acervulina[J]. Avian Dis, 1999, 43(1): 89-97.
[5]夏丽英.现代中药毒理学[M].天津:天津科技翻译出版公司,2005:712-713.
[6]张雅,李春,雷国莲.常山化学成分研究[J].中国实验方剂学杂志,2010,16(5):40-42.
[7]南京中医药大学.中药大辞典下册(第2版)[M].上海:上海科学技术出版社,2006: 2942-2944.
[8]宋世荣,李良荣,高学军,等.常山粉对鸡球虫病的防治效果[J].中国兽医科技,2000,30(5):37-38.
[9]颜正华.中药学(第2版)[M].北京:人民卫生出版社,2006:1100-1101.
[10] Deng Yonghong, Xu Rensheng, Ye Yang. A New Quinazolone Alkaloid from Leaves of Dichroa febrifuga [J]. Journal of Chinese Pharmaceutical Sciences, 2000, 9(3): 116.
[11]江苏新医学院.中药大辞典(下册)[M].上海:上海人民出版社,1977:1201.
[12]孙红祥,吴军.中药常山炮制历史沿革[J].山东中医学院学报,1993,17(6):53.
[13]叶定江,丁安伟,蔡宝昌,等.常山炮制方法的研究[J].中成药研究,1981,(2):19-21.
[14]叶定江,赵蕴馥.常山饮片中常山碱含量差异初步研究[J].中成药研究,1985,(7):22- 23.
[15]陈国佩,刘华钢,覃丽秋.常山炮制的实验研究[J].中药材,1998,21(l):18-20.
[16]阴健.中药现代研究与临床应用[M].北京:学苑出版社,1993:244-248.
[17]张昌绍.现代的中药研究[M].北京:中国科学图书仪器公司,1953:139.
[18]福建医学院药物研究室.八仙花的抗鸡瘧作用及其初步药理观察[J].福建医学院学报, 1959,(l):93.
[19]王进英.甜菜抗疟药之效能[J].医药学,1950,3(l0):255.
[20]唐汝愚.常山碱乙与槟榔碱的抗疟试验[J].上海中医药杂志,1958,(2):37.
[21]赵灿熙.常山提取物对氯喹敏感株和抗氯喹株鼠疟原虫的效应观察[J].同济医科大学报,1986,15(2):129.
[22]赵灿熙.宿主和耐氯喹株疟原虫在常山治疗中的酶学变化[J].同济医科大学学报,1987 ,16(5):334.
[23]张覃沐,吕富华.几种抗疟药(常山碱乙,氯胍和环氯胍)及黄芩素等的抗阿米巴作用[J].武汉医学院学报,1958,(l):11.
[24]李广勋.中药药理毒理与临床[M].天津:天津科技翻译出版公司,1992:135-136.
[25]张昌绍,黄琪章.常山碱的药理[J].生理学报,1956,20(l):30.
[26]郭晓庄.有毒中草药大辞典[M].天津:天津科技翻译出版公司,1992:186.
[27]江文德,张昌绍,杨藻宸.常山碱催吐作用的研究-Ⅰ.常山碱对鸽催吐作用的机制[J].上海第一医学院学报,1957,(3):253-258.
[28]江文德.常山碱对狗催吐作用的机制[J].生理学报,1961,(3-4):180.
[29]王本祥.现代中药药理学[M].天津:天津科技出版社,1997:764-765.
[30]吴葆杰.中草药药理学[M].北京:人民卫生出版社,1983:278.
[31]徐小洲.“常山合剂”治疗小儿上呼吸道感染63例疗效观察[J].新医药学杂志,1979, (5):33.
[32]夏斌.梅核参常滚痰汤治梅核气60例[J].陕西中医,1958,(l):11.
[33]丁书文,焦华琛,尹柱汉.青蒿、常山对大鼠急性心肌缺血所致心律失常的影响[J].山东中医杂志,2003,22(12):742-743.
[34]丁书文,焦华琛,解砚英,等.青蒿常山对冠脉结扎所诱发犬急性心肌缺血所致心律失常的保护作用[J].中华中医药学刊,2008,26(8):1613-1614.
[35]徐景藩.常山治疗蓝氏贾第虫病的体会[J].江苏中医中医,1962,(l0):17.
[36]孔荣顺.常山治寒热效佳[J].甘肃中医学院学报,1962,(l0):17.
[37]晋爱兰,陈秀真,张供领,等.中药常山及其组方与西药对鸡球虫病的疗效对比试验[J].中国动物检疫,2009,26(8):56-57.
[38]郭永和.中草药体外抗阴道毛滴虫试验及临床应用研究[J].济宁医学院学报,2000,23 (l):58.
[39]褚建军.高速逆流色谱分离中药中的生物碱[D].浙江工业大学硕士学位文,2004,5:1.
[40]玄玉实.高效制备色谱法制备菱角壳中生物碱的研究[D].东北师范大学硕士学位论文, 2005,5:1.
[41]肖崇厚.中药化学[M].上海:上海科学技术出版社,1987:71.
[42]陈晓青,蒋新宇,刘佳佳.中草药成分分离分析技术与方法化学[M].北京:化工工业出版社,2006:1-3.
[43]肖崇厚,杨松松,洪筱坤.中药化学[M].上海:上海科学技术出版社,1997:84-85.
[44]刘成梅,游海.天然产物有效成分的分离与应用[M].北京:化学工业出版社,2003:148- 150.
[45]张德华,黄仁术,左露,等.生物碱的提取方法研究进展[J].中国野生植物资源,2010,29 (5):15-20.
[46]周贤春,何春霞,苏力坦·阿巴白克力.生物碱的研究进展[J].生物技术通讯,2006,17(3):476-479.
[47]李倩霞,赵青,蒋林,等.酶法提取岩黄连总生物碱的研究[J].现代中药研究与实践, 2009,23(4):47-49.
[48]何际婵,董志超,王建荣,等.纤维素酶在提取长春花碱中的应用[J].时珍国医国药, 2010,21(3):544-545.
[49]魏金莹,朱宏吉,等.酶法提取贝母中总生物碱的工艺研究[J].中草药,2007,38(9): 1344-1346.
[50]邹建国,刘飞,徐小龙,等.微波辅助提取马钱子中总生物碱工艺[J].食品科学,2011,31(18):116-119.
[51]赵华,李会鹏,廖克俭.微波辅助提取汉防己总生物碱及其中汉防己碱含量的测定[J].化学与生物工程,2011,28(3):91-94.
[52]莫肖云,黄丽秀,田芳年,等.超声波提取两面针中的总碱[J].中国野生植物源,2009, 28(5):58-59.
[53]田明,朴胜华,栗德林.超临界萃取法提取草乌总生物碱有效成分的研究[J].中医药信息,2004,21(5):66.
[54]赵宋亮,陶春元,谢宝华.超临界CO2萃取菊三七生物碱的工艺研究[J].中药材,2008,31 (11):1749-1751.
[55]张良,袁瑜,李玉锋.CO2超临界萃取川贝母游离生物碱工艺研究[J].西华大学学报(自然科学版),2008,27(1):39-41.
[56]刘军红,廖国玲.生物碱提取、分离和纯化的研究进展[J].时珍国医国药,2007,18(5) :1230-1231.
[57]张兆旺,孙秀梅,尉小慧,等.川乌2种方法提取液的成分比较[J].中成药,1999,21(1):5 .
[58]卢艳花.中药有效成分提取分离技术[M].北京,化工工业出版社,2007:136.
[59]李霞,熊远福,文祝友,等.大孔吸附树脂纯化石蒜中石蒜碱[J].应用化学,2009,26(3): 325-328.
[60]王震,常金花,刘翠哲.离子交换树脂法提取野罂粟总生物碱的工艺优化[J].承德医学院学报,2010,27(1):14-15.
[61]霍建新.膜分离法的应用[J].安徽农学通报林产化学与工业,2007,13(14):41-42.
[62]陈立娜,都述虎,马坤芳,等.分子印记技术在葛根素分离中的应用及溶剂对聚合物识别能力的影响[J].林产化学与工业,2008,28(3):18-22.
[63] Xu S L, Wang J W, Xu S M,et al. Purif ication of octacosano by agitated short- path distillation[J]. Chinese Journal of Chemical Engineering, 2003, 11(4): 480-482.
[64]郑来丽,田平芳,李永生,等.分光光度法测定附桂骨痛复方中总生物碱的含量[J].北京化工大学学报(自然科学版),2009,36(4):80-83.
[65]何蓉,马惠芬,杨卫,等.高效液相色谱法测定大黄藤中生物碱的研究[J].安徽农业科学,2009,37(21):9980-9981.
[66]罗忠勇,曾建国,黄敬,等.RP-HPLC法测定小果博落回中7种异喹啉类生物碱[J].中草药,2010,41(7):1188-1190.
[67]许燕娟,白长敏,钟科军,等.气相色谱/质谱分析烟草中的主要生物碱[J].分析化学, 2006,34(3):382-384.
[68]张素梅,李耀维.交流示波极谱滴定法测定三黄片中盐酸小檗碱的含量[J].中国中药杂志,1997,22(4):224-226.
[69]薛燕,陈世智.乌头炮制品中酯型生物碱含量的红外光谱定量分析研究[J].中医药刊, 2005,23(6):987-988.
[70]庞志功,汪宝琪,王翔,等.用荧光猝灭法测定苦参碱和氧化苦参碱[J].药物分析杂志, 1998,18(6):378-380.
[71]金阳.薄层荧光扫描法测定黄柏胶囊中小蘗碱的含量[J].沈阳药科大学学报,2009,7: 113.
[72]姜秀娟,孟庆艳,余明星,等.双波长薄层扫描法测定刺山柑不同部位盐酸水苏碱的含量[J].光谱实验室,2010,27(5):1959-1963.
[73]庞志功,汪宝琪,范春.薄层-胶束荧光法测定北豆根中的两种生物碱[J].分析化学, 1995,23(5):539-542.
[74]蔡梅超,周洪雷,王真,等.高效毛细管电泳法测定黄柏干皮和枝皮中盐酸小檗碱的含量[J].西北药学杂志,2010,25(4):270-271.
[75]许雪琴,黄李彦,陈国南.化学发光法测定枳壳中生物碱含量[J].光谱实验室,2008,25 (5):955-958.
[76]鹿尘,曾华金,杨冉,等.流动注射化学发光法测定药品中青藤碱[J].现代预防医学, 2007,37(15):2883-2884.
[77]陈焕文,郑健,王伟萍,等.电喷雾解吸电离质谱快速测定吴茱萸中生物碱[J].分析化学,2009,37(2):237-241.
[78] El-Abasy M., M. Motobu, et al. Protective effects of sugar cane extracts (SCE) on Eimeria tenella infection in chickens[J]. J Vet Med Sci, 2003, 65(8): 65-71.
[79] Guo F C, K wakkel R P, Williams B A.et, al. Effects of mushroom and herb polysaccharides on cellular and humoral immune responses of Eimeria tenella infected chickens[J]. Poultry Science, 2004, 83(7): 1124-1132.
[80] Allen PC, Lydon J, Danforth HD. Effects of components of Artemisia annua on coccidian infections in chickens[J].Poultry Science, 1997, 76: 1156-1163.
[81]陈蒲丹,刘娟.青篙及其提取物对鸡柔嫩艾美耳球虫的抑制作用[J].中兽医医药杂志, 2008,2:24-26.
[82]马廷法,孟祥民,宋宪波,等.青篙素分散体在畜禽球虫病防治中的应用[J].当代畜牧, 2004,(8):32.
[83] Giannenas 1. Efect of dietary supplementation with oregano essential oil on performance broilers after experimental infection with Eimeria tenella[J]. Arch Tierernahr, 2003, 57(2): 99-106.
[84]刘国华,陈犬国,曾荣,等.牛至油对肉鸡球虫的防治效果研究[J].畜禽业,2006(204): 8-10.
[85]韩春来,郑明学,范瑞文.甜菜碱在防治鸡球虫病中的作用[J].中国家禽,2001,23(8): 32-33.
[86]张伟武,谢正,金明升,等.应用甜菜碱及球虫疫苗防治肉鸡球虫病的效果试验[J].浙江畜牧兽医,2006,5:30-31.
[87]李宝春,俞浩.白头翁水提液预防鸡球虫病作用的研究[J].安徽科技学院学报,2006,20(3):1-4.
[88]李禄先,区嘉伟,谢淑敏.常山碱的比色测定法[J].化学学报,1965,31(6):482-485.
[89]徐叔云,卞如濂,陈修.药理实验方法学(第3版)[M].北京:人民卫生出版社,2002:227-231.
[90]沈建忠.动物毒理学[M].北京:中国农业出版社,2002:83-97.
[91] Morehouse N F, Baron D R. Coccidiosis evaluation of cocidiostats by mortality, weight gains and fecal score[J]. Exp Parasital, 1970, (28): 5-29
[92] Johnson J, Reid W M. Anticoccidia drugs:lesin scoring techniques in battery and floor-pan experiments[J]. Exp Parasitol, 1970, (28): 30-36.
[93]索勋,李国清.鸡球虫病学[M].北京:中国农业大学出版社,1998:158-167.
[94]角田清,陈谊,明如镜译.鸡球虫病[M].上海:上海科学技术文献出版社,1986,101-103.
[95] Merk sharp and Dohome Laboratory, Anticoccidial Index,1976.
[96]刘其容.常山防治雏鸡球虫病[[J].中兽医医药杂志,1985,(1):17.
[97]孙浴东.常山酮对火鸡球虫病的效果观察[[J].家禽辑要,1989,(3):33.