【作者】 刘会会；

【导师】 付玉彬； 郑纪勇；

【作者基本信息】 中国海洋大学 ， 材料物理与化学， 2011， 硕士

【摘要】 生物污损是一直困扰航海界的棘手问题,全世界每年由污损生物造成的损失无法估计。污损生物增大了船体的自重和摩擦阻力,使航行速度降低,严重影响了船舶的机动性。同时也使燃料消耗增加,费用上升,增加船舶的维修费用,缩短船舶的使用寿命,海生物的代谢介质还会造成船体的腐蚀,影响船舶的安全性。所以防止海生物的污损成为必须要解决的一个重要问题。对船舶而言,外壳水下部位最有效的防污措施是涂装防污涂料,传统防污涂料却以有机锡、氧化亚铜等有毒防污剂的毒杀方式达到防污目的,因此对海洋环境造成严重污染。随着人们的环保意识的增强,新型环保防污涂料成为研究热点。海洋天然产物防污研究因其独特的防污作用机理和高效的防污活性重新引起人们注意。孔石莼作为我国黄渤海沿海常见的大型藻类植物,其自身具有防止其它污损生物附着的能力。本论文即以孔石莼为研究对象,借助现代色谱分离和分析技术,从孔石莼中提取、分离防污活性物质,然后借助红外光谱,核磁共振谱,气相色谱-质谱联用技术等对孔石莼提取物的结构进行分析鉴定,为后续新型防污剂开发提供重要的依据。首先孔石莼乙酸乙酯提取物UPAFS-A经二次硅胶柱层析分离后,得到防污能力最强的UPAFS-A52,UPAFS-A52分别经乙醇、氯仿/乙腈、盐酸/乙酸乙酯处理后,得到四种产物UPAFS-A521、UPAFS-A522、UPAFS-A523、UPAFS-A524。本文利用羽状舟形藻、紫贻贝、石莼孢子和白脊藤壶幼虫等典型污损生物检测了孔石莼提取物及各段分离和纯化产物的防污活性。结果显示:(1)UPAFS-A对羽状舟形藻附着完全抑制的最小浓度为0.7 mg/mL,而完全抑制紫贻贝足丝的附着的浓度为0.4 mg/mL,完全抑制石莼孢子附着的浓度为0.4 mg/mL。(2)UPAFS-A经柱层析分离得到的六段产物中,对羽状舟形藻抑制最好的部分是UPAFS-A3段,0.3 mg/mL时抑制率就达到90 %以上,其次是UPAFS-A5,0.6 mg/mL时抑制率可达到90 %以上;紫贻贝足丝附着实验中,UPAFS-A3在0.3 mg/mL能完全抑制紫贻贝足丝的附着,UPAFS-A5在0.5 mg/mL能完全抑制紫贻贝足丝的附着,而UPAFS-A6在0.4 mg/mL能完全抑制紫贻贝足丝的附着,且对紫贻贝有毒杀作用;对石莼孢子的抑制试验结果表明,UPAFS-A3对孢子分裂生长的抑制率达到90 %的浓度为0.7 mg/mL,UPAFS-A5为0.2 mg/mL,UPAFS-A6为0.3 mg/mL;白脊藤壶幼虫实验结果表明:UPAFS-A3药效最强,对白脊藤壶幼虫的全部致死浓度≤0.1 mg/mL。UPAFS-A5在0.4 mg/mL时所有白脊藤壶幼虫死亡,其LC50为0.287 mg/mL。UPAFS-A6在0.4 mg/mL时可使所有白脊藤壶幼虫死亡,其LC50为0.311 mg/mL。(3)UPAFS-A3、UPAFS-A5进行硅胶柱二次层析得到的八段产物的防污评价显示:防污活性最强的部分是UPAFS-A52,0.3 mg/mL时完全抑制羽状舟形藻、紫贻贝足丝附着,同时也可以完全抑制石莼孢子分裂生长;白脊藤壶幼虫实验中,UPAFS-A52毒性最弱,0.8 mg/mL时仍有白脊藤壶幼虫存活,其LC50为0.550 mg/mL,而UPAFS-A51在0.4 mg/mL时白脊藤壶幼虫已全部死亡,其LC50为0.217 mg/mL;UPAFS-A31在0.2 mg/mL时白脊藤壶幼虫死亡个数大大增加,死亡率高达86.7 %,其LC50为0.158 mg/mL。(4)UPAFS-A52纯化后产物UPAFS-A521、UPAFS-A522、UPAFS-A523、UPAFS-A524对羽状舟形藻抑制实验显示,这四种产物均促进羽状舟形藻附着,防污能力显著降低。UPAFS-A521~A524经气相色谱-质谱联用、红外波谱、核磁共振波谱检测发现,UPAFS-A521和UPAFS-A522中含有饱和十六酸,UPAFS-A523可能是盐晶体,UPAFS-A524中含有饱和十六酸和饱和十六酸乙酯。结合羽状舟形藻防污评价实验,发现饱和十六酸、饱和十六酸乙酯不具有防污能力,或它们需与UPAFS-A52中其他物质协同作用才能起到较好的防污效果。更多还原

【Abstract】 Marine biological fouling is a difficult problem in the sailing industry, every year the losses caused by the fouling organisms can not be estimated. Marine fouling organisms live in the marine environment or attach to the ship and various underwater artificial facilities, have the negative impact to human activities, give investors a negative benefit of general, including some large-scale algae, hydra, external anal animal, dragon crustaceans, bivalves, barnacles and sea squirts. It increases the weight and friction of the hull, so that the sailing speed is reduced, and the ship’s maneuverability is seriously affected. While the fuel consumption is increased, costs is rose, and the hulls’corrosion will be caused by the marine biological’s metabolic media, maintenance costs of ships will be increased, the life of the ships will be shorten, and the ship’s safety will be affected critically. Therefore, prevention of marine fouling organisms has become an important issue to be addressed. As we know, painting antifouling coating is a most effective measure for the underwater parts of boats. The traditional anti-fouling paints, including organic tin, cuprous oxide antifouling agents, cause serious pollution of the marine environment. Therefore, the toxic antifouling paints will gradually be replaced with the strength of people’s awareness of environmental protection and the investigation of new antifouling paint, especially finding the right natural anti-fouling agents, prevent the attachment without destroying the environment. Until recently, Marine natural products have been attracted by people due to its unique antifouling mechanism and efficient antifouling activities to be noticed. Ulva pertusa is a common large-scale algae plant along the coast of Yellow Sea and Bohai Sea, China, which has the capacity to prevent its surface from other fouling organism attachment. In this thesis, ulva pertusa was based as the research object, and the antifouling active substances were extracted and separated by the modern chromatographic separation and analysis technology, then the molecular formulas and structures were identified by IR, NMR, GC-MS, et al. It will provide important evidences for the follow-up development of new anti-fouling agents.In this article, the extraction substance UPAFS-A was isolated by silica gel column chromatography for twice, the most powerful antifouling production UPAFS-A52 was obtained, then UPAFS-A52 was elected to further purification processing. After being treated by ethanol, chloroform/acetonitrile, hydrochloric acid/ethyl acetate, UPAFS-A52 got four products UPAFS-A521, UPAFS-A522, UPAFS-A523, UPAFS-A524, respectively.We used the N.pinna sp.nov., Mytilus galloprovincialis, Ulva linza zoospores, Balanus albicostatus larvaes as typical fouling organisms to test the antifouling activities of the Ulva pertusa extractions and the paragraphs of separation and purification. The results showed that:(1) The smallest concentration of UPAFS-A against N.pinna sp.nov. complete attachment was 0.7 mg/mL, there was no mussel to reattach at the concentration of 0.4 mg/mL, and there was no spore growth on the suface of glass plate at the concentration of 0.4 mg/mL.(2) After column chromatography separation, six productions of UPAFS-A were obtained, the best inhibition part to N.pinna sp.nov. was UPAFS-A3, the inhibition rate was more than 90 % at the concentration of 0.3 mg/mL, the following was UPAFS-A5, its inhibition rate was more than 90 % at the concentration of 0.6 mg/mL; The Mytilus galloprovincialis test indicated that, UPAFS-A3 could inhibit Mytilus galloprovincialis to reattach at the concentration of 0.3 mg/mL. For UPAFS-A5, there was no Mytilus galloprovincialis to reattach at the concentration of 0.5 mg/mL. For UPAFS-A6, there was no Mytilus galloprovincialis to reattach at the concentration of 0.4 mg/mL, but it was toxic; The Ulva linza zoospores inhibition test results indicated that the inhibition rate of UPAFS-A3 was more than 90 % at the concentration of 0.7 mg/mL. For UPAFS-A5, the inhibition rate to Ulva linza zoospores was more than 90 % at the concentration of 0.2 mg/mL, and for production UPAFS-A6, the inhibition rate of growth of Ulva linza zoospores split up to 90 % at the concentration of 0.3 mg/mL; Balanus albicostatus larvaes tests revealed that UPAFS-A3 had the strongest effect, because all the larvaes were dead at the concentration less than 0.1 mg/mL. When UPAFS-A5 was at the concentration of 0.4 mg/mL, there was no larva to live, and its LC50 was 0.287 mg/mL. For UPAFS-A6, there was no larva to live at the concentration of 0.4 mg/mL, its LC50 was 0.311 mg/mL.(3) The bioassays of eight productions from UPAFS-A3 and UPAFS-A5 indicated that: UPAFS-A52 had the strongest antifouling activities, the attachment of the N.pinna sp.nov. and Mytilus galloprovincialis could be completely inhibited at the concentration of 0.3 mg/mL, and there was no Ulva linza zoospore to split growth at the same concentration. Balanus albicostatus larvaes experimental results indicated that, UPAFS-A52 had the weakest toxicity, there were larvaes still alive at the concentration of 0.8 mg/mL, and its LC50 was 0.550 mg/mL. For UPAFS-A51, there was no larva alive at 0.4 mg/mL, its LC50 was 0.217 mg/mL. For UPAFS-A31, at the concentration of 0.2 mg/mL, the death greatly increased and the mortality rate was as high as 86.7 %, and its LC50 was 0.158 mg/mL.(4) The N.pinna sp.nov. inhibition experiments of purification products UPAFS-A521, UPAFS-A522, UPAFS-A523, UPAFS-A524 had shown that this four kinds of products promoted the N.pinna sp.nov. attachment, and the antifouling ability reduced significantly.In this article, UPAFS-A521, UPAFS-A522, UPAFS-A523, UPAFS-A524 after being detected by GC-MS, IR, NMR indicated that, UPAFS-A521 and UPAFS-A522 contained the saturated sixteen acid, UPAFS-A523 might be the salt crystals, UPAFS-A524 contained the saturated sixteen acid and saturated sixteen acid ethyl ester. Combined with the N.pinna sp.nov. inhibition experiment, it was found that saturated sixteen acid and saturated sixteen acid ethyl ester do not have antifouling ability, or they might play better antifouling effect just with other substances in UPAFS-A52.更多还原