150 resultados para FINGERPRINT CHROMATOGRAPHIC
Resumo:
课题组在不断地创制新的同源四倍体材料的同时,连续多年以提高结实率为目的培育、筛选自交系材料,已获得自交繁殖十二年的高代自交系材料。相对于诱导创制初期,材料表现出的结实率低,同种系单株间的差异较大;高代材料已表现出较显著的结实率提升和较一致的农艺性状表型。 本实验选取课题组多年培育的同源四倍体水稻高代自交系材料,通过形态学、农艺性状和细胞遗传学比较,研究水稻同源四倍体与二倍体之间的异同。结果显示,所有同源四倍体材料的染色体组成均为2N=4X=48,花粉母细胞(PMC)减数分裂行为较正常,99%以上的染色体都能在减数分裂中期I(MI)发生联会、配对,形成四价体和二价体,这与理论染色体组构成相符。在减数分裂过程中,结实率较高的材料染色体异常现象较少而结实率较低的材料染色体异常现象较严重。统计分析表明,二价体和四价体的比例对结实率没有显著影响,但是三价体的数目对结实率有一定影响。这一结果表明了结实率和细胞减数分裂行为可能存在相关性,同源四倍体的减数分裂行为为筛选高结实率的同源四倍体种系提供了依据。 然后,对同源四倍体水稻高代自交系材料进行农艺性状和品质性状的统计与分析。主要针对结实率、每穗实粒数、有效分蘖和穗长等主要农艺性状,以及直链淀粉含量这一重要的品质性状进行统计。将统计结果与1996年诱导加倍的初代材料的数据相对比分析,结果显示所有材料经过多代选育培养,其农艺性状已经有了较显著的提高,同时同源四倍体材料的农艺性状稳定性也有了较显著的提升。如结实率的提高幅度较大,所有材料的平均结实率均显著高于加倍初代,而同种材料不同单株间的结实率差异也显著地减少,变异系数(CV)的平均值由1996年的41.15%减少到了2008年的28.81%。其他重要农艺性状也有不同程度的改良,种内变异系数也相应地降低。此外,实验测量了同源四倍体材料和来源二倍体材料的直链淀粉含量。分析结果显示,部分材料的直链淀粉含量与二倍体亲本产生了较显著的差异,这可能是诱导加倍过程中的遗传变异造成的;同源四倍体材料的种内变异系数(CV)平均值由1996年的6%下降到了2008年的3.88%,显示出在品质性状方面,同源四倍体材料的遗传稳定性也有较显著的增加。同源四倍体材料农艺性状经过多年的选育,表现出一定的提升,同时,经过多年自交纯化,所有材料种系内的性状差异逐渐缩小,说明同源四倍体水稻的遗传稳定性随着自交纯化而增强,这为同源四倍体水稻的进一步选育打下了良好的基础。 最后,通过测量连续两年的自交系材料的遗传多态性,分析材料间遗传差异和种群遗传结构,深入研究连续两代材料间的遗传差异,研究同源四倍体水稻与二倍体材料遗传稳定性之间的差异。实验采用18对SSR微卫星标记对连续两代15个材料,共94份样本进行差异分析。通过扩增条带长度多态性分析,计算不同材料以及同种材料不同世代间的遗传距离,构建同源四倍体和二倍体水稻的分子指纹库,并绘制聚类图。结果显示,同源四倍体和二倍体不同材料间的遗传差异比较大,遗传距离处于0.4757至0.2816之间;而相同品种不同世代材料间的遗传差异较小,但也表现出一定的遗传差异。同种同源四倍体材料不同世代间的遗传差异比二倍体材料更大,两代四倍体材料间遗传距离处于0.1359至0.0485之间;而两代二倍体材料间的遗传距离处于均小于0.0388。结果表明,同源四倍体水稻高代材料具有一定的遗传稳定性,但与来源二倍体材料相比,其世代间的遗传变异性仍然较强。这种结果说明,经过多代的自交纯化培育,同源四倍体水稻材料能够建立起相对稳定的遗传结构,同时,其强于二倍体亲本的变异性有能够为新品种的选育,农艺性状、品质性状的改良提供一定的遗传基础。此外,分析结果表明通过分子标记辅助检验,水稻材料间的遗传多态性能够有效地区分不同的品种,这为水稻品种的分子鉴定提供了一定的依据。 本研究从细胞学鉴定,农艺性状统计分析以及分子标记辅助聚类分析多方面地对同源四倍体水稻高代系进行了研究,对探究同源四倍体水稻的遗传规律,进一步揭示其遗传特性、农艺性状的遗传构成,为进一步选育优质的多倍体水稻提供了一定的理论依据。 This group insists on creating new Autotetraploid Rice (Oryza sativa L.) materials, while improving the seed-setting of them for many years, cultivated and selected the inbred line materials, has obtained the high generation inbred lines after twelve years cultivation. Compared to the early induced materials, which shown the low seed setting, and the large difference between the different plants in the same germ-line; the high generation materials have shown significant improvement in seed setting and more uniform phenotype agronomic traits. The autotetraploid rice high generation inbred lines material, which has been cultivated for more than 12 years, was chose in this experiment. The similarities and differences between autotetraploid and diploid rice was studied through morphological, agronomic and cytogenetic ways. The results showed that all the chromosome of autotetraploid materials are composed of 2N=4X=48, the pollen mother cells (PMC) meiosis behavior is normal, more than 99% chromosomes in metaphase I(MI) were federated and paired to form tetravalents or bivalents, which constitutes a consistent theory of genome. In the meiosis process, the material with a higher seed setting showed less chromosome abnormal than the material whose seed setting is lower. However, statistical analysis showed that the bivalent and tetravalent rate had no significant impact on seed setting, but the number of trivalent had a certain impact on seed setting. The result shows that the seed setting may be related to the meiosis behavior, which provides a basis to cultivate new autotetraploid germ line with high seed setting through the meiotic behavior. Furthermore, the agronomic and quality traits of autotetraploid rice high generation inbred material were statistically analyzed. The statistically analysis was focused on major agronomic traits such as: seed setting, grains per panicle, effective tillers and panicle length, as well as the important quality trait amylose content. The statistic data was compared with the data in 1996, when the first induced generation of autotetraploid material, and the result shows that after a multi-generation breeding, the agronomic traits has been significantly improved in all the materials, while the stability of agronomic traits also significant upgraded. For instant, the seed setting increased significantly, the average seed setting of all materials was significantly higher than the first induced generation, and the differences between different plants in the same species also significantly reduced, the average of the coefficient of variation (CV) was reduced from 41.15% in 1996 to 28.81% in 2008. Other important agronomic traits had improved in different degrees; the coefficient of variation within species is also reduced accordingly. In addition, the amylose content of autotetraploid and diploid materials was measured in this experiment. The results shows that the amylose content of some of the material differed from diploid parents significantly, it may caused by the genetic change during the inducing, autotetraploid materials intra-specific coefficient of variation (CV) average reduced from 6% in 1996 to 3.88% in 2008, shows that this is a significant increase of quality traits stability in autotetraploid rice. Agronomic traits of autotetraploid material shows some improvement after years of breeding, at the same time, after years of purification, all material within the germ-line gradually narrow the differences in traits indicates that autotetraploid rice genetic stability was enhanced, which laid a good foundation for the further autotetraploid rice breeding. Finally, this experiment studied the genetic differences between materials of two generations and researched the difference of genetic stability between diploid and autotetraploid rice materials through investigating the genetic polymorphism, genetic differences between materials and population genetic structure of inbred line materials of two consecutive years.18 pairs of SSR microsatellite markers for 15 materials of two generations were used in this experiment, and the total of 94 samples were analyzed. Through the amplification length polymorphism analysis of different materials and materials in different generations, the genetic distance between materials and generations was analyzed, a diploid and autotetraploid rice molecular fingerprint database and map rendering cluster were constructed. The result shows that the genetic distance is between 0.4757 to 0.2816 among different autotetraploid and diploid materials; the genetic distance between different generations of same species was less, but also shows a certain degree of genetic differences. The inter-generational genetic differences of autotetraploid materials were greater than of the diploid materials, which are 0.1359 to 0.0485 as the genetic distance; comparing with the 0.0388 of diploid materials. The result shows that high generation inbred autotetraploid rice material has a certain genetic stability, but the genetic variation between generations is still strong comparing with the source diploid materials. It indicates that, after many generations of purification cultivation, autotetraploid rice materials established a relatively stable genetic structure, at the same time, stronger variability than its diploid parents are useful in the breeding of new varieties, provides a genetic foundation to the agronomic and quality traits improvement. In addition, the analysis result shows that the through the molecular marker-assisted testing, rice genetic polymorphism between materials can effectively distinguish different species, provides a certain basis for molecular identification of varieties of rice. A series of investigation such as cytological identification, statistical analysis of agronomic traits, molecular marker-assisted cluster analysis was applied in this experiment to research genetic pattern of autotetraploid rice high generation inbred lines, revealed the genetic characteristics and the genetic composition of agronomic traits, provides a theoretical basis for the further selection of high quality autotetraploid rice.
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In this paper, bioconversion of trans-cinnamic acid(t-Ca)to L-phenylalanine (L-phe) has been investigated by using immobilized yeast cells with induced L-phe Ammonia-lyase(PAL, EC.4.3.1.5) as biocatalysts. The contents are the following. (1) Thirty strains of yeasts, including two genera (Rhodotorula, Sporobolomyces), six species (R. glutinis R. minuta,R.rubra,R.sineses,R.roseus and S.salmonicolor)were screened for their ability to converse the substrates, t-Ca and ammonia, to the product, L-phe, by using yeast cells as biocatalyst, and primary evaluation for PAL activity of the selected strains was investigated. From the results of the screening experiments, it was found that 22 strains were able to produce L-phe from t-Ca with the range of conversion yield from 2% to 67%. Studies on PAL formation time course during cultivation show that the maximum PAL activity of several different strains ranges from 2.3 to 14.4×10-3U/mg cell dry weight. The biomass of tested strains at their maximum enzyme activity is also greatly varied. (2)One of the selected strains, R. rubra as 2.166, was used for immobilized cells as biocatalysts to produce L-phe. The optimum conversion conditions and effective stablization agents were investigated. The results shown that polyacrylamide gel was chosen as a suitable matrix for immobilization of the yeast cells, and it can retain 88% of the PAL activity in the reverse direction at the following reactive conditions: [t-Ca]: 34mM. [NH4OH]: 6.OM.PH10.00, temperature: 30℃. (3) The effects of various kinds of effectors on the production of L-phe were also examined. Membrane permeabilizing agents can stimulate L-phe synthesis, but make the stability of PAL decline greatly. Polyalchoholic agents and glutamic acid were very effective for the stabilization of PAL. At the presence of glutamic acid (5%), the half life of L-phe productivity with the immobilized cells was extended to 192 hours, which was much higher than most of that having been reproted, while the half life of resting cells was only about 15 hours. (4) Use of initial velocity studies on the kinetics of enzyme-catalized reaction indicated that the apparent Km value was 13.0mM for the immobilized cells, and 4.8mM for the resting cells. Thermostability of the immobilized cells was better than the resting cells. Fluid bed bioreactor is more effective than batch bioreator in prolonging the thermostability of the biocatalysts. (5) CGA- 688 resin column chromatographic procedure was employed in the isolation and purification of L-phe, t-Ca and other substances from the reactire mixture. (6) Preparative-scale production of L-phe on a level of gram amount by immobilized cells from the culture broth of R. rubra AS2.166 allowed for the conversion yield with 30%. The characteristic physico-chemical criteria (including melting point, optical activity, elements analysis, IR, NMR) are the same with the standard L-phe. 本文报告了利用诱导的苯丙氨酸解氨酶 (PAL.EC.4.3.1.5)催化反式肉桂酸(t-Ca)氨加 成制备L-苯丙氨酸(L-phe)的研究,主要内容为:(1) 我们搜集了三十株酵母菌株,利用全细胞转化t-Ca生成L-phe的能力进行了直 接筛选,并对其PAL活性水平进行了初步评估研究。研究结果表明,其中22株酵母具有转化t-Ca生产L-phe的能力,它们包括 Rhodotorula glutinis,R.rubra, R.sineses 和Sporobolomyces roseus 的菌株,转化率在2-67%。细胞生长和PAL形成过程的研究 表明,不同菌株PAL最大活力在2.3-14.4×10-3U/mg 细胞干重,达到最大PAL活性时各株酵母的生长情况也极不一致。(2) 利用筛 选出的一株深红酵母R.rubra AS2.166 作为供试菌株,研究了细胞固定化条件下生物转化的最适条件及PAL在固定化条件下的稳定 性。结果表明以聚丙烯酰胺凝胶包埋法较为理想,能使细胞合成L-phe活力保持88%,最适t-Ca浓度为34mM,最适NH4OH浓度为6M,最 适PH10.0,最适温度45℃。(3) 多种效应物对L-phe 合成的影响研究表明:表面活性剂能刺激L-phe的合成,但使PAL稳定性下降。 多羟基化合物及Glu对PAL的稳定十分有效在有Glu存在下,能使固定化细胞合成L-phe的半寿期达192小时左右,高于大部分现已报 导的固定化结果。(4) 用初速度法研究了深红酵母AS2.166中PAL的酶促反应特征,测得固定化细胞对t-Ca的表观米氏常数Km为 13.0mM,全细胞为4.8mM,细胞固定后热稳定性提高。(5) 建立了适合低浓度分离纯化产物与底物的聚苯乙烯大孔树脂柱层析技术 ,能使L-phe与t-Ca及产物混合物中其它成分有效分开。(6) 利用固定化的R.rubra AS2.166细胞所做的制备实验能够使L-phe的产 率达到30%左右,其主要的理化指标(包括熔点、比旋光度、元素分析、IR、NMR等)与标准L-phe一致。
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Based on Monte Carlo method the microscopic gas-phase adsorption chromatographic behaviors of volatile chemical compounds were simulated,and the migration process of those compounds followed by the carrier gases along the chromatographic surface and the process of adsorption-desorption were also described.The programs have been compiled to simulate the processes of thermochromatography and isothermal chromatography with different experimental conditions and properties.The calculated results agree well with ...中文摘要:用蒙特卡罗方法模拟了易挥发化合物的气相色谱微观动力学行为,描述了化合物随气流在固体色谱柱表面的迁移以及吸附-解吸等过程。按气相色谱微观动力学模型编译的计算机程序模拟了超重元素化合物在等温色谱及热色谱的动力学过程,并根据不同实验条件进行了大量计算。计算结果与实验数据较为吻合。讨论了超重元素的半衰期、吸附态的周期、载气流量以及化合物的质量密度等对实验结果的影响及该理论模型的优点和待改进的地方。
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An algorithm for the enhancement of fingerprints was presented;an algorithm for realizing the foreground/background segmentation was also discussed,based on a new mathematical transform,namely, the second generation curvelet transform.The result shows that the algorithm is effective and the transform is attractive.分析了现有的一些增强、分割算法的不足,提出了基于第二代Curvelet变换的指纹图像预处理算法.实验结果表明提出的算法简单有效,能够实现指纹图像增强的要求.
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In order to study the gas-phase chemical behavior of transactinides, an on-line isothermal chromatography apparatus has been developed and applied to separate short-lived technetium isotopes in the form of TcO3 from fission products. The fission products from a Cf-252 source were continuously and rapidly transported through the capillary to the isothermal chromatography apparatus using the N-2/KBr gas-jet techniques. Volatile oxide molecules were formed at the reaction zone kept at 900 degrees C since a trace amount of oxygen existed in the N-2 carrier gas. With the new developed isothermal chromatography apparatus, a selective separation of Tc from fission products was achieved. After isothermal chromatographic separation, Tc-101,Tc-103,Tc-104,Tc-105,Tc-106,Tc-107,Tc-108 were dominantly observed together with their Ru daughters in the gamma-spectrum, The chemical yields of Tc-101, and Tc-104 and Tc-105 isotopes with longer half-lives are about 55-57%, and those of Tc-103, Tc-106 and Tc-108 isotopes with shorter half-lives dropped down to 25-28%. The adsorption enthalpy of the investigated compounds on quartz surfaces was determined to be -150 +/- 5 kJ/mol by fitting the measured retention curves with a Monte Carlo model. The observed species of technetium oxide is attributed to TcO3, which is in good agreement with previous experimental results. That means our system worked properly and it can be used to investigate the gas-phase chemical behavior of transactinides.
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本论文系统地描述了指纹图像自动识别系统的理论、算法及进行了相应的讨论,对批纹图像预处理中的各种技术进行了分析。在遵循指纹图像处理流程:预处理、分类及特征比对的基础上,本论文重点讨论了指纹图像预处理技术,这也是本文的主要目的。在指纹图像的增强处理当中,本文首先实验了几种常用的图像增强技术。直方图修改技术,对比度扩展技术、图像平滑及图像锐化技术,其中图像平滑技术分别实验了邻预平均法、中值滤波法以及低通滤波法;图像锐化技术也分别实验了拉普拉斯算子锐化、理想高通滤波器、梯度算子锐化。在比较了各种图像增强的实际效果后,提出了在以图像平滑及图像锐化两种方法联合的基础上进行指纹图像的增强。在指纹图像的修饰及二值化处理中,本文实验了固定阈值法、直方图法、微分直方图法、多阈值法以及李硕同学的研究成果,给出了一种较好的指纹图像修饰及二值化方法。作为指纹图像预处理以及数字图像处理技术的一种探索,本论文实验了各种图像处理技术,包括边缘检测、图像平滑、图像锐化、对比度扩展等。并比较了在指纹图像处理这一特定的领域中各自的优劣,以及在总结李硕同学的研究结果的同时,提出了一种比较好的指纹图像预处理技术,并在实验中得到了验证。最后,本文对指纹图像的分类以及特征比对也给出了一些探讨。
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To avoid the limitation of the widely used prediction methods of soil organic carbon partition coefficients (K-OC) from hydrophobic parameters, e.g., the n-octanol/water partition coefficients (K-OW) and the reversed phase high performance liquid chromatographic (RP-HPLC) retention factors, the soil column liquid chromatographic (SCLC) method was developed for K-OC prediction. The real soils were used as the packing materials of RP-HPLC columns, and the correlations between the retention factors of organic compounds on soil columns (k(soil)) and K-OC measured by batch equilibrium method were studied. Good correlations were achieved between k(soil) and K-OC for three types of soils with different properties. All the square of the correlation coefficients (R-2) of the linear regression between log k(soi) and log K-OC were higher than 0.89 with standard deviations of less than 0.21. In addition, the prediction of K-OC from K-OW and the RP-HPLC retention factors on cyanopropyl (CN) stationary phase (k(CN)) was comparatively evaluated for the three types of soils. The results show that the prediction of K-OC from k(CN) and K-OW is only applicable to some specific types of soils. The results obtained in the present study proved that the SCLC method is appropriate for the K-OC prediction for different types of soils, however the applicability of using hydrophobic parameters to predict K-OC largely depends on the properties of soil concerned. (C) 2004 Elsevier B.V. All rights reserved.
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The dynamic flow profiles and separation performances in conically shaped preparative liquid chromatographic columns (inlet i.d. larger than outlet i.d.) with three different angles (7, 10 and 15degrees) were studied and compared with cylindrical column of the same length and internal volume. The shapes of dynamic flow profiles were studied by on-column visualization method. The transparent chromatographic columns made of polymethyl methacrylate (PMMA), packed with C-18 bonded silica, were immerged into a cubic pool filled with glycerol to eliminate the cylindrical and conical lens effect. The flow profiles of colored iodine solution in the columns were observed clearly using cyclohexane as mobile phase since the refractive indices of C-18, column wall and the mobile phase are very close. In the conical column of 15degrees (20-7 mm i.d.) the mobile phase in the central region migrated slower than in wall region as it moved toward the column outlet, while in the conical column of 7degrees (17-11 mm i.d.) the mobile phase in the central region migrated faster than in wall region just like in cylindrical column. We found that a plug-like flow profile was generated in the conical column of 10degrees (18-9 mm i.d.) during the whole migration process. A carmine and brilliant blue mixture was used as a probe to test the separation ability of the columns. The resolutions of the two compounds on the conical column of 7, 10, 15degrees and on the cylindrical column were 0.6, 1.57, 1.29 and 0.8, respectively. (C) 2003 Elsevier B.V. All rights reserved.
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A comprehensive two-dimensional liquid chromatographic separation system based on the combination of a CN column and an ODS column is developed for the separation of components in a traditional Chinese medicine (TCM) Rhizoma chuanxiong. Two columns are coupled by a two-position, eight-port valve equipped with two storage loops and controlled by a computer. The effluent is detected by both the diode array detector and atmospheric pressure chemical ionization (APCI) mass spectrometer. More than 52 components in the methanol extract of R. chuanxiong were resolved and 11 of them were preliminary identified according to their UV and mass spectra. (C) 2004 Elsevier B.V. All rights reserved.
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Three molecularly imprinted monolithic columns with different length but almost identical column volume had been prepared. It was observed that the separation factors of diastereomers and enantiomers were almost unaffected by column length. However, the short column with dimension of 38 mm x 8 mm W. showed much lower resistance to flow rate so that it could be operated at much higher flow rates. By combining stepwise gradient elution with elevated flow rate, the diastereomers of cinchonine and cinchonidine and the enantiomers of Cbz-DL-Trp and Fmoc-DL-Trp were successfully separated within 3 min on the short column with dimension of 38 mm. x 8 mm i.d.. Based on the above results, a cinchonine imprinted monolithic disk with dimension of 10 mm x 16 mm W. was further developed. The SEM image and the pore size distribution profile showed that large flow-through pores are present on the prepared monolith, which allowed mobile phase to flow through the disk with very low resistance. Chromatographic performances on the monolithic disk were almost unchanged compared with the long columns. A rapid separation of cinchonine and cinchonidine was achieved in 2.5 min at the flow rate of 9.0 ml/min. Furthermore, it was observed that there was almost no effect of the flow rate on the dynamic binding capacity at high flow rates. In addition, the effect of the loading concentration of analytes on the dynamic binding capacity, namely adsorption isotherm, was also investigated. A non-linear adsorption isotherm of cinchonine was observed on the molecularly imprinted monolith with cinchonine as template, which might be a main reason to result in the peak tailing of template molecule. (C) 2004 Elsevier B.V. All rights reserved.
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The dynamic flow profiles and column efficiencies in conically shaped semi-preparative liquid chromatographic columns (inlet ID larger than outlet ID) with two different conical angles (7degrees and 15degrees) were studied. The dynamic flow profiles were studied by an on-column visualization method. Conical columns were compared with cylindrical column of the same length and internal volume. The results showed that the flow profile of a sample band in the conical column of 7degrees (50 mm x 17 mm --> 11 mm ID) was parabolic in shape. The sample band migrated slower in the wall region than in the central region, as in the cylindrical column (50 mm x 14 mm ID). However, the sample band in the conical column of 15degrees (50 mm x 20 mm --> 7 mm ID) migrated slower in the central region than in the wall region, resulting in a reverse parabolic flow profile, in contrast to that in cylindrical column. This indicated that a flat flow profile might be realized in a conical column with a conical angle between 7degrees and 15degrees. The conical column of 15degrees had the highest column efficiency among the three columns under the same conditions. Compared with the cylindrical column packed with identical packing material, the conical column of 15degrees had 22%-45% higher column efficiency and 11%-27% higher peak height.
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An electroosmotic pump (EOP) capable of generating pressure above 3 MPa and mul/min flow rate with reverse phase mobile phases of HPLC was constructed and evaluated. The pump consisted of three parallel connected fused silica capillary columns (25 cm x 320 mum I.D.) packed with 2 mum silica materials, hollow electrodes, a high voltage DC power supply, and. a liquid pressure transducer. The EOP was applied in a capillary liquid chromatographic system for mobile phase delivery instead of a mechanical pump. Standard samples containing thiourea, naphthalene, anthracene, phenanthrene and acetonitrile were separated on a 15 cm x 320 mum I.D. 5 mum Chromasil C-18 packed capillary column with acetonitrile/water as mobile phase. (C) 2003 Elsevier Science B.V. All rights reserved.
Resumo:
The transport processes of components in capillary electrochromatographic column was investigated based on the basic model of relaxation theory. A principal transport equation of chromatographic relaxation theory was established and mathematical expressions for eluting curves were obtained under the situations of both capillary electrophoresis and chromatography. Characteristics of peak symmetry and its effecting factors are discussed. Tailing peaks, symmetrical peaks and fronting peaks would be observed simultaneously, which was further proved with reversed capillary electrochromatographic experiments.