硅胶基质高效液相色谱填料研究进展

高效液相色谱(HPLC)不仅是一种有效的分析分离手段,也是一种重要的高效制备分离技术。色谱柱是HPLC系统的核心,不同性能的填料是HPLC广泛应用的基础。硅胶是开发最早、研究最为深入、应用最为广泛的HPLC固定相基质,其制备方法主要有喷雾干燥法、溶胶-凝胶法、聚合诱导胶体凝聚法及模板法等。近年来,亚2μm小粒径硅胶、核-壳型硅胶、双孔径硅胶、介孔性硅胶、有机杂化硅胶等新型硅胶应用于HPLC并取得了色谱分离技术的飞速发展,例如基于亚2μm填料的超高压液相色谱技术、基于核-壳型填料的快速分离技术、基于杂化硅胶填料的高温液相色谱技术等。硅胶经表面化学键合、聚合物包覆等有机改性可制得先进的大分子限进填料、温敏性填料、手性填料等,大大扩展了HPLC的应用范围。本文对液相色谱用硅胶的制备方法、改性与修饰方法以及硅胶基质固定相的评价方法加以系统综述,概述了新型硅胶在HPLC中的应用进展,并对硅胶基质填料的发展方向与应用前景进行了展望。     

中药物质基础的高效液相色谱分离分析方法研究

高效液相色谱方法已成为中药物质基础研究的重要手段．在中药质量控制、中药标准品制备、活性化合物发现等方面发挥着不可替代的作用．然而我们的实验数据表明,一个中药材可能包含上万个化合物．中药物质基础的复杂性对高效液相色谱方法提出了巨大挑战．本文针对液相色谱方法在中药物质基础分离分析中存在的问题与难点,结合红花、黄连、姜黄三味药材样品,从亲水色谱分离模式、新型色谱固定相、二维液相色谱分离系统和液相色谱质谱联用技术等方面讨论了液相色谱的分离分析方法和发展方向．结果表明,高效液相色谱新技术新方法在中药复杂体系的分离分析中具有很大的发展潜力和应用前景．     

填充型纳流液相色谱柱技术的研究进展

纳流液相色谱是高效液相色谱微型化的重要形式,由于其具有溶剂消耗少、样品需求量小、检测灵敏度高以及与质谱联用兼容性好等特点,在生命科学、生物医药以及微纳分析等领域得到越来越广泛的应用。填充柱是最常用的纳流色谱柱类型,通常由球形色谱填料颗粒填充入毛细管内制成。样品组分是在色谱柱上实现分离,色谱柱的性能对色谱分析的选择性和分辨率至关重要,因此制备高质量的纳流色谱填充柱对获得高效的纳流液相色谱分离尤为关键。纳流色谱柱的制备包含两个重要技术环节：柱塞技术和填充工艺。近年来,随着柱塞与填充技术的进步,填充型纳流液相色谱柱的稳定性和分离性能得到了显著提升。本文对近十年来纳流液相色谱柱技术的研究进展,主要是柱塞技术和填充技术的研究和应用进展进行了概述,介绍了其在组学研究和生物制药领域中的应用,并对其发展前景进行了展望。     

中国科学院兰州化物所色谱技术研究开发中心

中国科学院兰州化物所色谱技术研究开发中心是我国唯一从事研制、生产高效气相色谱柱、高效液相色谱柱、液相色谱填料以及相关产品的高科技企业。中心致力于我国色谱柱产业核心技术体系的发展,独创出国际领先的液相沉渍。原位一体化合成制备高效PLOT色谱柱技术:拥有先进的超净-静态制备高效WCOT色谱柱技术:在国内率先完成了球形硅胶色谱填料制备研究开发工作,研制开发出多种液相手性分析填料以及国内急需的专用色谱柱,创造出多项     

中国科学院兰州化物所色谱技术研究开发中心

<正>中国科学院兰州化物所色谱技术研究开发中心是我国唯一从事研制,生产高效气相色谱柱.高效液相色谱柱.液相色谱填料以及相关产品的高科技企业.中心致力于我国色谱柱产业核心技术体系的发展.独创出国际领先的液相沉渍。原位一体化合成制备高效PLOT色谱柱技术.拥有先进的超净——静态制备高效WCOT色谱柱技术.在国内率先完成了球形硅胶色谱填料制备研究开发工作.研制开发出多种液相手性分析填料以及国内急需的专用     

中国科学院兰州化物所色谱技术研究开发中心

<正>中国科学院兰州化物所色谱技术研究开发中心是我国唯一从事研制、生产高效气相色谱柱、高效液相色谱柱、液相色谱填料以及相关产品的高科技企业。中心致力于我国色谱柱产业核心技术体系的发展,独创出国际领先的液相沉溃、原位一体化合成制备高效PLOT色谱柱技术;拥有先进的超净-静态制备高效WCOT色谱柱技术;在国内率先完成了球形硅胶色谱填料制备研究开发工作;研制开发出多种液相手性分析填料以及国内急需的专用色谱柱;创造出多项     

制备型高效液相色谱在天然产物分离中的应用

制备型高效液相色谱法在天然产物的分离纯化中得到广泛应用。从制备型高效液相色谱法的技术分类及其在天然产物研究中的实际应用方面对目前国内外制备型高效液相色谱技术研究现状进行了综述,对其发展动态进行了展望。     

高效制备液相色谱柱技术的研究进展

本文概述了高效制备液相色谱柱的柱型结构、填料以及柱填充方法等研究的最新进展,讨论了制备柱与分析柱的不同特征,对目前普遍使用的压缩型制备柱的类型、结构及填充方法作了较为全面的评述,总结比较了工业化制备色谱填料不同于分析色谱填料的特点,探讨了高效制备液相色谱柱技术的应用和发展前景.     

基于大孔树脂与制备液相色谱技术快速分离野地瓜茎中的绿原酸

建立了基于大孔吸附树脂快速富集,制备高效液相色谱高效分离野地瓜茎中绿原酸的制备方法:AB-8,D101,HPD600,CN206和NKA-Ⅱ5种树脂中,经静态吸附-解吸附试验发现NKA-Ⅱ型大孔树脂对目标化合物具有较好的吸附率和解析率;采用NKA-Ⅱ型大孔树脂,经4倍柱体积(bed volume,BV)5%(V/V)乙醇除杂后,用7 BV 10%(V/V)乙醇洗脱得到目标化合物组分,HPLC分析目标化合物的峰面积比达到80.8%;由制备高效液相色谱对目标化合物做进一步纯化并开发了重复进样分离模式,提高了分离效率,经纯化后目标化合物纯度达到98.6%;1H NM R和13C NM R鉴定目标化合物为绿原酸。该方法适合于野地瓜中绿原酸化合物的大规模制备。     

中国科学院兰州化物所色谱技术研究开发中心

中国科学院兰州化物所色谱技术研究开发中心是我国唯一从事研制,生产高效气相色谱柱,高效液相色谱柱,液相色谱填料以及相关产品的高科技企业。中心致力于我国色谱柱产业核心技术体系的发展,独创出国际领先的液相沉渍、原位一体化合成制备高效PLOT色谱柱技术;拥有先进的超净-静态制备高效WCOT色谱柱技术;在国内率先完成了球形硅胶色谱填料制备研究开发工作;研制开发出多种液相手性分析填料以及国内急需的专用色谱柱,创造出多项"中国第一"。早在1988年,由中国科学院推荐,国家机械工业委员会将我中心生产的系列色谱柱列为"推荐替代进口产品",中心     

Separation of Cinchona alkaloids on a novel strong cation-exchange-type chiral stationary phase—comparison with commercially available strong cation exchanger and reversed-phase packing materials

A recently reported chiral strong cation exchanger (cSCX) type stationary phase was investigated for the LC separation of a series of Cinchona alkaloids and synthetic derivatives thereof to test its usefulness as alternative methodology for the separation of those important pharmaceuticals. The cSCX column-packing material was qualitatively compared on the one hand against a commercially available non-enantioselective SCX-material, PolySulfoethyl-A, and, on the other hand, against a modern C18 reversed-phase stationary phase which is commonly employed for Cinchona alkaloid analysis. Both SCX columns showed no pronounced peak-tailing phenomena which typically hamper Cinchona alkaloid RP analysis and require specific optimization. Thus, the cSCX-based assay provided new feasibilities for the separation of the Cinchona alkaloids in polar organic mode as opposed to conventional reversed-phase methodologies. In particular, a method for the simultaneous determination of eight Cinchona alkaloids (quinine, quinidine, cinchonine, cinchonidine, and their corresponding dihydro analogs) using the cSCX column in HPLC has been developed and exemplarily applied to impurity profiling of a commercial alkaloid sample. Furthermore, both SCX materials allowed successful separation of C9-epi and 10,11-didehydro derivatives from their respective educts in an application in synthetic Cinchona alkaloid chemistry. Figure An alternative separation principle - HPLC separation of strongly basic natural Cinchona alkaloids and synthetic derivatives thereof by means of a strong cation-exchanger type chiral stationary phase     

An Application of Capillary Electrophoresis for the Analysis of Algal Toxins from the Aquatic Environment

In this work capillary electrophoresis (CE) with UV detection has been applied to the analysis of different natural toxins produced in the aquatic environment. This technique is presented as an alternative to other chemical techniques such as HPLC, and the optimisation of analytical methodologies was carried out for diverse marine toxins including Paralytic and Amnesic and some polyether toxins, such as Yessotoxins, as well as for certain microcystin toxins produced by cyanobacteria present in freshwaters. Sample preparation steps were optimised and adequate electrophoretic conditions developed for achieving a complete separation of compounds with similar structures involved in such contamination. The influence of the biological matrices where they are involved has also been studied and the potential use of CE-UV as a tool for monitoring these aquatic toxins is also discussed.     

Profiling of different bioactive compounds in functional drinks by high-performance liquid chromatography

In the present work, an HPLC method is proposed to simultaneously detect and quantify water- and fat-soluble vitamins, phenolic compounds, carotenoids and chlorophylls in a single run, by using an ultradeactivated C18 column and gradient separation using trifluoroacetic acid, water and methanol. It is shown that the HPLC method provides baseline separation of all these compounds with good resolution values in 40 min. Moreover, other figures of merit of the method show a good linear response and low detection limits for all the compounds considered in the present study. Furthermore, the usefulness of this method is demonstrated via its successful application to the analysis of different beverages from different natural origin (orange, strawberry, apple, peach pineapple, plum and blackcurrant juices, soybean milk, beers) without the need of any previous sample preparation. A good correlation is also found by comparing the total phenol content (measured by Folin-Ciocalteu method) with the sum of total phenolic compounds obtained using the proposed HPLC method. By using statistical tools, the main compounds associated with antioxidant activity of the extracts (measured by 1,1-diphenyl-2-picrylhydrazyl radical scavenging) were assessed.     

Screening of Catharanthus roseus secondary metabolites by high-performance liquid chromatography

Two direct HPLC analytical methods for the screening of the major indole alkaloids of Catharanthus roseus hairy roots and their iridoid precursors have been developed. Photodiode array and fluorescence detection were performed. The separation was achieved on a reversed-phase C18 column. The first method allowed the separation of catharanthine, serpentine, tabersonine, vindoline, vinblastine, and vincristine in 20 min. Ajmalicine, tryptophan, tryptamine and secologanine were separated using the second method in 13 min. The identification of the compounds was based on the retention time and the comparison of UV spectra with those of authentic standards. A simplified alkaloid extraction method was developed in order to accelerate sample preparation. The assays were successfully used to quantify major compounds of the secondary metabolism of hairy root cultures of C. roseus, thus providing a reliable tool for rapid screening of C. roseus secondary metabolite samples. In these cultures, ajmalicine, serpentine, catharanthine, tabersonine, and tryptamine were detected, but tryptophan, vindoline, vinblastine and vincristine were not.     

手性固定相AD、AS和OD的拆分性能

手性固定相-高效液相色谱法在手性药物、手性农药等的分离分析中应用广泛。本文采用3种多糖衍生物的手性固定相(即EnantioPak AD、AS和OD)对20种手性化合物开展手性分离研究,进而探讨样品分子结构、多糖骨架和衍生基团对手性分离的影响。结果表明,除化合物13外,其余化合物在EnantioPak AD上均实现基线分离,分离度多在2.0以上,在正己烷-醇流动相中加入酸碱添加剂可改善和优化酸性或碱性化合物的分离效果;芳香醇(化合物13~16)随着侧链碳数增加在色谱柱上的保留减弱,其分离度呈现增加的趋势;对比8种化合物在3种手性固定相上的分离结果可知,EnantioPak AD表现出更优的分离性能。这为深入研究和了解多糖手性固定相、拓展其手性分离应用提供了参考。     

Liquid chromatographic analysis of the anticancer alkaloid luotonin A and some new derivatives in human serum samples

The quantitation of the natural cytotoxic and anti‐inflammatory alkaloid luotonin A and five recently synthesized derivatives is described, constituting the first report of a HPLC method for the analysis of these compounds in human serum samples. The conditions for the chromatographic separation were optimized and the method was validated for the analysis of these compounds in biological samples according to international guidelines. An RP‐HPLC method with fluorimetric detection and a C₁₈ stationary phase was applied. Different ACN/water mobile phases were assayed, including 0–4% of a mobile phase modifier such as tetrahydrofuran, dioxane or tert‐butyl methyl ether. Isocratic and gradient elution conditions are compared. The influence of pH on the efficiency and resolution of the separation was also considered. The developed method was applied to the determination of luotonins in pooled human serum samples by gradient elution RP‐HPLC using a simple cleanup procedure. The proposed chromatographic method exhibits satisfactory analytical figures of merit, with LOD from 1.0×10^?10 to 2.0×10^?10 M, intraday and interday precision below 6% except for the concentration level closest to LOD, and good agreement between experimental and theoretical concentrations. Therefore, the developed method is suitable, reliable, rapid, and simple.     

超临界流体色谱与高效液相色谱分离手性化合物的比较

超临界流体色谱(SFC)分离具有速度快、分离效率高、溶剂消耗少等优点,近年来在手性化合物的分离分析中得到诸多应用。本文对比研究了涂覆型多糖手性色谱柱在SFC和高效液相色谱(HPLC)上拆分24种手性化合物的差异。通过比较这些化合物在色谱柱上的保留时间和选择因子等发现多数化合物在SFC上的分离效率要高于其在HPLC上的分离效率,但HPLC对轴手性化合物的分离效率要优于SFC。SFC和HPLC的分离表现出一定的互补性,随着苯环侧链烷基的碳数增加,化合物在SFC上的保留逐渐增强,而在HPLC的保留却逐渐减弱。叶菌唑在使用SFC和HPLC分析时出现了洗脱顺序反转的现象。这些结果为SFC手性拆分提供了参考。     

Total synthesis of (±)-tangutorine and chiral HPLC separation of enantiomers

The first total synthesis of racemic tangutorine, a novel indole alkaloid, was performed in 7 steps. The key reactions, dithionite reduction and acidic cyclization provided easy access with good yields to the tangutorine skeleton. Comprehensive NMR spectroscopic data of new compounds are given. Chiral HPLC separation of enantiomers is reported.     

Improved chromatographic separation of thio-arsenic compounds by reversed-phase high performance liquid chromatography-inductively coupled plasma mass spectrometry

A new group of arsenic species, thio-arsenicals, have recently been reported in several natural samples such as molluscs, algae, and urine. These compounds are the sulfur analogues of oxo-arsenicals, a large group of naturally-occurring compounds, whereby the arsinoyl (As=O) group is substituted by an arsinothioyl group (As=S). The most common separation technique for oxo-arsenicals is anion-exchange HPLC with polymer-based columns, but under these conditions the thio-arsenicals show strong retention, resulting in unacceptably long analysis times and broad peaks. We report the development of a reversed-phase HPLC method, with ICPMS detection, which allows separation of the known thio-arsenicals within 15 min with significantly improved peak shapes. The detection limit is about 0.6 microg As/L based on 10 microL injection volume. Further, we have applied the method to the identification and quantification of thio-arsenic species in two standard reference materials, BCR 710 oyster tissue and NIES 18 human urine.