不同水平的聚合草碱、甜菜碱及蓖麻碱对小鼠毒性和生长影响的研究
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摘要
论文研究了三种常见的生物碱--聚合草碱、甜菜碱、蓖麻碱的基本提取、
    制备、测定方法,并以小白鼠为试验动物模型探讨了三种不同生物碱不同添加
    剂量对小白鼠毒性及生长作用的影响。
     三种生物碱测定方法的探讨:本试验以聚合草碱、甜菜碱、蓖麻碱为例,试验中,聚合草碱采用质量法测得根中含生物碱总量为0.05克/100克,叶中含量为0.030克/100克,并得到聚合草碱与生物碱试剂的颜色反应和沉淀反应;蓖麻碱采用有机溶剂萃取及分光光度法测定得到去壳蓖麻籽、脱毒饼粕、未脱毒饼粕中蓖麻碱的平均含量分别为0.077%、0.224%和0.333%,同时得到蓖麻碱与生物碱显色剂的显色反应;甜菜碱的测定采用分光光度法得到甜菜碱盐酸盐浓度在0.2~1.0mg/ml范围内与其吸收度呈线性关系。
    三种生物碱对小鼠毒性及生长影响的试验研究:分别选择断奶后雄性小白鼠(体重为15~21克)72只,分4组(一组为对照组),每组18只,试验组为添加不同生物碱不同水平(高剂量HD、中剂量MD、低剂量LD)的日粮。聚合草碱的添加剂量分别为0.25g/kg、0.375g/kg、0.5g/kg;甜菜碱的添加剂量分别为2.5g/kg、3.75g/kg、5g/kg;蓖麻碱的添加剂量分别为0.05g/kg、0.10g/kg、0.30g/kg日粮。试验结果表明,添加聚合草碱组的小鼠四周总增重LD组(18.30±1.75)比对照组(16.96±2.67)增加8%(P<0.05),且在第一周体增重最为明显,但随着周数的增加呈下降趋势;总采食量HD组(143.76±1.22)、MD组(166.71±2.32)、LD组(169.24±4.66)分别比对照组(177.17±2.02)减少19%(P<0.05),6%和4%。小鼠的血液指标GOT(谷草转氨酶)值HD组(216.78±16.74)比对照组(100.33±9.22)增加116%(P<0.01);GPT(谷丙转氨酶)值LD组(30.11±5.19)、MD组(41.94±6.30)、HD组(112.22±9.08)分别比对照组(28.72±6.80)增加5%(P<0.05),46%(P<0.01),291%(P<0.01)。
    添加蓖麻碱组的小鼠四周总增重LD组(17.17±1.80)比对照组(16.96±2.67)增加1%(p>0.05),而HD(16.33±1.60)却比对照组减少4%(p<0.05);采食量方面LD组(163.05±1.93)、MD组(152.25±1.64)、HD组(147.75±2.16)分别比对照组(177.17±2.02)减少8%,14%(p<0.05),17%(p<0.05)。其血液指标GOT值LD组(119.56±9.73)、MD组(156.00±
    
    15.39)、HD组(163.11±9.16)分别比对照组(100.33±9.22)增加19%(p<0.05),55%(p<0.01),63%(p<0.01);GPT值LD组(31.78±3.19)、MD组(47.94±4.12)、HD组(60.39±5.23)分别比对照组(28.72±6.80)增加11%(p<0.05),67%(p<0.01)和110%(p<0.01)。
    添加甜菜碱组小鼠的四周体增重HD组(15.10±2.46)比对照组(16.96±2.67)减少了11%(p<0.05),采食量HD组(170.87±1.97)比对照组(177.17±2.02)减少了4%(p>0.05)。
    在做病理切片时,发现添加聚合草碱HD组40%的小鼠其肝脏边缘有肿胀现象;添加蓖麻碱的HD组60%的小鼠其肝脏边缘及肾小管上皮细胞均有肿胀现象;添加甜菜碱组的小鼠四周体增重及采食量虽也出现下降趋势,但GPT和GOT值却并无明显上升,病理解剖时肝、肾组织也无病理变化。
    以上试验结果表明:(1)就三种生物碱的提取、测定方法而言,聚合草碱的提取采用质量法简单可行;甜菜碱和蓖麻碱的测定采用分光光度法可取得理想的结果。(2)就三种生物碱不同添加剂量对小鼠的毒性及生长影响而言,聚合草碱低剂量组(LD组)对小鼠具有促生长作用,而剂量达到0.375g/kg日粮时小鼠出现中毒症状;蓖麻碱低剂量和中剂量对小鼠体增重及采食量均呈现负作用,当蓖麻碱添加剂量达到0.30g/kg日粮时,对小鼠肝、肾有毒害作用;而甜菜碱对小鼠几乎是无毒性的。
This paper studied extraction, prepartion and determination methods of three kinds of common alkaloids(sympthine, betaine, ricinine)and determinated the effect of three alkaloids with different level on the toxicity and growth of mouse.
     Detemination's explortion of three kinds of common alkaloids. The experiment took sympthine ,betaine and recinine as example. The results showed gross alkaloid in sympthine root and leaf were 0.05g/100g and 0.030g/100g respectively by the way of mass methods . At the same time , we obtained results about colour and sediment reactions of alkaloid agent and sympthine. We also acquired recinine in husking castor bean, detoxic cake dregs and toxic cake dregs being 0.077%,0.224%,and 0.33% respectively by the way of organic solvent extraction and spectrophotemetry and got colour reactions of alkaloid agent and recinine . Meanwhile, we found betaine hydrochloric acid salt gotten by spectrophotemetry had liner relationship to do with absorptivity in concentration scope of 0.2~1.0mg/ml.
    Study about the effect of three alkaloids on the toxicity and growth of mouse. Mouse(waned) was choosed as object( body weigh range from 15 to 21g) and parted into four groups (n=18,per group). The experiment groups were fed different alkaloids with different level of ration(HD,MD,LD) .Sympthine were fed into the experiment groups at the doses of 0.25g/kg, 3.75g/kg and 5g/kg,betaine were 2.5g/kg,3.75g/kg and 5g/kg and recinine 0.05g/kg, 0.1g/kg and 0.3g/kg. The results showed within four week there was significant difference of gross body weigh gains between the controlled class and HD group(p<0.01) and feed intake and feed efficiency of mouse showed decreasing tendency in betaine group and sympthine group. At the same time, with the increase of adding dose of these two alkaloids, GPT and GOT of all experimental groups showed increasing tendency and had significant differencebetween all experimental groups and the controlled class(p<0.05),especially between HD group and the controlled class. There were tumid phenomena in pathologico-anatomic liver margin and epithelial cell of renal tubles in these two alkaloids groups. However, there was no pathological change in betaine groups.
    The above results suggested : (1) As to extraction and determination methods of the three alkaloids, mass method, spectrophotometry and spectrophotometry were
    
    simple and fesible respectively. (2) As to toxicity and growth effect of three alkaloids with different level, sympthine at the lower dose level could promote the growth of mouse. However, when its proportion in fodder was more than 0.375g/kg,there was toxicity sign in mouse; ricinine at the middle and low dose level had a negative effect on the body weigh gains and feed intake of mouse. When its proportion in fodder was more than 0.3g/kg ,there was damage effect on the liver and renal of mouse. However ,betaine was not found toxicity on mouse.
引文
1. 马养民 ,傅建熙 ,生物碱研究概况, 陕西林业科技, No.1,1997,pp.75-79
    2. 韩博,史言,尹军利 ,尹长安,苦豆子草渣和籽渣急性毒性试验,黑龙江畜牧兽医,1997(1)
    3. 张建华,乌云,苦豆子中生物碱含量测定方法新探,中草药,1997,28—8
    4. 胡宏敏,有机化学,高等教育出版社,1995,6
    5. 于炎湖 ,饲料毒物学附毒物分析,农业出版社,1990
    6. 朱蓓蕾,动物毒理学,上海:上海科学技术出版社,1989,37—40
    7. 袁森泉,羽扇豆——高蛋白饲料原料运用前景,国外畜牧学,1997,05:18-20译自《Feed international》.June,1997,8-13
    8. 中草药化学,全国中等卫生学校统编教材,南京,江苏科学技术出版社,1980,44-45
    9. 林隆泽,赵志远,徐任生等,抗癌植物喜树化学成分的研究,化学学报,1997,35(3-4):193-200
    10. 周韵丽,黄鹂英,周清等,美登木中美登素和美登普林的分离和鉴定,科学通报,1980,25(9):427-429
    11. 郑锡文,赵守训,蝙蝠葛中的新生物碱--蝙蝠葛苏林碱,科学通报,1979,24(6):285-288
    12. 龚培力,杜佐华等,蝙蝠葛酚性碱的抗心律失常作用,中药新药与临床药理,1995,6(3):13
    13. 李建科,中国棘豆属植物开发利用研究现状及前景评述,草与畜杂志,1997,(4)
    14. 李建科,甘肃棘豆饲喂小白鼠的亚慢性毒性试验的血液学分析,中兽医医药杂志,1996,(3):6—9
    15. 曹光荣,张守信等,关于宁夏黄花棘豆中毒的调查报告,畜牧兽医杂志,1988,(3):14—19
    16. 陈友梅,中药化学,济南:山东科学技术出版社,1988,53—57
    17. 黄丽瑛,周韵丽等,二杈狗牙花中生物碱的研究,中草药,1997,28(8)
    18. 中科院上海药物研究所编,中草药有效成分提取与分离,上海科学技术出版社,1983,263-269
    19. 张辉等,饲料中有毒有害物质的研究与测定,北京农业出版社,1994
    20. 张佐玉,南天竹根茎总生物碱含量测定 , 贵州林业科技, 1995(2)
    
    
    21. 汪儆,丁伯良等,朝鲜聚合草生物碱毒性的研究, 中国草原, 1984(2)
    22. 王秉栋,许益民等,聚合草喂猪的毒性试验,江苏农学院学报,1982年第3卷第3期
    23. 翟永信,现代食品分析手册,北京大学出版社,1988
    24. 邱行飞 张鸿钧,实用畜禽中毒手册 ,四川大学出版社 ,1996(1)
    25. 于建国,现代实用仪器分析方法,中国林业出版社,1993
    26. 史志诚,动物毒物学,中国农业出版社, 2001
    27. 王庭喜,甜菜碱及其应用,饲料研究,1987年第7期
    28. 吴万福等,甜菜碱的营养作用及机理研究进展,中国饲料,1999,(4)
    29. 曾端,肉碱和甜菜碱营养生理作用的比较,饲料研究,2001,(3)
    30. 方希修,王冬梅,李继秋,甜菜碱的营养生理功能与应用,中国饲料,2001,(6)
    31. 胡彩虹, 夏牧生等, 甜菜碱分析方法进展,中国饲料,1999,(7)
    32. 刘文彬 ,钟华, 甜菜碱的快速测定法,饲料工业,1998,19(7)
    33. 王茹菊,甜菜碱的生化作用及检验要点,中国甜菜糖业,1999,(6):29—30
    34. 梁冬生,陈益众, 甜菜碱,饲料工业,1997年第18卷第3期
    35. 包文芳,高顺国,范知宇等,糖蜜中甜菜碱的分离、鉴定与含量测定,沈阳药学院学报
    36. 沙世炎,徐礼,严如敏等,中草药有效成分分析法,北京人民卫生出版社,1985,
     101-1043
    37. 张永忠,徐雅琴,甜菜碱简介,饲料博览,1999,11(1):33
    38. 谢仲权,牛树琦,刘凤华主编,天然物中草药饲料添加剂研究方法,中国农业科技出版
     社,2001,6
    39. 赵 兴,赵立超等,两种纯化蓖麻毒素方法的比较,生物化学杂志,1994,10(5):636
    40. 屈健,甜菜碱,兽药与饲料添加剂,2000,5
    41. 李尚波,麦波,李兆仁,畜禽十大高效饲料添加剂,辽宁科学技术出版社,2002,6
    42. 高宝岩,霍建中等,蓖麻碱的提取及光谱研究,光谱试验室,2000,17(1):880
    43. 刘立群,毒物分析,上海科学技术出版社,1963,6
    44. 徐文清,陈艾等,紫外分光光度法测定蓖麻籽饼中蓖麻碱的含量,中国油脂,2001, 26
    45. 许秀举,李丰等,脱脂蓖麻饼的饲料价值,粮食与饲料工业,1989,4:45
    46. 许秀举,肖梦兰等,蓖麻饼制取分离蛋白的研究,食品科学,1990(12):3
    47. 张建华,倪培德,蓖麻饼粕毒性成分分析及取度的研究,中国油脂,1996,21(4):22
    48. 赵英,蓖麻毒素毒性作用机理研究进展,中华预防医学杂志,1999,33(1):54
    
    
    49. 上海第一医院中国医学科学院卫生研究所,食品毒理,人民卫生出版社,1978.12
    50. 钟品仁,哺乳类实验动物,人民卫生出版社,1983.6
    51. 曙明,张辉,饲料中有毒有害物质的控制与测定,北京农业大学出版社,1994.6
    52. 温玉麟,药物与化学物质毒性数据,天津科技出版社,1989
    53. 王洪章,段得贤主编,家畜中毒学,农业出版社,1985
    54. 李祚煌主编,家畜中毒及毒物检验,农业出版社,1992,9
    55. 史志诚,洪子鹂,丁伯良,王建华 ,李绍君,植物有毒成分的分析与检验1983,6
    56. H.butler and D.M.creasy 1996 F. Food and Chemical Toxicology 531-536
    57. kovatsis and F.tsafaris 1996 J. F Food and Chemical Toxicology 31:11,841-845;
    58. Artyukova and Yuldashe, et al, Pilot plant deactivation kf castor meal antigen lime process ,
     J.AGR.FOOD CHEM.1968,16(5):727
    59. Full, G.andWalker,J.R.et al,potential for detoxified Castor Meal, J.A.O.C.S., 1971, 48:616
    60. Simmons B.M. and Russell J.H.,A single affinity colnmn step method for the purificati
     on of ricin toxin from castor beans(ricinus communis),Analytical Biochemistry,
     1985,146:206
    61. plit andsgar,Some physico chemical and nutritional properties of caster bean(Ricinus
     communis L.)meal with rats,J. FS. SC. Technol 1987,24:33
    62. Gorog,Set al,Analyst,94;89:282
    63. Richard F'keeler et at 1987 Effects of poisionous plants on Livetock chapter New yourk, Academic press inc
    64. Furga'T. and K.Araki,1986:studies on constuent of crude drugs Alkoloids of Symphtum
     officinale Linn'chem'pharm Bull 16(12):5212
    65. I.Hirono et at’1978:the cancer activity of Symphtum officinale Linn Journal of the National cancer institnte’61:865
    66. Mossoba-MM; LinH-S; Andrzejewski-D; Sphon-JA; Betz-JM; Application of gas
     chromatography/matrix isolation/Fourid spectroscopy to the identification of pyrrolizidine
     alkaloid(Symphytum officinale L.);Journal of Aoac Internation.
    67. Furuya,T. And K.Araki,1968: studies on constituent of crued drugs. I.Alkaloids of
     symphytum officinale Linn.Chem.Pharm.Bull., 16(12):2512.
    68. Furuya, T. And.And.M.Hikichi, 1971: Alkaloids and tyiterpenoids of symphytum
    
    
     officale .Phytochemistry,10:2217.
    69. Bull.L.B.,C.C.J.Culvenor.and.A.T.Dick.The.Pyrrolizidine.Alkaloids,Wiley,NewYork,1968
    70. I. Hironlo et at'1978 : The cancer acitivity of symphtum officinle Linn Journal of the National cancer Institute'61:865
    71. W.H.BuTLER, G.P.FoRD and D.M.CREASY, A 90-day Feeding study of Lupin Flour
     Spiked.with Lupin Alkaloids in the Rat Food and Chemical Toxicology, 1996.
    72. Harada T. Maronpot R.R. Morris R.W. and Boormin G.A.(1989) obervatiois studies in rats .Toxicologic Pathology 17,690-708.
    73. LUQUE Marquez R. Gutierrez-Rave M. and infante Miranda F.(1991) Acute Poisoning by Luoine sed debitotering water ,Veterinary and Humin Toxicology 33, 265-267.
    74. John R k. Aust J Chem,1975,28:1813
    75. G.Takken,M,T. Fleotcher and B.J.Blanui,Isoauinoline Alkaloids and Reto-Fatty Acids of Argemone ochroleucn and A.mexican seed,Australian Journal of Agriculcural Research, 1993, 44: 277-85
    76. Norton,J,H, and O'Roure,P.k.(1980):Oedemn disease In chickens caused by mexican poppy seed, Aust,VET, J.56, 187-9
    77. Jonathin Arzt DVM,PVM,Hepatotoxicity Assoicated with Pyrrolizidine Alkaloids ingestion in a Horse on E aster Island,Vet Human Toxical 41(2) April 1999
    78. Wlker,K.H.,1981,Aus.Vet.J.57~Jan
    78. Errol Zeiger,Ph.D, Raymond Tice,Ph.D, Comfrey and one of Its Constituent Alkaloids Review of Toxicological Literature, 1997.
    80. Jubb, K.V.E., 1970,Pathology of Domestic Animals Sec.edi.

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