溢油及表面活性劑成分對刺參幼參的毒理效應(yīng)研究
發(fā)布時間：2019-05-21 19:28
【摘要】：隨著經(jīng)濟社會的發(fā)展，表面活性劑成分的應(yīng)用延伸到生產(chǎn)和生活的各個方面，用途和用量也不斷增加，并伴隨生產(chǎn)和生活污水的排放進入環(huán)境中；除此之外，規(guī)模較大的突發(fā)性溢油事故不斷頻發(fā)，也給環(huán)境帶來嚴(yán)重危害。本文重點研究常見的表面活性劑成分十二烷基磺酸鈉（SDS）、十二烷基苯磺酸鈉（SDBS）及消油劑處理海上溢油事故對海水養(yǎng)殖刺參（Stichopus japonicas）的影響，運用毒理學(xué)研究手段進行相應(yīng)的毒理效應(yīng)研究，開展了如下研究： 為探究表面活性劑和消油劑處理溢油對海洋珍品刺參的毒理學(xué)效應(yīng)，SDS、SDBS、消油劑、機械分散原油（water accommodated fraction,WAF）和化學(xué)分散原油（chemical enhanced water accommodated fraction,CEWAF）對刺參幼參的急性毒性。結(jié)果顯示，SDS對4月齡刺參的72h-LC50和96h-LC50分別為15.03和10.89mg·L-1；SDBS對4月齡刺參的72h-LC50和96h-LC50分別為2.50和1.71mg·L-1；消油劑對4月齡刺參的96h-LC50為7498.94mg·L-1。由此可見，試驗中所用消油劑屬微毒性，毒性遠(yuǎn)遠(yuǎn)小于SDBS和SDS。原油、消油劑的96h大劑量表面涂抹并未使刺參幼參出現(xiàn)死亡，僅消油劑對刺參幼參產(chǎn)生較小的刺激，表現(xiàn)為軀體短時間的收縮。原油CEWAF對3月齡和4月齡刺參幼參的96h-LC50分別為246.09mg·L-1和399.15mg·L-1。 為了優(yōu)化原油WAF和CEWAF毒性試驗中的暴露過程，探究了紫外分光光度法測定總石油烴濃度、實驗室條件下的風(fēng)化過程和組分的分析。結(jié)果顯示，為了更準(zhǔn)確地的測定試驗原油CEWAF的TPH質(zhì)量濃度，確定采用原油+消油劑的濃度-吸光度曲線作為原油CEWAF的總石油烴質(zhì)量濃度標(biāo)準(zhǔn)曲線，消油劑在正己烷中有白色絮狀物生成，在制備標(biāo)準(zhǔn)儲備液后須充分靜置（或以2000r/min離心5min），吸取上清液進行稀釋，制備標(biāo)準(zhǔn)使用液，以免帶入絮狀物而干擾吸光度讀數(shù)；在實驗室條件下，80h內(nèi)，機械分散的原油中總石油烴呈現(xiàn)線性風(fēng)化的趨勢，隨著時間的推移風(fēng)化速率減小。實驗室條件下，化學(xué)分散的原油在120h內(nèi)的風(fēng)化趨勢可以看出，開始的12h內(nèi)總石油烴含量基本沒有變化，但是12h到36h內(nèi)出現(xiàn)了急劇的風(fēng)化，36h后基本穩(wěn)定下來，但仍存在波動�？赏ㄟ^一些方法減小誤差，例如可以考慮在保證對受試生物無傷害的情況下頻繁的更換試驗液，WAF建議2～3小時換液一次，CEWAF建議10～12小時換液一次，也可測定試驗起始時總石油烴含量，并做相關(guān)說明。 試驗原油WAF中的多環(huán)芳烴類物質(zhì)豐度較高，而原油和原油CEWAF中正構(gòu)烷烴的豐度較高，原油WAF中的烷烴所占比例雖遠(yuǎn)小于原油和原油CEWAF中的比例，試驗原油CEWAF中除C11，C12和C15以外其它正構(gòu)烷烴相對比例均低于試驗原油WAF，特別是姥鮫烷在試驗原油CEWAF中的相對比例明顯低于在試驗原油WAF中的相對比例，多環(huán)芳烴總比例在試驗原油WAF中高達44.44%，而試驗原油CEWAF中僅占10.35%，烷烴類總比例占試驗原油WAF中的16.16%，消油劑大大增加了烷烴類在水相中的比例，而機械攪拌溶解的多是原油中的芳烴類物質(zhì)，萘和菲及其取代物在試驗原油WAF和CEWAF中所占比例較高，試驗原油WAF中的萘含量達到13.36%。 為探討表面活性劑成分對刺參幼參的亞急性毒性，以超氧化物歧化酶（SOD）和過氧化氫酶（CAT）為生物標(biāo)志物，實驗室模擬刺參幼參污染暴露于十二烷基磺酸鈉（SDS）和消油劑7天，隨后再進行7天的免疫恢復(fù)，采用羥胺法和可見光分光光度法測定了4月齡刺參體壁肌和內(nèi)臟中SOD、CAT活性。結(jié)果表明，在SDS暴露期間比較SOD和CAT的活力變化趨勢大體相近，這也反映了在抗氧化反應(yīng)中兩者合作進行活性氧（ROS）氧化分解的關(guān)系。SDS脅迫下對比刺參幼參體壁肌中的SOD和CAT活性變化發(fā)現(xiàn)，SDS污染暴露對刺參幼參體壁肌中SOD和CAT活性的誘導(dǎo)效應(yīng)存在時間依賴性，對內(nèi)臟中SOD活性的誘導(dǎo)效應(yīng)存在時間依賴性，與SDS暴露濃度無關(guān)，內(nèi)臟中CAT活性則沒有這樣的表現(xiàn)。消油劑脅迫下對比刺參幼參體壁肌中的SOD和CAT活性變化發(fā)現(xiàn)，消油劑對刺參幼參體壁肌中的SOD和CAT活性的影響沒有明顯的時間和劑量依賴性，這樣的現(xiàn)象比較異常。考慮到試驗中SDS和消油劑的濃度并不在一個量級上，推測消油劑對刺參幼參SOD和CAT活性的影響較SDS小，但消油劑在處理溢油事故中一般是大量使用，，且可導(dǎo)致大量刺參幼參“排臟”，因此似乎具有較深遠(yuǎn)的影響，這一點值得關(guān)注。 為探討消油劑處理溢油事故的安全性，以SOD和CAT作為生物標(biāo)志物，實驗室模擬刺參幼參污染暴露于原油CEWAF和原油WAF7天，隨后再進行7天的免疫恢復(fù)，測定刺參幼參體壁肌和內(nèi)臟中的SOD和CAT活性。結(jié)果顯示，原油WAF和CEWAF暴露對刺參幼參體壁肌和內(nèi)臟中的抗氧化酶活性均有影響，但原油WAF免疫恢復(fù)后，無論是體壁肌還是內(nèi)臟中的抗氧化酶活性均未能恢復(fù)正常，而原油CEWAF免疫恢復(fù)后有所恢復(fù)，表明原油WAF污染暴露對刺參幼參的長期影響較原油CEWAF嚴(yán)重，盡管原油CEWAF中的總石油烴濃度高于原油WAF中總石油烴濃度，結(jié)合對原油WAF和CEWAF的GC-MS分析，發(fā)現(xiàn)原油WAF和原油CEWAF中組分存在差異，多環(huán)芳烴總比例在試驗原油WAF中高達44.44%，而試驗原油CEWAF中僅占10.35%，相反烷烴類總比例占試驗原油WAF中的16.16%，而占試驗原油CEWAF中的39.37%，有研究認(rèn)為多環(huán)芳烴的生物毒性較烷烴類大，這可能是導(dǎo)致原油WAF污染暴露對刺參幼參的長期影響較原油CEWAF嚴(yán)重的原因。通過比較我們發(fā)現(xiàn)，內(nèi)臟中CAT活性明顯比體壁肌中高，對于刺參這樣較為低等的生物，以內(nèi)臟作為抗氧化酶測定的組織是比較合適的。
[Abstract]:With the development of the economic and social development, the application of the surfactant component extends to various aspects of production and life, and the application and the dosage are also continuously increased, and the emission of production and domestic sewage enters the environment; besides, the large-scale sudden oil spill accidents are frequent, And also brings serious harm to the environment. In this paper, the effects of common surfactant component sodium dodecyl sulfonate (SDS), sodium dodecyl benzene sulfonate (SDBS) and oil-removing agent on the sea oil spill were studied. The following studies have been carried out: In order to study the toxicological effects of surfactants and oil-removing agents on the marine treasures, such as the toxicological effects of the oil spill on the marine treasures, SDS, SDBS, oil-removing agent, water-dispersed crude oil (WAF) and chemical-dispersed crude oil (CEWAF), the acute toxicity of the sea-oil-oil-oil spill to the juvenile sea cucumber (Apostichopus japonicus) was studied. The results showed that the 72 h-LC50 and 96 h-LC50 of the four-month-old Apostichopus japonicus were 15.03 and 10.89 mg 路 L-1, respectively. The 72h-LC50 and 96h-LC50 of the four-month-old stichopus japonicus were 2.50 and 1.71 mg 路 L-1, respectively. The 96h-LC50 of the four-month-old Apostichopus japonicus was 7498.94 mg 路 L-1. 1. It can be seen that the deoiling agent used in the test is of micro-toxicity, and the toxicity is much smaller than that of SDBS and SD. S. The application of 96-h large-dose surface of crude oil and oil-removing agent does not cause the young ginseng of the stichopus japonicus to die, and only the oil-removing agent can generate small stimulation to the young ginseng of the stichopus japonicus, and the oil-removing agent can be collected for a short period of time. The 96 h-LC50 of the crude oil CEWAF for the 3-month-old and 4-month-old stichopus japonicus was 246.09 mg 路 L-1 and 399.15 mg 路 L-, respectively. 1. In order to optimize the exposure process of WAF and CEWAF toxicity test of crude oil, an ultraviolet spectrophotometry was used to determine the total petroleum hydrocarbon concentration and the weathering process and composition under laboratory conditions. The results show that, in order to determine the TPH mass concentration of the crude oil CEWAF more accurately, it is determined that the concentration-absorbance curve of the crude oil + deoiling agent is used as the standard curve of the total petroleum hydrocarbon mass concentration of the crude oil CEWAF, and the deoiling agent has a white floccus in n-hexane. The product is generated. After the standard stock solution is prepared, it shall be allowed to stand sufficiently (or be centrifuged at 2000r/ min for 5 min), the supernatant is extracted for dilution, and the standard solution is prepared so as not to bring the floccule to interfere with the absorbance reading; in laboratory conditions, the total petroleum hydrocarbon in the mechanically dispersed crude oil exhibits linear weathering. The trend of the weathering rate over time The rate is reduced. Under the laboratory conditions, the weathering tendency of the chemically dispersed crude oil in 120 h can be seen, the total petroleum hydrocarbon content in the beginning of the 12 h is basically unchanged, but there is a sharp weathering within 12 h to 36 h, which is basically stabilized after 36 h, but still remains The error can be reduced by some methods, such as frequent replacement of the test solution in the event of no harm to the subject to be tested, WAF recommended for 2 to 3 hours of change, the CEWAF is recommended for 10 to 12 hours to be changed once, and the total petroleum hydrocarbon content at the start of the test can also be determined, and the phase can be made The results show that the abundance of the polycyclic aromatic hydrocarbons in the WAF of the crude oil and the crude oil is high, while the proportion of the paraffin in the crude oil WAF is much smaller than that of the crude oil and the crude oil CEWAF, and the test crude oil CEWAF The relative proportion of the other n-alkanes except C11, C12 and C15 is lower than that of the test crude oil WAF, especially the relative proportion of the basestalk in the test crude oil CEWAF. The total proportion of the polycyclic aromatic hydrocarbons is as high as 44.44% in the experimental crude oil WAF, and only 1 of the test crude oil CEWAF is 1. The total proportion of alkane is 16.16% in WAF of the test crude oil, and the oil-removing agent greatly increases the proportion of the alkane in the water phase, while the mechanical stirring and dissolution are the aromatic substances in the crude oil, and the amount of the oil and the phenanthrene and the substitute thereof in the experimental crude oil WAF and CEWAF The proportion is high, and the oxygen content in the WAF of the test crude oil reaches 1. 3.36%. In order to study the sub-acute toxicity of the surfactant component to the young ginseng of the stichopus japonicus, the superoxide dismutase (SOD) and catalase (CAT) were used as biomarkers, and the laboratory simulated stichopus japonicus was exposed to sodium dodecyl sulfonate (SDS). And 7 days after the deoiling agent,7 days of immune recovery is carried out, and a hydroxylamine method and a visible light spectrophotometry are adopted to determine the SO in the body wall and the internal organs of the body wall of the stichopus japonicus at the age of 4 months. The results showed that the changes of the activity of SOD and CAT were similar in the course of SDS exposure, which also reflected the active oxygen species (ROS) in the anti-oxidation reaction. The changes of SOD and CAT activity in the body wall of the stichopus japonicus were found by SDS under the stress of SDS. The time-dependent effect of SDS-contaminated exposure on the activity of SOD and CAT in the body wall of the stichopus japonicus was found. The S-exposure concentration is independent, and the CAT activity in the viscera The activity of SOD and CAT in the body wall of the stichopus japonicus was found to have no significant time and dose-dependent effect on the activity of SOD and CAT in the body wall of the stichopus japonicus. The results show that the concentration of the SDS and the deoiling agent in the test is not on the order of one order, the effect of the deoiling agent on the SOD and CAT activity of the stichopus japonicus is less than that of the SDS, but the oil-removing agent is generally used in a large amount in the treatment of the oil-spill accident, and can lead to a large number of stichopus japonicus. ", so it seems to have a far-reaching impact, In order to study the safety of oil spill accident, SOD and CAT were used as biomarkers, and the laboratory simulated stichopus japonicus was exposed to the crude oil CEWAF and the crude oil WAF for 7 days, and then the immune recovery was carried out for 7 days, and the body wall and the internal organs of the stichopus japonicus were determined. The activities of SOD and CAT showed that the exposure of WAF and CEWAF of crude oil had an effect on the activities of antioxidant enzymes in the body wall and the internal organs of the stichopus japonicus, but the activity of the antioxidant enzymes in the body wall and the internal organs of the crude oil did not return to normal after the recovery of the WAF of the crude oil, and the CEWAF of the crude oil The recovery of the immune recovery indicates that the long-term effect of WAF contamination exposure on the crude oil WAF is more severe than that of the crude oil CEWAF, although the total petroleum hydrocarbon concentration in the crude oil CEWAF is higher than the total petroleum hydrocarbon concentration in the crude oil WAF, the crude oil WAF and the crude oil CEW are found in combination with the GC-MS analysis of the crude oil WAF and CEWAF. The total proportion of the polycyclic aromatic hydrocarbons in the AF was 44.44% in the pilot crude WAF, and only 10.35% of the test crude oil CEWAF, while the total proportion of the opposite alkanes was 16.16% in the experimental crude oil WAF, accounting for 39.37% of the test crude oil CEWAF, and it was considered that the polycyclic aromatic hydrocarbons Biotoxicity is large, which may result in a long-term effect of WAF contamination exposure of crude oil on the juvenile of the stichopus japonicus compared to the crude oil C EWAF is a serious cause. By comparison we have found that the activity of CAT in the viscera is significantly higher than that in the body wall muscle, and for the lower-grade organisms such as the stichopus japonicus, the internal organs are determined as the antioxidant enzymes.
【學(xué)位授予單位】：上海海洋大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：X55;X171.5
本文編號：2482332
[Abstract]:With the development of the economic and social development, the application of the surfactant component extends to various aspects of production and life, and the application and the dosage are also continuously increased, and the emission of production and domestic sewage enters the environment; besides, the large-scale sudden oil spill accidents are frequent, And also brings serious harm to the environment. In this paper, the effects of common surfactant component sodium dodecyl sulfonate (SDS), sodium dodecyl benzene sulfonate (SDBS) and oil-removing agent on the sea oil spill were studied. The following studies have been carried out: In order to study the toxicological effects of surfactants and oil-removing agents on the marine treasures, such as the toxicological effects of the oil spill on the marine treasures, SDS, SDBS, oil-removing agent, water-dispersed crude oil (WAF) and chemical-dispersed crude oil (CEWAF), the acute toxicity of the sea-oil-oil-oil spill to the juvenile sea cucumber (Apostichopus japonicus) was studied. The results showed that the 72 h-LC50 and 96 h-LC50 of the four-month-old Apostichopus japonicus were 15.03 and 10.89 mg 路 L-1, respectively. The 72h-LC50 and 96h-LC50 of the four-month-old stichopus japonicus were 2.50 and 1.71 mg 路 L-1, respectively. The 96h-LC50 of the four-month-old Apostichopus japonicus was 7498.94 mg 路 L-1. 1. It can be seen that the deoiling agent used in the test is of micro-toxicity, and the toxicity is much smaller than that of SDBS and SD. S. The application of 96-h large-dose surface of crude oil and oil-removing agent does not cause the young ginseng of the stichopus japonicus to die, and only the oil-removing agent can generate small stimulation to the young ginseng of the stichopus japonicus, and the oil-removing agent can be collected for a short period of time. The 96 h-LC50 of the crude oil CEWAF for the 3-month-old and 4-month-old stichopus japonicus was 246.09 mg 路 L-1 and 399.15 mg 路 L-, respectively. 1. In order to optimize the exposure process of WAF and CEWAF toxicity test of crude oil, an ultraviolet spectrophotometry was used to determine the total petroleum hydrocarbon concentration and the weathering process and composition under laboratory conditions. The results show that, in order to determine the TPH mass concentration of the crude oil CEWAF more accurately, it is determined that the concentration-absorbance curve of the crude oil + deoiling agent is used as the standard curve of the total petroleum hydrocarbon mass concentration of the crude oil CEWAF, and the deoiling agent has a white floccus in n-hexane. The product is generated. After the standard stock solution is prepared, it shall be allowed to stand sufficiently (or be centrifuged at 2000r/ min for 5 min), the supernatant is extracted for dilution, and the standard solution is prepared so as not to bring the floccule to interfere with the absorbance reading; in laboratory conditions, the total petroleum hydrocarbon in the mechanically dispersed crude oil exhibits linear weathering. The trend of the weathering rate over time The rate is reduced. Under the laboratory conditions, the weathering tendency of the chemically dispersed crude oil in 120 h can be seen, the total petroleum hydrocarbon content in the beginning of the 12 h is basically unchanged, but there is a sharp weathering within 12 h to 36 h, which is basically stabilized after 36 h, but still remains The error can be reduced by some methods, such as frequent replacement of the test solution in the event of no harm to the subject to be tested, WAF recommended for 2 to 3 hours of change, the CEWAF is recommended for 10 to 12 hours to be changed once, and the total petroleum hydrocarbon content at the start of the test can also be determined, and the phase can be made The results show that the abundance of the polycyclic aromatic hydrocarbons in the WAF of the crude oil and the crude oil is high, while the proportion of the paraffin in the crude oil WAF is much smaller than that of the crude oil and the crude oil CEWAF, and the test crude oil CEWAF The relative proportion of the other n-alkanes except C11, C12 and C15 is lower than that of the test crude oil WAF, especially the relative proportion of the basestalk in the test crude oil CEWAF. The total proportion of the polycyclic aromatic hydrocarbons is as high as 44.44% in the experimental crude oil WAF, and only 1 of the test crude oil CEWAF is 1. The total proportion of alkane is 16.16% in WAF of the test crude oil, and the oil-removing agent greatly increases the proportion of the alkane in the water phase, while the mechanical stirring and dissolution are the aromatic substances in the crude oil, and the amount of the oil and the phenanthrene and the substitute thereof in the experimental crude oil WAF and CEWAF The proportion is high, and the oxygen content in the WAF of the test crude oil reaches 1. 3.36%. In order to study the sub-acute toxicity of the surfactant component to the young ginseng of the stichopus japonicus, the superoxide dismutase (SOD) and catalase (CAT) were used as biomarkers, and the laboratory simulated stichopus japonicus was exposed to sodium dodecyl sulfonate (SDS). And 7 days after the deoiling agent,7 days of immune recovery is carried out, and a hydroxylamine method and a visible light spectrophotometry are adopted to determine the SO in the body wall and the internal organs of the body wall of the stichopus japonicus at the age of 4 months. The results showed that the changes of the activity of SOD and CAT were similar in the course of SDS exposure, which also reflected the active oxygen species (ROS) in the anti-oxidation reaction. The changes of SOD and CAT activity in the body wall of the stichopus japonicus were found by SDS under the stress of SDS. The time-dependent effect of SDS-contaminated exposure on the activity of SOD and CAT in the body wall of the stichopus japonicus was found. The S-exposure concentration is independent, and the CAT activity in the viscera The activity of SOD and CAT in the body wall of the stichopus japonicus was found to have no significant time and dose-dependent effect on the activity of SOD and CAT in the body wall of the stichopus japonicus. The results show that the concentration of the SDS and the deoiling agent in the test is not on the order of one order, the effect of the deoiling agent on the SOD and CAT activity of the stichopus japonicus is less than that of the SDS, but the oil-removing agent is generally used in a large amount in the treatment of the oil-spill accident, and can lead to a large number of stichopus japonicus. ", so it seems to have a far-reaching impact, In order to study the safety of oil spill accident, SOD and CAT were used as biomarkers, and the laboratory simulated stichopus japonicus was exposed to the crude oil CEWAF and the crude oil WAF for 7 days, and then the immune recovery was carried out for 7 days, and the body wall and the internal organs of the stichopus japonicus were determined. The activities of SOD and CAT showed that the exposure of WAF and CEWAF of crude oil had an effect on the activities of antioxidant enzymes in the body wall and the internal organs of the stichopus japonicus, but the activity of the antioxidant enzymes in the body wall and the internal organs of the crude oil did not return to normal after the recovery of the WAF of the crude oil, and the CEWAF of the crude oil The recovery of the immune recovery indicates that the long-term effect of WAF contamination exposure on the crude oil WAF is more severe than that of the crude oil CEWAF, although the total petroleum hydrocarbon concentration in the crude oil CEWAF is higher than the total petroleum hydrocarbon concentration in the crude oil WAF, the crude oil WAF and the crude oil CEW are found in combination with the GC-MS analysis of the crude oil WAF and CEWAF. The total proportion of the polycyclic aromatic hydrocarbons in the AF was 44.44% in the pilot crude WAF, and only 10.35% of the test crude oil CEWAF, while the total proportion of the opposite alkanes was 16.16% in the experimental crude oil WAF, accounting for 39.37% of the test crude oil CEWAF, and it was considered that the polycyclic aromatic hydrocarbons Biotoxicity is large, which may result in a long-term effect of WAF contamination exposure of crude oil on the juvenile of the stichopus japonicus compared to the crude oil C EWAF is a serious cause. By comparison we have found that the activity of CAT in the viscera is significantly higher than that in the body wall muscle, and for the lower-grade organisms such as the stichopus japonicus, the internal organs are determined as the antioxidant enzymes.
【學(xué)位授予單位】：上海海洋大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：X55;X171.5
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相關(guān)期刊論文 前10條
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