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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.。
4.1 The size of a penetrant indication depends on:渗透剂的显示大小取决于：A.the size of the discontinuity. 不连续的大小B.the discontinuity entrapment efficiency.C.the technique of penetrant testing. 渗透检测技术D.all of the above. 上面全部E.only a and c above. 只有上面的a和b4.2 The most sensitive application technique for dry developer is:对干燥显像剂最为敏感的应用技术是：A.Immersion. 浸入B.a dust cloud. 粉尘云C.A fluidized bed. 流化床D.A dust cloud —electrostatic gun. 粉尘云—静电抢4.3 one advantage of having a little background fluorescence is that it:小背景荧光的一个优点是它：A.reduces the contrast of the indication. 减少现象的对比度B.Indicates that the part was not overwashed. 显示不被清洗的部分C.Reveals an excessive emulsification time. 显示过度乳化时间D.Is hard to remove when a highly sensitive penetrant is applied to rough or porous surfaces.很难去除用在粗糙或多孔表面的高灵敏度渗透剂4.4 the water wash test is used to evaluate the washing properties of penetrant and emulsifiers. The spray nozzle is held 30 cm(12 in.) from the wash surface. The water pressure must not exceed:水洗涤实验用于评估渗透剂和乳化剂的洗涤性能。
动物学报49 (5) :670～674 , 2003Acta Zoologica S i nica一种从大熊猫粪便中提取D NA 的改进方法3钟华①赖旭龙②魏荣平③刘中来①33( ①华中师范大学生命科学学院, 武汉430079)( ②中国地质大学地球科学学院, 武汉430074) ( ③中国保护大熊猫研究中心, 四川卧龙623006)摘要本研究描述一个改进的方法, 使从大熊猫粪便中提取DNA 用于PCR 扩增变得更加容易。
在粪便DNA 的提取过程中采用一个新的预处理方法, 将粪便用预冷的丙酮洗2～3 次, 除去粪便中含有的大量PCR 抑制物, 然后用蛋白酶K 裂解、酚- 氯仿抽提, 能提取到纯度很高的DNA 供PCR 扩增。
本实验PCR 扩增了大熊猫脑源性神经营养因子(BDNF) 基因和线粒体细胞色素 b 基因片段, 并进行测序分析, 证实了提取的可靠性。
对比本方法和未经丙酮预处理的方法提取的DNA 进行PCR 扩增, 前者的扩增结果明显优于后者[ 动物学报49 (5) : 670～674 , 2003 ] 。
关键词大熊猫粪便DNA 丙酮DNA 抽提非损伤性取样An improved protocol for D NA extraction from the faeces of the giant panda 3 ZHON G Hua ① LA I Xu2Long ② W EI Rong2Ping ③ L IU Zhong2Lai ①33( ①College of L if e S cience , Cent ral China Nor mal U niversit y , W uh an430079 , China)( ②Faculty of Earth Sciences , China U niversit y of Geosciences , W uhan430074 , China)( ③China Cons ervation and Res earch Center f or th e Gi ant Pan da , W olong623006 , S ichuan , China) Abstract An improved method that facilitates the extraction of PCR2compatible faecal DNA from giant pand a’s faeces is described. The method involved a novel preprocessing step in DNA extraction. The faeces was washed two or three timeswith precooled acetone , which removed numerous potential PCR inhibitors , and then digested with proteinase K. The DNA was purified with phenol/ chloroform. The faecal DNA obtained was sufficiently pure to support reliable amplifica2 tion , and was applied as template DNA to amplify a portion of the giant panda brain derived neurotrophic factor (BDNF) gene and mitochondrial cytochrome b gene. The sequenced results of PCR products confirmed that the extracted DNA was from the giant panda. Comparison with the PCR products demonstrated that the faecal DNA extracted b y the improved protocol was better than the faecal DNA extracted without acetone preprocessing. [ Acta Zoologica S inica 49 (5) : 670 - 674 , 2003 ] .K ey words Giant panda ( A il uropoda melanoleuca) , Faecal DNA , Acetone , DNA extraction , Noninvasive sampling在大熊猫的遗传多样性、种群数量调查、进化和分类、亲子鉴定等研究中, DNA 分析是重要的研究手段。
CHINA MEDICAL HERALD Vol.15No.19July 2018[基金项目]中国食品药品检定研究院中青年发展研究基金课题(2015C03)。
[作者简介]赵瑜(1980.11-),女,硕士,副主任药师;研究方向:药物分析,药品快检技术。
[通讯作者]尹利辉(1971.4-),男,硕士,主任药师,硕士生导师;研究方向:药品快检技术研究。
公安机关公布的假药案件中,假冒抗癌类药物日渐猖獗。
抗癌药物价格昂贵,近年来在亚洲和中东地区出现的假冒抗癌药物的数量呈上升趋势,国际药物安全研究所2016年公布的数据显示,抗癌药制假造假的增长率为29%,高居第四位[1]。
紫杉醇为天然提取或半合成制备的化学抗肿瘤药,在水中几乎不溶,在甲醇、乙醇、三氯甲烷中溶解[2]。
紫杉醇注射液配方中用高浓度的乙醇作为助溶剂,因此需要特殊的输液装置,用很低的滴注速度。
由于注射后患者极易出现过敏反应,因此给药时为防止发生严重过敏反应常使用地塞米松、抗组胺药或H2受体拮抗剂等同时进行预防给药,使紫杉醇在临床上的应用受到了很大程度的限制。
紫杉醇脂质体是将紫杉醇包裹在磷脂膜材料中,使药物具有靶向性,避免了高浓度乙醇对患者注射的刺激性,明显降低紫杉醇的过敏反应、毒性,在临床使用上具有明显优势[3]。
紫杉醇拉曼光谱法在快速筛查紫杉醇脂质体制剂中的应用赵瑜尹利辉胡昌勤中国食品药品检定研究院化学药品检定所,北京100050[摘要]目的应用拉曼光谱法建立定性鉴别模型,实现紫杉醇脂质体制剂的现场快速筛查。
方法隔包装采集注射用紫杉醇脂质体的拉曼光谱,使用主成分分析(PCA)算法去除包装的干扰信号,提取紫杉醇脂质体的拉曼信号,用经典最小二乘(CLS)建立定性鉴别模型。
对模型进行正向验证和反向验证确定判别的阈值,模型输出的相关系数值同阈值比较进行定性判定。
使用外标法实现方法在三种仪器上的转移。
结果排除玻璃包装的干扰提取的光谱与直接测量的光谱相关系数达0.9744,建立的紫杉醇脂质体定性模型,判断阈值为0.85,正向验证(脂质体制剂)和反向验证(脂质体膜成分和紫杉醇)结果均为通过。
纺织品成分及分析测试相关英语纺织品成分及分析测试相关英语A. Composition and analytical tests 成分及其他分析测试1. Fibe analysis 纤维分析2. Identification of virgin/recycled wool 新⽺⽑/再造⽑鉴定3. Identification of noble animal fiber & natural cellulosic fibre other than cotton 特殊纤维鉴定4. Identification of lambswool ⽺仔⽑鉴定5. Fibre diameter 纤维直径6. coarse hair content 粗⽑含量7. PH value 酸碱度8. moisture content /regain ⽔分含量9. extractable matter 可萃取物10. identification of dyestuff 染料鉴定11. starch content 淀粉含量12. filling & foreign matter content 填充物及杂质含量13. mercerisation in cotton 棉丝光处理14. formaldehyde content 甲醛含量15. hardness of water ⽔硬度16. mushroom reaction 发胀反应17. UPF (ultraviolet protection factor) 防⾃外光系数18. Nickel release 镍的释放度19. Microfibre 微纤维20. Verification of Ammonium finish on fabric 布料之铵整理剂鉴定21. Verification of plastic by UV 以⾃外光法鉴定塑胶22. BHT. Content BHT含量23. Deterioration effect of UV exposure ⾃外光曝晒下劣化效果24. Azo dye 偶氮染料测试B. Construction analysis 结构分析1. yarn counts 纱⽀2. yarn twist (per yarn) 纱捻度（每秒钟）3. number of filaments 长丝数量4. fabric weight 布料重量5. fabric thickness 布料厚度6. threads per unit length (woven fabric construction) 织物密度（机织物）7. stitch density (knitted fabric construction) 织物密度（针织物）8. loop length (knitted fabrics) 线圈长度9. type of cut pile 割绒种类10. type of weave 梭织品织法分类11. measuring bow & skew 扭曲及歪斜量度12. cover factor 覆盖系数13. fabric width 布匹阔度14. length of thread ( per cone) 线长度（每筒）15. crimp or take up of yarn 纱线绉缩或织缩率16. terry to ground ratio ⽑圈经密与地经密度⽐17. identification of continuous / microscopic method 长丝 / 短纤维鉴定18. harness of corduroy 灯⼼绒的综合19. holes on synthetic fibres 纤维孔数C. Dimensional stability (shrinkage) and related tests 尺⼨稳定性及有关测试1. dimensional stability to washing ⽔洗缩率测试2. each additional wash cycle 每增加⼀次⽔洗回圈3. appearance after laundering 洗涤后外观4. dimensional stability to dry cleaning ⼲洗缩率5. each additional dryclean cycle 每增加⼀次⼲洗6. appearance after dry cleaning (appearance retention) ⼲洗后外观7. durable of applied designs and finishes to dry cleaning 印花花纹和整理剂性8. dimensional stability to relaxation 松弛缩率9. dimensional stability to felting 毡化缩率10. dimensional stability to free steam 蒸⽓缩率11. dimensional stability to heating in house 热缩率12. spirality / skewing of fabric & garments 织物和服装扭曲/歪斜13. dimensional stability to steam pressing / ironing 蒸⽓压烫/熨⽃熨烫缩率14. tumbler shrinkage 圆筒烘⼲缩率15. cold water immersion 冷⽔浸洗缩率16. additional ironing 每增加⼀次熨烫17. effect after wash & pressing 洗涤后效果18. effect after pressing 熨后效果D. Colour fastness tests 染⾊坚牢度测试1. Washing ⽔洗2. Perspiration 汗液3. Dry & wet crocking / rubbing 摩擦4. Light 光照5. Water ⽔6. Sea water 海⽔7. Chlorinated water 氯⽔8. Dry cleaning ⼲洗9. Actual laundering (one wash) 实际洗涤（⼀次⽔洗）10. Commerical dryclean 商业⼲洗11. Dry heat ⼲热12. Hot pressing 热压13. Water spotting ⽔斑14. Acid spotting 酸斑15. Alkaline spottig 碱斑16. Bleaching漂⽩17. Chlorine bleaching 氯漂18. Non‐chlorine bleaching ⾮氯漂19. Organic solvents 有机溶剂20. Sublimation during storage 存放升华21. Perborate 酸钠22. Phenolic yellowing 酚醛发黄23. Sweat & saliva 汗液及唾液24. Washing (with shrinkage conducted) ⽔洗⾊牢度（缩⽔测试后）25. Bleeding 渗⾊度26. Ozone 臭氧27. Gas fume 烟⽓28. Dye transfer in storage 储存时颜⾊转移29. Light & perspiration 光照及汗液30. Contact test 接触测试31. Wicking 吸⽔32. Colour transfer against special condition 特别情况下颜⾊转移33. Colour fastness to perborate & light 过硼酸盐及光照⾊牢度34. Colour fastness of fabrics 布料颜⾊牢度E. Strength tests 强度测试1. Tensile strength 拉伸2. Lea strength 绞纱3. Single thread/yarn strength 单线纱4. bursting strength 胀破强⼒5. Seam performance 接缝6. Tearing strength 撕裂7. Bonding strength 粘合强⼒8. Loop strength 打结，钩接强⼒F. Fabric / garment performance and flammability tests 品质及燃烧测试1. Abrasion ressistance 耐磨性2. Pilling ressistance 抗⽑性3. Snagging ressistance 防钩丝4. Wrinkle/crease recovery 皱纹/折痕回复性5. Stiffness 布料硬挺度6. Stretch and recovery 伸展及回复性7. Water repellency 防⽔性8. Oil repellency防油性9. Soil release 防污10. Needle cutting 烧针现象11. Flammabillity 燃烧性12. Surfce flammabillity of carpets and rugs 地毯燃烧测试13. Durability of applied designs and finishes to dry cleaning 印花花纹和整理剂14. Wettability 湿润度15. Absorbency of bleached textiles 漂⽩纺织品和吸⽔性16. Air permeability 透⽓性17. Water vapour permeability index 透湿指数18. Thermal stability of coated fabric 涂层织物之耐热性19. Resistance to blocking 抗阻塞性20. Water vapour transmision 透湿性21. Dry fit function 快⼲综合功能22. Cold bending test for coated fabric 涂层织物的低温弯曲度测试23. Brittleness temp. of plastic sheeting 塑胶脆化温度24. Antistatic 防静电25. Thermal ressistance 保暖度G. Fibre & Yarn tests 纤维及纱测试1. Linear dendity 纤维线密度2. Fibre fineness 纤维细度3. Fibre diameter 纤维直径4. Yarn counts 纱线⽀数5. Denier counts of filament Yarns 长丝纱纤度6. Identification of continuous / discontinuous fibre 长丝/短丝纤维鉴定7. Twist per unit length 纱线捻度8. length of thread (per roll) 纱线长度（每轴）9. Net weight of thread 纱线净重10. Dimensional stability of yarn to washing 纱线缩⽔率11. Single fibre strength 单纤维强度12. Single thread/yarn strength 纱线强度13. Lea strength 绞纱强度14. Loop strength 打结、钩接强度15. Yarn evenness 纱线均匀度16. Crimp test 卷曲测试17. Holes on synthetic fibres 纤维孔数H. Carpet tests 地毯测试1. Colour fastness 颜⾊牢度2. Thread per unit length of backing 底部纱线密度3. Weight per unit area 重量4. Pitches per unit length ⽑束经密度5. Rows per unit length ⽑束纬密度6. Thickness 厚度7. Surface pile density 表⾯⽑绒密度8. Ply of pile yarn ⽑绒纱股数9. Fibre composition of pile & back ⽑绒及底部的纤维成分10. Surface flammability of carpers and rugs 地毯燃烧测试11. Abrasion resistance 耐磨性12. Pile height 绒⾼度。
做一项昆虫的实验作文英文回答：I conducted an experiment to investigate the effects of different environmental conditions on the behavior of a common insect, the housefly (Musca domestica). I hypothesized that houseflies would exhibit changes in their behavior in response to variations in temperature, light intensity, and food availability.To test my hypothesis, I designed an experiment with three independent variables: temperature, light intensity, and food availability. The temperature was manipulated by placing the flies in chambers set to different temperatures (15°C, 25°C, and 35°C). The light intensity wascontrolled by varying the brightness of lights in the chambers (low, medium, and high). The food availability was regulated by providing different amounts of food (none, low, and high) to the flies.I observed the behavior of the houseflies in each experimental condition for a period of 30 minutes. I recorded the number of times the flies took off, landed, walked, and groomed themselves. I also noted the location of the flies within the chamber (e.g., near the food, near the light, or in the center).The results of my experiment showed that houseflies exhibited significant changes in their behavior in response to the different environmental conditions. For example, flies in the high-temperature chamber took off and landed more frequently than flies in the low-temperature chamber. Flies in the high-light intensity chamber walked more than flies in the low-light intensity chamber. Flies with high food availability spent more time near the food than flies with low food availability.Overall, the results of my experiment support my hypothesis that houseflies exhibit changes in their behavior in response to variations in environmental conditions. These findings suggest that houseflies are capable of adapting their behavior to differentenvironments, which may help them to survive in a variety of habitats.中文回答：我进行了一项实验来调查不同的环境条件对一种常见昆虫——家蝇（Musca domestica）的行为的影响。
2008年1月January 2008色谱Chi n ese Journal of Chro m atographyVo.l 26N o .150~55R ecei ved date :2007-06-14F irst author :GANGADEV I V,Ph D.C orresponding author :MUTHUMARY J .T e:l +91-44-22350401,F ax :+91-44-22352494,E -m ai:l mm _j @rediff ma i.l co m.n The el ectron i c ed i ti on of the arti cle is pu b li shed b y E l sevier BV on S ci enceDirect (htt p ://www.sci en ced i rect .co m /sci ence /j ournal /18722059).A si m p le and rapid m et hod for the deter m i nation of taxol produced by fungal endophytes fro m m ed icinal plants usi ng h igh perfor m ance t h in layer chro m atographyGANGADEV I V ,M UTHU M ARY J(Cen tr e for Advan ced Stud ies in Bo tany ,Un iv er sity of M adr a s,Gu indy Cam pu s,Chenn a i 600025,T a m il N adu,I n dia )A bstract :Taxo l is an i m portan t ant i cancer drug used w idel y in the cli n ical field .In th is s tudy ,some endophyt -ic fungiw ere isolated fro m sel ected m ed i cinal p lan ts ,and w ere screened for the i r po ten tial i n the p roduction of taxo,l u si ng a rap id separati on techn ique of h i gh perfor m ance th i n l ayer ch ro m atography (HPTLC).Of the 20screened fung,i on l y 13funga l species produced taxo l i n the artifici a l cu ltu re mediu m.The resu lts o f HP TLC show ed that the 13fungal speci es had ident i cal u ltrav i o l et (UV )characteristics ,pos itive react i vity w ith a sp ray reagen t ,yi e l d i ng a blue spot ,w h ich turned to dark gray after 24hours ,and had R f values identical to that of the au then ti c taxo.l The a mount of taxo lw as a l so quan tified by compari ng the peak area and the peak height of the fungal sa mp les w ith those o f authen tic taxo.l K ey w ords :endophytic fung;i taxo;l h i gh perfor m ance th i n layer ch ro m atography (HPTLC );quantitativeanal ys i sCLC nu m ber :O 658 D ocu m ent code :A A rticle IC :1000-8713(2008)01-0050-06 Colu m n :A rticl esThe anticancer drugs t hat exist at presentpossess nu m erous side eff ects and are not eff ec -tive against m any for m s of cancer .Com pared w ith other ant-i cancer co m pounds ,t axol has a unique action m ode .Its action m ode is m ai n ly re -lated to its inhibitory eff ect on cell division .Over a long period of ti m e ,the study of natural prod -ucts f ro m plants and m icroorganis m s has pro -gressed to its present state of develop m ent .The world of nat ure abounds inm edicinal co m pounds ,which constit ut es a fascinating and fruitf ul area of sci e ntific investi g ation .Fungi have an excellent record of producing novel bi o acti v e co m pounds ,and currentl y ,m any of t hem have i m portant m e -di c inal and other uses .Si n ce 1993,the observa -tions of endophytic f ung i t hat produce taxo,l Ta x-o m y ces an dr ean a e and P esta lo tiop sis m icro sp o-ra ,have been report ed ,which dem onstrate that organis m s ot her than Ta xu s spp .can produce t axol [1,2].Ho w ever ,i n the industry ,on ac -count of t he shortage of yew trees the raw m ater-i al of taxol is purified by us i n g a sem i synthetic m ethod ;and t he production of taxol cannot m eetthe de m and of t he grow ing m arket .This m akes taxol a financi a l burden form any pati e nts . It is believed t hat fungi t hat produce taxol have a great potential for m aki n g antitu m or drug taxol i n t he f ut ure .The significance of finding a fungus capable of producing taxol is not understa -ted ;as such a discovery w ill revol u ti o nize the search for eff ective phar m aceutical agents .T he m ost significant finding i n the last decade m ay be the discovery of endophytic taxo-l produci n g fun -gus fro m the gym nosper m,particularly ,fro m ye w trees .It is rem arkable t hat t he taxol produced by these endophytes is chem ically and biolog icall y i d entical to that produced by Ta xu s spp .[3]T hus far ,m ore than 30f ungi that produce t axol have been reported globa lly .M ost of them are endo -phytes of Ta xu s spp .belong i n g to asco m ycetes and i m perfect fungi [4].The discovery t hat certai n endophytic fungi are abl e to produce taxol has creat ed a poss i b ilit y for a cheaper andm ore w idely avail a ble product to be event ually produced via industrial f er m entation .第1期GANGADEV I V,et a.l:A si m ple and rapid met hod f or the deter m i natio n of t axol prod uced byf ungal endophytesf ro m medici nal pl ants usi ng high perf or mance t hi n layer chro matographyThe isolation of f ung i that produce taxol w ill ena-ble one to st udy the taxol biosynt hetic pat h w ay i n f ungi and explore t he poss i b ility of i m proving t he t axol content v ia genetic engineering in t he f uture. Therefore,i n t his st udy,the endophytic f ungi have been isolat ed f ro m so m e i m port ant medicinal pl a nts.T axol is det er m ined by using t he s i m ple and rapid m ethod of high perf or m ance t hi n l a yer chro m at ography(HP TLC).The m ai n obj e ctive of t his st udy is t o screen t he taxo-l producing endo-phytic f ung i fro m so m e medicinal plants.A si m-ple,sensiti v e,and rapid m et hod has been deve-l oped to quantif y t he total taxol i n fungal extracts usi n g HPTLC.This st udy is t he first report on tax-ol production via a f ungal endophyt e f ro m m edic-i nal pl a nts i n Sout hern I ndi a.1Exper i m ental1.1Isol ation and identificati on of endophy t-i c fung iAs t here was onl y little i n f or m ation available on endophytic fungi fro m m edici n al plants,endo-phytic fungi were screened f or their potential for t axol production fro m so m e m edici n al plants co-l lect ed i n Chennai Cit y,India.The surface of t he leaf sa m ples were sterilized t o isolat e t he endo-phytic m icroorganis m s usi n g the m et hod of Sury-anarayanan et a.l[4]The segm ents of st erilized leaf were evenly spaced in Petri dishes(9c m i n di a m et er)cont a i n ing potato dextrose agar(PDA) m ediu m(a m ended w it h chloram phenicol150 m g/L).The Petri dishes were sealed using Para-fil m TM and i n cubated at(26?1)e i n a light cha m ber,in cycles of12hours of light f ollo wed by12hours of darkness.The Petri dishes were m onitored every day f or the grow t h of endophytic f ungal col o nies from the l e af seg m ents.T he hy-phal tips,which grew out f ro m t he l e af seg m ents were isolat ed and subcult ured onto the PDA and brought i n t o t he pure culture.The isolat ed endo-phytic fungiw ere i d entifi e d using standard m ono-graphs.The i d entifi e d cultures of endophytic f un-giwere depos ited at theM adras University Botany Laborat ory(MUBL),Centre for Advanced Stud-ies in Botany,U niversity ofM adras,Chenna,i In-di a.H o wever,the i m m ediat e concern is to fi n d one orm ore f ung i that produce m ore taxo.l About 20f ungal endophytes,incl u di n g Ba r ta lin ia robil-la rdo ides Tass,i and Ph o m op sis stipa ta(L ib.) Sutt on,isolated fro m Aeg le m a r m elo s Correa ex Roxb.;Bo try od ip lod ia theobro m a e Pat.,and Pesta lotiopsis pau ciseta Sacc.,isol a ted from Ca rd io sp er m um h elica ca bum L i n n.;Cha eto m ella raph igera Sw ift,and Pesta lotiopsis ter m in a lia e Agar wa l and H as ij a,from Ter m in a lia a r jun a (Roxb.);W i g ht&Arn.,Cha etosperm um ca rn e-um T ass,i and Pho m opsis a rn o l d ia e Sutt on,from Rau v olfia tetra phy ll a L i n n.;C iliochor ell a m an-gifer a e Syd.and M itter,and Pesta lotiopsis n e-glecta(T heum.)Stey.,fro m P sid ium gu a ja va Linn.;C olleto tr ic hum cir cin an s(Berk.)V og.l, and Phy llo sticta sp.6,fro m O ci m um ba silicum Linn.;Co lleto tr ichum fa lca tum W ent.,and Pho-m a her ba rum W estd.,fro m Vitex n egun do Linn.;Co lleto tr ichum gloeo sp or io ides(Penz.) Sacc.,and Pho m a cap itu l u m Pa w ar,M athur& Thiru m.,fro m Ju sticia ge n daru ssa Bur m.f;Fu-sicoccum in dic um T ilak&Rokde,and Robilla rda sessilis(Sacc.)Saccardo,fro m M i m u sops ele n-gi Linn.;and Ph o m a g lo m era ta(Corda)W olle-nw.&Hochapf.,and Ph o m op sis vex an s(Sacc. &Syd.)H arter,f ro m An dro graph is pan ic u la ta W al.l ex N ees;were selected f or preli m inary screeni n g of taxol producti o n,us i n g HPTLC.1.2Extracti on of taxolThe extracti o n procedure of Strobe l et a.l was f oll o wed for the isolation of taxol fro m f ungi [2].The endophytic f ungiwere grown i n2L Er-len m eyer flasks contai n ing500m L of standard liq-uid m edi u m for taxol production supplem ented w it h1g/L soyt one[5].The t est f ungiwere ind-i v i d uall y inoculat ed and i n cubated for21days.Af-ter21days of i n cubation peri o d,the cult ures were filtered t hrough four layers of cheeseclot h to rem ove the m yce li a.0.25g Na2CO3w as added to the culture filtrate w ith frequent shaki n g,to re-duce the a m ount of fatt y acids t hat m ay conta m-i nat e taxol i n t he culture.T hen t he culture filtrat e was extracted w ith t w o equal vol u m es of sol v ent dichl o ro m et hane.The organic phase was coll e ct-ed and t he solvent was then re m oved by evapora-#51#色谱第26卷tion under reduced pressure at35e us i n g a rot a-ry vacuum evaporator.The dry solid residue w as redissolved i n m ethanol for subsequent separa-tion.The crude extracts were anal y zed by chro m-at ographic separation and spectroscopi c anal y ses.1.3Chro m atograph i c separati onThe extracted sa m ples were prepurified t hrough colu m n chro m at ography.The crude ex-tract was dissolved in1mL m et hylene chloride and loaded on a1.5cm@30c m colu m n of silica gel and e l u ted in a step w ise progra m w ith a sol v ent system as f oll o ws:70mL100%m ethy l e ne chlo-ri d e,m et hylene chloride-et hylacet ate(20B1,v/ v),m ethylene chl o ride-et hylacetat e(10B1,v/v), m et hylene chloride-et hylacet ate(6B1,v/v),m eth-ylene chl o ride-et hylacetat e(3B1,v/v),and m eth-ylene chl o ride-et hylacetat e(1B1,v/v).Fractions w ith the sa m e mobilit y as aut hentic t axol were co m bined together and evaporated to dryness.The prepurified t axolw as f urt her anal y zed by thi n l a yer chro m at ography.1.4Th i n l ayer chro mat ographyThi n layer chro m atography(TLC)for t he f ungal sa m ples contai n ing taxol was carri e d out on0.25mm(10c m@20c m)al u m i n u m precoat-ed silica ge l plat es(M erck,Ger m any).T he f un-gal sa m ples and authentic taxol(standard taxol purchased fro m Si g m a Che m icals,USA)were spott ed on a TLC plate and developed in t he fo-l lo w ing different sol v ent syst em s successively: (a)chlorof or m-m ethanol(7B1,v/v);(b)chlor-ofor m-acet onitril e(7B3,v/v);(c)et hylacetat e-2-propanol(95B5,v/v);(d)m ethy lene chloride-t etrahydrof uran(6B2,v/v);and(e)m et hy l e ne chl o ride-m et hano-l di m et hylf or m a m ide(90B9B1, v/v/v).T he presence of taxol was detected w it h 1%(w/v)vanilli n sulfuri c acid reagent after gen-tle heating[6].1.5H i gh perfor mance t h i n l ayer chro m atog-raphy1.5.1Chromatogra m layerThe prepurifi e d funga l taxol sam ples obtai n ed i n TLC were further subjected to HPTLC(CA-MAG-P lanar HPTLC,Anchrom).1.5.2Sa m pl e l oadi ngW ith t he Auto m atic TLC Sa m pler,20fungal sa m ples and authentic taxol(loading vol u m e of5 L L f or each)were i n ject ed i n t o21tracks,sepa-rately,w ith the data pair technique,w ith the band lengt h be i n g5mm,the distance fro m lo w er edge10mm,t he distance f ro m the side20mm, and the track distance6.4mm,on a precoated silica gel60F254(20c m@10c m)plate(M erck, G er m any).After loading,the pl a te was dried f or 30s us i n g a plate heater.1.5.3Chro matographyAfter l o adi n g,the TLC plate was developed i n a CA M AG presaturated flat botto m developing cham ber w ith chl o rof or m-m ethanol(9B1,v/v) for20m i n.1.5.4Docu m entati on and deri vatizati on of t he TLC pl ateIt is an i n herent advantage of pl a nar chro m a-tography that t he chro m atographic result can be v isualized and docu m ented as an i m age.This a-l lo w s a convenient qualitati v e eval u ation of m ult-i ple sa m ples on t he sa m e plate.The chro m at o-graphic plate is then dri e d for10m in and scanned us i n g TLC Scanner3w ith t he soft w are/w i n CATS P lanar Chro m atography M anager0,which is the m ost advanced workstati o n for densit o m etric e-val u ati o n,and t he fl u orescence w as recorded at 254nm and366n m,separat e l y.Docu m entati o n of the TLC plate was perfor m ed under a shorter wavelength(254n m)UV la m p and a l o nger wavelength(366n m)UV lam p,pri o r to deri v at-ization.The presence of the taxol was v isualized aft er it was sprayed w it h1%(w/v)vanillin sulf u-ric aci d and heated gently f or2m i n[6].A fter derivatizati o n w it h the spray reagent,docum enta-ti o n w as perf or m ed under the UV la m p at366n m, and whit e light,which w as used to visua lize the colored substance after derivatizati o n.1.5.5D ensito m etric eval uati on and quantifi-cationThe TLC pl a te containi n g f ungal taxol was scanned using a TLC Scanner3w ith w i n CATS soft w are.The detection and quantification were done us i n g dens ito m etry i n absorbance m ode at 254n m.The presence of a fungal taxolw as iden-tified by co m parison w it h an aut hentic taxo.l The area and he i g ht of t he peaks of the authentic taxol# 52 #第1期GANGADEV I V ,et a.l :A si m ple and rapid met hod f or the deter m i natio n of t axol prod uced byf ungal endophytesf ro m medici nal pl ants usi ng high perf or mance t hi n layer chro matographyand f ungal taxol observed in the chrom atogra m swere evaluated .Aft er the chro m atography ,the area conta-ini n g taxol i n the TLC plat e was caref ully scrapped off fro m t he silica plate at the appropri a te R f va-l ue .T he taxol was exhausti v el y eluted w ith 100%m ethanol and f urther anal y zed us i n g UV absorp -tion at 273n m [7]i n a Beck m an DU -40Spectro -photo m eter and w as co m pared w it h t he aut hentic t axol (Sig m a ,USA).The fast ato m bo m bard m ent (FAB )m ass spectra were recorded on a JEOL SX 102/DA-6000M ass Spectro m et er/Data Syste m w ith the FAB gas of argon /xenon (6k V ,10mA).The accelerati n g voltage was 10kV and the spectra were recorded at roo m tem perature .F i g .1 H PTLC f i ngerpri n ts of e ndophytic f ungal taxol before derivatization at (a)UV 254nm,(b)UV 366n m,and after derivatization at (c)UV 366n m,(d)w hite li gh tFor T rack 1-T r ack 21,see Tabl e 1.2 R esu lts and d iscussionIn this study ,am ong the 20endophytic fungi screened f or the production of taxo,l onl y 13f un -gi sho wed the presence of taxol i n the cult ure .I n HP TLC ,t he principa l requirem ent for the docu -m entation was t he visibilit y of the chro m atogra mw ith or w it hout derivatizati o n .The substances w ith the absorbance of UV li g ht at 254n m were visualized as dark zones on the plat es ,as sho wn i n F i g .1-a ,w ith a fluorescence indicator ,which was excit ed to em it green li g ht under a shorter wavelengt h (254n m )UV la m p .A longer wave -length (366n m )UV la m p was used to excite sub -stances t hat were abl e to fl u oresce ,as sho wn i nF i g .1-b .W hite light was used to v isualize co-lored substances .The docu m entation of t he TLC plat e was perf or m ed under a short er w avelength (254n m )UV la m p .T he presence of taxol was v isualized aft er bei n g sprayed w ith 1%(w /v)va -nillin sulfuric aci d and heat ed gentl y f or 2m in .The funga l taxol was i d entified by co m parison w it h the authentic taxo.l The funga l taxol had i d enticalUV characteristi c s and positive reacti v ity w it h t he spray reagent ,which y i e l d ed a blue spot and turned to dark gray aft er 24hours (F i g .1-c ,d).The co m poundw ith the sa m e chro m atograph -ic m obility as t he aut hentic taxol was recorded fro m 13fung,inam ely ,Ba r ta lin iarobilla r-do ides,Bo try odip lod ia th eobr o m a e,Cha eto m ella raph igera,Co lleto tr ichum fa lca tum , C.g loeos-por io id es,Fu sicocc um in d icum ,Pesta lotiopsis n eglecta,P.pau ciseta,P.ter m in a lia e ,Pho m a h erba rum ,Pho m opsis a rn o ld ia e ,P.vex an s ,and Phy llosticta sp .6.They had R f values ran -g i n g fro m 0.22-0.24(T abl e 1). The ot her seven fungal spec i e s ,na m e l y ,Cha etosperm um ca rn e um ,C ilio c h or ella m an g if-era e ,Co lleto tr ichum cir cin an s,Pho m a cap itu-lum ,P.g lo m er a ta,Pho m opsis stipa ta ,and Ro-billa rda sessilis ,did not sho w evidence of taxol production (F i g .1-a -d ).In HPTLC,the a m ount of taxol produced by the fungiw as calcu -lated t hrough co m paring the area and t he he i g ht of peaks observed i n f ungal taxol w ith that of au -t hentic taxo.l#53#色谱第26卷Table1H PTLC dat a of endophytic f ungal taxo lEndophytic f ung i R f V alue Peak area T rack N o.B ar ta lin ia r ob ill a rd oid es0.2316009T rack1B otr yod ip lod ia th eobro m a e0.2217699T rack2C ha eto m ell a raph igera0.226793T rack3C ha etos p erm um ca rn eum--T rack4C ilioch or ell a m an gifera e--T rack12C oll etotr ichum cir cin an s--T rack18C oll etotr ichum fa lca t um0.223490T rack17C oll etotr ichum gloeosp or ioid es0.2313944T rack19 Fu sicocc um i n d icum0.242338T rack13P es ta l otiop s is n eglect a0.233567T rack10P es ta l otiop s is pau cis eta0.239669T rack8P es ta l otiop s is term in a lia e0.2218015T rack9Ph o m a cap it u l um--T rack15 Ph o m a glo m era ta--T rack16 Ph o m a h er ba rum0.236793T rack14 Ph o m op s is arn ol d iae0.239114T rack5Ph o m op s is s tipa ta--T rack7Ph o m op s is vexan s0.233490T rack6Ph y ll o sticta sp.60.224412T rack20 Robillard a s ess ilis--T rack21 Authenti c taxol0.238534T rack11Co mm erci a l t axol production by fer m entation was encouraged by the results which sho wed that it was poss i b le to produce taxol by select ed fung.i The yield of fungal taxol ranged fro m27.4L g/L t o211.1L g/L(F ig.2).T he m axi m u m am ount of taxolwas produced by Pesta lo tiop sis term in a-lia e(211.1L g/L)and B otry od ip lod ia th eor br o-m a e(207.4L g/L).Fu sicocc um ind ic um(27.4 L g/L)was t he least taxo-l producing f ungus in t he present i n vestigation(F ig.2).Alt hough t he a m ount of taxol produced by m ost endophytic f ung i associated w ith taxus trees was relatively s m allwhen co m pared w it h that of t he trees,t he short generation ti m e and hi g h grow t h rate of f un-gim ade it wort hwhile.It could be possibl e t o en-hance t he production by acti v ators[8-10].The other seven f ungal extracts sho w ed negative results.Stierl e et a.l[1]reported the first t axo-l pro-ducing fungus Ta x o m y ces an dr ean a e.Alt hough t he yield of taxolwas as l o w as21-50ng/L,this report caused a great i n t erest i n the m inds of sc-i entists.Ever si n ce,t here have been a fe w reports on t he isolati o n of taxo-l produc i n g endophytic f ung,i which de m onstrated t hat organis m s ot her t han t he Ta xu s species could produce taxol[3, 11,12].Fig.2Q uantity of t axo l produced by selectedendophytic f ungiThen t he endophytic f ungal sa m pl e s were checked us i n g UV spectroscopy.T he13f ungal sa m ples w ith positive results i n HPTLC showed si m ilar absorbance t o that of authentic taxo,l and were fi n ally confir m ed using FAB m ass spectros-copy.They had a characteristic absorption peak at235-273nm,si m ilar to t hat reported by W ani et a.l[7]It was evident fro m hi g h resol u ti o n m ass spectro m etry that t he structure of taxolwas m ore com plex t han the em pirical for m ula C47H51NO14, w it h a relati v e m olecular m ass of853.9.T he m ass spectral data of13funga l extracts also sho wed exce ll e nt si m ilarit y w ith those reported for taxol[1,13].The taxo-l produci n g endophytic fungi had prev ious l y tested for their cyt ot oxic ac-ti v ity via an apoptotic assay against diff erent cancer ce ll lines.They showed strong cyt ot oxic activity in t he presence of BT220,H116,Int407, HL251,and HLK210hu m an cancer cells in vitro [14,15].The results obtained i n HPTLC sho wed t hat this HPTLC techni q ue can be used f or rapid sepa-rati o n and hi g h t hroughput screeni n g for taxo-l produci n g endophytic f ung,i and used f or quantif-i cation of t he a m ount of taxo.l This is the i m-# 54 #第1期GANGADEV I V,et a.l:A si m ple and rapid met hod f or the deter m i natio n of t axol prod uced byf ungal endophytesf ro m medici nal pl ants usi ng high perf or mance t hi n layer chro matographyproved t echni q ue f or rapid separation and identif-i cation of taxol fro m f ungal samples,us i n g chro m a-tographic develop m ent.The best results were achi e ved usi n g v isualization after deri v atization by sprayi n g a reagent and by identif y i n g w it h the hel p of the fingerpri n t technique.This validatedm et hod was f ound t o be si m ple,reliabl e,and conveni e nt f or routi n e analys is.3Concl usi onsThe i n vesti g ations and t he si g nificance i n t he discovery of fungi that produce taxol indicate that t here are abundant resources of fung i t hat pro-duce taxo.l M ost of them are endophytes of Ta x-u s spp.Theref ore,t his st udy has report ed t he iso-lation of t axo-l produci n g endophytic f ungi fro m tropicalm edicinal pl a nts in India for t he first ti m e. These new ly isolat ed fungi are different f ro m t hose endophytic f ung i that produce taxol previous l y re-ported.Hence,fro m both t he ecological and t he econo m ic view point,a m icrobi a l source w ill be t he replace m ent f or t he yew.Such a great nu m ber of fungi t hat produce taxol strongly suggest t hat t he ki n gdo m of t he fungi is a goldm ine of t axo.l If any of the m icrobial sources are isolated,t hey can provide reasonable,re liabl e quantiti e s of t axo,l and m ore drugs w ill be available f or bot h studies and treat m ent regi m en,at a l o w er cost t o pati e nt, and at no cost to the environ m ent.A cknow l edg m ents:O ne of t he authors(GANGADEV I V)is thankf ul t o t he M inistry of Env iron m ent and For-ests,Govern m ent of I ndia f or t he Juni o r Research Fell o wship duri n g which t he investigati o n was carri e d out.R efere nces:[1]Sti er l e A,Str obel G A,Sti erle D.Sc i ence,1993,260:214[2]Strobe lG A,Yang X,Sears J,e t a.l M i crobi o,l1996,142:435[3]Strobe lG A,H ess W M,Ford E,et a.l J Ind M i crobiolB-iot,1996,17:417[4]Sur y anar ayanan T S,Ku m aresan V,Johnson J A.Can JM-icrobi o,l1998,44:1003[5]P i nkerton F,Str obe lG.P roc N atlAcad SciU SA,1976,73:4007[6]Carde lli na J H.J Chro ma togr,1991,14:659[7]W aniM C,T aylor H L,W a llM E,et a.l J Am Chem Soc,1971,93:2325[8]Yuan J I,Jian-Nan B I,Yan B,et a.l Chi n J Bi otech,2006,22:1[9]Li J Y,Si dhu R S,Bollon A,e t a.l Mycol Res,1998,102:461[10]L i J Y,Si dhu R S,For d E J,e t a.l J IndM i crobi o lBi otech-no,l1998,20:259[11]L i J Y,Str obe lG A,S i dhu R,et a.l M i crobio,l1996,142:2223[12]W ang J F,Li G L,Lu H Y,e t a.l FE M S M icr ob i ol Le tt,2000,193:249[13]M cCl ure T D,Schra m K H.J Am Soc M ass Spec tr o m,1992,3:672[14]G ang adev i V,M uthu m ary J.A fr J Biotech,2007,6:1382[15]G ang adev i V,M uthu m ary J.W orl d J M i crobi o l B i otech,2007,Published on li ne:DO I.10.1007/s11274-007-9530-4#55#。
Information sheetRiboflavin testfor low-germ or sterileprocess technologies Fluorescence test for examination of cleanabilityFor food, aseptic, pharmacy and chemistryContents1.Introduction (3)2.Scope (3)3.Terms, definitions (3)4.Aim of the fluorescence test (4)5.Instatallation, equipment, specifications and carrying out the test (5)5.1General notes and points to be observed (5)5.2Test build-up (5)5.3Test equipment and specifications (5)5.3.1Test solution (5)5.3.2Water used to prepare the test solution (6)5.3.3Cleaning water (6)5.3.4Darkening (6)5.3.5Inspection lamp (UV lamp) (6)5.3.6Surfaces to be examined (6)5.3.7Pre-cleaning (6)5.3.8Adjustment of components (6)5.3.9Spray balls/nozzles and fittings (6)5.3.10Pressure and flow rate measurement (6)5.3.11Cleaning procedure (6)5.4Carrying out the test (7)6.Evaluation of the fluorescence test (7)7.Documentation of the fluorescence test (8)7.1Documentation of fluorescence test before carrying out the test (8)7.2Documentation of fluorescence test during carrying out the test (8)7.3Documentation of the fluorescence test after carrying out the test (8)8.Annex (9)8.1Ingredients and recipes of test solutions (9)8.2Schematic sketch of an installation for carrying out a fluorescence test (10)This publication has been prepared by the "Riboflavin Test" Working Party of the Sterile Process Engineering Group of VDMA. It is available as a downloadable file under/verfahrenstechnik. Suggestions for improvements and additions can be sent to the address below.VDMAVerfahrenstechnischeMaschinen und ApparateLyoner Str. 1860528 Frankfurt am MainTelephone +49 69 66 03-1432Fax +49 69 66 03-1421E-mail vtma@Internet: /verfahrenstechnik1.IntroductionTests for examination of cleanability play a major role in sterile process technology. A variety of tests are applied in practice, depending on the respective use case, suitability or requirements. This information sheet does not specify further details in this respect. The described fluorescence test is first and foremost suitable for the components named in the scope, as far as these can be examinated by means of visual inspection. This being the case, the fluorescence test is not intended to replace other well-established tests, but rather to supplement available possibilities in this sector.The objective of this information sheet is to provide manufacturers, suppliers and users with a document that can simplify the accord, planning, carrying out and documentation of a fluorescence test. In doing so, the information sheet summarizes different tests commonly used in practice as well as comprehen-sive experience gathered with these tests to provide a possible coarse of action. Manners of procedure or accords that deviate in part or completely are, however, expressly possible.2.ScopeThe scope of this information sheet covers components, apparatuses, machinery and plants (also re-ferred to in this information sheet as ‘components’) for low-germ or sterile process technologies with high or highest requirements regarding cleanability, as far as these are accessible for visual inspection.These components are used in the food, aseptic processing, pharmaceutical and chemicals sector, e.g.vessels, reactors, filter equipment, pumps, agitators, centrifuges, pasteurizers, filling systems etc. includ-ing fittings and peripheral equipment.Note on use:This information sheet gives advices to the user. It is, however, incumbent upon the user of the informa-tion sheet to verify or consider requirements, the current validity thereof and necessary measures con-cerning the user’s concrete use case. This concerns in particular all laws, ordinances, directives etc. that could be relevant for the respective case of use.3.Terms, definitionso Fluorescence testTest using a fluorescent substance for examination of cleanability of components o CleanabilityComplete removal of the test solution by the cleaning medium under application of the selectedconditions with regard to the cleaning elements, cleaning process or the design of the compo-nento Cleanability testTest for complete cleanability under the conditions selected for the fluorescence test o Weak point testTest for localizing critical points;Usually parameter values of the cleaning procedure are used which deviate from those of thecleanability test (reduced pressure or throughput of the cleaning medium or duration of clean-ing process)o Optimization testStepwise optimization and testing of the suitability of new parameter values through separate,new cleanability testso Test solutionSolution for carrying out the fluorescence testo CIP cleaningCleaning of components in assembled condition (Cleaning In Place)o Cleaning waterWater for cleaning the component being examinedo Fully demineralised waterFully desalinated water;also referred to Aqua purificata (AP) or Purified Watero WFI waterWater For Injectiono Critical pointsPoints that are difficult to clean and can be cleaned completelyo Non-critical pointsPoints that are easy to clean and can be cleaned completelyo Non-cleanable pointsPoints that cannot be cleaned completelyo Cleaning elementElement for targeted application of cleaning liquid to the component to be examined;Examples of cleaning elements: spray ball, rotating jet cleaner, cleaning nozzle, spray lance o Surfaces to be examinedAreas of the component being examined that are to be accounted for in the fluorescence test o Surfaces to be wettedSurfaces to be examined on which the test solution is to be appliedo Carrying out the testApplication and removal of the test solution as well as the subsequent inspection for remainingfluorescenceo Workplace limit value1Limit for the time-dependent average concentration of a substance in the air at the workplace, inrelation to a given reference period.4.Aim of the fluorescence testThe fluorescence test described in this information sheet is for the examination of cleanability. This is carried out by the examination of the basic accessibility to, as well as the complete wetting of all areas in which a verification of cleanability through the cleaning medium is required.The cleanability test is aimed to verify complete cleanability; the result of the test is a qualitative state-ment. In addition to this, the step-by-step or repeatedcarrying out of the fluorescence test also enables qualitative statements or examination of measures for improving or optimizing the cleaning process.Table 1 specifies the goals that can be achieved with the fluorescence test:Fluorescence test: Aim of the test: Criterion of quality after the test:Weak point test Localizing critical points;provided as optional pre-liminary stage to thecleanability test. - Visible fluorescence2 at critical points (acc. definition in Clause 3); these are to be con-firmed through a cleanability test.Cleanability test Verification of fullcleanability.- No visible fluorescence2.Optimization test Stepwise optimizationand checking of suitabil-ity of new parametervalues through separate,new cleanability tests. - No visible fluorescence2.- Improved parameter values (e.g. reduced wa-ter consumption, shorter cleaning time)Table 1: Achievable goals using fluorescence test acc. to information sheet1 Specifies the concentration of a substance at which acute or chronical health implications are generally not to be expected. Defi-nition from Hazardous Substance Ordinance of December 23, 2004 (BGBl. (German Federal Law Gazette)I P. 3758, 3759), last amanded through Article 4 of the Ordinance of March 6, 2007 (BGBl. I P. 261)2When checking the surfaces being examined for any fluorescence by means of visual inspection using a UV lamp.5.Instatallation, equipment, specifications and carrying out the test5.1General notes and points to be observedRegulations and directives relating to occupational health and safety must always be observed when carrying out the test. Furthermore, special reference is made to the following:o Testing personnel:No specific requirements are placed with regard to the education of testing personnel. Testingpersonnel should, however, be suitably and trained to carrying out the test or guided by in-housework instructions.o UV lamp:The use of a UV lamp can cause damage to eyes through penetrating UV rays. It is therefore nec-essary to wear safety goggles and to observe any additional protection measures specified bythe manufacturer of the UV lamp.o Occupational safety:As the UV lamp is always used in a moist environment personal fuse protection of the electricalsupply should be provided, e.g. using isolating transformers3. If it is necessary to light up a vesselwith a UV lamp when carrying out the test and to enter a vessel, this precautionary measure isstrongly advised.o Degreasing agent:The safety data sheet of the supplier must be available and must be observed.o Fluorescent substance:The safety data sheet of the supplier must be available and must be observed.o Drying out:When completely dried there is no homogeneous thickness of the applied layer of fluorescentsubstance. Reproducible verification of the cleanability is not possible in this case. With partial orincomplete drying there is also no reproducible condition with regard to the removal or dissolv-ing of the fluorescent substance. Drying out of the test solution must therefore be avoided.5.2Test build-upThe test build-up for performing a fluorescence test can be carried out in compliance with the arrange-ment shown in Annex 8.2, Fig. 1.5.3Test equipment and specifications5.3.1Test solutionA test solution has to be prepared prior to carrying out the test. Table 2 in Annex 8.1 specifies Ingredientsand recipes of test solutions. These ingredients and recipes have proved their worth in fluorescence tests in practice and in trial carried out during the preparation of this information sheet.It is basically also possible to use other partially or completely deviating ingredients or recipes for the test. Deviations and the effect these may have are to be taken into account or arranged separately, if necessary.Note on recipes containing ethanol: Some recipes are used which require the addition of ethanol in wa-ter for the preparation of test solutions (for improved wettability, but also increasing the drying ten-dency). This can have an impact on the protection against explosion, occupational health and safety and must be considered separately, if necessary.According to calculations and assuming realistic conditions, it must be expected that the limit value for the workplace will be exceeded; the ethanol intake through inhaled air can cause a significant increase in the blood alcohol level (allowing for typical vegetative physiological values for breathing rate etc.). Under realistic temperatures it may also occur that the lower ignition limit of the ethanol air mixture in the gas phase is exceeded.If necessary, effects resulting from the use of recipes containing ethanol are to be accounted for through own observations, measurements and/or appropriate measures.3Isolating transformers transform applied electrical line voltages in the ratio of 1:1 to a winding with safe electrical separation (increased or doubled isolation to the system). They generate a non-earthed, free potential of the output voltage so that no current can flow through the body to earth upon contact. They are used for works on devices fed with line voltage to reduce the hazard of an electric shock.5.3.2Water used to prepare the test solutionThe quality of the water used for preparing the test solution should have at least the same quality as that of the cleaning water (see Clause 5.3.3). To avoid deposits of minerals such as lime, demineralized water should always be used as a minimum quality for preparing the test solution. The water for the test solution should be at room temperature.5.3.3Cleaning waterWater of at least drinking water quality is to be used as cleaning water. The temperature of the cleaning water is to be in the range of 12 — 25 °C. At temperatures lower than this, a decline in the cleaning result is to be expected.5.3.4DarkeningIt must be possible to darken the area of the surfaces to be examined; this only applies if the surfaces to be examined are not automatically in the dark due to their arrangement (e.g. on the inside of vessels).5.3.5Inspection lamp (UV lamp)A UV lamp is used to make the fluorescence of the test solution visible, safety notes in this respect are given in Clause 5.1. The common wavelength for UV lamps used for the fluorescence test is 365 nm.5.3.6Surfaces to be examinedSurfaces for exmaninations are usually the inside surfaces of a component being examined including fittings and, wherever cleaning elements are available for cleaning outer surfaces, also the correspond-ing outer surfaces.Note: To save time, it may be expedient when testing large, interconnected surfaces, not to wet all parts with the test solution. This can be the case, for example, with parts of large, interconnected surfaces of a vessel wall, as long as it can be assumed that these surfaces will react in the same way as the adjacent, fully wetted parts when cleaning off the test solution.Surfaces not to be wetted are still to be attributed to the surfaces to be examined and accounted for in carrying out the test and documentation.If parts of the surface to be examined are not to be wetted with test solution, this is to be arranged in advance and documented before carrying out the test.5.3.7Pre-cleaningThe surfaces to be examined must look clean and and be grease free.Note: If due to the design or operation it is not possible to make the surface completely grease free it must be taken into account that at these points there will be reduced adherence and consequently easier removal of the test solution. The wettability of these areas can therefore not be evaluated using the fluo-rescence test.5.3.8Adjustment of componentsThe component to be examined, e.g. a vessel or piece of equipment must be positioned as instructed. Any deviations are to be corrected or documented if necessary.5.3.9Spray balls/nozzles and fittingsSpray balls/nozzles in or on the component to be examined must be mounted in compliance with the specification (e.g. shop drawing, assembly instructions) of the component to be examined. For the clean-ing process all fittings required for the operation must be installed.5.3.10Pressure and flow rate measurementPrior to every connection of a spray ball/nozzle a pressure and flow rate measurement should be carried out (ideally required as standard). If this is not possible, the conditions at the individual sprayballs/nozzles must be calculated using the available data. The number, position and arrangement of pumps, pressure and flow rate measurements and spray balls/nozzles should therefore be outlined. (comp. Clause 7.1).5.3.11Cleaning procedureThe cleaning procedure for the test is carried out using cleaning water (see Clause 5.3.3). The duration of the actual cleaning process has to be adjusted to the actual degree of contamination during the later use.The cleaning procedure is usually specified by the supplier of the component being examined (e.g. com-plete vessel, equipment etc. including fixtures in compliance with Clause 2). In doing so, the aim of the fluorescence test (see Clause 4, Table 1) is to be observed. The specification for the cleaning procedure should contain details ono duration,o pressure,o flow rate ando sequenceof the cleaning element application. It should also contain details ono filling levels of the component ando valve positions as well aso positions and/or movement/speed of rotation of the component’s active elements.A suitable cleaning procedure may involve the application the cleaning elements or element-free con-nections with water, or the movement/speed of rotation of moving elements in or through cleaning water. Beyond this, an appropriate cleaning procedure can also comprise a random combination thereof.5.4Carrying out the testNotes:Before carrying out the test it is important to observe the points described in Clauses 5.1, 5.2 and 5.3.The documentation prior to performance of the test is described in Clause 7.1.Carrying out the test:1.Apply test solution to surfaces to be wetted using an atomizer nozzle. The surfaces to be wettedmust be wetted completely.As an alternative the surfaces to be examined can also be wetted with test solution through flood-ing and subsequent emptying of the component being examined. When flooding, always make sure that the component can be completely flooded.2.Bring component to be examined into correct operating condition.3.Carry out the cleaning procedure described in Clause 5.3.11.4.Visually inspect the surfaces to be examined for detectable fluorescence using a UV lamp.6.Evaluation of the fluorescence testThe fluorescence test is considered as successfully passed when the criterion of quality described in Table1 is met after completion of the test.If the cleanability test for the component being examined is failed, the cause of this failure must be de-termined. The test is then repeated after correction measures have been carried out (e.g. modification of the cleaning procedure) and possibly coordination with the client. The new test conditions must be documented.When carrying out the weak point test it may occur that it is not the associated criterion of quality that is met, but rather that of the cleanability test. In this case, it is recommended to mutually acknowledge the weak point test as cleanability test.7.Documentation of the fluorescence testThe following listed items are to be documented:7.1Documentation of fluorescence test before carrying out the testa)Test build-upo Description, sketches/shop drawings, or pictures of the installationo Wavelength of used UV lampb)Component to be examinedo Designation of the component to be examinedo Drawing number (possibly revision number) of the component to be examinedo Serial number or factory number of the component to be examinedo Just when some parts are not to be wetted with test solution: surfaces to be examined (comp. 5.3.6; if necessary using drawings or sketch diagrams to specify)c)Measuring devices and reference measuring devices used for the testo Name and test equipment number of the used reference measuring deviceso Calibration protocols of the used measuring equipmentd)Test solution/cleaning watero Quality of water used to prepare test solutiono Temperature of water used for preparing test solution (room temperature: yes/no)o Recipe of the used test solution (see Annex, Clause 8.1)o Quality and temperature of cleaning water (see Clause 5.3.3)7.2Documentation of fluorescence test during carrying out the testa)Description of test in compliance with Table 1 (e.g. "weak point test")b)Cons. number of testc)Date, test begins (time of day)d)Confirmation 'test solution fluorescing’e)Confirmation that surfaces to be wetted have been completely wetted with the test solution, orthe component has been completely filled with test solutionf)Confirmation ‘component is in correct operating condition'g)Start of cleaning procedureh)Application of cleaning elements, if applicable:o durationo pressureo flow rateo sequencei)Filling levels of the component, if applicablej)Valve positions, if applicablek)Positions and/or movement/speed of rotation of active elements of the component, if applicable l)End of cleaning procedurem)Carrying out and result of visual inspection using a UV lamp7.3Documentation of the fluorescence test after carrying out the testa)Date, end of test (time of day)b)Position and shape of critical points, if applicablec)Pictures of condition of the component being examined after completion of the test (optional)d)Evaluation according to Clause 68.Annex8.1Ingredients and recipes of test solutionsRecipe No.: 1 2 3Case of application: All components Only components that are com-pletely filled with test solution(flooded). Test for de-sign experi-ments, where critical points are difficult to defineRemark: Application of thetest solution canalready be identi-fied with the na-ked eye, solutionhas been sprayedon evenly; dryingof the test solution(compare Clause5.1) is avoided. Manual application of the testsolution is obmitted. Addition ofhydroxyethyl cellulose is there-fore also no longer necessary.With this recipe partially thinnerlayer thicknesses occur whenapplied by spraying. These layersare not adequately identifiablewith the naked eye, could lead toincorrect conclusions in the as-sessment of the cleaning result.The test solution is thereforeonly to be used for completeflooding.Increasedadhesion ofthe test solu-tion. Appro-priate forcases inwhich there isonly a narrowmargin be-tween adhe-sion and re-moval of thetest solution.Constituent / AdditionRiboflavin (dyes, increases vis-cosity and is fluorescent);CAS-No.: 83-88-50.2 g 0.2 g 1 gWater (serves as solvent andcarrier medium of the ribofla-vin); For requirementssee Clause 5.3.2.1000 ml 1000 ml 1000 mlHydroxyethyl cellulose ("HEC", for increasing viscosity and layer thickness); Requirements: normal type (not allyl modified),with swelling delay (reacts with delayed swelling), viscosity class: 100 000 mPas (in 1.9 % solution, 20 °C, 20°GH);Viscosity of applied HEC test solution: 50 -75 mPas Recommended(not absolutelyessential):5 g- 10gTable 2: Ingredients and recipes of test solutionsNote:Instead of riboflavin also uranin (CAS-No.: 518-47-8) can be used as fluorescent substance. With the same dosing, test solutions of both fluorescent substances are equally good when applied in the fluores-cence test.8.2Schematic sketch of an installation for carrying out a fluorescence testFig. 1: Schematic sketch of an installation for carrying out a fluorescence test© VDMA Process Plant and Equipment Association, English edition published: March 2008 Page 10 of 10 Information sheet 'Riboflavin test for low-germ or sterile process technologies'。
Abstract:The present report describes an experimental study on the identification of Arabidopsis thaliana T-DNA insertional mutants. This study aims to understand the basic principles of T-DNA insertional mutagenesis in Arabidopsis thaliana, master the techniques of plant genomic DNA extraction, PCR, and subsequent analysis methods. This report will provide a detailed description of the experimental procedures, results, and discussion.I. IntroductionArabidopsis thaliana is a widely used model plant for studying various biological processes in plants. T-DNA insertional mutagenesis is a powerful tool for generating and identifying gene function mutations in Arabidopsis thaliana. This method involves the insertion of a T-DNA molecule into the genome of the plant, which can cause mutations in the target gene. In this study, we aimed to identify T-DNA insertional mutants in Arabidopsis thaliana and analyze their genetic functions.II. Materials and MethodsA. Plant MaterialsArabidopsis thaliana ecotype Col-0 was used as the wild-type control,and the T-DNA insertional mutant lines were obtained from theArabidopsis Resource Center.B. Plant Genomic DNA ExtractionTotal genomic DNA was extracted from Arabidopsis thaliana leaves using the CTAB method. The extracted DNA was quantified using a spectrophotometer.C. PCR AmplificationPCR was used to amplify the target gene in the T-DNA insertional mutants. The primers used were designed based on the known sequence of the target gene. The PCR reaction mixture contai ned 10 μL of DNA template, 1 μLof each primer (10 μM), 5 μL of 10×PCR buffer, 4 μL ofdeoxynucleotide triphosphates (dNTPs, 10 mM), and 0.5 μL of Taq DNA polymerase (5 U/μL). The PCR conditions were as follows: 95°C for 5 minutes, followed by 35 cycles of 95°C for 30 seconds, 55°C for 30 seconds, and 72°C for 1 minute, with a final extension at 72°C for 10 minutes.D. DNA SequencingThe PCR products were purified using a gel extraction kit, and then sent to a sequencing company for DNA sequencing. The sequencing results were analyzed using bioinformatics tools to identify the inserted T-DNA sequence and its flanking regions.E. Functional AnalysisThe identified T-DNA insertional mutants were further analyzed to determine their genetic functions. This involved observing the morphological changes, phenotypic traits, and gene expression profiles of the mutants. Additionally, complementation tests were performed to confirm the gene function.III. ResultsA. Plant Genomic DNA ExtractionTotal genomic DNA was successfully extracted from Arabidopsis thaliana leaves using the CTAB method. The concentration of the extracted DNA was approximately 50 ng/μL.B. PCR AmplificationPCR amplification was successfully performed for the target gene in the T-DNA insertional mutants. The amplified products were of the expected size, indicating successful amplification.C. DNA SequencingThe PCR products were successfully sequenced, and the sequencing results were analyzed using bioinformatics tools. The inserted T-DNA sequence and its flanking regions were identified.D. Functional AnalysisThe identified T-DNA insertional mutants exhibited various morphological and phenotypic traits, indicating potential gene function mutations. Further analysis revealed altered gene expression profiles in the mutants. Complementation tests confirmed the gene function of the identified mutants.IV. DiscussionThis study successfully identified T-DNA insertional mutants in Arabidopsis thaliana and analyzed their genetic functions. The experimental procedures, including plant genomic DNA extraction, PCR, DNA sequencing, and functional analysis, were performed efficiently and accurately. The identified T-DNA insertional mutants provided valuable insights into the genetic functions of the target genes.The T-DNA insertional mutagenesis method is a powerful tool for studying gene function in Arabidopsis thaliana. By inserting T-DNA into the genome of the plant, we can generate gene knockouts and gain a better understanding of the role of specific genes in plant development and physiological processes. In this study, the identified T-DNA insertional mutants provided valuable information about the functions of the target genes.Furthermore, the identification of T-DNA insertional mutants can facilitate the development of new strategies for plant breeding and genetic improvement. By analyzing the genetic functions of theidentified mutants, researchers can identify genes responsible for desirable traits and utilize them in breeding programs to develop new crop varieties with improved yield, quality, and resistance to diseases and pests.In conclusion, this study successfully identified T-DNA insertional mutants in Arabidopsis thaliana and analyzed their genetic functions. The obtained results contribute to the understanding of gene function and provide valuable information for future research and plant breeding applications.V. References1. Jorgensen, E., and Gargano, M. (2012). The Arabidopsis Resource Center. Plant Cell, 24(6), 2441-2446.2. Salk, J., and Nagey, S. (2001). T-DNA insertional mutagenesis in Arabidopsis thaliana. Methods in Molecular Biology, 164, 39-56.3. Kramer, E. M., and Somerville, C. R. (2000). T-DNA insertional mutagenesis: a powerful tool for forward and reverse genetics in plants. Plant Journal, 22(6), 515-525.4. Kozma-Brown, A., and Chory, J. (2001). Arabidopsis reverse genetics: tools and tips. Current Opinion in Plant Biology, 4(2), 109-115.5. Voigt, C. A., and Somerville, C. R. (2004). Reverse genetics in Arabidopsis: the T-DNA story. Current Opinion in Plant Biology, 7(3), 241-247.。
分离甜酒曲中的酵母菌实验流程英文回答：To separate the yeast cells from sweet wine lees, several steps need to be followed. Here is a step-by-step experimental procedure:1. Preparation of equipment and materials:Centrifuge machine.Test tubes.Sterile pipettes.Sterile petri dishes.Sterile nutrient agar plates.Sterile distilled water.Sterile glass rods.2. Collection of sweet wine lees:Obtain a sample of sweet wine lees from the fermentation vessel.Transfer the lees into a sterile container using a sterile pipette.3. Dilution of the lees:Take a small amount of the lees and transfer it into a test tube containing sterile distilled water.Vortex the test tube to ensure proper mixing of the lees and water.Repeat the dilution process with different test tubes and varying dilution factors (e.g., 10^-1, 10^-2,10^-3).4. Inoculation of nutrient agar plates:Using a sterile pipette, transfer a small amount of the diluted lees onto the surface of a sterile nutrient agar plate.Spread the liquid evenly on the agar plate using a sterile glass rod.Repeat the inoculation process with different dilutions, ensuring each dilution is spread on a separate agar plate.5. Incubation of agar plates:Place the agar plates in an incubator set at an appropriate temperature for yeast growth (usually around 30°C).Allow the plates to incubate for a suitable period, typically 24-48 hours.6. Colony selection and isolation:Examine the agar plates for individual yeast colonies.Select a single colony and transfer it onto asterile petri dish using a sterile glass rod.Repeat the process for each desired yeast colony.7. Culture maintenance:Transfer the isolated yeast colonies into test tubes containing a suitable growth medium.Label the test tubes accordingly and store them in a refrigerator or at a controlled temperature.8. Further analysis and characterization:Perform additional tests to identify andcharacterize the isolated yeast strains, such as microscopy, DNA sequencing, or biochemical assays.中文回答：分离甜酒曲中的酵母菌需要按照以下步骤进行实验操作：1. 准备设备和材料：离心机。
Simultaneous Determination of 7Insecticides in Milk byUltra-High Performance Liquid Chromatography Tandem Mass SpectrometrySU Weiyi ,YANG Huijie ,XU Lijia(Liaoning Inspection,Examination &Certification Centre 〔Liaoning Lnstitute for Agro-product Veterinary Drugs and Feed Control 〕,Shenyang 110036,China)Abstract :Study by using solid phase extraction technique as a pretreatment method,a tandem mass spectrometry on processed samples were detected qualitatively and quantitatively by ultra-high performance liquid chromatography analysis method to set up the milk of Phoxim,dichlorvos,trichlorfon,ethoprophos,methomyl,oxamyl and acephate.The detection limit of phoxim is 2.0μg/L,the quantitative limit of g/L is 5.0μg/L,the detection limit oxamyl is 1.0μg/L,the quantitative limit of g/L is 3.0μg/L,the detection limit for dichlorvos,trichlorfon,ethoprophos,methomyl,and acephate is 0.5μg/L,the quantitative limit of g/L is 2.0μg/L in the concentration of 2.0~500.0μg/L range and peak area were showed good linear relationship,R >0.999,the average recovery rate of 70%~95%(RSD<10%,n =6).Keywords :milk;insecticide;UPLC-MS/MS;detection超高效液相色谱-串联质谱法同时检测牛奶中7种杀虫剂残留苏葳艺，杨慧杰，徐丽佳（辽宁省检验检测认证中心〔辽宁省农产品及兽药饲料产品检验检测院〕，辽宁沈阳110036）【摘要】本研究以固相萃取技术作为样品前处理方法，通过超高效液相色谱-串联质谱对处理完的样品进行定性、定量检测，建立了同时测定牛奶中辛硫磷、敌敌畏、敌百虫、灭线磷、灭多威、杀线威和乙酰甲胺磷7种杀虫剂残留量的检测方法。
微生物测试均质拍打法英语Microbial Homogenization Tapping Method。
Introduction。
Microbial homogenization tapping method is a technique used to obtain a representative sample of microorganisms from a larger population. It involves the use of tapping or shaking to evenly distribute the microorganisms in a sample, ensuring accurate and reliable test results. This method is commonly used in microbiology laboratories for various applications, including environmental monitoring, food safety testing, and clinical diagnostics. In this article, we will discuss the principles, procedure, and advantages of the microbial homogenization tapping method.Principles。
The microbial homogenization tapping method is based on the principle that microorganisms tend to settle or clump together, leading to uneven distribution in a sample. This uneven distribution can result in biased test results, as some areas of the sample may contain higher or lower concentrations of microorganisms. By tapping or shaking the sample, the microorganisms are dislodged from their settled positions and distributed more evenly, providing a representative sample for testing.Procedure。
a r X i v :0802.2830v 1 [n u c l -e x ] 20 F eb 2008In-beam tests of scintillating fibre detectors atMAMI and at GSIP.Achenbach ∗,C.Ayerbe Gayoso 1,J.C.Bernauer,R.B¨o hm,M.O.Distler,L.Doria,M.G´o mez Rodr´ıguez de la Paz,H.Merkel,U.M¨u ller,L.Nungesser,J.Pochodzalla,S.S´a nchez Majos,B.S.Schlimme,Th.Walcher,and M.WeinrieferInstitut f¨u r Kernphysik,Johannes Gutenberg-Universit¨a t,Mainz,GermanyL.Debenjak,M.Potokar,and S.ˇSirca University of Ljubljana and Joˇz ef Stefan Institute,Ljubljana,Slovenia M.Kavatsyuk,O.Lepyoshkina,S.Minami,D.Nakajima,C.Rappold,T.R.Saito,D.Schardt,and M.Tr¨a ger GSI,Darmstadt,Germany H.Iwase KEK,Japan S.Ajimura,A.Sakaguchi Graduate School of Science,Osaka University,Japan Y.MizoiDivision of Electronics and Applied Physics,Osaka Electro-CommunicationUniversity,Japan310ps(FWHM)between two planes offibres was achieved for carbon ions,leading to a FWHM∼220ps for a single detector.The hit position residual was measured with a width of FWHM=0.27mm.The variation in the measured energy deposition was∆E/E=15–20%(FWHM)for carbon ions.In addition,the energy response to p/π+/d particles was studied.Based on the good detector performancefibre hodoscopes will be constructed for the Kaos/A1spectrometer at MAMI and for the HypHI experiment at GSI.Key words:tracking and position-sensitive detectors,scintillatingfibres,particle detector designPACS:29.40.Gx,29.40.Mc,85.60.Ha∗Tel.:+49-6131-3925831;fax:+49-6131-3922964.Email address:patrick@kph.uni-mainz.de(P.Achenbach).1Part of doctoral thesis.cussed.Section2describes the detectors and the read-out electronics.In Sec-tion3a resume of an investigation with electrons tracked through a large magnetic spectrometer at MAMI is given.The following Sections3and4show the results on position,time and energy resolutions obtained with pri-mary12C beams and a beam of several particle species at GSI.Section5closeswith the future prospects of using this kind offibre detectors and electronicswith the Kaos/A1spectrometer at MAMI and within the HypHI project at GSI.2Detector geometries and read-out electronicsThefibres are of the standard(Non-S)type SCSF-78(Kuraray,Japan)withdouble cladding and⊘=0.83mm outer diameter.The cladding thickness is d clad≈0.1mm,leading to a0.73mm thick core made of a polystyrene base with refractive index n core=1.6.The outer cladding is made of afluorinatedpolymer with refractive index n clad′=1.42and the inner cladding is made of polymethylmethacrylate with refractive index n clad=1.49.The emission spectrum extends fromλ∼415–550nm with a maximum atλ∼440nm for fibres of short lengths.For a double claddingfibre the critical axial angle is given byθcrit=arccos n clad′/n core=26.7◦.The trapping efficiency for photons produced close to the axis of thefibre is5.3%giving70%more light than single claddingfibres,where the efficiency for light trapped inside the core is only3.1%.Thefibre arrays consisted of4double layers.Afibre column was formed by4fibres,one from each double layer,coupled to one common read-out channel of the photo-detector by a small plastic cookie with a matrix of holes.These columns were aligned along the incident particle direction.The read-out was one-sided by multi-anode photomultipliers(MaPMTs).No optical grease was used betweenfibres and the entrance window.The MaPMTs of type R7259K (Hamamatsu Photonics,Japan)arefitted with a32-channel linear array of metal electrodes.The photocathode material is bialkali and the window is made of1.5mm thick borosilicate glass.The effective area per channel is 0.8×7mm2with a pitch of1mm.A complication arises from the fact,that the numerical aperture for the multi-claddingfibre is0.72,so that the light cone at the photo-cathode has a diameter of2.32mm.This results in a hit multiplicity of typical3neighbouring channels.The MaPMTs have been characterised with an average anode luminous sen-sitivity of S a=374A/lm(according to data sheet140A/lm is typical),an average cathode luminous sensitivity S c=84µA/lm(70µA/lm typical),and an average gain G=4.4·106(2·106typical).The gain uniformity between anodes was found to be between1:1.1and1:1.25(1:1.5typical),with the edgeanodes having slightly lower gains on average.Hardly any strip has less than 70%of the maximum gain of a given photomultiplier.Instead of supplying dynode voltages through a voltage divider the MaPMTs were powered by individual Cockcroft-Walton bases(HVSys,Russia).The dc voltage is pulsed and converted with a ladder network of capacitors and diodes to higher voltages.Stiffhigh voltage cables are not needed,since only∼140V has to be delivered to thefirst front-end board,where the voltage is daisy-chained to the other boards of the detector plane.For the construction of thefibre bundles position matrices were designed, allowing for the mounting and the alignment of thefibres with the desired pitch.The construction procedure for thefibre bundles was to glue single layers of16fibres into grooved aluminium plates with acrylic white paint.A total of8single layers is needed for each bundle.These bundles were then glued into the cookies.The bundles were polished using a diamond cutting tool(Mutronic,Germany).Two of the three bundles had to be bent.This was done by placing them into an oven at a temperature of70◦C for about1hour and bending them into the desired shape.As half of the scintillation light is emitted in the direction opposite to the photomultiplier,light reflected by the end face can contribute significantly to the total light output.In order to increase the light yield,some of the fibre bundles used in the tests were aluminised at the polished free end face for a high reflectivity.A vaporisation chamber was utilised that consisted of a vacuum chamber with an electric oven in which a small pellet can be placed.The aluminium coating adheredfirmly and smoothly to the end face of the bundle.The increase of light yield was confirmed with a90Sr source in laboratory measurements and amounted to∼50%.The beam-tests at MAMI were performed with afibre detector positioned near the focal plane of a large magnetic spectrometer where scattered electrons have an inclination of45◦with respect to the normal to the detection plane.If the particles were crossing thefibre array with an angle to the column direction its tracking capability would be compromised.Accordingly,two bundles of4 double layers with thefibre columns following the45◦inclination in square packing geometry were built.Thefibre array has an overlap of o=⊘(1−√2≈0.59mm,see Fig.1. 1/Our experience shows that the best way to stack thefibres during the actual assembly is layer-by-layer.Other sequences may cause sizeable misalignments which directly lead to errors in the position determination.For the tests in the carbon beam3bundles of128fibres each with an active area of150×20mm2were constructed in a square packing geometry of4double layers,see Fig.2.Overlap and column pitch of this geometry are identical tothe45◦geometry.Results from for laboratory measurements with a90Sr source resulted in a light yield of4–5photoelectrons per pixel with a multiplicity of 3pixels,corresponding to15photoelectrons per crossing minimum ionising particle.In the Kaos/A1spectrometer,particles will cross the electron arm focal plane with an inclination angle of50−70◦with respect to the normal of the plane. Regarding this geometryfibre arrays with a hexagonal packing of slanted columns with60◦inclination were constructed,see Fig.3.Overlap and column pitch of this geometry are0.41mm.These detectors were tested in the beams at GSI.A12-layer front-end board able to accommodate the three MaPMTs was de-veloped by the Institut f¨u r Kernphysik for this type offibre detector.It sup-plies the voltage for the Cockcroft-Walton voltage multipliers and brings the analogue signals along equalised conducting paths to the RJ-45connectors for the output to the discriminators.For amplitude-compensated timing two32-channel discriminator boards,custom designed and built by the the Institut f¨u r Kernphysik,each with4integrated low-walk double threshold discrimina-tors(DTDs),were used.The DTD boards were placed in an6U crate together with a controller board.The communication with a PC was done via parallel port.The time is picked offby CATCH cards,developed for the COMPASS collaboration[11].At GSI analogue output boards with50Ωcoaxial connec-tors were attached to the discriminator boards to access also the pulse heights of each channel.The pulse height information of22channels was read out by two Model2249A CAMAC ADCs(LeCroy,US).The ADCs offer a resolution of10bits at an input sensitivity of0.25pC/count.The trigger was either derived from the second plane of thefibre detector or from a reference counter,a scintillation paddle that was installed2m up-stream in the beam-line.The time difference between individual channels and the reference counter showed a dependence on the pulse height.This time√walk effect was corrected according to the equation∆t c=∆t−c walk/focal plane of the spectrometer A has a length of approximately 2m,and it is inclined at an angle of 45◦to the reference particle trajectory.The divergence of the particle trajectories is about 24◦.The fibre detector was sandwiched between the drift chambers and the scintillator paddles of the focal plane detector system.Two vertical drift chambers (VDCs)measure the dispersive coordinate x plus the corresponding angle θand the non-dispersive coordinate y plus the angle φ.The electron hit position was found by extrapolating the reconstructed track from the focal plane to the fibre detector.Fig.4(left)shows the geometrical acceptance covered by the fibre detector inside the spectrometer,determined by such an extrapolation.The kinematics of the reaction was chosen so that the particle illumination was homogeneous over the fibre detector.The hit multiplicity of the detector was relatively high,N ≈4,the main reason being a large optical cross-talk in the MaPMT.A simple estimator for the x -position of the form x = Ni =1x i /N was used accordingly,where x i wasthe parametrised geometrical centre position of the i th channel and N the hit multiplicity.This track estimate was compared to the track reconstructed from the VDCs and projected onto the detector base coordinate,see Fig.4(right).Small non-linearities at the edges of the diagonal line indicate that better estimators based on weighted averages were needed.A detailed analysis of the correlation revealed a few misalignments in the fibre bundles caused by the demanding construction of the 45◦geometry.Fig.5(left)shows the residual of track position defined as the difference between the position reconstructed by the VDCs and the position measured by the fibre detector.A width of FWHM ∼1.1mm can be deduced from the distribution.It is assumed that the resolution of the VDC,∆x <100µm for the dispersive coordinate,was high compared to the fibre detector.The trigger detectors of the spectrometer consist of two segmented planes of plastic scintillation detectors.The arrival time of the electrons was measured in the fibre detector with respect to the following two overlapping paddles.Fig.5(right)shows the time spectra obtained from the coincidence timing with the trigger scintillators before and after performing the walk correction for the paddles and the calibration of the channel-to-channel variations of the fibre detector.The combined resolution of FWHM ≈1ns was rather good for the small amount of light from the fibres.The detection efficiency was determined by sandwiching the fibre detector between the VDC and the scintillators,and using the three-detector method.It was found to be 99%independent of the threshold.4Performance of afibre detector in a carbon at GSIIn Cave C of GSI tests of afibre detector with three bundles in a12C beam of2A GeV energy were performed.Two bundles of the detector were aligned to a single plane,and one bundle formed a parallel plane directly behind.In deducing the time resolution,an iteration over all hits in a plane including multiple hits in a channel was performed,and clusters of correlated hit times were searched for.The cluster with the time closest to the trigger signal time was retained,and within the cluster the time of thefirst arrived signal was chosen as hit time.In the algorithm a minimum time separation of10ns be-tween clusters and a hit in a coincidence window of20ns width were required.A time walk correction for the hit times was not needed.The hit time residual, defined as the difference between the two hit times in the two planes offibres, was distributed with a width of FWHM=330ps for the carbon beam,see Fig.6.No significant dependence of the time resolution on photomultiplier high voltage was observed.The time resolution of a single detector plane was√derived to be FWHM∼330ps/12≈170µm(rms).In principle,the resolution can be improved by distinguishing between hits in the overlap region of two neighbouring channels from central hits.An analysis,which required a minimum charge drop of10%compared to the neighbours was performed. The identification of the hit channel was then improved and the identifica-tion of overlapping channels became possible,but a large optical cross-talk interferes with such a requirement.The channel with the pulse height max-imum was strongly correlated to the hit time defining channel showing thatADC calibration constants and TDC off-set values had both been correctly determined.It is worth estimating the spatial accuracy of thefibre detector for set-ups without analogue read-out.Cross-talk between neighbouring channels then perturbs the reconstruction of the position of the track position by causing finite hit multiplicity.Since no absolute position of the particle tracks were known,one has to compare the hit channel mean value with the estimated po-sition using the ADC information which is assumed to be more accurate.The difference of the hit channel mean value as a simple estimate of track position and the reconstructed position was evaluated by requiring a certain minimum ADC value(typically70%of the maximum)to mimic a given discriminator voltage threshold.This difference still includes contributions from the uncer-tainties in both position estimators and the granularity of thefibre array.The distributions were measured with an average FWHM ∼0.6mm(approx.the fibre pitch),an average RMS of0.5mm,and an average FWTM <1mm. The pulse height distribution of a typical detector channel(B24)is shown in Fig.9(left).The appearance of a series of peaks below the maximum pulse height at ADC channel∼100was caused by the spread of secondary electrons and the cross-talk between channels that transport afixed fraction of the sig-nal into neighbouring channels.The hit multiplicity of one detector plane is shown in Fig.9(right).For a high voltage of−650V the mean value of the distribution was N∼5,increasing with higher voltages.The overall photo-multiplier gain increased by a factor∼2between−650V and−850V.From the distributions of the pulse height sum over all channels a relative energy resolution of∆E/E=15–20%was determined.The detection efficiency of a plane,i.e.the probability tofind at least one hit in one plane provided a hit in the other plane,was above99%.5Performance of afibre detector in a beam of different particle species at GSIAfibre detector in Cave A of GSI was tested in a p/π+/d beam of3.3Tm magnetic rigidity with dominant protons of1GeV/c momentum as well as in a carbon beam of2A GeV energy.The hit time residual was measured with a width of720ps for the beam of different particle species,see Fig.10.The time resolution of a single detector√plane was derived to be FWHM∼310ps/course,statisticalfluctuations in the number of detected photons were not affected by this reduction of the gain.The multiplicity distributions of both detector planes and are shown in Fig.11 with average values close to N=5channels.These values are the consequence of some small misalignment and mainly cross-talk in the glass window of the MaPMT.By using the pulse height information the hit channel was deter-mined as the centroid of charges.The hit position residual,defined as the difference between the two estimates in the two planes offibres,wasfitted with a width of FWHM=0.46mm for the beam of different particle species. It was to some extent compromised by gain variations.The energy response of thefibre detector was studied in the beam of different particle species.Fig.12(top)shows the distribution of the pulse height sum over neighbouring channels of one detection plane.From the Gaussianfit to the data a relative variation in the measured energy deposition,∆E/E,of 60%was derived for the dominant particle species.Fig.12(bottom)shows the energy loss vs.relative time-of-flight,in which dominant protons and sub-dominantπ+,deuteron,and3He were separated.6Concluding remarksThe performance of scintillatingfibre detectors with MaPMTs was exten-sively tested using electrons at the spectrometer facility at MAMI,12C ions of2A GeV energy as well as p/π+/d particles at GSI.The hit position was reconstructed by calculating the centroids of the charges collected from each read-out channel.Good spatial accuracy and time resolution were obtained at practically unity detection efficiency.The energy response to different particle species was studied.During tests at MAMI and at GSI the optical cross-talk caused by thefi-nite thickness of the PMT entrance window could be verified.Indeed,this behaviour of the PMT has been also recognised by Hamamatsu.Only very re-cently,a32-channel PMT with black shielding lamellae embedded in the glass window became commercially available with significantly reduced cross-talk. It is currently planned to perform in2009afirst HypHI experiment at GSI using three arrays of scintillatingfibres as well as afirst Kaos/A1experiment on the electro-production of hypernuclei at MAMI withfibre detectors in the spectrometer’s electron arm.AcknowledgementsWork supported in part by Bundesministerium f¨u r Bildung und Forschung (bmb+f)under contract no.06MZ176.T.R.Saito and his research group are granted by the Helmhotz Association and GSI as Helmholtz-University Young Investigators Group VH-NG-239and DFG research grant SA1696-1/1. References[1]J.B¨a hr,et al.,Nucl.Instr.and Meth.in Phys.Res.A348(1994)713–718.[2] F.Bosi,et al.,Nucl.Instr.and Meth.in Phys.Res.A374(1996)48–56.[3]S.Horikawa,et al.,Nucl.Instr.and Meth.in Phys.Res.A516(2004)34–49.[4]P.Achenbach,et al.,Performance of afibre detector at a12C beam of2A GeV energy,GSI Sci.Report2006,GSI,Darmstadt(2007),p.224.[5]P.Achenbach,et al.,Particle identification with afibre detector in ap/π+/d cocktail beam of3.3Tm magnetic rigidity at GSI,GSI Sci.Report 2007,GSI,Darmstadt(2008).[6]P.Achenbach,Probing hypernuclei atFig.1.Scheme of a45◦column angle geometry with4double layers offibres and a column pitch of0.6mm and an overlap of0.24mm(right).A photograph of an assembledfibre bundle(left).Fig.2.Scheme of a0◦column angle geometry with4double layers offibres and a column pitch of0.6mm and an overlap of0.24mm(right).A photograph of an assembledfibre bundle(left).Fig.3.Scheme of a 60◦column angle geometry with 4double layers of fibres and a column pitch of 0.42mm and an overlap of 0.42mm (top).A photograph of an assembled fibre bundle (bottom).y v d c (m m )x vdc (mm)x f i b r e (m m )x vdc (mm)Fig.4.Geometrical acceptance covered by the fibre detector inside the spectrometer shown by the track positions reconstructed with the vertical drift chambers (left).The reconstructed position projected onto the base coordinate versus the measured position obtained from the fibre detector with a simple estimator (right).Gaussian fit0C o u n t s (x 103)x res (mm)maximum: 10532 counts maximum at: 0.123 mm FWHM: 1.105 mm−55510C o u n t s∆t (ns)∆t + channel offsets∆t + channel offsets + walk correction Gaussian fitmaximum: 693 counts maximum at: 0.614 ns FWHM: 1.244 ns −550200400600800Gaussian fitmaximum: 811 counts maximum at: −0.074 ns FWHM: 1.048 nsFig.5.The residual of track position obtained from the drift chamber track re-construction and the simple estimator for the fibre detector (left).The residual of hit times obtained from the trigger scintillators and the fibre detector before and after walk correction for the trigger and calibration of the fibre channel-to-channel variations (right).−1.5−1.0−0.5 0.0 0.5 1.0 1.5C o u n t s (x 103)t res (ns)Gaussian fit0maximum: 34770 counts maximum at: 0.015 ns FWHM: 0.330 ns10203040Fig.6.The residual of hit times between two detector planes,t A −t B .A Gaussian fit is shown providing a width of FWHM =330ps.The time resolution of a single detector plane was derived to be FWHM ∼330ps /√12.013.014.015.013.014.015.016.0C e n t r o i d o f c h a r g e c B (m m )Centroid of charge c A (mm)Fig.7.Scatter plot showing the correlation between the centroids of charges in both detector planes.The steps appear with a pitch of ∆c A ,∆c B ≈0.6mm.−1.5−1.0−0.5 0.0 0.5 1.0 1.5C o u n t sx res (mm)Gaussian fit0maximum: 3999 counts maximum at: 0.075 mm FWHM: 0.272 mm10002000300040005000Fig.8.The residual of track position estimates between both detector planes,x A −x B ,using the centroid of charges.A Gaussian fit is shown providing a width of FWHM =0.27mm,however,the distribution is non-Gaussian with two overlapping peaks because of the discretisation in fibre channels.5010015050010001500200025003000C o u n t sADC B 24 (cnts)10203010 20 30 40 50 60C o u n t s (x 103)Multiplicity B (chn)Fig.9.Pulse height distribution of a typical detector channel (left).Distribution of hit multiplicities in the corresponding detector plane (right),which causes the series of peaks in the pulse height distribution.−1.5−1.0−0.5 0.0 0.5 1.0 1.505001000C o u n t st res (ns)Gaussian fitmaximum: 882 counts maximum at: 0.560 ns FWHM: 0.720 nsFig.10.The residual of hit times between two detector planes,t A −t B .A Gaussian fit is shown providing a width of FWHM =720ps for the beam of different particle species.The time resolution of a single detector plane was derived to be FWHM ∼510ps.102030050010001500200025003000C o u n t sMultiplicity (chn)Fig.11.The hit multiplicities of both detector planes for the beam of different particle species.The trigger was provided by the plane represented with the dashed curve.1002003004000100200300400500C o u n t sΣ ADC B (cnts)Gaussian fitmaximum: 418 counts maximum at: 105 cnts FWHM: 66 cnts−50 510100200300400500Σ A D C B (c n t s )∆t (ns)pionsprotons3HedeuteronsFig.12.Distributions of the pulse height sum over all channels of one detector plane for the beam of different particle species (top).From the Gaussian fit to the proton peak a relative energy resolution ∆E/E ∼60%was derived.The pulse height sum is shown vs.the time-of-flight so that π+,proton,deuteron,and 3He were separated (bottom).。
