羧甲基壳聚糖银络合及活性研究
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摘要
壳聚糖在自然界储量极其丰富,但是由于壳聚糖不溶于水,制约了应用范围的扩大,因此合成水溶性好的壳聚糖衍生物是壳聚糖改性研究中最引人注目的方向之一。
本文在反应时间、温度等条件不变的情况下,改变反应物之间量的配比关系,得出碱化氯乙酸法最佳反应比例关系-壳聚糖:30%NaOH为1g: 120ml,NaOH:氯乙酸=1:1.05(质量比)。取1g壳聚糖通过碱化氯乙酸法修饰得到成品984mg收率98.4%;用胶体滴定法测定羧甲基壳聚糖取代度为0.98,其中O位取代度:0.78,N位取代度:0.2。
对羧甲基壳聚糖与银离子吸附过程进行考察,结果表明:羧甲基壳聚糖吸附银离子的行为符合庄国顺等根据质量作用定律和单分子层吸附机理提出的的动力学方程:t/Qt=t/Qeq+M/KCM(0) ,吸附能力取决于活性基团的数目,吸附量Q随着-COOH取代度的增大而增大;在强酸性条件下(pH < 2) , -COOH的离解受到抑制,对银离子的络合能力大大减弱,随着pH值增大,与银离子的络合能力逐渐增大,当溶液pH值达到5～7时,羧甲基壳聚糖与Ag+络合程度趋于最大。当pH > 7时, Ag+和OH-先形成AgOH然后迅速形成氧化银沉淀,影响测定结果;不同温度下吸附试验的结果表明,吸附量Q随温度升高而略有增大,这说明吸附过程是吸热过程,升温有利于吸附。
对银离子进行吸附能力比较,结果表明:羧甲基壳聚糖由于引进了羧基,对银离子的络合能力大大增强,是壳聚糖的大约6倍,是水溶性低聚壳聚糖的4.5倍左右。
目前临床上常用的有抗生素和化学合成药物,容易破坏口腔的微生态平衡,造成过敏反应,因此,开发安全有效的药物用于牙周炎的治疗具有重要意义。本文测定羧甲基壳聚糖银对口腔常见致病菌-牙龈卟啉单胞菌的抑菌活性,结果表明:羧甲基壳聚糖银抑菌圈直径随着溶液的浓度的增大而变大,当浓度为40mg/ml时,抑菌圈直径为9.5 mm(﹥9 mm) ,表现出较好的抑菌效果;羧甲基壳聚糖银最低抑菌浓度是1.25mg/ml。动物实验炎症指标分析结果,药物组与牙周炎组相比BI值有显著性差异(P﹤0.05),PLI值接近显著(P=0.07),替硝唑组、健康组、药物组之间没有显著性差异,羧甲基壳聚糖银在治疗牙周炎方面有一定效果,但还不能完全阻止菌斑形成,和成品药替硝唑相比,疗效还有一定差距。
Chitosan having most reserves in nature doesn’t dissolve in water, which limits its application. Therefore the pop study way is to synthesize the new characteristic derivative of chitosan which could have the better water-solubility.
Under the circumstances of reaction time, temperature and other conditions unchanged, changed the ratio between reagents, come to the optimum method of chloroacetic acid alkaline proportion of the reaction-chitosan : 30%NaOH to 1g : 120ml, NaOH : chloroacetic acid = 1:1.05 (mass ratio). 1g chitosan was modified to be finished product of 894mg by the method of alkalified chloroacetic acid, yield ratio was 98.4%. By the method of colloid titration, the degree of substitution of CMCTS(carboxymethyl chitosan) is 0.98, O-substitution : 0.78, N-substitution : 0.2.
By the inspection adsorption process of CMCTS and silver ion, the results showed : CMCTS silver ion adsorption behavior accorded with quality role under the law and single molecular layer adsorption mechanism of kinetic equations advanced by Guoshun Zhang : t/Qt=t/Qeq+M/KCM(0) , adsorption capacity rest with the number of active groups, Q adsorption 2COOH replaced with the larger increases; In strongly acidic conditions (pH<2), 2COOH dissociation to be restrained on the silver ion complexation capacity greatly weakened, with the pH value increases, with silver ion complexation capacity increased gradually. when the pH reached 5~7, CMCTS Ag complex with the greatest degree of convergence. When the pH >7, first Ag+ and OH- form AgOH, then quickly emerged silver oxide precipitation, impacted measurement results. Assay results under different temperature showed that Q adsorption increased with temperature slightly increased, This indicates adsorption process is endothermic process, and warming is in favor of adsorption.
The strong adsorption capacity to silver ion showed that CMCTS with the introduction of carboxyl, silver ion complexation capacity has been greatly enhanced, is about six times than chitosan, and is about 4.5 times than water-soluble oligomeric chitosan.
At present, antibiotic and chemosynthesis druggery is commonly used, but it will destroy the microecosystem, bring hypersusceptibility. Therefore it is important to open out the efficiency druggery. In our study, we determine inhibition of CMCTS silver on . The activity results showed : CMCTS silver inhibition zone diameter become laeger with the increase of solution concentration, and When the concentration is at 40mg/ml, the inhibition zone diameter is 9.5 mm ( >9 mm), displayed preferable antibacterial effects. CMCTS silver minimum inhibitory concentration is at 1.25mg/ml. Animal experimental inflammation indicator analysis indicated: B3 drug group compared with periodontitis group, BI value presenced significant difference (P<0.05), PLI value is nearly significant (P= 0.07). Group C tinidazole, health group D, drug group B3 did not presenced significant difference, in the treatment of periodontitis CMCTS silver have certain effects, but cann’t completely prevent the formation of dental plaque, and the therapeutic effect of CMCTS silver is inferior to finished products tinidazole.
本文在反应时间、温度等条件不变的情况下,改变反应物之间量的配比关系,得出碱化氯乙酸法最佳反应比例关系-壳聚糖:30%NaOH为1g: 120ml,NaOH:氯乙酸=1:1.05(质量比)。取1g壳聚糖通过碱化氯乙酸法修饰得到成品984mg收率98.4%;用胶体滴定法测定羧甲基壳聚糖取代度为0.98,其中O位取代度:0.78,N位取代度:0.2。
对羧甲基壳聚糖与银离子吸附过程进行考察,结果表明:羧甲基壳聚糖吸附银离子的行为符合庄国顺等根据质量作用定律和单分子层吸附机理提出的的动力学方程:t/Qt=t/Qeq+M/KCM(0) ,吸附能力取决于活性基团的数目,吸附量Q随着-COOH取代度的增大而增大;在强酸性条件下(pH < 2) , -COOH的离解受到抑制,对银离子的络合能力大大减弱,随着pH值增大,与银离子的络合能力逐渐增大,当溶液pH值达到5～7时,羧甲基壳聚糖与Ag+络合程度趋于最大。当pH > 7时, Ag+和OH-先形成AgOH然后迅速形成氧化银沉淀,影响测定结果;不同温度下吸附试验的结果表明,吸附量Q随温度升高而略有增大,这说明吸附过程是吸热过程,升温有利于吸附。
对银离子进行吸附能力比较,结果表明:羧甲基壳聚糖由于引进了羧基,对银离子的络合能力大大增强,是壳聚糖的大约6倍,是水溶性低聚壳聚糖的4.5倍左右。
目前临床上常用的有抗生素和化学合成药物,容易破坏口腔的微生态平衡,造成过敏反应,因此,开发安全有效的药物用于牙周炎的治疗具有重要意义。本文测定羧甲基壳聚糖银对口腔常见致病菌-牙龈卟啉单胞菌的抑菌活性,结果表明:羧甲基壳聚糖银抑菌圈直径随着溶液的浓度的增大而变大,当浓度为40mg/ml时,抑菌圈直径为9.5 mm(﹥9 mm) ,表现出较好的抑菌效果;羧甲基壳聚糖银最低抑菌浓度是1.25mg/ml。动物实验炎症指标分析结果,药物组与牙周炎组相比BI值有显著性差异(P﹤0.05),PLI值接近显著(P=0.07),替硝唑组、健康组、药物组之间没有显著性差异,羧甲基壳聚糖银在治疗牙周炎方面有一定效果,但还不能完全阻止菌斑形成,和成品药替硝唑相比,疗效还有一定差距。
Chitosan having most reserves in nature doesn’t dissolve in water, which limits its application. Therefore the pop study way is to synthesize the new characteristic derivative of chitosan which could have the better water-solubility.
Under the circumstances of reaction time, temperature and other conditions unchanged, changed the ratio between reagents, come to the optimum method of chloroacetic acid alkaline proportion of the reaction-chitosan : 30%NaOH to 1g : 120ml, NaOH : chloroacetic acid = 1:1.05 (mass ratio). 1g chitosan was modified to be finished product of 894mg by the method of alkalified chloroacetic acid, yield ratio was 98.4%. By the method of colloid titration, the degree of substitution of CMCTS(carboxymethyl chitosan) is 0.98, O-substitution : 0.78, N-substitution : 0.2.
By the inspection adsorption process of CMCTS and silver ion, the results showed : CMCTS silver ion adsorption behavior accorded with quality role under the law and single molecular layer adsorption mechanism of kinetic equations advanced by Guoshun Zhang : t/Qt=t/Qeq+M/KCM(0) , adsorption capacity rest with the number of active groups, Q adsorption 2COOH replaced with the larger increases; In strongly acidic conditions (pH<2), 2COOH dissociation to be restrained on the silver ion complexation capacity greatly weakened, with the pH value increases, with silver ion complexation capacity increased gradually. when the pH reached 5~7, CMCTS Ag complex with the greatest degree of convergence. When the pH >7, first Ag+ and OH- form AgOH, then quickly emerged silver oxide precipitation, impacted measurement results. Assay results under different temperature showed that Q adsorption increased with temperature slightly increased, This indicates adsorption process is endothermic process, and warming is in favor of adsorption.
The strong adsorption capacity to silver ion showed that CMCTS with the introduction of carboxyl, silver ion complexation capacity has been greatly enhanced, is about six times than chitosan, and is about 4.5 times than water-soluble oligomeric chitosan.
At present, antibiotic and chemosynthesis druggery is commonly used, but it will destroy the microecosystem, bring hypersusceptibility. Therefore it is important to open out the efficiency druggery. In our study, we determine inhibition of CMCTS silver on . The activity results showed : CMCTS silver inhibition zone diameter become laeger with the increase of solution concentration, and When the concentration is at 40mg/ml, the inhibition zone diameter is 9.5 mm ( >9 mm), displayed preferable antibacterial effects. CMCTS silver minimum inhibitory concentration is at 1.25mg/ml. Animal experimental inflammation indicator analysis indicated: B3 drug group compared with periodontitis group, BI value presenced significant difference (P<0.05), PLI value is nearly significant (P= 0.07). Group C tinidazole, health group D, drug group B3 did not presenced significant difference, in the treatment of periodontitis CMCTS silver have certain effects, but cann’t completely prevent the formation of dental plaque, and the therapeutic effect of CMCTS silver is inferior to finished products tinidazole.
引文
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[1] 王正国,付培蓉,程天民. 创伤愈合与组织修复[ M] . 济南,山东科学出版社,第一版,219.
[2] 熊远珍,湛学军,柳 羧甲基壳聚糖银、锌、铈的合成及抑菌实验的研究药物生物技术2004 ,11 (6) :361～363
[3] 夏金兰,王 春,聂珍媛,刘新星 羧甲基壳聚糖银噻苯咪唑的制备及其抑菌性能 中南大学学报(自然科学版) 2005 年2 月 第36 卷第1 期
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[1] 王爱勤,阐志宏,贾宝全,等. 羧甲基壳聚糖的制备与质量分析[ J] . 中国生化药物杂志,1996 ,17 (4) :147.
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[1]林友文,陈伟等羧甲基壳聚糖对锌离子的吸附性能研究.海峡药学,2000,12(3):69-72
[2]许才利,唐星华.羧甲基壳聚糖对Fe2 + 的络合及光谱分析.江西化工2005 年第3 期
[3]Ernest R Hayes. N ,O 2Carboxymethylch ito san [P ]. U SA4619995, 19861
[4] M ast riM S, Randall V G. Removal of metallic ions bypart ially cro sslinked1po lyam ine po lymers [ J ]1Po lymerP rep rints. 1978, 19 (1) : 483- 4871
[5]汪玉庭, 彭长宏, 唐玉蓉等. 交联壳聚糖冠醚的合成及其对金属离子的吸附性能[J ]. 环境化学, 1998, 17 (4) : 349
[6]程发, 李厚萍, 魏玉萍, 冯建新, 李桂风交联羧甲基壳聚糖制备及对Cu 的螯合性能.天津大学学报.第35 卷第3 期2002 年5 月
[7]韦进宝, 钱沙华. 环境分析化学. 北京化学工业出版社, 2002 , 4
[8]宋吉英,侯明.羧甲基壳聚糖吸附痕量汞研究.分析试验室,2006 年5 月第25 卷第5 期
[9]刘艳如等:水溶性壳聚糖与锌的络合,福建师范大学学报(自然科学版),1998,14(2):58～61.
[10]庄国顺等:国液界面吸附活性能的测定及其原理,化学学报,1984,42:1085～1088.
[1] 王正国,付培蓉,程天民. 创伤愈合与组织修复[ M] . 济南,山东科学出版社,第一版,219.
[2] 熊远珍,湛学军,柳 羧甲基壳聚糖银、锌、铈的合成及抑菌实验的研究药物生物技术2004 ,11 (6) :361～363
[3] 夏金兰,王 春,聂珍媛,刘新星 羧甲基壳聚糖银噻苯咪唑的制备及其抑菌性能 中南大学学报(自然科学版) 2005 年2 月 第36 卷第1 期
[4] Ashimoto A, Chen C,Bakker I, et al. Polymerase chain reaction de2 tection of 8 putative periodontal pathogens in subgingival p laque ofgingivitis and advanced periodontitis lesions[ J ]. OralMicrobiol Im2munol, 1996, 11 (4) : 2662273.
[5] 任蕾,杨圣辉,刘颖. 牙周炎常见菌对抗菌药物的筛选及活性测定. 现代口腔医学杂志. 2000 ;14 (4) :256
[6] 韩宁,任蕾,杨圣辉等. 大黄滴剂对金黄地鼠实验性牙周炎的作用. 北京口腔医学. 1999 ;7 (4) :170
[7] 卞金有主编. 口腔预防医学. 第三版. 人民卫生出版社.2000 ;33 - 36
[8] 陈新谦,金有豫主编. 新编药物学. 第14 版. 人民卫生出版社. 1997 ;74 - 76
[9]曹采方主编. 牙周病学. 人民卫生出版社. 2001 ;183 - 184
[10]罗明生,高天惠主编. 药物辅料大全. 成都. 四川科技出版社. 1993 ;126 - 319
[11] Choi BK, Kim KY, Yoo YJ et al. In vitro antimicrobial ac2tivity of achitooligosaccharide mixture againstactinobacillusactinomycetemcomitans and streotococcus mutans. Int J An2timicrob Agents. 2001 ; 18(6) : 553
[12] Ikinci G,Senel S ,Akincibay H et al. Effect of chiosan on aperiodontal pathogen porphyromna gingivalis. Int J Pharm
[13] Holt SC , Kesavalu L ,Walker S , et al . Virulence factors of Porphy2 romonas gingivalis[J ] . Periodontol ,2000 , 1999 ,20 :168 - 238.
[14] Fives2Taylor PM,Meyer DM,Mintz KP , et al . Virulence factors of Acti2 noba2cillus actinomycetemcomitans [J ] . Periodontol 2000 ,1999 (20) : 136 - 167.
[15] Listgarten MA. Nature of periodontal disease : Pathogenic mechanisms