Molecular properties of (U)LIRGs CO, HCN, HNC and HCO+
2021 年度诺贝尔化学奖:大道至简
大 学 化 学Univ. Chem. 2022, 37 (1), 2112014 (1 of 5)收稿:2021-12-07;录用:2021-12-17;网络发表:2021-12-22*通讯作者,Email:******************.cn基金资助:国家自然科学基金(21825108)•今日化学• doi: 10.3866/PKU.DXHX202112014 2021年度诺贝尔化学奖:大道至简冯向青1,2,杜海峰1,2,*1中国科学院化学研究所分子识别与功能院重点实验室,北京 1001902中国科学院大学,北京 100049摘要:有机小分子成为继酶和金属催化剂之后发展的一类新型催化剂,被称为第三类催化。
有机小分子催化作为一种精确的分子构建新工具,对手性新药研发产生了巨大影响,在药物、农药、化工、材料等领域都得到了广泛的应用。
2021年的诺贝尔化学奖授予了德国化学家本杰明·利斯特和美国化学家大卫·迈克米伦,以表彰他们在这一领域做出的开创性重要贡献。
本文简述了手性现象和不对称催化,有机小分子催化的发展历程及其催化优势和未来前景。
关键词:手性;不对称催化;有机小分子催化;诺贝尔化学奖中图分类号:G64;O6The 2021 Nobel Prize in Chemistry: The Simpler the BetterXiangqing Feng 1,2, Haifeng Du 1,2,*1 CAS Key Laboratory of Molecular Recognition and Function, Institute for Chemistry, Chinese Academy of Sciences, Beijing 100190, China.2 University of Chinese Academy of Sciences, Beijing 100049, China.Abstract: Organic molecules have become one novel type of catalysts developed after enzymes and metal catalysts, which are named as organocalysis, the third type of catalysis. As a new tool toward the precise construction of molecules, organocatalysis has a huge impact on the development of chiral new drugs, which has been used in the fields of pharmacy, pesticides, chemicals, materials, and so on. The 2021 Nobel Prize in Chemistry was awarded to German chemist Benjamin List and American chemist David W. C. MacMillan for their pioneering and important contributions to this field. This article will briefly describe chirality and asymmetric catalysis, especially, the history of organocatalysis development, its advantages and future prospects.Key Words: Chirality; Asymmetric catalysis; Organic small molecule catalysis; Nobel prize in chemistry1 2021年诺贝尔化学奖获得者简介2021年10月6日,长期被戏称为“理综奖”的诺贝尔化学奖被授予“对于有机小分子不对称催化[1]的重要贡献”的两位化学家,分别是德国化学家本杰明∙利斯特(Benjamin List)和美国化学家戴维∙麦克米伦(David W. C. MacMillan)。
微生物屏障试验 DIN 58953-6_2010 Test report
Interlaboratory T est …Microbial barrier testing of packa ging materials for medical devices which are tobe ster ili ze d“according to DIN 58953-6:2010Test re portJanuary 2013Author: Daniel ZahnISEGA Forschungs- und Untersuchungsgesellschaft mbHTest report Page 2 / 15Table of contentsSeite1.General information on the Interlaboratory Test (3)1.1 Organization (3)1.2 Occasion and Objective (3)1.3 Time Schedule (3)1.4 Participants (4)2.Sample material (4)2.1 Sample Description and Execution of the Test (4)2.1.1 Materials for the Analysis of the Germ Proofness under Humidityaccording to DIN 58953-6, section 3 (5)2.1.2 Materials for the Analysis of the Germ Proofness with Air Permeanceaccording to DIN 58953-6, section 4 (5)2.2 Sample Preparation and Despatch (5)2.3 Additional Sample and Re-examination (6)3.Results (6)3.1 Preliminary Remark (6)3.2 Note on the Record of Test Results (6)3.3 Comment on the Statistical Evaluation (6)3.4 Outlier tests (7)3.5 Record of Test Results (7)3.5.1 Record of Test Results Sample F1 (8)3.5.2 Record of Test Results Sample F2 (9)3.5.3 Record of Test Results Sample F3 (10)3.5.4 Record of Test Results Sample L1 (11)3.5.5 Record of Test Results Sample L2 (12)3.5.6 Record of Test Results Sample L3 (13)3.5.7 Record of Test Results Sample L4 (14)4.Overview and Summary (15)Test report Page 3 / 15 1. General Information on the Interlaboratory Test1.1 OrganizationOrganizer of the Interlaboratory Test:Sterile Barrier Association (SBA)Mr. David Harding (director.general@)Pennygate House, St WeonardsHerfordshire HR2 8PT / Great BritainRealization of the Interlaboratory Test:Verein zur Förderung der Forschung und Ausbildung fürFaserstoff- und Verpackungschemie e. V. (VFV)vfv@isega.dePostfach 10 11 0963707 Aschaffenburg / GermanyTechnical support:ISEGA Forschungs- u. Untersuchungsgesellschaft mbHDr. Julia Riedlinger / Mr. Daniel Zahn (info@isega.de)Zeppelinstraße 3 – 563741 Aschaffenburg / Germany1.2 Occasion and ObjectiveIn order to demonstrate compliance with the requirements of the ISO 11607-1:2006 …Packaging for terminally sterilized medical devices -- Part 1: Requirements for materials, sterile barrier systems and packaging systems“ validated test methods are to be preferably utilized.For the confirmation of the microbial barrier properties of porous materials demanded in the ISO 11607-1, the DIN 58953-6:2010 …Sterilization – Sterile supply – Part 6: Microbial barrier testing of packaging materials for medical devices which are to be sterilized“ represents a conclusive method which can be performed without the need for extensive equipment.However, since momentarily no validation data on DIN 58953-6 is at hand concerns emerged that the method may lose importance against validated methods in a revision of the ISO 11607-1 or may even not be considered at all.Within the framework of this interlaboratory test, data on the reproducibility of the results obtained by means of the analysis according to DIN 58953-6 shall be gathered.1.3 Time ScheduleSeptember 2010:The Sterile Barrier Association queried ISEGA Forschungs- und Unter-suchungsgesellschaft about the technical support for the interlaboratory test.For the realization, the Verein zur Förderung der Forschung und Ausbildungfür Faserstoff- und Verpackungschemie e. V. (VFV) was won over.November 2010: Preliminary announcement of the interlaboratory test / Seach for interested laboratoriesTest report Page 4 / 15 January toDecember 2011: Search for suitable sample material / Carrying out of numerous pre-trials on various materialsJanuary 2012:Renewed contact or search for additional interested laboratories, respectively February 2012: Sending out of registration forms / preparation of sample materialMarch 2012: Registration deadline / sample despatchMay / June 2012: Results come in / statistical evaluationJuly 2012: Despatch of samples for the re-examinationSeptember 2012: Results of the re-examination come in / statistical evaluationNovember 2012: Results are sent to the participantsDecember 2012/January 2013: Compilation of the test report1.4 ParticipantsFive different German laboratories participated in the interlaboratory test. In one laboratory, the analyses were performed by two testers working independently so that six valid results overall were received which can be taken into consideration in the evaluation.To ensure an anonymous evaluation of the results, each participant was assigned a laboratory number (laboratory 1 to laboratory 6) in random order, which was disclosed only to the laboratory in question. The complete laboratory number breakdown was known solely by the ISEGA staff supporting the proficiency test.2. Sample Material2.1 Sample Description and Execution of the TestUtmost care in the selection of suitable sample material was taken to include different materials used in the manufacture of packaging for terminally sterilized medical devices.With the help of numerous pre-trials the materials were chosen covering a wide range of results from mostly germ-proof samples to germ permeable materials.Test report Page 5 / 15 2.1.1 Materials for the Analysis of Germ Proofness under Humidity according to DIN 58953-6, section 3:The participants were advised to perform the analysis on the samples according to DIN 58953-6, section 3, and to protocol their findings on the provided result sheets.The only deviation from the norm was that in case of the growth of 1 -5 colony-forming units (in the following abbreviated as CFU) per sample, no re-examination 20 test pieces was performed.2.1.2 Materials for the Analysis of Germ Proofness with Air Permeance according to DIN 58953-6, section 4:The participants were advised to perform the analysis on the samples according to DIN 58953-6, section 4, and to protocol their findings on the provided result sheets.2.2 Sample Preparation and DespatchFor the analysis of the germ proofness under humidity, 10 test pieces in the size of 50 x 50 mm were cut out of each sample and heat-sealed into a sterilization pouch with the side to be tested up.Out of the 10 test pieces, 5 were intended for the testing and one each for the two controls according to DIN 58953-6, sections 3.6.2 and 3.6.3. The rest should remain as replacements (e.g. in case of the dropping of a test piece on the floor etc.).For the analysis of the germ proofness with air permeance, 15 circular test pieces with a diameter of 40 mm were punched out of each sample and heat-sealed into a sterilization pouch with the side to be tested up.Test report Page 6 / 15 Out of the 15 test pieces, 10 were intended for the testing and one each for the two controls according to DIN 58953-6, section 4.9. The rest should remain as replacements (e.g. in case of the dropping of a test piece on the floor etc.).The sterilization pouches with the test pieces were steam-sterilized in an autoclave for 15 minutes at 121 °C and stored in an climatic room at 23 °C and 50 % relative humidity until despatch.2.3 Additional Sample and Re-examinationFor the analysis of the germ proofness under humidity another test round was performed in July / August 2012. For this, an additional sample (sample L4) was sent to the laboratories and analysed (see 2.1.2). The results were considered in the evaluation.For validation or confirmation of non-plausible results, occasional samples for re-examination were sent out to the laboratories. The results of these re-examinations (July / August 2012) were not taken into consideration in the evaluation.3. Results3.1 Preliminary RemarkSince the analysis of germ proofness is designed to be a pass / fail – test, the statistical values and precision data were meant only to serve informative purposes.The evaluation of the materials according to DIN58953-6,sections 3.7and 4.7.6by the laboratories should be the most decisive criterion for the evaluation of reproducibility of the interlaboratory test results. Based on this, the classification of a sample as “sufficiently germ-proof” or “not sufficiently germ-proof” is carried out.3.2 Note on the Record of Test Results:The exact counting of individual CFUs is not possible with the required precision if the values turn out to be very high. Thus, an upper limit of 100 CFU per agar plate or per test pieces, respectively, was defined. Individual values above this limit and values which were stated with “> 100” by the laboratories, are listed as 100 CFU per agar plate or per test piece, respectively, in the evaluation.Test report Page 7 / 153.3 Comment on the Statistical EvaluationThe statistical evaluation was done based on the series of standards DIN ISO 5725-1ff.The arithmetic laboratory mean X i and the laboratory standard deviation s i were calculated from the individual measurement values obtained by the laboratories.The overall mean X of the laboratory means as well as the precision data of the method (reproducibility and repeatability) were determined for each sample3.4 Outlier testsThe Mandel's h-statistics test was utilised as outlier test for differences between the laboratory means of the participants.A laboratory was identified as a “statistical outlier” as soon as an exceedance of Mandel's h test statistic at the 1 % significance level was detected.The respective results of the laboratories identified as outliers were not considered in the statistical evaluation.3.5 Record of Test ResultsOn the following pages, the records of the test results for each interlaboratory test sample with the statistical evaluation and the evaluation according to DIN 58953-6 are compiled.Test report Page 8 / 153.5.1 Record of Test Results Sample F1Individual Measurement values:Statistical Evaluation:Comment:Laboratory 4, as an outlier, has not been taken into consideration in the statistical Evaluation.Outlier criterion: Mandel's h-statistics (1 % level of significance)Overall mean X:91.0CFU / agar plateRepeatability standard deviation s r:17.9CFU / agar plateReproducibility standard deviation s R:19.8CFU / agar plateRepeatability r:50.0CFU / agar plateRepeatability coefficient of variation:19.6%Reproducibility R:55.5CFU / agar plateReproducibility coefficient of variation:21.8%Evaluation according to DIN 58953-6, Section 3.7:Lab. 1 - 6:Number of CFU > 5, i.e. the material is classified as not sufficiently germ-proof.Conclusion:All of the participants, even the Laboratory 4 which was identified as an outlier, came to the same results and would classify the sample material as “not sufficiently germ-proof”Test report Page 9 / 153.5.2 Record of Test Results Sample F2Individual Measurement values:Statistical Evaluation:Comment:Laboratory 4, as an outlier, has not been taken into consideration in the statistical Evaluation.Outlier criterion: Mandel's h-statistics (1 % level of significance)Overall mean X:0CFU / agar plateRepeatability standard deviation s r:0CFU / agar plateReproducibility standard deviation s R:0CFU / agar plateRepeatability r:0CFU / agar plateRepeatability coefficient of variation:0%Reproducibility R:0CFU / agar plateReproducibility coefficient of variation:0%Evaluation according to DIN 58953-6, Section 3.7:Lab. 1 – 3:Number of CFU = 0, i.e. the material is classified as sufficiently germ-proofLab. 4:Number of CFU ≤ 5, i.e. a re-examination on 20 test pieces would have to be done Lab. 5 – 6:Number of CFU = 0, i.e. the material is classified as sufficiently germ-proofConclusion:All of the participants, except for the Laboratory 4 which was identified as an outlier, came to the same results and would classify the sample material as “sufficiently germ-proof”.Test report Page 10 / 153.5.3 Record of Test Results Sample F3Individual Measurement values:Statistical Evaluation:Overall mean X:30.1CFU / agar plateRepeatability standard deviation s r:17.2CFU / agar plateReproducibility standard deviation s R:30.9CFU / agar plateRepeatability r:48.2CFU / agar plateRepeatability coefficient of variation:57.1%Reproducibility R:86.5CFU / agar plateReproducibility coefficient of variation:103%Evaluation according to DIN 58953-6, Section 3.7:Lab. 1 - 4:Number of CFU > 5, i.e. the material is classified as not sufficiently germ-proof. Lab. 5:Number of CFU = 0, i.e. the material is classified as sufficiently germ-proof. Lab. 6:Number of CFU > 5, i.e. the material is classified as not sufficiently germ-proof.Conclusion:Five of the six participants came to the same result and would classify the sample as “not sufficiently germ-proof”. Only laboratory 5 would classify the sample material as “sufficiently germ-proof”.Test report Page 11 / 153.5.4 Record of Test Results Sample L1Individual Measurement values:Statistical Evaluation:Overall mean X:0.09CFU / test pieceRepeatability standard deviation s r:0.32CFU / test pieceReproducibility standard deviation s R:0.33CFU / test pieceRepeatability r:0.91CFU / test pieceRepeatability coefficient of variation:357%Reproducibility R:0.93CFU / test pieceReproducibility coefficient of variation:366%Evaluation according to DIN 58953-6, Section 4.7:Lab. 1 - 6:Number of CFU < 15, i.e. the material is classified as sufficiently germ-proof.Conclusion:All participants came to the same result and would classify the sample as “sufficiently germ-proof”.Test report Page 12 / 153.5.5 Record of Test Results Sample L2Individual Measurement values:Statistical Evaluation:Overall mean X:0.73CFU / test pieceRepeatability standard deviation s r: 1.10CFU / test pieceReproducibility standard deviation s R: 1.18CFU / test pieceRepeatability r: 3.07CFU / test pieceRepeatability coefficient of variation:151%Reproducibility R: 3.32CFU / test pieceReproducibility coefficient of variation:163%Evaluation according to DIN 58953-6, Section 4.7:Lab. 1:Number of CFU > 15, i.e. the material is classified as not sufficiently germ-proof. Lab. 2 - 6:Number of CFU < 15, i.e. the material is classified as sufficiently germ-proof.Conclusion:Five of the six participants came to the same result and would classify the sample as “sufficiently germ-proof”. Only laboratory 1 exceeds the limit value slightly by 1 CFU, so that the sample would be classified as “not sufficiently germ-proof”.Test report Page 13 / 153.5.6 Record of Test Results Sample L3Individual Measurement values:Statistical Evaluation:Overall mean X:0.36CFU / test pieceRepeatability standard deviation s r: 1.00CFU / test pieceReproducibility standard deviation s R: 1.06CFU / test pieceRepeatability r: 2.79CFU / test pieceRepeatability coefficient of variation:274%Reproducibility R: 2.98CFU / test pieceReproducibility coefficient of variation:293%Evaluation according to DIN 58953-6, Section 4.7:Lab. 1 - 6:Number of CFU < 15, i.e. the material is classified as sufficiently germ-proof.Conclusion:All participants came to the same result and would classify the sample as “sufficiently germ-proof”.Test report Page 14 / 153.5.7 Record of Test Results Sample L4Individual Measurement values:Statistical Evaluation:Overall mean X:35.1CFU / test pieceRepeatability standard deviation s r:18.8CFU / test pieceReproducibility standard deviation s R:42.6CFU / test pieceRepeatability r:52.7CFU / test pieceRepeatability coefficient of variation:53.7%Reproducibility R:119CFU / test pieceReproducibility coefficient of variation:122%Evaluation according to DIN 58953-6, Section 4.7:Lab. 1 - 3:Number of CFU > 15, i.e. the material is classified as not sufficiently germ-proof. Lab. 4:Number of CFU < 15, i.e. the material is classified as sufficiently germ-proof. Lab. 5 - 6:Number of CFU > 15, i.e. the material is classified as not sufficiently germ-proof.Conclusion:Five of the six participants came to the same result and would classify the sample as“not sufficiently germ-proof”.Test report Page 15 / 15 4. Overview and SummarySummary:In case of four of the overall seven tested materials, a 100 % consensus was reached regarding the evaluation as“sufficiently germ-proof”and“not sufficiently germ-proof”according to DIN 58 953-6.As for the other three tested materials, there were always 5 concurrent participants out of 6 (83 %). In each case, only one laboratory would have evaluated the sample differently.It is noteworthy that the materials about the evaluation of which a 100 % consensus was reached were the smooth sterilization papers. The differences with one deviating laboratory each occurred with the slightly less homogeneous materials, such as with the creped paper and the nonwoven materials.。
MolecularThermodynamicsPDF:分子热力学的PDF
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阳离子脂质体介导日本血吸虫组织蛋白酶L1基因的真核表达
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阳离子脂质体介导 日本血吸 虫组织 蛋 白酶 L 基 因的真核表达 1
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关键词 : 日本 血 吸 虫 ; 织 蛋 白酶 L;阳 离 子 脂质 体 ;基 因转 染 ;基 因表 达 组
中 图分 类 号 : 3 3 2 文献 标 识 码 : R 8 . A
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维普资讯
26458035_酪蛋白水解物碳源SBR_生物除磷系统启动
酪蛋白水解物碳源SBR 生物除磷系统启动杨鹤1,2,王少坡1,2,张铁凡3,4,于静洁1,2,李亚静1,2,王晨晨1,2,邱春生1,2,孙力平1,2(1.天津城建大学环境与市政工程学院,天津300384;2.天津市水质科学与技术重点实验室,天津300384;3.天津创业环保集团股份有限公司,天津300384;4.天津凯英科技发展股份有限公司,天津300384)[摘要]为研究酪蛋白水解物为碳源的强化生物除磷系统的启动及其性能,采用序批式反应器对系统启动过程中污染物的去除效果、聚磷菌的富集以及不同温度和厌氧时间下系统内菌群结构变化进行了考察。
结果表明:常温(20℃)下,增加厌氧时间,除磷率无明显变化,PO 43-去除率均在40%以下;增加厌氧时间后降低温度至15℃,除磷率提高,PO 43-去除率均在65%以上,且污泥增殖迅速,沉降性能较好。
高通量测序结果表明,9种聚磷菌的相对丰度能随运行条件的改变而不断变化。
[关键词]生物除磷;酪蛋白水解物;聚磷菌;菌群结构[中图分类号]X703[文献标识码]A[文章编号]1005-829X (2021)02-0052-06Start ⁃up of sodium casein hydrolysate carbon sourceSBR biological phosphorus removal systemYang He 1,2,Wang Shaopo 1,2,Zhang Tiefan 3,4,Yu Jingjie 1,2,Li Yajing 1,2,Wang Chenchen 1,2,Qiu Chunsheng 1,2,Sun Liping 1,2(1.School of Environment and Municipal Engineering ,Tianjin Chengjian University ,Tianjin 300384,China ;2.Tianjin Key Laboratory of Water Quality Science and Technology ,Tianjin 300384,China ;3.Tianjin Captial Environmental Protection Group Co.,Ltd.,Tianjin 300384,China ;4.Tianjin Caring Technology Development Co.,Ltd.,Tianjin 300384,China )Abstract :In order to study the start ⁃up and performance of the enhanced biological phosphorus removal system us ⁃ing sodium casein hydrolysate as the sole carbon source ,a sequencing batch reactor was used to investigate the effect of pollutant removal and the enrichment of polyphosphate accumulating organisms during the start ⁃up of the system ,and the changes in the microflora structure of the system under different temperatures and anaerobic times.The resu ⁃lts showed that at room temperature (20℃),increasing anaerobic time ,the phosphorus removal rate did not changeobviously ,and the removal rate of PO 43-were below 40%.After increasing the anaerobic time ,the temperature redu ⁃ced to 15℃,the phosphorus removal rate increased ,the removal rate of PO 43-were above 65%,and the sludge mul ⁃tiplied rapidly ,the sedimentation performance was better.The results of high ⁃throughput sequencing showed that the relative abundance of 9types of polyphosphate accumulating organisms changed with the operating condition.Key word :biological phosphorus removal ;sodium casein hydrolysate ;polyphosphate accumulating organisms ;mic ⁃roflora structure[基金项目]国家自然科学基金资助项目(51678388)水体富营养化已经成为全球面临的重大水环境问题,其产生的主要原因是人类活动导致的氮、磷等营养元素在水体中的积累。
liquid–liquid phase separation
Biophysical Chemistry 109(2004)105–1120301-4622/04/$-see front matter ᮊ2003Elsevier B.V .All rights reserved.doi:10.1016/j.bpc.2003.10.021Cloud-point temperature and liquid–liquid phase separation ofsupersaturated lysozyme solutionJie Lu *,Keith Carpenter ,Rui-Jiang Li ,Xiu-Juan Wang ,Chi-Bun Ching a ,a a b bInstitute of Chemical and Engineering Sciences,Ayer Rajah Crescent 28,࠻02-08,Singapore 139959,Singapore aChemical and Process Engineering Center,National University of Singapore,Singapore 117576,SingaporebReceived 31July 2003;received in revised form 8October 2003;accepted 16October 2003AbstractThe detailed understanding of the structure of biological macromolecules reveals their functions,and is thus important in the design of new medicines and for engineering molecules with improved properties for industrial applications.Although techniques used for protein crystallization have been progressing greatly,protein crystallization may still be considered an art rather than a science,and successful crystallization remains largely empirical and operator-dependent.In this work,a microcalorimetric technique has been utilized to investigate liquid–liquid phase separation through measuring cloud-point temperature T for supersaturated lysozyme solution.The effects of cloud ionic strength and glycerol on the cloud-point temperature are studied in detail.Over the entire range of salt concentrations studied,the cloud-point temperature increases monotonically with the concentration of sodium chloride.When glycerol is added as additive,the solubility of lysozyme is increased,whereas the cloud-point temperature is decreased.ᮊ2003Elsevier B.V .All rights reserved.Keywords:Biocrystallization;Microcalorimetry;Cloud-point temperature;Liquid–liquid phase separation1.IntroductionKnowledge of detailed protein structure is essen-tial for protein engineering and the design of pharmaceuticals.Production of high-quality pro-tein crystals is required for molecular structure determination by X-ray crystallography.Although considerable effort has been made in recent years,obtaining such crystals is still difficult in general,and predicting the solution conditions where pro-*Corresponding author.Tel.:q 65-6874-4218;fax:q 65-6873-4805.E-mail address:lujie@.sg (J.Lu ).teins successfully crystallize remains a significant obstacle in the advancement of structural molecu-lar biology w 1x .The parameters affecting protein crystallization are typically reagent concentration,pH,tempera-ture,additive,etc.A phase diagram can provide the method for quantifying the influence of solu-tion parameters on the production of crystals w 2,3x .To characterize protein crystallization,it is neces-sary to first obtain detailed information on protein solution phase behavior and phase diagram.Recently physics shows that there is a direct relationship between colloidal interaction energy106J.Lu et al./Biophysical Chemistry109(2004)105–112and phase diagram.Gast and Lekkerkerker w4,5x have indicated that the range of attraction between colloid particles has a significant effect on the qualitative features of phase diagram.A similar relationship should hold for biomacromolecules, i.e.the corresponding interaction potentials govern the macromolecular distribution in solution,the shape of the phase diagram and the crystallization process w6x.Many macromolecular crystallizations appear to be driven by the strength of the attractive interactions,and occur in,or close to,attractive regimes w7,8x.Recent intensive investigation has revealed that protein or colloidal solution possesses a peculiar phase diagram,i.e.liquid–liquid phase separation and sol–gel transition exists in general in addition to crystallization w9,10x.The potential responsible for the liquid–liquid phase separation is a rather short range,possibly van der Waals,attractive potential w11,12x.The measurement of cloud-point temperature T can provide useful informationcloudon the net attractive interaction between protein molecules,namely,the higher the cloud-point tem-perature,the greater the net attractive interaction. Herein Taratuta et al.w13x studied the effects of salts and pH on the cloud-point temperature of lysozyme.Broide et al.w14x subsequently meas-ured the cloud-point temperature and crystalliza-tion temperature for lysozyme as a function of salt type and concentration.From these works the cloud-point temperature was found to be typically 15–458C below the crystallization temperature. Furthermore,Muschol and Rosenberger w15x deter-mined the metastable coexistence curves for lyso-zyme through cloud-point measurements,and suggested a systematic approach to promote pro-tein crystallization.In general,an effective way to determine the strength of protein interactions is to study temperature-induced phase transitions that occur in concentrated protein solutions.Liquid–liquid phase separation can be divided into two stages w11x:(1)the local separation stage at which the separation proceeds in small regions and local equilibrium is achieved rapidly;and(2) the coarsening stage at which condensation of these small domains proceeds slowly to reduce the loss of interface free energy w16x.The coexisting liquid phases both remain supersaturated but differ widely in protein concentration.The effect of a metastable liquid–liquid phase separation on crystallization remains ambiguous w17x.Molecular dynamics simulations and analyt-ical theory predict that the phase separation will affect the kinetics and the mechanisms of protein crystal nucleation w18x.tenWolde and Frenkel w19x have demonstrated that the free energy barrier for crystal nucleation is remarkably reduced at the critical point of liquid–liquid phase separation, thus in general,after liquid–liquid phase separa-tion,crystallization occurs much more rapidly than in the initial solution,which is typically too rapid for the growth of single crystal with low defect densities w15x.The determination of the location of liquid–liquid phase separation curve is thus crucial for efficiently identifying the optimum solution conditions for growing protein crystals. Microcalorimetry has the potential to be a useful tool for determining:(1)the metastable-labile zone boundary;(2)the temperature-dependence of pro-tein solubility in a given solvent;and(3)the crystal-growth rates as a function of supersatura-tion w20x.Microcalorimeters can detect a power signal as low as a few microwatts whereas standard calorimeters detect signals in the milliwatt range. Because of this greater sensitivity,samples with small heat effects can be analyzed.In addition, microcalorimetry has the advantage of being fast, non-destructive to the protein and requiring a relatively small amount of material.The present work is concerned with the analysis of the transient heat signal from microcalorimeter to yield liquid–liquid phase separation information for lysozyme solutions at pH4.8.To further examine the role of salt and additive on interprotein interactions, cloud-point temperature T has been determinedcloudexperimentally as a function of the concentrations of salt,protein and glycerol.2.Materials and methods2.1.MaterialsSix times crystallized lysozyme was purchased from Seikagaku Kogyo,and used without further107J.Lu et al./Biophysical Chemistry 109(2004)105–112purification.All other chemicals used were of reagent grade,from Sigma Chemical Co.2.2.Preparation of solutionsSodium acetate buffer (0.1M )at pH 4.8was prepared with ultrafiltered,deionized water.Sodi-um azide,at a concentration of 0.05%(w y v ),was added to the buffer solution as an antimicrobial agent.Protein stock solution was prepared by dissolving protein powder into buffer.To remove undissolved particles,the solution was centrifuged in a Sigma centrifuge at 12000rev.y min for 5–10min,then filtered through 0.22-m m filters (Mil-lex-VV )into a clean sample vial and stored at 48C for further experiments.The concentration of protein solution was determined by measuring the absorbance at 280nm of UV spectroscopy (Shi-madzu UV-2550),with an extinction coefficient of 2.64ml y (mg cm )w 21x .Precipitant stock solution was prepared by dissolving the required amount of sodium chloride together with additive glycerol into buffer.The pH of solutions was measured by a digital pH meter (Mettler Toledo 320)and adjusted by the addition of small volumes of NaOH or HAc solution.2.3.Measurement of solubilitySolubility of lysozyme at various temperatures and precipitant y additive concentrations was meas-ured at pH 4.8in 0.1M acetate buffer.Solid–liquid equilibrium was approached through both crystallization and dissolution.Dissolving lasted 3days,while the period of crystallization was over 2weeks.The supernatant in equilibrium with a macroscopically observable solid was then filtered through 0.1-m m filters (Millex-VV ).The concen-tration of diluted supernatant was determined spec-troscopically and verified by refractive meter(Kruss)until refractive index remained unchanged ¨at equilibrium state.Solubility of each sample was measured in duplicate.2.4.Differential scanning microcalorimetry Calorimetric experiments were performed with a micro-differential scanning calorimeter with anultra sensitivity,micro-DSC III,from Setaram SA,France.The micro-DSC recorded heat flow in microwatts vs.temperature,thus can detect the heat associated with phase transition during a temperature scan.The sample made up of equal volumes of protein solution and precipitant solu-tion was filtered through 0.1-m m filters to remove dust particles further.To remove the dissolved air,the sample was placed under vacuum for 3min while stirring at 500rev.y min by a magnetic stirrer.The degassed sample was placed into the sample cell of 1.0ml,and a same concentration NaCl solution was placed into the reference cell.The solutions in the micro-DSC were then cooled at the rate of 0.28C y min.After every run,the cells were cleaned by sonicating for 10–15min in several solutions in the following order:deionized water,methanol,ethanol,acetone,1M KOH and finally copious amounts of deionized water.This protocol ensured that lysozyme was completely removed from the cells.The cells were then placed in a drying oven for several hours.The rubber gaskets were cleaned in a similar manner except acetone and 1M KOH were omitted and they were allowed to dry at low temperature.3.Results and discussionA typical micro-DSC scanning experiment is shown in Fig.1.The onset of the clouding phe-nomenon is very dramatic and easily detected.The sharp increase in the heat flow is indicative of a liquid–liquid phase separation process producing a latent heat.This is much consistent with many recent investigations of the liquid–liquid phase separation of lysozyme from solution w 22,23x .In fact,such a liquid–liquid phase separation is a phase transition with an associated latent heat of demixing.In this work,the cloud-point tempera-tures at a variety of lysozyme,NaCl and glycerol concentrations are determined by the micro-DSC at the scan rate of 128C y h.3.1.Effect of protein concentrationIn semilogarithmic Fig.2we plot the solid–liquid and liquid–liquid phase boundaries for lyso-108J.Lu et al./Biophysical Chemistry 109(2004)105–112Fig.1.Heat flow of a typical micro-DSC scan of lysozyme solution,50mg y ml,0.1M acetate buffer,pH 4.8,3%NaCl.The scan rate 128C y h is chosen referenced to the experimental results of Darcy and Wiencek w 23x .Note the large deflection in the curve at approximately 4.38C indicating a latent heat resulting from demixing (i.e.liquid–liquid phase separation )process.Fig.2.Cloud-point temperature and solubility determination for lysozyme in 0.1M acetate buffer,pH 4.8:solubility (5%NaCl )(s );T (5%NaCl,this work )(d );T (5%cloud cloud NaCl,the work of Darcy and Wiencek w 23x )(*);solubility (3%NaCl )(h );T (3%NaCl )(j ).cloud Fig.3.Cloud-point temperature determination for lysozyme as a function of the concentration of sodium chloride,50mg y ml,0.1M acetate buffer,pH 4.8.zyme in 0.1M acetate buffer,pH 4.8,for a range of protein concentrations.It is worth noting that,at 5%NaCl,our experimental data of T from cloud micro-DSC are quite consistent with those from laser light scattering and DSC by Darcy and Wiencek w 23x ,with difference averaging at approx-imately 0.88C.This figure demonstrates that liquid–liquid phase boundary is far below solid–liquid phase boundary,which implies that the liquid–liquid phase separation normally takes place in a highly metastable solution.In addition,cloud-point temperature T increases with the cloud concentration of protein.3.2.Effect of salt concentrationFig.3shows how cloud-point temperature changes as the concentration of NaCl is varied from 2.5to 7%(w y v ).The buffer is 0.1M acetate (pH 4.8);the protein concentration is fixed at 50mg y ml.Over the entire range of salt concentrations studied,the cloud-point temperature strongly depends on the ionic strength and increases monotonically with the concentration of NaCl.Crystallization is driven by the difference in chemical potential of the solute in solution and in the crystal.The driving force can be simplified as w 24xf sy Dm s kT ln C y C (1)Ž.eq109J.Lu et al./Biophysical Chemistry 109(2004)105–112Fig.4.The driving force required by liquid–liquid phase sep-aration as a function of the concentration of sodium chloride,50mg y ml lysozyme solution,0.1M acetate buffer,pH 4.8.In the same way,we plot the driving force,f ,required by liquid–liquid phase separation as a function of the concentration of sodium chloride in Fig.4.At the moderate concentration of sodium chloride,the driving force required by liquid–liquid phase separation is higher than that at low or high salt concentration.As shown in Fig.3,with NaCl concentration increasing,the cloud-point temperature increases,which is in accord with the results of Broide et al.w 14x and Grigsby et al.w 25x .It is known that protein interaction is the sum of different potentials like electrostatic,van der Waals,hydrophobic,hydration,etc.The liquid–liquid phase separation is driven by a net attraction between protein molecules,and the stronger the attraction,the higher the cloud-point temperature.Ionic strength is found to have an effect on the intermolecular forces:attractions increase with ionic strength,solubility decreases with ionic strength,resulting in the cloud-point temperature increases with ionic strength.It is worth noting that,the effect of ionic strength on cloud-point temperature depends strongly on the specific nature of the ions w 13x .Kosmotropic ions bind adjacent water molecules more strongly than water binds itself.When akosmotropic ion is introduced into water,the entro-py of the system decreases due to increased water structuring around the ion.In contrast,chaotropes bind adjacent water molecules less strongly than water binds itself.When a chaotrope is introduced into water,the entropy of the system increases because the water structuring around the ion is less than that in salt-free water.This classification is related to the size and charge of the ion.At high salt concentration ()0.3M ),the specific nature of the ions is much more important w 25x .The charges on a protein are due to discrete positively and negatively charged surface groups.In lysozyme,the average distance between thesecharges is approximately 10Aw 26x .As to the salt ˚NaCl used as precipitant,Na is weakly kosmo-q tropic and Cl is weakly chaotropic w 27x .At low y NaCl concentrations,as the concentration of NaCl increases,the repulsive electrostatic charge–charge interactions between protein molecules decrease because of screening,resulting in the increase of cloud-point temperature.While at high NaCl con-centrations,protein molecules experience an attrac-tion,in which differences can be attributed to repulsive hydration forces w 14,25x .That is,as the ionic strength increases,repulsive electrostatic or hydration forces decrease,protein molecules appear more and more attractive,leading to higher cloud-point temperature.At various salt concentra-tions,the predominant potentials reflecting the driving force for liquid–liquid phase separation are different.Fig.4shows that the driving force,f ,is parabolic with ionic strength,while Grigsby et al.w 25x have reported that f y kT is linear with ionic strength for monovalent salts.The possible reasons for that difference include,their model is based on a fixed protein concentration of 87mg y ml,which is higher than that used in our study,yet f y kT is probably dependent on protein concentration,besides the solutions at high protein and salt concentrations are far from ideal solutions.3.3.Effect of glycerolFig.5compares cloud-point temperature data for 50mg y ml lysozyme solutions in absence of glycerol and in presence of 5%glycerol,respec-110J.Lu et al./Biophysical Chemistry109(2004)105–112parison of cloud-point temperatures for lysozyme at different glycerol concentrations as a function of the con-centration of sodium chloride,50mg y ml,0.1M acetate buffer, pH4.8:0%glycerol(s);5%glycerol(j).Fig.6.Cloud-point temperatures for lysozyme at different glycerol concentrations,50mg y ml lysozyme,5%NaCl,0.1M acetate buffer,pH4.8.Fig.7.Cloud-point temperature and solubility determination for lysozyme at different concentrations of glycerol in0.1M acetate buffer,5%NaCl,pH4.8:solubility(0%glycerol)(s); T(0%glycerol)(d);solubility(5%glycerol)(h);cloudT(5%glycerol)(j).cloudtively.Fig.6shows the cloud-point temperature as a function of the concentration of glycerol.The cloud-point temperature is decreased as the addi-tion of glycerol.In semilogarithmic Fig.7we plot the solid–liquid and liquid–liquid phase boundaries at dif-ferent glycerol concentrations for lysozyme in0.1 M acetate buffer,5%NaCl,pH4.8,for a range of protein concentration.This figure demonstrates that liquid–liquid and solid–liquid phase bounda-ries in the presence of glycerol are below those in absence of glycerol,and the region for growing crystals is narrowed when glycerol is added. Glycerol has the property of stabilizing protein structure.As a result,if crystallization occurs over a long period of time,glycerol is a useful candidate to be part of the crystallization solvent and is often included for this purpose w28x.In addition,glycerol is found to have an effect on the intermolecular forces:repulsions increase with glycerol concentra-tion w29x.Our experiment results of solubility and cloud-point temperature can also confirm the finding.The increased repulsions induced by glycerol can be explained by a number of possible mecha-nisms,all of which require small changes in the protein or the solvent in its immediate vicinity.The addition of glycerol decreases the volume of protein core w30x,increases hydration and the size of hydration layer at the particle surface w31,32x. In this work,we confirm that glycerol shifts the solid–liquid and liquid–liquid phase boundaries. The effect of glycerol on the phase diagram strong-111 J.Lu et al./Biophysical Chemistry109(2004)105–112ly depends on its concentration and this canprovide opportunities for further tuning of nuclea-tion rates.4.ConclusionsGrowing evidence suggests protein crystalliza-tion can be understood in terms of an order ydisorder phase transition between weakly attractiveparticles.Control of these attractions is thus keyto growing crystals.The study of phase transitionsin concentrated protein solutions provides one witha simple means of assessing the effect of solutionconditions on the strength of protein interactions.The cloud-point temperature and solubility datapresented in this paper demonstrate that salt andglycerol have remarkable effects on phase transi-tions.The solid–liquid and liquid–liquid bounda-ries can be shifted to higher or lower temperaturesby varying ionic strength or adding additives.Ourinvestigation provides further information upon therole of glycerol used in protein crystallization.Glycerol can increase the solubility,and decreasethe cloud-point temperature,which is of benefit totuning nucleation and crystal growth.In continuingstudies,we will explore the effects of other kindsof additives like nonionic polymers on phasetransitions and nucleation rates.Much more theo-retical work will be done to fully interpret ourexperimental results.AcknowledgmentsThis work is supported by the grant from theNational Natural Science Foundation of China(No.20106010).The authors also thank Professor J.M.Wiencek(The University of Iowa)for kinddiscussion with us about the thermal phenomenaof liquid–liquid phase separation.Referencesw1x A.McPherson,Current approaches to macromolecular crystallization,Eur.J.Biochem.189(1990)1–23.w2x A.M.Kulkarni, C.F.Zukoski,Nanoparticle crystal nucleation:influence of solution conditions,Langmuir18(2002)3090–3099.w3x E.E.G.Saridakis,P.D.S.Stewart,L.F.Lloyd,et al., Phase diagram and dilution experiments in the crystal-lization of carboxypeptidase G2,Acta Cryst.D50(1994)293–297.w4x A.P.Gast, C.K.Hall,W.B.Russel,Polymer-induced phase separations in non-aqueous colloidal suspensions,J.Colloid Interf.Sci.96(1983)251–267.w5x H.N.W.Lekkerkerker,W.C.K.Poon,P.N.Pusey,et al., Phase-behavior of colloid plus polymer mixtures,Euro-phys.Lett.20(1992)559–564.w6x A.Tardieu,S.Finet,F.Bonnete,Structure of the´macromolecular solutions that generate crystals,J.Cryst.Growth232(2001)1–9.w7x D.Rosenbaum,C.F.Zukoski,Protein interactions and crystallization,J.Cryst.Growth169(1996)752–758.w8x A.George,W.W.Wilson,Predicting protein crystalli-zation from a dilute solution property,Acta Cryst.D50(1994)361–365.w9x D.Rosenbaum,P.C.Zamora, C.F.Zukoski,Phase-behavior of small attractive colloidal particles,Phys.Rev.Lett.76(1996)150–153.w10x V.J.Anderson,H.N.W.Lekkerkerker,Insights into phase transition kinetics from colloid science,Nature416(2002)811–815.w11x S.Tanaka,K.Ito,R.Hayakawa,Size and number density of precrystalline aggregates in lysozyme crys-tallization process,J.Chem.Phys.111(1999)10330–10337.w12x D.W.Liu,A.Lomakin,G.M.Thurston,et al.,Phase-separation in multicomponent aqueous-protein solutions,J.Phys.Chem.99(1995)454–461.w13x V.G.Taratuta,A.Holschbach,G.M.Thurston,et al., Liquid–liquid phase separation of aqueous lysozymesolutions:effects of pH and salt identity,J.Phys.Chem.94(1990)2140–2144.w14x M.L.Broide,T.M.Tominc,M.D.Saxowsky,Using phase transitions to investigate the effect of salts onprotein interactions,Phys.Rev.E53(1996)6325–6335. w15x M.Muschol,F.Rosenberger,Liquid–liquid phase sep-aration in supersaturated lysozyme solutions and asso-ciated precipitate formation y crystallization,J.Chem.Phys.107(1997)1953–1962.w16x C.Domb,J.H.Lebowitz,Phase Separation and Critical Phenomena,Academic,London,1983.w17x D.F.Rosenbaum,A.Kulkarni,S.Ramakrishnan,C.F.Zukoski,Protein interactions and phase behavior:sen-sitivity to the form of the pair potential,J.Chem.Phys.111(1999)9882–9890.w18x O.Galkin,P.G.Vekilov,Nucleation of protein crystals: critical nuclei,phase behavior and control pathways,J.Cryst.Growth232(2001)63–76.w19x P.R.tenWolde, D.Frenkel,Enhancement of protein crystal nucleation by critical density fluctuations,Sci-ence277(1997)1975–1978.w20x P.A.Darcy,J.M.Wiencek,Estimating lysozyme crystal-lization growth rates and solubility from isothermalmicrocalorimetry,Acta Cryst.D54(1998)1387–1394.112J.Lu et al./Biophysical Chemistry109(2004)105–112w21x A.J.Sophianopoulos,C.K.Rhodes,D.N.Holcomb,K.E.vanHolde,Physical studies of lysozyme.I.Characteri-zation,J.Biol.Chem.237(1962)1107–1112.w22x Y.Georgalis,P.Umbach, A.Zielenkiewicz,et al., Microcalorimetric and small-angle light scattering stud-ies on nucleating lysozyme solutions,J.Am.Chem.Soc.119(1997)11959–11965.w23x P.A.Darcy,J.M.Wiencek,Identifying nucleation tem-peratures for lysozyme via differential scanning calorim-etry,J.Cryst.Growth196(1999)243–249.w24x M.L.Grant,Effects of thermodynamics nonideality in protein crystal growth,J.Cryst.Growth209(2000)130–137.w25x J.J.Grigsby,H.W.Blanch,J.M.Prausnitz,Cloud-point temperatures for lysozyme in electrolyte solutions:effectof salt type,salt concentration and pH,Biophys.Chem.91(2001)231–243.w26x D.Voet,J.Voet,Biochemistry,Wiley,New Y ork,1990. w27x K.D.Collins,Charge density-dependent strength of hydration and biological structure,Biophys.J.72(1997)65–76.w28x R.Sousa,Use of glycerol and other protein structure stabilizing agents in protein crystallization,Acta Cryst.D51(1995)271–277.w29x M.Farnum, C.F.Zukoski,Effect of glycerol on the interactions and solubility of bovine pancreatic trypsininhibitor,Biophys.J.76(1999)2716–2726.w30x A.Priev,A.Almagor,S.Yedgar,B.Gavish,Glycerol decreases the volume and compressibility of proteininterior,Biochemistry35(1996)2061–2066.w31x S.N.Timasheff,T.Arakawa,Mechanism of protein precipitation and stabilization by co-solvents,J.Cryst.Growth90(1988)39–46.w32x C.S.Miner,N.N.Dalton,Glycerol,Reinhold Publishing, New Y ork,1953.。
导电聚合物
聚对苯撑乙烯 (PPV)
聚对苯(PPP)
聚芴(PF)
R1
R2
常见的共轭聚合物的分子链结构
聚乙炔的链结构
反式聚乙炔的链结构和结构缺陷
(a) (b) (c) (d)
基态简并共轭链 孤子结构缺陷 极化子结构缺陷
聚苯胺的主链结构
H N H N
x
N
N
1-x n
x = 1时为全还原态(leucoemeraldine,PAn-I) x = 0.5时为半氧化态(emeraldine, PAn-II)、碱式聚苯胺(emeraldine base, EB) x = 0时为全氧化态(pernigraniline, PAn-III)。
碱式聚苯胺(EB)的质子酸掺杂
H N H N N N
-2HCl
+2HCl
Cl -
H N
+
H N
Cl -
H+ N
H N
聚吡咯的两种掺杂结构
A
-
AH N
(
H N
+ N H
)n
(
H N
H2 + N H
H N
)n
载流子与相关的结构缺陷
孤子(Soliton),用S表示。这是基态简并的反式聚乙炔所 特有的一种结构缺陷,这种孤子是离域在很长的共轭链段上, 并且其在共轭链上可自由迁移。孤子的自旋为1/2 中性孤子被氧化失去电子形成电荷载流子正孤子S+,被还原 得到电子形成电荷载流子负孤子S-。正孤子和负孤子则没有 自旋(自旋为0)。
扩展到包括本征态共轭聚合物。
电子聚合物 Electronic polymers
指具有光电性能的共轭聚合物,包括掺杂态导电聚 合物和具有半导体特性的本征态共轭聚合物(主要 是发光聚合物和用于聚合物太阳能电池的共轭聚合
不同酶消化法提取猪原代肝细胞的效果比较
532024.4·试验研究0 引言猪圆环病毒(PCV )是Circoviridae 科Circovirus 属的一种无囊膜的单链环状DNA 病毒。
在已知的4个血清型中,PCV2为猪易感的致病性病毒[1]。
PCV2感染会诱导宿主免疫抑制引起猪圆环病毒病(PCVD ),包括断奶仔猪多系统衰竭综合征、新生仔猪先天性脑震颤、皮炎与肾病综合征、猪呼吸道病综合征、母猪繁殖障碍等,给全世界养猪业带来较大的经济损失,是世界各国的兽医与养猪业者公认的造成重大影响的猪传染病[2]。
PCV2的感染在猪生长发育的不同阶段有不同的组织嗜性。
但无论是胎儿阶段还是出生后,肝细胞都是PCV2感染和复制的靶细胞。
因此,PCV2也被视为一种能够诱导猪肝炎的病毒[3]。
且PCV2诱导的肝细胞凋亡在PCV2引发的相关病变和疾病的发病机制中具有关键性作用[4]。
因此,方便、快捷地获取大量有活性的猪肝细胞对于研究PCVD 的致病机制具有重大意义。
目前获取肝细胞常用的方法主要包括机械分离细胞法、非酶分离细胞法、离体酶消化法和酶灌流法等[5]。
因此,本试验采用简便、经济、无需特殊设备、仅需部分肝组织的离体酶消化法,比较不同酶消化分离猪原代肝细胞的效果,为一般实验室提取分离大量有活性的猪肝细胞提供参考。
1 材料与方法1.1 材料1.1.1 主要试剂新鲜猪肝组织,Hank's 平衡盐溶液(HBSS ),磷酸盐缓冲液(无菌PBS ),4%多聚甲醛(PFA ),收稿日期:2024-01-27基金项目:国家自然科学基金项目:复杂器官与组织在脾脏内的功能性再生(32230056)作者简介:周徐倩(1999-),女,汉族,浙江温州人,硕士在读,研究方向:组织工程与再生医学。
*通信作者简介:董磊(1978-),男,汉族,安徽阜阳人,博士,教授,研究方向:组织工程与再生医学、生物材料。
周徐倩,董磊.不同酶消化法提取猪原代肝细胞的效果比较[J].现代畜牧科技,2024,107(4):53-55. doi :10.19369/ki.2095-9737.2024.04.014. ZHOU Xuqian ,DONG Lei .Comparison of the Effect of Different Enzyme Digestion Methods on Extraction of Porcine Primary Hepatocytes[J].Modern Animal Husbandry Science & Technology ,2024,107(4):53-55.不同酶消化法提取猪原代肝细胞的效果比较周徐倩,董磊*(南京大学,江苏 南京 210023)摘要:猪肝细胞是猪圆环病毒的靶细胞,简单快速地提取猪原代肝细胞对于研究猪圆环病毒病的致病机制具有重要意义。
丛柳珊瑚Euplexaura rhipidalis中细胞毒活性二萜成分分析
正 反 相 硅胶 柱 层 析 、 S e p h a d e x L H一 2 0凝 胶 柱 层 析 和半 制 备 HP L C等方法进行分离和纯化 , 再利用 N MR、 MS等 波谱方法 , 并 与 文 献 对 照 确 定 了 3个 高 氧 化 度 的 西 松 烷 二 萜 类 化 合 物 , 分别 为 c o r u l l o i d o l i d e A( 1 ) , ( 1 R, 3 S, 4 S, 7 E, 1 1 E) 一 3 , 4 - e p o x y c e mb r a 一 7 , 1 1 , 1 5 一 t r i e n e( 2 ) ,( 一) 一 c o r u l l o i d o l i d e E( 3 ) ; 细胞毒活性模型评 价结果发现 ,
An a l y s i s o f Cy t o t o x i c Di t e r p e n o i d Co m po s i t i o n f r o m t he
Go r g o ni a n Co r a l Eu p l e xau r a r h i pi dal i s
广 西科 学 Gu a n g x i S c i e n c e s 2 0 1 3 , 2 0 ( 4 ) : 2 6 9  ̄2 7 1 , 2 7 5
The Properties and Applications of Biochemistry
The Properties and Applications ofBiochemistry生物化学的性质与应用生物化学是研究生命活动的化学基础的学科。
它涉及许多重要的生物大分子,如蛋白质、核酸、多糖和脂质等。
这些生物大分子具有各自独特的性质和广泛的应用。
本文将介绍这些生物大分子的性质和应用。
蛋白质蛋白质是一种拥有复杂结构的生物大分子,是组成人体的重要物质之一。
蛋白质的结构可分为四级,分别是原位结构、二级结构、三级结构和四级结构。
蛋白质的性质包括酶促活性、免疫性、分子识别性和构象可变性等。
蛋白质的应用非常广泛,如生物制药、食品添加剂、工业酶、生物传感器等。
核酸核酸是生物体内遗传信息的主要存储和传递分子,包括脱氧核糖核酸(DNA)和核糖核酸(RNA)两种。
DNA是双链结构,RNA是单链结构。
核酸的主要功能是编码蛋白质的氨基酸序列。
核酸包括许多重要的结构和功能性元素,如基序、剪接位点、启动子和转录因子结合位点等。
核酸的应用领域涉及多方面,如基因工程、药物研究、生物信息学等。
多糖多糖是一类由单糖分子组成的高分子化合物,是植物和动物细胞壁、软组织、骨骼等的主要组成部分。
多糖分子结构单一,但其结构与含量却具有很大的变异性。
多糖具有多种生物学功能,如细胞识别、浸润和保护、免疫调节等。
多糖也被广泛应用在食品工业、药物研究、生物传感器等领域中。
脂质脂质是一类极为广泛的生物大分子,包括脂肪酸、甘油三酯、磷脂、鞘磷脂、皂质和固醇等。
脂质具有多种生命活动的功能,如细胞膜形成、激素合成、能量储存等。
脂质的应用也非常广泛,如人体营养、洗涤剂、润滑剂、药物传递等。
总体来说,生物化学是非常重要的研究领域,其研究内容和应用都非常广泛。
各种生物大分子都有其独特的性质和应用,深入研究这些生物大分子能为广大科学工作者提供更多的理论基础和实践应用价值。
原子力显微镜观察虫草多糖分子的结构形貌
第18卷第1期1999年2月电 子 显 微 学 报Journa l of Ch i nese Electron M icroscopy Soc ietyVol-18,No111999-02文章编号:100026281(1999)01201032105原子力显微镜观察虫草多糖分子的结构形貌蔡林涛 李 萍3 陆祖宏(东南大学分子与生物分子电子学开放研究实验室,南京210096)(3中国医科大学生药学教研室,南京210009)摘 要:本文通过原子力显微镜直接观察虫草多糖分子的三维结构形貌。
虫草多糖分子的稀溶液铺展在N i2+处理的云母片上,经干燥,乙醇固定后,获得稳定、重复的图像。
结果表明虫草多糖分子具有高度分枝的结构,并且糖链间形成小环或螺旋结构。
关键词:虫草;多糖分子;生物样品;原子力显微镜分类号:TH74219;Q949132718;O629112;Q336 文献标识码:A原子力显微镜(A FM)[1]是观察物质表面细微形貌和表面加工的强有力工具。
它在生物形态学[2—3]上的应用引起了人们广泛的关注。
由于生物样品或大分子表面往往很脆弱且易被扫描针尖破坏,为获得重现性好、分辨率高的图像,必须改进制样方法和减小针尖和样品间的相互作用力,采用轻敲(T app ing M ode)模式或在液体池中成像,有助于提高图像质量。
多糖是一种很重要的生物大分子,目前,采用A FM对多糖分子[4]的研究,还远远落后于核酸分子(DNA)、蛋白质和磷脂生物膜等生物材料。
多糖在体内不仅参与能量的储存和传递,而且也作为细胞骨架材料和参与细胞识别等多种生命功能活动。
本研究通过改进制样方法,采用A FM的轻敲模式,观察从中华医药瑰宝虫草中提取出来的多糖分子的形貌结构,获得了清晰、稳定的图像。
为进一步研究其立体结构及其构效关系,以及在分子水平上阐述虫草多糖分子的药效和作用机制作准备。
材料与方法11虫草多糖的分离和纯化 虫草菌体培养液经浓缩后,去除残渣,乙醇沉淀,脱蛋白、脱色,透析、浓缩、醇沉,得粗多糖。
反相高效液相色谱法分离制备银杏内酯
24反相高效液相色谱法分离制备银杏内酯唐晓丹1,丛景香1,潘明宽2,林炳昌11.辽宁科技大学分离技术中心 (鞍山 114051);2.辽宁科技大学化工学院 (鞍山 114051)摘要采用高效液相色谱技术,分离制备高纯度银杏内酯。
通过单因素及正交试验,对银杏内酯的浸提条件进行优化,再通过反相高效液相色谱分离系统,采用浓度梯度洗脱的方式,分离出高纯度银杏内酯。
结果显示最佳浸提条件:提取剂为乙酸乙酯,浸提时间4 h,温度65℃,固液比1∶5,银杏内酯含量由原料中的6%,提高到29%。
通过制备色谱柱层析,得到产品中银杏内酯纯度≥90%,回收率为70%。
关键词银杏内酯;高效液相色谱(HPLC);分离制备Separation and Preparation of Ginkgolides by Reverse Phase-high-performance Liquid ChromatographyTang Xiao-dan 1, Cong Jing-xiang 1, Pan Ming-kuan 2, Lin Bing-chang 11.Center of Separation Technology, University of Science and Technology Liaoning (Anshan 114051);2.School of Chemical Engineering, University of Science and Technology Liaoning (Anshan 114051)Abstract Separation and preparation of high purity of ginkgolides from Ginkgo biloba leaves was investigated by high-performance liquid chromatography (HPLC). The ef fi cient factors of extraction of ginkgolides by single factor tests and orthogonal experiment were optimized. RP-HPLC using gradient elution method was utilized. And then the high purity ginkgolides was achieved from the crude extract of Ginkgolide. Results showed: the optimum conditions of extraction as follows: the extractant of ethyl acetate, extraction temperature 65℃, extraction time 4 h, ratio of solid to liquid extraction 1:5. The content of ginkgolides was increased from 6% to 29%. The product of ginkgolides was obtained by RP-HPLC, product purity was about 90%, product yield was 70%.Keywords ginkgolides ;high performance liquid chromatography(HPLC);separation and preparation 基金项目:中国高教部高等学校博士学科点专项科研基金项目(20070146001),辽宁省教育厅团队项目(2008T093)银杏叶中主要含有银杏黄酮和银杏内酯两大类生物活性物质。
博士复试英文PPT
3. PTBP1 enhances exon11a skipping of Mena premRNA in lung cancer cells
Results
1. PTBP1 is highly expressed in lung adenocarcinoma (LUAD) tissues and 95-D cells and upregulation of PTBP1 is associated with EMT progress
2. PTBP1 promotes migration and invasion of lung cancer cells
Master Research
PTBP1 enhances exon11a skipping in Mena premRNA to promote migration and invasion in lung
carcinoma cells
Background Objectives Technology Methods Results Conclusions
5. PTBP1-mediated migration and invasion of 95-D cells are partially dependent on MenaINV
Results
2.1. Overexpressed PTBP1 promotes levels of EMT-related proteins in lung cancer cells
Technology Methods
Results
1. PTBP1 is highly expressed in lung adenocarcinoma (LUAD) tissues and 95-D cells and upregulation of PTBP1 is associated with EMT progress
常压酸化法从磷钾伴生矿酸浸液中制备硫酸钙晶须
2017年7月第28卷第4期化学研究C H E M I C A L R E S E A R C H432http :// 常压酸化法从磷钾伴生矿酸浸液中制备硫酸钙晶须刘存成\贺方杰\孙顺平2,吕仁亮\覃远航\王为国\王存文1S(1.武汉工程大学化工与制药学院,湖北武汉430073; 2.中国天辰工程有限公司,天津300400)摘要:以磷钾伴生矿的盐酸浸出液为介质,研究常压酸化法制备硫酸钙晶须,分别考察了不同硫酸浓度、加热 温度、™(S〇4_)/™(Ca2+)、滴加速率以及搅拌转速等操作条件对硫酸钙晶须长径比的影响.实验结果表明:在硫酸 浓度为 0.87 mol/L、加热温度为 105 T;、n>(S〇4_)/n(Ca2+)= 1.2、滴加速率为 2.5 m L/m i n、搅拌转速为 166 l'/min 较佳的条件下,可制得长径比为32的二水硫酸钙晶须.关键词:磷钾伴生矿;酸浸液;硫酸;常压酸化法;硫酸钙晶须中图分类号:TB32;TD985 文献标志码:A文章编号:1008-1011(2017)04-0432-07Preparation of calcium sulfate whisker from acid-leaching solution of phosphate-potassium associated oreby atmospheric acidification methodL I U C u n c h e n g1,H E Fangjie1,S U N Shunping2,L U Renliang1,Q I N Y u a n h a n g1,W A N G W e i g u o1,W A N G C u n w e n1*(1.School o f Chemical Engineering and Pharmacy, Wuhan Institute o f Technology, Wuhan430073, Hubei, China;2.China Tianchen Engineering Corporation, Tianjin300400, China)Abstract:Calcium sulfate whisker was prepared by atmospheric acidification method using the hydrochloric acid-leaching solution ol phosphate-potassium associated ores. T h e effects ol different concentrations ol sulfuric acid,heating temperatures,the molar ratios ol ore S〇4 /C a2+,the dropping rates ol sulfuric acid and stirring speeds on the aspect ratio ol calcium sulfate whisker were investigated in detail. T h e results ol experiment showed that the calcium sulfate whisker with an aspect ratio ol 32 could be obtained under the o p t i m u m conditions ol the sulfuric acid concentration 0.87 m o l/L,heating t emperature 105 T,the molar ratio ol sulfate ion to calcium ion 1.2,the dropping rate ol sulfuric acid 2.5 m L/m i n and the stirring speed 166 r/min.Keywords:phosphate-potassium associated ore; acid-leaching solution;sulfuric acid; atmospheric a- cidilication method ;calcium sulfate whisker硫酸钙晶须是硫酸钙的纤维状单晶体,有二水、半水以及无水三种类别.外观上均呈白色蓬松状. 硫酸钙晶须具有短纤维状填料的长径比,颗粒状填收稿日期:2017-01-17.基金项目:国家自然科学基金资助项目(51274153);湖北省自然科 学基金重点项目(2011CDA120);湖北省自然科学基金资助项目(2015CFB523).作者简介:刘存成(1990-),男,硕士生,主要从事矿产综合利用方 面的研究.* 通讯联系人,E-mail: wangcw0120@.料的粒度.此外,它还具有韧性好、耐高温、强度高、尺寸稳定、抗化学腐蚀等特性,是一种价格较低、性 能优良的绿色环保材料,广泛应用在塑料、胶黏剂以 及造纸等领域[|-2].水热法和常压酸化法是制备硫 酸钙晶须的主要方法[3-4].常压酸化法是向酸性溶 液中加人浓度较高的生石膏悬浮液,生石膏可向纤 维状的硫酸钙晶须转变.常压酸化法由于制备工艺 简单,不存在高温高压操作,因而受到了广泛的应 用[5-7]第4期刘存成等:常压酸化法从磷钾伴生矿酸浸液中制备硫酸钙晶须433〇1--------------------------1--------------------------'--------------------------1--------------------------0 12 34硫酸浓度/(mol/L)图1硫酸浓度对晶须的长径比的影响Fig.l Effect o f sulfuric acid concentrationon the aspect ratio of whiskers由图1和图2可以看出:在硫酸浓度为0.21〜3.48 mol /L范围内,随着滴人硫酸浓度增大,晶须的丈后保温陈化3 h 得到晶须产物,过滤,滤饼用95丈热水充分洗涤三次,60丈下干燥4 h 后密封保存于干燥器中.采用偏光显微镜对所得晶须产物进行 观察,获得其形貌及长径比.2结果与讨论2.1硫酸浓度的影响向100 m L 磷钾伴生矿浸出液(盐酸浓度约为 2.7 m o L /L 、C a 2+浓度为0.87 m o l /L )中滴加硫酸,硫 酸浓度的变化将对酸浸液中硫酸钙的溶解度产生影 响,进而影响溶液的过饱和度.在加热温度85丈、 n_(S〇4 )/n_(Ca 2+) = 1、滴加速率为 2.5 m L /m i n 以及我国湖北宜昌地区拥有储量丰富的磷钾伴生矿 资源,综合利用该地区磷钾伴生矿资源,对于提升磷 资源综合利用率以及减少钾资源对外依存度有着重 大意义[8—9].课题组前期以盐酸处理磷钾伴生矿, 钾、钙浸出率均达到90%以上,酸浸液中含有大量 的钾、钙资源[10].钾用来制备NPK 复合肥,大量的 钙如不加以利用,不但是资源的极大浪费,钙离子的 存在还会给后续工艺带来麻烦.因此,酸浸液中的 钙急需分离利用,可考虑制备硫酸钙晶须.以磷钾伴生矿的盐酸浸出液为介质,直接向酸浸液中滴加硫酸,米用常压酸化法制备硫酸钙晶须.考察硫酸浓度、加热温度、几(S 〇4_ )/ 4 Ca 2+)、滴加速率以及搅拌转速等操作条件对 硫酸钙晶须长径比的影响,从而获得较佳的硫酸 钙晶须制备工艺条件.1实验部分1.1试剂和仪器实验所用试剂和仪器见表1和表2.表1实验所用试剂Table 1 Reagents for the experiment名称化学式级别生产厂家浓盐酸HC1A R 开封东大化工甲基红c,5h ,5N3〇2A R 国药集团化学试剂无水乙醇C A 〇A R 天津博迪氢氧化钾K O H A R 国药集团化学试剂三乙醇胺C 6H i 5N 〇3A R 南京化学试剂钙试剂C 20H,3N 2N a O 5SA R 国药集团化学试剂乙二胺四乙酸二钠C 10H 14N 2Na2O 8A R国药集团化学试剂表2实验所用仪器Table 2 Instruments for the experiment仪器与设备名称型号生产厂家电子天平PL203梅特勒-托利多仪器上海有限公司偏光显微镜XP-330C上海蔡康光学仪器有限公司1.2实验方法采用文献[11]的方法制备磷钾伴生矿酸浸液,取100 mL 酸浸液加热到一定温度,用恒压滴液漏斗 将不同浓度的硫酸滴加到酸浸液中,保温1.5 h ,然 后趁热抽滤,将滤液快速转移到三颈烧瓶中,在一定 的搅拌速率下关闭油浴加热装置自然降温,降至20搅拌转速为166 r /min 的条件下,研究硫酸浓度对 晶须长径比的影响,实验结果见图1和图2.10i ^434化学研究2017 年(a) 3.48 mol/L,(b) 2.61 mol/L,(c) 1.74 moL/L,(d) 0.87 mol/L,(e) 0.43 mol/L.图2不同硫酸浓度下硫酸钙晶须的光学显微图像Fig.2 Optical microscope images o f whiskers obtained a tdifferent H2S O4concentrations长径比先增大后减小.这是因为当按《(8〇4_)/ra(C a2+)= 1滴加硫酸时,硫酸的滴人将造成酸浸液 体积与体系中盐酸浓度的变化,这将对酸浸液中硫 酸钙的溶解度产生影响,进而影响溶液的过饱和度.前期研究结果表明[12]:向磷钾伴生矿酸浸液中滴人 0.43〜3.48 m o l/L硫酸,酸浸液中盐酸浓度的变化范 围为0.9 ~2.16 m o l/L.在该盐酸浓度范围内,硫酸钙 的溶解度随盐酸浓度的变化范围为22〜29 g/100 L (见图3),酸浸液体积的变化范围为300〜125 m L.因此,酸浸液体系的过饱和度随着滴人硫酸浓度的 减小而减小.当滴人较大浓度的硫酸时,晶体呈现粗大且不 规则的短柱状,并团聚在一起,产品颗粒夹杂也十分 严重(如图2(a)).这是由于滴人较大浓度的硫酸 时,生成的硫酸钙在酸浸液中溶解不完全,体系的过 饱和度过大.当过饱和度过大时,将很难维持整个 晶面过饱和度的恒定,体系中有大量不定型的硫酸 钙,部分硫酸钙还未按晶状生长就被掺杂进人晶体 中[13—14].此外,过饱和度越大,晶体的生长速度也就 越快,较快的生长速度使酸浸液中的金属离子很容图3 25尤和80尤下硫酸钙在不同浓度盐酸中的溶解度 Fig.3 Solubility o f calcium sulfate in different concentrations in hydrochloric acid at 25 ^ and 80 ^易吸附在晶体表面或者进人晶体,晶体生长的均勻 性遭到破坏,导致被破坏的晶面生长速率总是比光 滑面的生长速率大,造成相对生长速率的改变,这样 就影响到晶体的生长形态[|5].另外,磷钾伴生矿的 浸出液中存在大量的金属离子,这些金属离子在酸 浸液中成为结晶中心,促使了产物的聚集生长[16].当滴人较小浓度的硫酸时,形成的晶体长径比 较小.这是由于滴人的硫酸浓度较小时,体系的过 饱和度过小,不足以促使晶体生长.因此,适宜的硫 酸浓度为0.87 mol/L.2.2加热温度的影响加热温度对硫酸钙的溶解度有重要影响,进而 直接影响到溶液的过饱和度.在硫酸浓度为0.87 m o l/L、ra(S〇4_)/ra(Ca2+)=1、滴加速率为 2.5 m L/ m i n以及搅拌转速为166 r/m i n的条件下,研究在较 低加热温度范围内(65〜105丈),温度对晶须长径 比的影响,实验结果见图4和图5.Fig.4 Effect o f heating temperatureon the aspect ratio o f whiskers第4期刘存成等:常压酸化法从磷钾伴生矿酸浸液中制备硫酸钙晶须4351.01.21.4SO^与Ca2_的物质的量之比图6 «•( S〇4_ )/«•( Ca2+ )对晶须长径比的影响Fig.6 Effect o f the molar r a tio o f sulfate ion andcalcium ion on the aspect rat i o o f whiskers(a) 1.0,(b) l.l,(c) 1.2,(d) 1.3,(e) 1.4.图 7 不同 f t ( S〇4_ ) /ft ( Ca2+ )的晶须光学显微图像Fig.7 Optical microscope images o f whiskers obtained a t different molar ratios of sulfate ion and calcium ion着低能面不断的生长[IS],但s 〇4—的浓度过高时,影 响程度逐渐减小[|7].因此,适宜的f t (s 〇4-)/ft(Ca 2+)为 1.2.2.4滴加速率的影响在硫酸浓度为0.87 m o l /L 、加热温度105丈、ft(S〇24_)/ft(Ca 2+)= 1.2 以及搅拌转速为 166 r/min 的条件下,研究滴加速率对晶须长径比的影响,实验\>、!//\丨A,¥[.]f l O O ^m1、11100 urn(a) 65 ^,(b)75 ^,(c)85 ^,(d)95 ^,(e)105 图5不同加热温度下硫酸钙晶须的光学显微图像Fig.5 Optical microscope images of whiskers obtainedat different heating temperatures由图4和图5可以看出:加热温度在65〜105丈范围内,随着温度的升高,晶须的长径比逐渐增大.这是因为:当体系中硫酸钙的含量一定时,溶液 的过饱和度随温度的升高而逐渐下降.当体系温度 较低时,硫酸钙的溶解度较小,导致溶液的过饱和度过大.此时溶液中有大量的无定型的硫酸钙粒子, 黏度急剧增大,酸浸液浓度和温度的均勻性不能得 到保证,增大了二次成核的可能,造成产品的均勻性 和长径比下降[13].因此,适宜的加热温度为105丈. 2.3 ra (S 〇4_)/«(Ca 2+)的影响在硫酸浓度0.87 mol /L 、滴加速率2.5 m L /m i n 、 加热温度105丈以及搅拌转速为166 r /m i n 的条件 下,研究f t( S 〇4_ )/ft( C a 2+ )对晶须长径比的影响,实 验结果见图6和图7.由图6和图7可以看出:在ft(S〇4_)/ft(Ca2+) 为1.0〜1.4的范围内,随着ft(S〇4—)/ft(Ca2+)的增 大,晶须的长径比先增大后基本保持不变.这是因 为:酸浸液中H2SO 4浓度对晶体的形貌有重要影响,在一定范围内,过量的H2S O 4有利于增加体系中S 〇4-与C a 2+在晶核表面的结合率,有利于形成较大尺寸的晶体[17].此外,S O 4—离子的增加,促进晶体向^436化学研究2017 年246滴加速率/ (mL / min)图8滴加速率对晶须长径比的影响Fig.8 Effect o f the dropping rate on theaspect r a tio of whiskers(a) 1. 5 mL/min,( b) 2.5 mL/ m in,( c) 3.5 mL/ m in,(d) 4.5 mL/min,(e) 5.5 mL/min.图9不同滴加速率的晶须光学显微图像Fig.9 Optical microscope images o f whisker obtainedat different dropping rates由图8和图9可以看出:随着滴加速率的增大, 晶须的长径比逐渐下降.这可能是由于当滴加速率较大时,在较短的时间里体系中产生了大量的不定 型的硫酸钙沉淀,导致酸浸液的浓度过大,黏度也随 之增大,部分硫酸钙并没有完全溶解,未溶解的颗粒 在冷却结晶的过程中充当了晶种,体系中出现了二 次成核.考虑到生产效率,适宜的滴加速率为2.5结果见图8和图9.m L /min .2.5撿拌转速的影响搅拌转速对结晶过程有着重要影响.在硫酸浓 度为 0. 87 mol /L 、加热温度 105 丈、( SO 2,)/ r a (Ca 2+)= 1.2以及滴加速率为2.5 m L /min 的条件 下,研究搅拌转速对晶须长径比的影响,实验结果见 图10和图11.Fig.10 Effect o f the s t i rring speed on theaspect r at i o o f whiskers(a) 0 r/min,(b) 166 r/min,(c) 333 l'/min,(d) 498 r/min,(e) 664 r/min.图11不同搅拌转速下晶须的光学显微图像 Fig.11 Optical microscope images o f whisker obtainedat different s tirring speeds第4期刘存成等:常压酸化法从磷钾伴生矿酸浸液中制备硫酸钙晶须437由图10和图11可以看出:随着搅拌转速的增 大,晶须的长径比先增大后减小.这主要是因为在 没有搅拌时,硫酸钙晶须不能均勻悬浮于酸浸液中,导致酸浸液中局部浓度偏高,生成的晶须长短不一.此外,没有搅拌时,细碎的晶体具有较高的比表面 能,晶体容易发生团聚并黏附于容器壁上,不易过 滤[19].但搅拌转速过高时,体系中易发生二次成核,造成成核过多,晶体长不大.在保证混合均勻、尽量 避免过多晶核的产生的前提下,适宜的搅拌转速为166 r/min.通过以上单因素实验可获得硫酸钙晶须较佳的 制备工艺条件.在硫酸浓度为0.87 m〇l/L、加热温度 为 105 T;、ra(S〇4_)/ra(C a2+)= 1.2、滴加速率为 2.5 m L/m i n、搅拌速率为166 r/m i n的条件下,制备得到 的二水硫酸钙晶须长径比可达32.3结论以磷钾伴生矿的盐酸浸出液为原料,研究了直 接滴加硫酸,采用常压酸化法制备硫酸钙晶须,分析 了不同操作条件对晶须长径比的影响.硫酸钙晶须 制备的较佳工艺条件:硫酸浓度为0.87 m o l/L、加热 温度为105丈、ra(S〇4_)/ra(Ca2+)= 1.2、滴加速率为 2.5 m L/m i n、搅拌转速为166 r/m i n,此时可制得长 径比为32的二水硫酸钙晶须.参考文献:[1]马继红.硫酸钙晶须的制备及在道路改性沥青中的应用研究[D].青岛:山东科技大学,2005: 2-10.M A J H. 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a r X i v :0709.3423v 1 [a s t r o -p h ] 21 S e p 2007Proceedings Astrophysical Masers and their Environments Proceedings IAU Symposium No.242,2007J.M.Chapman &W.A.Baan,eds.c2007International Astronomical Union DOI:00.0000/X000000000000000XMolecular properties of (U)LIRGs:CO,HCN,HNC and HCO +A.F.Loenen 1,2,W.A.Baan 2and M.Spaans 11Kapteyn Astronomical Institute,University of Groningen,P.O.Box 800,9700AV Groningen,The Netherlands email:loenen@astro.rug.nl,spaans@astro.rug.nl2ASTRON,P.O.Box 2,7990AA Dwingeloo,The Netherlandsemail:baan@astron.nlAbstract.The observed molecular properties of a sample of FIR-luminous and OH megamaser (OH-MM)galaxies have been investigated.The ratio of high and low-density tracer lines is found to be determined by the progression of the star formation in the system.The HCO +/HCN and HCO +/HNC line ratios are good proxies for the density of the gas,and PDR and XDR sources can be distinguished using the HNC/HCN line ratio.The properties of the OH-MM sources in the sample can be explained by PDR chemistry in gas with densities higher than 105.5cm −3,confirming the classical OH-MM model of IR pumped amplification with (variable)low gains.Keywords.galaxies:nuclei,galaxies:ISM,galaxies:active,galaxies:starburst,ISM:molecules,ISM:evolution,radio lines:ISM2Loenen,Baan &Spaans02468108910111213time (T units)78910111289101112130.050.10.150.2Figure 1.left,from topto bottom:Integrated line ratios HCN(1-0)/CO(1-0),HNC(1-0)/CO(1-0),andHCO +(1-0)/CO(1-0)versus FIR luminosity.Squares represent reliable values and triangles upper or lower limits.Filled symbols are sources with OH-MM activ-ity.right,from top to bottom:Three FIR luminosity curves used in the simulations with different maximum lumi-nosities,the luminosity of a high-density component,and the variation of the high-versus low-density ratio during the outburst starting at the upper part of the curve.trace the high-density component (HD)which mostly resides in the cores of the galaxies,at the sites of the star-formation activity.On the left hand side of Fig.1,the relative contributions of these two density compo-nents are shown,by plotting the ratios of the high-density and CO(1-0)line strengths.The figure shows that the distribution of line ratios for all molecules increases with FIR luminosity,which gives an upwardly curved lower boundary for the distribution at higher L FIR .The highest values are found at L FIR 1010.5L ⊙.The figure also shows that in general the OH-MM sources have a much larger spread in HD/CO(1-0).This behavior can be explained as the result of ongoing star formation.The FIR luminosity of the ULIRG during the evolution of the outburst reflects energy generated by the star formation activity.The FIR luminosity integrated over the course of the outburst would reflect the amount of high-density molecular material consumed by the star formation process and destroyed or removed by feedback.In the following simplified scenario (see Baan et al.2007),we employ a model for the time-evolution of the high-density components in a galaxy during a FIR outburst.In the absence of a representative FIR light curve,we use a diffusion-like expression in time t as a response to a starburst starting at t =0defined as:L FIR (t )=1.35L FIR (0)tMolecular properties of(U)LIRGs3 of L FIR,but this curve does resemble the outcome of starburst-driven FIR evolution simulations(Loenen,Baan&Spaans2006).The high-density component HD can be defined asβLD,the low-density component. As a result,the high-low-density ratio varies with time during the FIR outburst as:HD(t)L FIR,int ,(2.2) whereγis the fraction of the initial HD component that is consumed during the whole outburst,and L FIR,int the FIR luminosity integrated over the whole course of the out-burst.The large-scale low-density component LD is assumed to remain unchanged. The results of these simulations have been presented on the right hand side in Fig.1. The top panel shows the FIR light curve of the outburst for three peak luminosities. The middle panel shows the luminosity of a representative high-density component for the three L FIR curves.The bottom panel shows the high-low-density ratio for these same FIR light bining the results of this simulation with the data shows that the HD/CO(1-0)data points in the left panels are a measure of the evolution of the starburst. This implies that the OH MM sources are galaxies in an early stage of star formation, which is consistent with OH MM sources being found in starburst-dominated galaxies (Genzel et al.1998;Baan&Kl¨o ckner2007).3.ChemistryThe model presented in the previous section does well in explaining the evolution of the different gas components,but it makes no distinction between the different high-density tracers.Even though the emission of the different molecules originates in the same regions,the line strengths are influenced by the environmental properties like the (column)density,temperature,and the type and strength of the prevailing radiation field.In order to study the effects of these parameters on the emission characteristics of the sources,we remove the intrinsic difference in line strength between the galaxies in our sample and use line ratios tofind diagnostic properties.Fig.2presents the ratios of the integrated lines of HCO+/HCN,HNC/HCO+and HNC/HCN against each other. In order to interpret the behavior of the sources in this diagram,we compare the data to the theoretical models that treat the chemistry and radiative transfer of molecular clouds, including all the relevant heating,cooling and chemical processes(see Meijerink&Spaans 2005;Meijerink et al.2007,and Spaans et al.in these proceedings).A large grid of mod-els was created by Meijerink et al.(2007),varying the strength of the radiationfield,its type(UV and X-ray),the gas density and the column density.These results are compared to our observational data in Fig.2(note:not all models are shown,some fall out of the range of ourfigure).3.1.PDR modelsThe results of the PDR models(UV radiationfield)are shown in Fig.2with heavy lines, where the line styles indicate different gas densities(solid:n=104.5,dashed:n=105.0,and striped:n=105.5cm−3).The tracks vary as a function of column density,which ranges from N=1022cm−2(the column density below which the strength of the emission lines decreases rapidly)to N=1023,N=1023.5and N=1024cm−2for the n=104.5,n=105.0, and n=105.5cm−3models,respectively(corresponding to a cloud size of1pc;indicated by the symbol at one end of the tracks).Two different radiationfield strengths are shown: F UV=1.6erg cm−2s−1,indicated by a plus symbol at the highest column density point, and F UV=160erg cm−2s−1,indicated with a circle.Molecular properties of(U)LIRGs5 affect the detection rate of XDR sources and could make hybrid sources look like PDR sources,even if the XDRs are energetically more important than the PDRs.Despite these drawbacks,the XDR models do provide diagnostics,since they clearly separate from the PDR models in their HNC/HCN line ratio.The PDR models approach, but never cross the HNC/HCN=1line and the XDR models all have HNC/HCN>1. Therefore,the HNC/HCN line ratio is an excellent way to determine whether a system’s chemistry is dominated by UV or X-ray photons.3.3.Terra IncognitaNot all our observational data in Fig.2is covered by the models.The few sources in our sample with known H2O MM activity(indicated by plot symbols surrounded by circles)are also located in this area,which is characterized by lower HNC and higher HCO+line strengths compared to HCN.The fact that this area is not covered by the models suggests that other processes influence the line ratios,such as strong shocks. Shocks are not treated in the models and can have profound effects on the chemistry in molecular clouds as they may selectively destroy HNC(Schilke et al.1992)and enhance HCO+relative to HCN(Dickinson et al.1980).A simultaneous increase in HCO+and a decrease in HNC shifts the PDR models to the uncovered region,implying that the H2O MM sources in our sample are UV driven systems with strong shocks.This would suggest that these H2O MM sources are similar to shock-induced Galactic H2O maser spots(see other contributions in these proceedings).4.ConclusionsMolecular line emissions of multiple species(and transitions)provide excellent diag-nostics for understanding the status of the nuclear gas in extra-galactic sources.The different molecules trace different(density)components and the ratio of high and low-density tracer lines follows the star formation activity in the paring different high-density tracers tells a lot about physical characteristics of the gas.The HCO+/HCN and HCO+/HNC line ratios are good proxies for the density of the gas,due to the differ-ent critical densities of the species.PDR and XDR sources can be distinguished using the HNC/HCN line ratio:PDR sources all have ratios lower than unity and XDRs have ra-tios larger than1.OH MM sources cluster in a particular part of the diagnostic diagram, which is only traced by PDR models with densities higher than105.5cm−3,confirming the classical OH MM model of FIR(UV radiation reprocessed by the surrounding gas and dust)pumped amplification with low but variable gains.ReferencesBaan,W.A.1989,ApJ338,804Baan,W.A.,Henkel,C.,Loenen,A.F.,Baudry,A.,Wiklind,T.2007,submitted to A&A Baan,W.A.,&Kl¨o ckner,H.-R.2006,A&A,449,559Dickinson,D.F.,Dinger,A.S.C.,Kuiper,T.B.H.,&Rodriguez Kuiper,E.N.1980,ApJ,237,L43 Genzel,R.,et al.1998,ApJ4498,579Loenen,A.F.,Baan,W.,A.,Spaans,M.2006,A&A458,89Loenen,A.F.,Baan,W.,A.,Spaans,M.2007,in preparationMeijerink,R.&Spaans,M.2005,A&A436,397Meijerink,R.,Spaans,M.,&Israel,F.P.2007,A&A461,793Schilke,P.,Walmsley,C.M.,Pineau Des Forets,G.et al.1992,A&A256,595。