Application of pharmacia automated FPLC System and PepRPC HR 5/5 bonded phase column for determination of dansylated polyamines

Summary The Pharmacia Automatized FPLC System equipped with a PepRPC HR 5/5 bonded-phase column was tested with dansyl polyamines, toluene, dimethyl- and dibutylphthalate, using either isocratic conditions or linear gradient-elution program of methanol in water as the mobile phase. A simple and reproducible method is described for the quantiation of dansylated natural polyamines, such as putrescine, spermidine and spermine originating from P388/S leukemia cells. Comparing the data from analyses performed in parallel by the Automated FPLC/PepRPC HR 5/5, and a Hewlett-Packard Model 1084B/LiChrosorb RP-8 systems a linear correlation has been found, with a regression coefficient of r=0.974.     

一种简单的制备反相电色谱柱的方法

 采用简便易行的方法自制内径为100 μm的电色谱柱,并考察了其性能,同时研究了磷酸盐缓冲液浓度、乙腈体积分数、电压、柱温对中性化合物保留行为的影响。     

开管毛细管亲和液相色谱初探

 提出了一种新的亲和色谱模式开管毛细管亲和液相色谱。在内径为 5 0 μm的毛细管内表面键合一层三嗪染料配体 ,利用毛细管电泳仪的压力系统进行亲和色谱实验。采用流动相切换洗脱技术 ,牛血清白蛋白和溶菌酶获得了有效的分离。连续运行 10次 ,溶菌酶的出峰时间、峰面积和峰高的相对标准偏差分别为0 1% ,4 3% ,3 7%。在 8 6ng～ 2 8 7ng范围内 ,溶菌酶的进样量与峰面积和峰高都呈线性关系 ,其相关系数分别为 0 9946和 0 9988。     

Dynamic Capacity Studies of CIM (Convective Interaction Media)® Monolithic Columns

The characterization of CIM^® DEAE monolithic columns in terms of dynamic binding capacity is presented in this paper. Breakthrough experiments were performed for capacity determination. Bovine serum albumin (BSA) was used as a model protein. It is shown that CIM^® monolithic columns have good batch‐to‐batch reproducibility as well as long‐term stability. The experiments performed under different linear velocities demonstrated that the dynamic capacity is unaffected at least up to a linear velocity of 2450 cm/h. Furthermore, the breakthrough curve slope is constant, indicating that the capacity would remain constant at even higher linear velocities. The adsorption isotherm of BSA dissolved in 20 mM Tris‐HCl buffer shows a constant capacity of around 30 mg/mL of support down to a concentration of 20 μg/mL. The capacity is substantially influenced by the ionic strength; however, 20% of the maximal capacity is still preserved at 0.3 M NaCl.     

Determination of fungicide residues in fruits by coupled-column liquid chromatography

Coupled-column liquid chromatography with fluorescence detection was applied to the determination of o-phenylphenol and bitertanol residues in orange and banana fruits. After extraction with a mixture of acetone, dichloromethane-petroleum ether, and ethyl acetate, an extract aliquot of 100 microL was injected directly without any additional clean-up into the chromatographic system using two reversed phase C18 coupled columns. The LC-LC approach allowed automated sample clean up of the vegetal extracts, leading to a simple and rapid analytical procedure, with limits of quantification between 0.01 and 0.05 mg kg(-1). Recovery experiments performed on orange and banana samples fortified at different concentrations (0.01 - 4 mg kg(-1)) gave average recoveries between 70 and 113% with relative standard deviations lower than 15%. The procedure developed was finally applied to orange and banana samples from different geographical locations and the results were confirmed by GC-MS.     

Rapid simultaneous determination of isoflavones in Radix puerariae using high-performance liquid chromatography-triple quadrupole mass spectrometry with novel shell-type column

A high-performance liquid chromatography (HPLC) coupled with triple quadrupole mass spectrometry (MS/MS) method was developed for rapid determination of 13 isoflavones in Radix puerariae. A novel shell-type column, namely Kinetex core-shell C(18) column (50 mm×2.1 mm id, 2.6 μm), and gradient elution were used during the analysis. The chromatographic peaks of 13 investigated compounds were identified by comparing their retention time and MS data with the related reference compounds. Multiple-reaction monitoring (MRM) was employed for the quantitative analysis with negative ionization mode. All calibration curves showed good linearity (r(2)>0.9990) within test ranges. The LOD and LOQ were lower than 0.017 and 0.873 μg/mL on column, respectively. The intra- and inter-day precisions for 13 analytes were <1.17 and 2.17%, respectively, and the recoveries were 93.1-104.4%. The validated method was applied for quantitative analysis of 13 isoflavones in 7 species of Radix puerariae. The result demonstrated that HPLC-MS/MS system with Kinetex column could be a promising analytical tool for the determination of isoflavones in traditional Chinese medicines, which is helpful for comprehensive evaluation of quality of R. puerariae.     

A study of the behaviour of some new column materials in the chromatographic analysis of Cinchona alkaloids

Summary A major problem in the HPLC analysis of alkaloids is the poor peak shape and consequently low resolution, due to the interactions of the basic alkaloids with the residual acidic silanol groups of most reversed phase materials. The performance of new packing materials specially designed for the separation of basic compounds has been studied using mobile phases without the special additives commonly applied in the analysis of alkaloids. Strongly basic Cinchona alkaloids were used as test compounds. Retention characteristics and selectivities of each material were studied, after mobile phase optimisation for the column. The influence of the major factors (nature and content of the organic modifier, pH value, salt concentration) affecting resolution was studied. The mobile phases were chosen so that they could be used in thermospray LC-MS. The addition of salts to the mobile phase improves separation but in general the modification of the mobile phase gave little change in selectivity. The performance of silicabased C₁₈ material proved superior to the polymer materials tested.     

毛细管气相色谱法快速测定羰基合成脂肪醇中的正构醇

The normal alcohols in fatty alcohols produced from hydroformylation unit were determined directly byOV-1 capillary GC method; This method is simple,rapid and accurate。The relative error is below 2%andcoefficient of variation is below 3%。     

高温毛细管气相色谱柱的制备及应用

研究了臭氧交联法制备毛细管气相色谱柱的方法,即用镀铝石英毛细管,涂渍ＳＥ３１,经臭氧交联,最高使用温度可达３６０℃。测试的结果表明其柱效高,吸附性小,热稳定性好,适用于烃类、醇类、多环芳烃及二硝基甲苯和三硝基甲苯的分离。     

气相色谱专家系统的进展

综述了国家色谱研究分析中心在气相色谱专家系统方面的进展,着重于色谱分离模式的推荐、气相色谱柱系统的推荐、气相色谱操作条件的最佳化、气相色谱在线定性定量分析四个方面;列举了在石化部门和环境监测中气相色谱专家系统应用的大量成功实例,以说明软件的实用性。