Development of a precolumn derivatization method for the determination of free amines in wastewater by high-performance liquid chromatography via fluorescent detection with 9-(2-hydroxyethyl)acridone

A simple, sensitive, and mild method for the determination of amino compounds based on a condensation reaction with fluorescence detection has been developed. 9-(2-Hydroxyethyl)acridone reacts with coupling agent N,N-carbonyldiimidazole at ambient temperature to form activated amide intermediate 9-(2-acridone)oxyethylcarbonylimidazole (AOCD). The amide intermediate (AOCD) preferably reacts with amino compounds under mild reactions in the presence of 4-(dimethylamino)pyridine (base catalyst) in acetonitrile to give the corresponding sensitively fluorescent derivatives with an excitation maximum lambda(ex) 404 mn and an emission maximum at lambda(em) 440 nm. The labeled derivatives exhibit high stability under reversed-phase conditions. The fluorescence intensities of derivatives in various solvents or at different temperatures were investigated. The method, in conjunction with a gradient elution, offers a baseline resolution of the common amine derivatives on a reversed-phase C-18 column. The LC separation for the derivatized amines shows good reproducibility with acetonitrile-water including 2.5% DMF as mobile phase. The relative standard deviations (n = 6) for each amine derivative are <4.5%. The detection limits (at a signal-to-noise ratio of 3) per injection were 0.16-12.8 ng/mL. Further research for the field of application, based on the AOCD amide intermediate as derivatization reagent, for the determination of free amines in real water samples is achieved.

Analysis of aromatic amines by high-performance liquid chromatography with pre-column derivatization by 2-(9-carbazole)-ethyl-chloroformate

2-(9-Carbazole)-ethyl-chloroformate (CEOC), a novel pre-column fluorescence derivatization reagent, has been developed for the analysis of aromatic amines. Taking five monocyclic aromatic amines (o-toluidine, aniline, 3,4-dimethylaniline, N-ethyl-p-toluidine, and p-phenylenediamine) as testing compounds, derivatization conditions such as pH of borate buffer, reaction time and fluorescent tagging reagent concentration have been investigated. By a one-step procedure, CEOC reacts readily with the aromatic amines to form stable derivatives with excitation and emission wavelengths, respectively, at 293 and 360 nm. This derivatization reaction could be finished within 20 min even at room temperature. The peak shapes of the derivatized aromatic amines can be improved greatly without any addition of competition amines into the mobile phase. Furthermore, this method can offer excellent quantitative precision with high tolerance of the matrix of samples. (C) 2003 Elsevier B.V. All rights reserved.

Determination of peptides and amino acids from wool and beer with sensitive fluorescent reagent 2-(9-carbazole)-ethyl chloroformate by reverse phase high-performance liquid chromotography and liquid chromotography mass spectrometry

A new method for the sensitive determination of amino acids and peptides using the tagging reagent 2-(9-carbazole)-ethyl chloroformate (CEOC) with fluorescence (FL) detection has been developed. Identification of derivatives was carried out by liquid chromotography mass spectrometry. The chromophore in the 2-(9-fluorenyl)-ethyl chloroformate (FMOC) reagent was replaced by carbazole, which resulted in a sensitive fluorescence lerivatizing agent CEOC. CEOC can easily and quickly label peptides and amino acids. Derivatives are stable enough to be efficiently analyzed by high-performance liquid chromatography. Studies on derivatization demonstrate excellent derivative yields over the pH range 8.8-10.0. Maximal yields close to 100% are observed with three- to fourfold molar reagent excess. Derivatives exhibit strong fluorescence and allow direct injection of the reaction mixture with no significant disturbance from the major fluorescent reagent degradation by-products, such as 2(9-carbazole)-ethanol and bis-(2-(9-carbazole)-ethyl) carbonate. In addition, the detection responses for CEOC derivatives are compared to those obtained with FMOC. The ratios AC(CEOC)/AC(FMOC) = 1.00-1.82 for fluorescence (FL) response and AC'(CEOC)/AC'(FMOC) = 1.00-1.21 for ultraviolet (UV) response are observed (here, AC and AC' are, respectively, FL and UV F response). Separation of the derivatized peptides and amino acids has been optimized on a Hypersil BDS C18 column. Excellent linear responses are observed. This method was used successfully to analyze protein hydrolysates from wool and from direct-derivatized beer. (C) 2003 Elsevier Science (USA). All rights reserved.

Dimerization of 1-butene via zirconium-based Ziegler-Natta catalyst

A zirconium-based Ziegler-Natta catalytic system has been tested in the dimerization of 1-butene. It was found that the concentration of Et2AlCl, Ph3P and PhONa as well as the reaction temperature had great influences on the activity and selectivity of the catalyst. Under the optimum reaction conditions, the conversion of 1-butene is 91.9%, and the selectivity of dimers is 76.7%. Basic ligands such as Ph3P and PhONa can inhibit isomerization of 1-butene to 2-butene effectively. In addition, the metal hydride mechanism was also suggested and some indirect evidence was obtained in favor of this mechanism.

科尔沁草甸草地放牧和割草条件下土壤种子库研究

研究了科尔沁沙地草甸草地在放牧、割草两种利用方式下的土壤种子库的大小、组成及其与地上植被的关系 .结果表明 ,在自由放牧下 ,土壤种子库密度为 6 15 8± 16 4 7粒·m-2 ,在割草利用下 ,土壤种子库密度为 8312± 2 5 4 0粒·m-2 .放牧干扰下 ,种子库组成以矮小、短命的一年生植物为主 ,其中一二年生植物占 81 6 6 % ,占比例最大的前 4种植物分别为虎尾草 (38 5 5 % )、灰绿藜 (15 4 2 % )、毛马唐 (14 95 % )和狗尾草 (9 83% ) ,多年生植物种子密度仅为 112 9± 30 2粒·m-2 ;割草干扰下 ,一二年生植物占6 8 0 8% ,其中狗尾草占 5 2 7% ;而割草地多年生植物种子密度为 2 6 5 3± 811粒·m-2 放牧地土壤种子库密度与地上植物相关不显著 ;割草地土壤种子库密度与地上植物多度显著相关 (r=0 76 ,P <0 0 1) .放牧地种子库的Shannon Wiener指数、丰富度指数分别为 2 96和 2 98,明显小于割草地的 3 10和 5 0 9,表明自由放牧更易使物种多样性下降 .

科尔沁沙地乌兰敖都地区降水特点分析

分析了科尔沁沙地西部乌兰敖都地区 2 1年的降水特点和降水趋势。年平均降水量为 31 1 .4mm ,降水集中在 5 - 9月份。夏季占全年降水量的 70 .1 % ,年平均降水日数约 76天。≥ 5 .0mm、≥ 1 0 .0mm的降雨日数全部集中于 3— 1 0月份。历年最长连续降水日数多发生于夏季 ,占 85 %。相对于年平均降水量 31 0mm而言 ,本地正常降水年份占 43%。异常降水年份占 5 7%。最长干旱持续期出现在冬春两季 ,约占 80 %。 2 1年平均降水量波动较大 ,并存在总体下降的趋势 ,可能与 1 999- 2 0 0 2年的年降雨量低于多年平均降雨量有关。

黄绵土不同形态有机氮径流流失规律

利用不同坡度径流小区 ,在自然降水条件下 ,研究侵蚀泥沙不同形态有机氮的流失规律 ,结果表明 :在 4次产流泥沙中 ,除氨基糖氮含量减少外 ,其它有机形态氮均存在增加趋势 ,以氨态氮含量增加最为显著。随坡度的下降 ,水解全氮、氨态氮和氨基酸氮含量呈增加趋势 ;泥沙中水解全氮、氨基酸氮、氨态氮、非酸水解氮均存在不同程度的富集 ,其富集率随坡度的下降而增大 ,以氨基酸氮富集程度高 ,泥沙中氨基糖态氮反而无富集现象存在 ;当坡度分别为 2 5°、2 0°、15°、10°和 5°时 ,水解全氮流失量分别为 1135 .0、76 3.7、485 .5、313.0和 15 4.4kg/ km2 ,而氨基酸氮流失量则为 2 34.8、182 .7、12 1.1、76 .8和 33.9kg/ km2 ,占水解全氮的 2 0 .6 %～ 2 4.9%。氨态氮流失量为 742 .5、5 0 3.3、32 2 .2、2 0 8.9、97.4kg/ km2 ,占水解全氮的 6 3.1%～ 6 6 .7%。氨基糖氮占酸水解氮 1.1%～ 2 .9%。非鉴别氮占水解全氮的 5 .8%～ 12 .6 %。而酸水解氮与非酸水解氮之比分别为 ...

水蚀条件下硝酸铵施用对黄绵土氮素流失的影响

研究结果表明不同坡度谷子地 ,高 N处理小区径流中铵态氮、硝态氮和有效氮浓度平均为 1.0 6、0 .76和1.82 mg/kg,低 N分别为 0 .6 4、1.2 9和 1.93mg/kg;高氮处理土壤铵态氮、硝态氮和有效氮平均流失量分别达到17.90、12 .93和 30 .84kg/(km2 · a) ,低 N流失量为 11.90、2 3.86和 35 .77kg/(km2 · a)。高氮处理小区泥沙中有机质和全氮浓度平均为 5 .2 1和 0 .5 36 g/kg,而低氮处理分别为 4.94和 0 .481g/kg;高氮和低氮处理土壤有机质流失量分别为 5 70 2和 5 743kg/(km2 · a) ,土壤全氮流失量为 498和 5 5 9kg/(km2 · a)

坡地集流梯田工程蓄水效果试验研究

为了探索在小流域土壤侵蚀治理基础上的雨水集蓄利用模式和技术,以丘陵半干旱区国家重点治理小流域为试验区,遵循坡地水量平衡和雨水叠加利用理论,将小流域综合治理与雨水集蓄利用技术有机集成,研究确定了坡地集流梯田工程的集流潜力、集流效果及其断面参数;并对该工程集蓄径流提高土壤水分和增产增收效果进行了研究。结果表明,该地区坡耕地适宜修建1∶1平坡比的集流梯田工程;与坡耕地相比,集流梯田土壤含水量提高1.96～3.12个百分点,特别是7,8,9三个月的差异更大,比坡耕地平均提高3.62个百分点,集流梯田产量提高58.28%～76.02%;产值增加70.98%～114.30%。

两种施氮水平下不同基因型冬小麦叶片光合特性与形态差异的研究

田间试验研究不施N和施N(90kg/hm2)条件下“NR9405”、“9430”、“偃师9号”、“小偃6号”、“陕229”、“西农2208”、“矮丰3号”和“商188”等8种不同基因型冬小麦中后期生理特性及其叶片形态的差异结果表明,抽穗期倒二叶和灌浆期旗叶的净光合速率、气孔导度、蒸腾速率及瞬时水分利用效率在不同基因型间存在显著差异,施N仅能显著降低抽穗期倒二叶的蒸腾速率,而对功能叶的其他生理指标无明显影响。小麦成熟期旗叶和倒二叶的长度、宽度及叶面积在不同基因型间也存在极显著差异,施N对这些叶片形态指标有极显著地促进作用,基因型和N肥同时影响灌浆期旗叶的SPAD值,而叶片衰老指数主要受基因型调控。总体上看,冬小麦叶片形态指标同时受施N和基因型影响,而生理指标主要受基因型影响,N肥的影响相对较小。

细菌二氯甲烷脱氯酶基因对拟南芥的转化

The dichloromethane dehalogenase gene (dcm) of Methylophilus was cloned into the plasmid pKYLX71-35S2 and fused with the 35S2 promotor,then transformed Arabidopsis by the infiltration mediated with Agrobacterium tumefaciens. Homozygous dcm seeds were obtained after several generations selection on the medium with 50 mg L -1 kanamycin.Northern blotting showed dcm mRNA was high in cytoplasm and the detection of DCM enzyme indicated that the lines containing high dcm mRNA expressed high DCM enzyme activities. Fig 4, Ref 9