沙枣止泻糖浆剂生产工艺及其止泻机制研究
摘要
目的:(1)优化沙枣中鞣质的提取工艺;(2)比较不同溶剂提取物的抗腹泻功效;(3)在药效学试验的基础上,进行抗腹泻的机制研究;(4)设计并制备沙枣糖浆剂型,建立质量标准;(5)观察糖浆剂的抗腹泻功效。方法:(1)以浸膏得率(浸膏量)和鞣质含量为指标,单因素筛选回流、煎煮、超声提取、冷浸等不同提取方法的提取效果,在单因素基础上采用正交试验设计,优化总鞣质提取工艺;(2)分别用番泻叶、蓖麻油致小鼠腹泻模型、墨汁胃肠推进运动实验、新斯的明致小肠运动亢进实验,以腹泻指数、稀便率、小肠推进率为指标,比较了不同提取物对以上指标模型的影响;(3)选择优选的提取物,通过对豚鼠离体回肠平滑肌运动的影响实验和对拮抗激动剂的作用方式观察,阐明其抗腹泻的作用机制;(4)以水提取物为原料制备成糖浆剂,建立糖浆剂的质量标准;(5)通过以上模型以及二甲苯致小鼠耳肿实验、醋酸诱发小鼠扭体实验、小鼠腹腔毛细管通透性试验、小鼠肠道水分吸收影响实验、对2,4-二硝基苯酚及干酵母片致大鼠发热影响实验、抗菌实验、观察沙枣糖浆剂的止泻、镇痛、抗炎、抗肠道水分、解热和抗菌等功效。结果:(1)优化的提取工艺为以70%丙酮为溶剂,超声提取3次,每次1.0h,料液比为1:10,在此工艺下,干浸膏中总鞣质含量为6.32%;②优化提取工艺下的丙酮提取物与水提物均可减少番泻叶、蓖麻油致小鼠腹泻的腹泻指数和稀便率,能抑制正常及小肠运动亢进的胃肠道推进运动(P<0.05),但水提物的效果比丙酮提取物好,可能与沙枣果实中的多种营养成分的协同作用;③水提物能够非竞争性地拮抗乙酰胆碱和氯化钡对离体豚鼠回肠平滑肌激动作用并随剂量加大而作用加强,而与组胺受体无关;④沙枣止泻糖浆剂的制备工艺为:取的沙枣水提物,浓缩成含0.7g(生药)/ml浓度流浸膏,加40%(g/ml)的蔗糖和0.25%(g/ml)的苯甲酸至规定的量。各项质量标准符合中国药典规定;⑤沙枣糖浆剂止泻药效结果同前,同时能抑制二甲苯致小鼠耳肿(P<0.05),交叉菜胶致小鼠足跖肿(P<0.05),小鼠腹腔毛细管炎性渗出(P<0.05),醋酸诱发小鼠扭体反应(P<0.05),还有轻微的解热作用和抗痢疾杆菌的作用,但对小鼠肠道水分几乎没有影响。结论:沙枣丙酮提取物、水提物和糖浆剂具有止泻作用;同时糖浆剂具有一定的镇痛、抗炎、解热和抗菌等功效。质量标准符合要求。
Objective: (1)To optimize the extracting process of tannins from Elaeagnus angustifolia L.;(2) Compare the anti-diarrheal effect of extracts, extracted with different solvent; (3) Study the anti-diarrhea mechanism on the base of pharmacodynamic result.;(4) Design and prepare the syrup-formulation, and set up the standards for quality control. (5)Observe the anti-diarrheal effect of the syrup .Methods: (1) The extracting efficiency was observed by the single factors such as a reflux extraction、decocted extraction、ultrasonic extraction and cold extraction with contents of tannin (amounts of extracts) as indexes,then optimized the extracting technology to total tannin using the orthogonal design. (2)The diarrhea models in mice were induced by senna and castor oil, and the ink gastrointestinal propulsive motility test, neostigmine-induced propellant augmentation of small intestine test were applied to compare the effects of different extracts, using the diarrhea exponent, loose stools rate, small intestine propulsive rate as indexes, to the models mentioned above ,respectively. (3) The mechanisms of extract selected were observed through the test of effectis on isolated guinea pig ileum smooth muscle and the reacting modes of excitant. (4) Prepared the water extract into a syrup ,and the standards set up for quality control. (5) The anti-diarrhea effects of syrup were investigated using the models mentioned above, as well as the anti-inflammation,analgesic,anti-intestinal water,pyretic and anti-bacterial effects were carried out by means of xylene-induced mice ear swelling test、cross plastic dishes-induced mice foot swollen test、mice peritoneal capillary permeability test、acetic acid-induced mice writhing response test、the impact of mice intestinal water absorption test、2,4-dinitrobenzene phenol and dry yest tablet-induced rat fever impact test、antibacterial test, respectively. Results: (1)The optimized process of extraction were : ultrasonic extracts 3 times ,each time for 1.0h with 70% acetone as solvent, the ratio of raw materials to solvent was 1:10, on this condation the contents of total tannin in the extracts reached to 6.32%.(2)Both extracts of acetone optimized and water could reduce the mice diarrhea exponent and loose stools rate of diarrhea mice induced by senna and castor oil ,could inhibit the normal and augmentated gastrointestinal movement of mice small intestine (P <0.05), the effects of water extracts were better than acetone ,it could be estimated as the synergy roles of other nutrients in fruts . (3) The water extracts could be used as the antagonist of the exciting role of isolated guinea pig ileum smooth muscle induced by acetylcholine and barium chloride , non-competitively, and enhanced with the increase of dosage. , not related with the histamine receptor. (4) The preparing process of syrup was :concentrated the water extracts to the concentration of 0.7g (crude drug)/ml ,add 40%(g/ml) sucrose and 0.25% (g/ml)of benzoic to prescriptive amounts.,all of the qualititive parameters reached the standards of chp. (5)The antidiarrheal effects of the syrup were as above, it could as well inhibit the xylene-induced mice ear swelling (P<0.05), cross plastic dishes-induced mice foot swelling (P<0.05), mice peritoneal capillary inflammatory exudation (P<0.05), acetic acid-induced writhing response (P <0.05). it also had the anti-dysentery bacilli effect and slight pyretic effect (P<0.05), but not any effect on intestinal water absorption. Conclusion: Both extracts of acetone and water from angustifolia showed the antidiarrheal effects;The syrup provided the effect of anti-inflammation,analgesic、pyretic and anti-bacterial effects. The parameters of quality were consistent with the standards.
Objective: (1)To optimize the extracting process of tannins from Elaeagnus angustifolia L.;(2) Compare the anti-diarrheal effect of extracts, extracted with different solvent; (3) Study the anti-diarrhea mechanism on the base of pharmacodynamic result.;(4) Design and prepare the syrup-formulation, and set up the standards for quality control. (5)Observe the anti-diarrheal effect of the syrup .Methods: (1) The extracting efficiency was observed by the single factors such as a reflux extraction、decocted extraction、ultrasonic extraction and cold extraction with contents of tannin (amounts of extracts) as indexes,then optimized the extracting technology to total tannin using the orthogonal design. (2)The diarrhea models in mice were induced by senna and castor oil, and the ink gastrointestinal propulsive motility test, neostigmine-induced propellant augmentation of small intestine test were applied to compare the effects of different extracts, using the diarrhea exponent, loose stools rate, small intestine propulsive rate as indexes, to the models mentioned above ,respectively. (3) The mechanisms of extract selected were observed through the test of effectis on isolated guinea pig ileum smooth muscle and the reacting modes of excitant. (4) Prepared the water extract into a syrup ,and the standards set up for quality control. (5) The anti-diarrhea effects of syrup were investigated using the models mentioned above, as well as the anti-inflammation,analgesic,anti-intestinal water,pyretic and anti-bacterial effects were carried out by means of xylene-induced mice ear swelling test、cross plastic dishes-induced mice foot swollen test、mice peritoneal capillary permeability test、acetic acid-induced mice writhing response test、the impact of mice intestinal water absorption test、2,4-dinitrobenzene phenol and dry yest tablet-induced rat fever impact test、antibacterial test, respectively. Results: (1)The optimized process of extraction were : ultrasonic extracts 3 times ,each time for 1.0h with 70% acetone as solvent, the ratio of raw materials to solvent was 1:10, on this condation the contents of total tannin in the extracts reached to 6.32%.(2)Both extracts of acetone optimized and water could reduce the mice diarrhea exponent and loose stools rate of diarrhea mice induced by senna and castor oil ,could inhibit the normal and augmentated gastrointestinal movement of mice small intestine (P <0.05), the effects of water extracts were better than acetone ,it could be estimated as the synergy roles of other nutrients in fruts . (3) The water extracts could be used as the antagonist of the exciting role of isolated guinea pig ileum smooth muscle induced by acetylcholine and barium chloride , non-competitively, and enhanced with the increase of dosage. , not related with the histamine receptor. (4) The preparing process of syrup was :concentrated the water extracts to the concentration of 0.7g (crude drug)/ml ,add 40%(g/ml) sucrose and 0.25% (g/ml)of benzoic to prescriptive amounts.,all of the qualititive parameters reached the standards of chp. (5)The antidiarrheal effects of the syrup were as above, it could as well inhibit the xylene-induced mice ear swelling (P<0.05), cross plastic dishes-induced mice foot swelling (P<0.05), mice peritoneal capillary inflammatory exudation (P<0.05), acetic acid-induced writhing response (P <0.05). it also had the anti-dysentery bacilli effect and slight pyretic effect (P<0.05), but not any effect on intestinal water absorption. Conclusion: Both extracts of acetone and water from angustifolia showed the antidiarrheal effects;The syrup provided the effect of anti-inflammation,analgesic、pyretic and anti-bacterial effects. The parameters of quality were consistent with the standards.
引文
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