高良姜DPH提取物凝胶剂的研究
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摘要
高良姜为姜科山姜属植物高良姜Alpinia officinarum Hance的干燥根茎。主要含有:挥发油类,黄酮类,二苯基庚烷类,苯丙素类,糖苷类及微量元素成分。具有温胃散寒,消食止痛作用,常用于脘腹冷痛,胃寒呕吐,嗳气吞酸的治疗。根据文献,高良姜中的二苯基庚烷类成分具有显著的镇痛作用,本实验据此研究一种新的用于局部镇痛的现代中药“高良姜DPH提取物凝胶剂”。
本实验以1-苯基-7-(3’-甲氧基-4’-羟基)苯基-5-醇-3-庚酮(以下简称二苯基庚烷A)为指标对高良姜药材进行了筛选。
采用超临界流体萃取技术对药材中有效部位进行提取,得到脂溶性成分高良姜超临界CO_2提取物(以下简称高良姜SFE提取物),采用分子蒸馏技术对有效部位高良姜SFE提取物进行富集,得到高良姜二苯基庚烷类提取物(以下简称高良姜DPH提取物),按单因素实验的方法,对富集工艺进行了优化,确定最佳工艺为:加热温度90℃时,真空度5Pa,将超临界萃取物预加热至60℃进料,转子刮膜的转速为260~280r·min~(-1),流速2ml·min~(-1)。
采用LC-MS分析技术对高良姜DPH提取物中的有效成分二苯基庚烷类进行确证,并对高良姜DPH提取物的质量控制方法进行了研究。
通过动物实验确定了高良姜SFE提取物和富集后的高良姜DPH提取物的LD50值,分别为:1.338g/kg和10.64g/kg。并采用醋酸扭体法验证了高良姜DPH提取物的镇痛药效,高良姜DPH提取物的量效呈线性关系,ED50为464.4mg/kg。
根据制剂原料的理化性质,结合凝胶剂本身的特点,优化了高良姜DPH提取物凝胶剂的制剂成型工艺。最佳工艺为:将卡波姆均匀撤于适量蒸馏水面上,静置,使之充分溶胀,配成3%的卡波姆水凝胶基质;将高良姜DPH提取物用无水乙醇溶解,依次加入氮酮、甘油、丙二醇、尼泊金乙酯混匀。将上述药液加入基质中,边加边搅拌,再缓慢滴加三乙醇胺,加水至足量,搅拌均匀,即得。
对制剂的质量控制方法进行了研究,采用紫外分光光度法对制剂中主要有效成分二苯基庚烷类进行了含量测定,建立了制剂的质量标准草案,并对制剂的稳定性进行了考察。
Rhizoma Alpiniae officinarum is drying plant rhizomes of Alpinia officinarum Hance. Mainly contains: Essential oils, flavonoids, diphenylheptane category, phenylpropanoid category, glycosides and trace constituents. With the effect of warming the middle warmer, cold-dispelling, promoting digestion, and alleviating pain, it commonly used for the treatment of abdominal psychroalgia, gastrofrigid vomiting,belching, etc. In recent years, both at home and abroad, studies of Galangal were mainly centered on flavonoids. In this experiment diphenylheptane constituents are studied, and a new modern medicine,“DPH Gel”for the treatment of pain is studied and prepared.
In this experiment, with diphenyl heptane A as indicator, Galangal medicines were screened.
Effective fraction in Chinese medicinal materials was extracted by supercritical fluid extraction technology and SFE extraction was obtained. Molecular distillation technology was used for further purification of SFE extraction from Rhizoma Alpiniae officinarum. By the method of single-factor experiment, purification process was optimized: temperature 90℃, the vacuum 5 Pa, super critical extraction pre-heated in 60℃, scraping membrane rotor speed 260 to 280 r·min~(-1), flow rate 2 mL·min~(-1).
Effective fraction in extracts by molecular distillation technology (shorted as“DPH extracts”)were as identified through LC-MS. Quality control methods for DPH extracts were studied.
Identified through animal experiments, the LD50 value of SFE extracts and DPH extracts were as follows: 1.3377 g / kg and 10.641 g / kg. And through acetic acid writhing test, the analgesic efficacy of Galangal extract was confirmed. A linear dose-effect relationship of Galangal extract was identified, which is ED50 464.39 mg·kg-1.
Based on the physical and chemical properties of raw materials and the characteristics of gel, DPH gel formulations process was optimized. Optimum: Carbomer was uniformly sprinkled on the surface of distilled water, and standing to the full swelling, then 3% Carbomer hydrogel matrix is prepared. Galangal extract was dissolved into ethanol,then Azone,glycerin,propyleneglycol, and ethyl Nipagin were joined and blended into uniform solution. The solution was added in the matrix, stirring, then triethanolamine was slowly added. Stirring, water was added to prepare DPH Gel.
Quality control methods for preparations were studied by ultraviolet spectrophotometry. Contents of main active ingredient in the preparation were determinated. Standards for quality control of the preparation were established.
本实验以1-苯基-7-(3’-甲氧基-4’-羟基)苯基-5-醇-3-庚酮(以下简称二苯基庚烷A)为指标对高良姜药材进行了筛选。
采用超临界流体萃取技术对药材中有效部位进行提取,得到脂溶性成分高良姜超临界CO_2提取物(以下简称高良姜SFE提取物),采用分子蒸馏技术对有效部位高良姜SFE提取物进行富集,得到高良姜二苯基庚烷类提取物(以下简称高良姜DPH提取物),按单因素实验的方法,对富集工艺进行了优化,确定最佳工艺为:加热温度90℃时,真空度5Pa,将超临界萃取物预加热至60℃进料,转子刮膜的转速为260~280r·min~(-1),流速2ml·min~(-1)。
采用LC-MS分析技术对高良姜DPH提取物中的有效成分二苯基庚烷类进行确证,并对高良姜DPH提取物的质量控制方法进行了研究。
通过动物实验确定了高良姜SFE提取物和富集后的高良姜DPH提取物的LD50值,分别为:1.338g/kg和10.64g/kg。并采用醋酸扭体法验证了高良姜DPH提取物的镇痛药效,高良姜DPH提取物的量效呈线性关系,ED50为464.4mg/kg。
根据制剂原料的理化性质,结合凝胶剂本身的特点,优化了高良姜DPH提取物凝胶剂的制剂成型工艺。最佳工艺为:将卡波姆均匀撤于适量蒸馏水面上,静置,使之充分溶胀,配成3%的卡波姆水凝胶基质;将高良姜DPH提取物用无水乙醇溶解,依次加入氮酮、甘油、丙二醇、尼泊金乙酯混匀。将上述药液加入基质中,边加边搅拌,再缓慢滴加三乙醇胺,加水至足量,搅拌均匀,即得。
对制剂的质量控制方法进行了研究,采用紫外分光光度法对制剂中主要有效成分二苯基庚烷类进行了含量测定,建立了制剂的质量标准草案,并对制剂的稳定性进行了考察。
Rhizoma Alpiniae officinarum is drying plant rhizomes of Alpinia officinarum Hance. Mainly contains: Essential oils, flavonoids, diphenylheptane category, phenylpropanoid category, glycosides and trace constituents. With the effect of warming the middle warmer, cold-dispelling, promoting digestion, and alleviating pain, it commonly used for the treatment of abdominal psychroalgia, gastrofrigid vomiting,belching, etc. In recent years, both at home and abroad, studies of Galangal were mainly centered on flavonoids. In this experiment diphenylheptane constituents are studied, and a new modern medicine,“DPH Gel”for the treatment of pain is studied and prepared.
In this experiment, with diphenyl heptane A as indicator, Galangal medicines were screened.
Effective fraction in Chinese medicinal materials was extracted by supercritical fluid extraction technology and SFE extraction was obtained. Molecular distillation technology was used for further purification of SFE extraction from Rhizoma Alpiniae officinarum. By the method of single-factor experiment, purification process was optimized: temperature 90℃, the vacuum 5 Pa, super critical extraction pre-heated in 60℃, scraping membrane rotor speed 260 to 280 r·min~(-1), flow rate 2 mL·min~(-1).
Effective fraction in extracts by molecular distillation technology (shorted as“DPH extracts”)were as identified through LC-MS. Quality control methods for DPH extracts were studied.
Identified through animal experiments, the LD50 value of SFE extracts and DPH extracts were as follows: 1.3377 g / kg and 10.641 g / kg. And through acetic acid writhing test, the analgesic efficacy of Galangal extract was confirmed. A linear dose-effect relationship of Galangal extract was identified, which is ED50 464.39 mg·kg-1.
Based on the physical and chemical properties of raw materials and the characteristics of gel, DPH gel formulations process was optimized. Optimum: Carbomer was uniformly sprinkled on the surface of distilled water, and standing to the full swelling, then 3% Carbomer hydrogel matrix is prepared. Galangal extract was dissolved into ethanol,then Azone,glycerin,propyleneglycol, and ethyl Nipagin were joined and blended into uniform solution. The solution was added in the matrix, stirring, then triethanolamine was slowly added. Stirring, water was added to prepare DPH Gel.
Quality control methods for preparations were studied by ultraviolet spectrophotometry. Contents of main active ingredient in the preparation were determinated. Standards for quality control of the preparation were established.
引文
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[2] 庄心良,曾因明,陈伯变. 现代麻醉学[M]. 北京:人民卫生出版社,2003:2505-2539.
[3] 韩济生. 神经科学原理[M]. 北京:北京医科大学出版社,1999.
[4] Wall PD,Melzack R. Textbook of pain[M]. 4th ed. London, Churchill Livingstone, 1999.
[5] Long-term Potentiation: a Central Mechanism in Plasticity of Nociceptive Transmission and Pain Perception[J]. Abstract viewer. 2005, 19.
[6] Cell signaling pathways in the dorsal horn in pain neuroplasticity[J]. Abstract Viewer. 2005, 372.
[7] Messenger molecule that transmits inflammatory signal fromthe periphery to the central nervous system[J]. Abstract Viewer. 2005, 29: 6.
[8] A novel form of synaptic plasticity in pain pathways mediates inflammatory pain[J]. Abstract Viewer. 2005, 405: 11.
[9] Sihvonen M J, Rvnp E. Advances in supercritical carbon dioxide technologies [J]. Trends in Food Science & Technology, 999, 10(6/7): 217-222.
[10] de Castro M D L,Jiménez-Carmona M M. Where is supercritical fluidextraction going[J]. Trends in Analytical Chemistry, 2000, 19(4): 223-227.
[11] 朱凯,杨光明,程康华. 岩蔷薇超临界二氧化碳萃取研究[J]. 香料香精化妆品,2004(3):14-16.
[12] Kopcak U,Mohamed R S[J]. Supercrit.Fluids, 2005, 34(2): 209-214.
[13] 朱自强.超临界流体技术——原理和应用[M]. 北京:化学工业出版社,2001.
[14] 许松林,应安国. 短程蒸馏技术提纯胡椒基丁醚的研究[J]. 农业工程学报,2005,21(2):169-171.
[15] 应安国,许松林,王淑华等. 应用分子蒸馏技术提纯胡椒基丁醚的工艺研究[J]. 化工进展,2005,24(1):57-60.
[16] 王军武,许松林,徐世民等. 分子蒸馏技术的应用现状[J]. 化工进展,2002,21(7):499-501.
[17] GreenbergDB.Theoretical andexperimental study of thecentrifugal molecular still[J]. AIChEJ, 1972, 18(2): 269-276.
[18] Xu S L,Xiang A S,Ying A G.Purification of 3-hydroxypropionitrilebywiped molecular distillation[J]. SciChina Series B-Chem, 2004, 47(6): 521-527.
[19] Bhandarker M, Ferron J R. Transport process in an agitated thin film evaporator[J]. Ind Eng Chem Res, 1988, 27: 1016-1024.
[20] Lutisan J, Cvengtos J, Micov M.Heat and mass transfer in theevaporating file of a molecular evaporator[J]. Chem Eng J, 2002, 85(2): 225-234.
[21] 中华人民共和国药典委员会编中华人民共和国药典[S]-部. 北京:化学工业出版社. 2000:236-237.
[22] 罗辉,蔡春,张建和等. 不同产地高良姜挥发油化学成分的比较[J]. 时珍国药研究,1998,9(1):29.
[23] 江苏医学院. 中药大辞典.下册. 上海人民出版社. 1997:1907
[24] 林敬明,刘双秀,贺巍等. 高良姜超临界 CO2 萃取物 GC-MS 分析[J]. 中药材,2000, 23(10);613.
[25] Ly TN, Yamauchi R, Shimoyamada M, et al. Isolution and structural elucidation of some glycosides from the rhizomes of smaller galanga (Alpinia officirzarnm Hance )[J]. J Agric Food Chem, 2002, 50(17): 4919.
[26] Ly TN, Shimoyamada M, Kato K,et al. Isolation and characterization of some antioxidative compounds from the rhizomes of smaller galanga (Alpinia officirzarum Hance)[J]. J Agric Food Chem, 2003; 51(17): 4924.
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