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Chinese Journal of Chromatography
2009, Vol. 27, No. 5
Online: 30 September 2009

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Developments and applications of multidimensional liquid chromatography in proteomics ZHU Guijie, LIANG Zhen, ZHANG Lihua, ZHANG Yukui 2009, 27 (5):  518-525.  Abstract ( 2573 )   [Full Text(HTML)] () PDF (314KB) ( 956 )   This review summarizes the recent developments of multidimensional liquid chromatography (MDLC) techniques, and their applications in proteomics study, including expression profiling, post-translational modification, quantitative analysis, and so forth. In addition, the prospect and further developments of MDLC are discussed.

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Recent developments in preparation of monolithic columns and their applications in proteomic analysis ZOU Hanfa, WU Minghuo, WANG Fangjun, WU Ren’an, YE Mingliang 2009, 27 (5):  526-536.  Abstract ( 3124 )   [Full Text(HTML)] () PDF (664KB) ( 1057 )   Monolithic column is a continuous unitary porous structure prepared by in-situ polymerization or consolidation inside the column tubing. Monolithic columns have been attracted great attention due to their simple preparation procedure, high permeability and high mass transfer rate. Big progress on the technology and methods for the preparation of monolithic columns and their applications to the highly efficient separation and pretreatment of biological samples have been made in last decade. This review summarizes the developments in the preparation of the monolithic columns and their applications to the proteomic analysis during the last five years.

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Advances of separation and analytical techniques in proteomic studies ZHANG Yangjun, ZHANG Wanjun, MA Yan, PENG Bo, QIAN Xiaohong 2009, 27 (5):  537-550.  Abstract ( 2184 )   [Full Text(HTML)] () PDF (437KB) ( 1078 )   The separation and analytical technique is one of the core technologies in proteomic studies. The advances in the applications of separation and analytical techniques in proteomic studies on protein expression profiling, protein modification profiling, protein complex and interaction network as well as proteome quantitation are reviewed, and 89 references are cited.

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Advances in multidimensional high performance liquid chromatography for separation technology in proteomic study GAO Mingxia, GUAN Xia, HONG Guangfeng, ZHANG Xiangmin 2009, 27 (5):  551-555.  Abstract ( 2224 )   [Full Text(HTML)] () PDF (215KB) ( 1026 )   With the developments of proteomic study, multidimensional high performance liquid chromatography technology has attracted increasing interests due to its obvious advantages, such as rapid analysis, high automation and easy combination with mass spectrometry and so on. This review emphasizes the advances of multidimensional high performance liquid chromatography technology, including classical bottom-up methods, top-down techniques and array-based two-dimensional liquid chromatography system, which was designed and set up by our lab to improve the throughput. These techniques showed promising potential applications in proteomics study.

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Recent advances in aptamer affinity chromatography ZHAO Qiang, LE X Chris 2009, 27 (5):  556-565.  Abstract ( 2842 )   [Full Text(HTML)] () PDF (334KB) ( 1397 )   Aptamer affinity chromatography makes the use of aptamers as affinity ligands to achieve chromatographic separation and concentration. Aptamers are single stranded oligonucleotides that can specifically bind to targets. They show unique advantages over antibodies in preparation, stability and applications. This article summarizes the applications of aptamer affinity chromatography to the separation and analysis of small molecules, proteins, and cells. By critically evaluating literature, this review describes the current status and discusses the future prospects of aptamer affinity chromatography.

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Applications of chromatography-mass spectrometry in metabonomics HUANG Qiang, YIN Peiyuan, LU Xin, KONG Hongwei, XU Guowang 2009, 27 (5):  566-572.  Abstract ( 2403 )   [Full Text(HTML)] () PDF (347KB) ( 1583 )   Metabonomics (or metabolomics) is one of platforms for function genomics and systems biology. Metabonomics focuses on the changes of low molecular mass metabolites in complex samples such as biofluids, cells and tissue/tissue extracts. Although it has not been possible for quantitative measurements of global metabolite profiling, the combination of chromatography-mass spectrometry and chemometrics has secured a central role in metabonomics research. Here, applications of chromatography-mass spectrometry in metabonomics are reviewed, the advantages and disadvantages of various separation techniques including gas chromatography, liquid chromatography and capillary electrophoresis are emphasized, their further developments in metabonomics field are prospected.

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Photonic crystals for analytical chemistry CHEN Yi, LI Jincheng 2009, 27 (5):  573-583.  Abstract ( 2298 )   [Full Text(HTML)] () PDF (640KB) ( 1130 )   Photonic crystals, originally created to control the transmission of light, have found their increasing value in the field of analytical chemistry and are probable to become a hot research area soon. This review is hence composed, focusing on their analytical chemistry-oriented applications, including especially their use in chromatography, capillary- and chip-based electrophoresis.

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Recent advances of gas chromatography FU Ruonong 2009, 27 (5):  584-591.  Abstract ( 2506 )   [Full Text(HTML)] () PDF (233KB) ( 1210 )   The recent advances and features of gas chromatography (GC) are briefly described. GC is a considered as a mature technique, and is applied in various fields. Now, in addition to stationary phases and columns, the investigations are focused on comprehensive two-dimensional gas chromatography (GC×GC), fast GC, portable GC, and micro GC (μGC). In the research of stationary phases, the room temperature ionic liquids continue its development, and modified cyclodextrins are other projects of GC stationary phases studied. Today a lot of chromatographers prefer to use commercial capillary GC columns rather than the home made one, and most of the stationary phases of commercial capillary GC columns used are 5% phenyl polydimethylsiloxane.

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Advances in micro gas chromatography GUAN Yafeng, WANG Jianwei, DUAN Chunfeng 2009, 27 (5):  592-597.  Abstract ( 2870 )   [Full Text(HTML)] () PDF (160KB) ( 1196 )   Miniaturization of analytical instruments has attracted great attentions both from scientists and industry because of their unique properties such as small volume, high performance, low power and material consumption. Gas chromatograph (GC) is one of the most widely used analytical instruments. Many efforts have been made to develop portable or micro GC for on-line, in-situ and fast analysis in field. In this review, the state of arts in the field of micro GC is summarized. The advances on micro GC including injectors, chromatographic separation columns and detectors are discussed separately. The applications of micro GC are also briefly reviewed. Finally, a prospect on the developments and applications of micro GC is outlined.

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Recent developments in capillary electrophoresis-mass spectrometry ZHOU Zhigui, LI Min, BAI Yu, LIU Huwei 2009, 27 (5):  598-608.  Abstract ( 2575 )   [Full Text(HTML)] () PDF (419KB) ( 1029 )   Capillary electrophoresis-mass spectrometry (CE-MS) has become a well-accepted analytical approach by combining high separation power, wide applicability of CE with high sensitivity, molecular information availability of MS. In this review, the new advances in CE-MS interface over the past few years are presented, and the recent applications are summarized in the fields of life sciences, food science and pharmaceutical analysis and so on.

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Recent advances and applications of capillary electrochro-matography and pressurized capillary electrochromatography WU Yi, ZHANG Xiaohui, WEI Juan, XUE Yunyun, BAHATIBIEKE Marjan, WANG Yan, YAN Chao 2009, 27 (5):  609-620.  Abstract ( 2467 )   [Full Text(HTML)] () PDF (326KB) ( 940 )   Capillary electrochromatography (CEC), in which electroosmotic flow (EOF) created from the electrical double layer is made to act as a pump to drive the mobile phase in a capillary column packed with micro-particulates or coated with stationary phase. Both neutral and charged species can be resolved by CEC. It has been demonstrated that the efficiency of a separation obtained by electroosmotic propulsions is superior to that obtained by pressure-driven flow (as is the case in HPLC). CEC combines the best features of CE and versatile selectivity and large sample capacity of HPLC, promising high efficiency, high resolution, high selectivity and high peak capacity. However, in practice, when CEC is used without pressure, often used on a commercial CE instrument, there are problems and difficulties associated with bubbles formation and column dry-out. These difficulties can be overcome by a pressurized CEC (pCEC) system, in which a supplementary pressure is applied to the column in addition to the EOF. In such a system, a pressure can be applied to the capillary column to suppress bubbles formation. Quantitative sample introduction in pCEC can be easily achieved through a rotary-type injector. Most importantly, it is amenable for a solvent gradient mode, similar to that in HPLC, by programming the composition of mobile phase. The article brings a comprehensive survey of recent development of CEC and pCEC, including the development of instrumentation, capillary columns and stationary phase as well as CEC and pCEC applications in life science, biotechnology, pharmaceutical analysis, food safety and environmental security. Prospects for CEC and pCEC development and application are also discussed.

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Recent applications of microemulsion electrokinetic chromatography PENG Zhenlei, LIN Jin-Ming 2009, 27 (5):  621-630.  Abstract ( 2156 )   [Full Text(HTML)] () PDF (397KB) ( 658 )   The separation principle of microemulsion electrokinetic chromatography (MEEKC) is similar to micellar electrokinetic chromatography (MEKC). Within the last eight years, a number of papers have appeared in the literature. An overview about the applications of MEEKC is given.

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Development of immunoaffinity capillary electrophoresis CHEN Hongxu, ZHANG Xinxiang 2009, 27 (5):  631-641.  Abstract ( 2279 )   [Full Text(HTML)] () PDF (649KB) ( 744 )   Immunoaffinity capillary electrophoresis is one of the important analytical technologies for complicated samples. It combines the advantages of both immunoassay and capillary electrophoresis, such as high specificity, high separation efficiency, rapid and low consumption. With the aid of laser-induced fluorescence detector and immuno-enrichment, the sensitivity of the method can meet the requirement for trace detection. In this paper, the research progress of homogenous capillary electrophoresis immunoassay (CEIA) and immunoaffinity capillary electrophoresis (IACE) is reviewed based on the research of the author’s group.

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Methodology of capillary electrophoresis-laser induced fluorescence immunoassay for highly sensitive detection of DNA adducts WANG Hailin, ZHANG Dapeng, WANG Zhixin, LI Tao, FENG Feng, WANG Chao, GAO Haiyan 2009, 27 (5):  642-647.  Abstract ( 2347 )   [Full Text(HTML)] () PDF (179KB) ( 832 )   DNA adducts is a very important class of biomarkers of human exposure to carcinogen, cancer risk assessment, and population susceptibility. There is a lack of a technology with a sufficient sensitivity to detect trace DNA adducts related to low environmental exposure levels. We attempt to develop a highly sensitive assay for the detection of DNA adducts by combining capillary electrophoresis-laser induced fluorescence (CE-LIF) with immunochemical recognition, or CE-LIF immunoassay. This review describes the recent research progress on CE-LIF instrument construction and the methodology of CE-LIF immunoassay for the detection of DNA adducts. The methodology study mainly involves the synthesis and characterization of the adduct containing DNA fluorescent probes, the interaction of DNA adducts and antibody, stabilization of trace DNA adducts-antibody complexes, and DNA-driven focusing.

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Immune algorithm: a new chemometrics tool for resolution of multicomponent overlapping chromatographic signals SHAO Xueguang, LIU Zhichao, CAI Wensheng 2009, 27 (5):  648-654.  Abstract ( 2181 )   [Full Text(HTML)] () PDF (345KB) ( 674 )   Chromatography and related hyphenated techniques have been one of the most important tools for complex sample analysis. Although advances in equipments and experimental techniques have been achieved, overlapping chromatographic peaks are still unavoidable when complex samples are analyzed. Chemometrics, which uses mathematical tools to deal with the overlapping analytical signals, provides alternative approaches for the resolution of overlapping chromatographic signals. In this review, the advances in the studies of immune algorithms (IAs) are summarized with the applications of the methods in the resolution of multicomponent overlapping chromatographic signals.

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Cell laboratory on a microfluidic chip QIN Jianhua, LIU Tingjiao, LIN Bingcheng 2009, 27 (5):  655-661.  Abstract ( 2343 )   [Full Text(HTML)] () PDF (408KB) ( 1530 )   In this paper, cell-based microfluidic laboratory as a useful platform for biomedical applications is reviewed. The characteristics of the microfluidic-based platform for cellular and organism-based research are discussed.

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Developments of high-throughput microfluidic sample introduction techniques YAO Bo, HE Qiaohong, DU Wenbin, SHI Xiaotong, FANG Qun 2009, 27 (5):  662-666.  Abstract ( 1876 )   [Full Text(HTML)] () PDF (250KB) ( 733 )   How to achieve the interfacing between macro world and microchip is still one of the important topics in the research of microfluidic chips. In this paper, the recent progress in high-throughput microfluidic sample introduction techniques is described, mainly based on the work achieved in the authors’group. Various high-throughput microfluidic sample introduction systems in three modes including reservoir, flow-through cell and sampling probe based on microchips or capillaries are described. The developing prospects of high-throughput sample introduction techniques are also forecasted.

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Analysis of sugars by high performance anion-exchange chromatography with pulsed amperometric detection MOU Shifen, YU Hong, CAI Yaqi 2009, 27 (5):  667-674.  Abstract ( 3108 )   [Full Text(HTML)] () PDF (332KB) ( 963 )   High performance anion-exchange chromatography （HPAEC） with pulsed amperometric detection （PAD） is a powerful method of recent development for the analysis of sugars. The separation of sugars by HPAEC and the detection of sugars using PAD, including stationary phase and mobile phase, and waveform of detection, are reviewed. Applications of the method are described in detail.

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Stationary phases for hydrophilic interaction liquidchromatography and their applications in separation of traditional Chinese medicines GUO Zhimou, ZHANG Xiuli, XU Qing, LIANG Xinmiao 2009, 27 (5):  675-681.  Abstract ( 2422 )   [Full Text(HTML)] () PDF (248KB) ( 1198 )   Hydrophilic interaction liquid chromatography (HILIC) is an alternative of reversed-phase liquid chromatography (RPLC) for the separation of polar compounds. The main characteristic of HILIC is the use of polar stationary phases and aqueous/organic (usually acetonitrile) mobile phases. In recent years, HILIC has attracted more and more attentions for the demanding of the separation of polar compounds in many research fields and the advantages of HILIC, such as the good retention of polar compounds, orthogonality to RPLC and compatibility with mass spectrometry (MS), etc. Stationary phases are the basis of the development and application of HILIC. The structure of the bonded phases, retention properties and applications of the HILIC stationary phases are reviewed in the present article. The conventional stationary phases for normal phase liquid chromatography used in HILIC and the stationary phases specially developed for HILIC are introduced. The advantages, drawbacks and the general situation of applications of the different stationary phases are commented. The applications in the separation of traditional Chinese medicines are also reviewed. The further development of HILIC stationary phases is looked ahead.

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Applications of fast and ultra performance liquid chromatography in the analysis of Chinese herbal medicines LIU Ying, ZHOU Jianliang, LI Ping 2009, 27 (5):  682-689.  Abstract ( 2188 )   [Full Text(HTML)] () PDF (224KB) ( 996 )   The analysis of chemical components of Chinese herbal medicines (CHMs) is one of the most critical issues not only for screening and analyzing the bioactive components but also for controlling their quality. However, due to the complexity of the chemical constituents of CHMs, it is difficult to separate them on column within a short time. In the recent, the fast and ultra performance liquid chromatography, including ultra high pressure liquid chromatography, high performance liquid chromatography based on the monolithic columns and high temperature liquid chromatography, are of particular interest because of the high resolution and fast analytical speed provided by these techniques. This overview covers the principle and separation characteristics of these techniques, as well as their applications in Chinese herbal medicines.

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Progress of on-line sample preparation techniques in chromatographic analysis ZHONG Qisheng, HU Yufei, LI Gongke, HU Yuling 2009, 27 (5):  690-699.  Abstract ( 2368 )   [Full Text(HTML)] () PDF (640KB) ( 1160 )   Sample preparation is very important in the chromatographic analysis due to its time consumed and deviations produced, therefore the problem of on-line sample preparation techniques is the front project in the field of analytical chemistry. In this paper, the progress of on-line sample preparation techniques in chromatographic analysis, including solid phase extraction, solid phase microextraction, liquid phase microextraction, membrane assisted extraction, field assisted extraction, gas phase extraction, thermal desorption and microfluidic chip is reviewed.

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Determination of pesticide residues in fugu, eel and prawn using gas chromatography-mass spectrometry with gel permeation chromatographic clean-up ZHENG Feng, PANG Guofang, LI Yan, WANG Minglin, FAN Chunlin 2009, 27 (5):  700-710.  Abstract ( 2269 )   [Full Text(HTML)] () PDF (470KB) ( 920 )   A multiresidue analytical method was developed for the determination of 191 pesticides in fugu, eel and prawn using gas chromatography-mass spectrometry (GC-MS). The samples were extracted with ethyl acetate and cyclohexane (1:1, v/v), and cleaned-up by gel permeation chromatography (GPC). The GPC eluant collected from 26 min to 44 min was concentrated to 1 mL, then analyzed using GC-MS. A DB-1701 column was used for the separation. The MS detection was performed in selected ion monitoring mode. The recoveries were determined at the two spiked levels of 1 LOQ and 4 LOQ (LOQ: limit of quantification). The overall recoveries were from 50.2% to 120%, and in which the recoveries of 89.5% pesticides were from 70% to 120%. The relative standard deviations (RSDs) of the recoveries were from 0.6% to 21.6%. The calibration curves of all pesticides showed good linearities in the respective ranges with the correlation coefficient above 0.97. The limits of detection and the limits of quantification were 0.002~0.3 mg/kg and 0.007~1.2 mg/kg, respectively. The sensitivity and accuracy of the method met the requirements of the multiple pesticide residues. This method was applicable to determine 191 multiple pesticide residues in fugu, eel, prawn and other fishes.

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Simultaneous speciation of arsenic and selenium by high performance liquid chromatography-double channel atomic fluorescence spectrometry WANG Zhenhua, HE Bin, SHI Jianbo, YIN Yongguang, JIANG Guibin 2009, 27 (5):  711-716.  Abstract ( 2293 )   [Full Text(HTML)] () PDF (325KB) ( 781 )   A comprehensive method for simultaneously detecting species of arsenic and selenium including arsenite (As(III)), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), arsenate (As(V)), selenocystine (SeCys), selenomethionine (SeMet) and selenate (Se(IV)) was developed with high performance liquid chromatography-hydride generation-double channel atomic fluorescence spectrometry (HPLC-HG-AFS). An anion-exchange column (PRP-X100) with eluent of 10 mmol/L NH4H2PO4 containing 2.5%(v/v) methanol was employed to separate these species within 12 min. The detection limits of As(III), DMA, MMA, As(V), SeCys, SeMet and Se(IV) were 1, 3, 2, 3, 4, 18 and 3 μg/L (200 μL of injection), respectively. The relative standard deviations in five independent determinations varied from 1.9% to 6.1% for arsenic and selenium species at the concentration levels of 100 and 300 μg/L. The proposed method was applied to analyze the selenium yeast tablet and human urine samples. The recoveries from spiked selenium yeast tablet and urine samples ranged from 88% to 105% and from 83% to 108%, respectively. The results showed that this method can be used for determining arsenic and selenium species in urinary metabolites and drug samples in daily analysis conveniently.

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Effect of dynamic factors on the resolution of intact protein separation by liquid chromatography MIN Yi, CHEN Gang, GENG Xindu 2009, 27 (5):  717-723.  Abstract ( 2138 )   [Full Text(HTML)] () PDF (324KB) ( 677 )   Based on the fact that the resolution of intact protein separation is almost independent of column length, the effect on the resolution for intact protein separation causing from dynamic factors in hydrophobic interaction chromatography (HIC) was investigated. A concept of “conditional plate height” (H) for protein separation is firstly suggested for characterizing this effect for protein separation under linear gradient elution. Standard proteins were separated with conventional chromatographic column and chromatographic cake, and the plot of the H vs the linear velocity of mobile phase (u) was made, respectively. It was found that the obtained plot is similar to the conventional van Deemter Plot but has some differences. The optimized u corresponding to the minimum H was determined to be approximate 0.2 mm/s for the chromatographic cake and 1~3 mm/s for the conventional column. Furthermore, in comparison with the latter, optimized u value for the former has much broader range. Based on this fact, the resolutions and speeds for standard protein separation between the chromatographic cake packed with silica-base HIC material and the conventional column packed with soft HIC media were compared. The chromatographic cake (10 mm×20 mm i.d.) was found to perform a complete separation of seven standard proteins in 10 min, while with the latter (55 mm×12 mm i. d.) only five standard proteins can be completely separated in 140 min, even though the sample load for the former having bed volume of 3.14 mL, five times of that of the latter. The HIC chromatographic cake was also employed for the renaturation with simultaneous purification of the recombinant human granulocyte colony stimulating factor. The obtained purity was ≥97%, mass recovery was 39%, specific bioactivity was 1×108 IU/mg with only one step HIC in 50 min. It would be expected that when a kind of packings having very small particle size is packed into a chromatographic cake with diameter to be greater than its thickness and is employed to separate, and/or renature proteins, a result of high speed and high resolution with simultaneous renaturation under high protein loading (“three H” target) could be obtained.

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Investigation on metabolism and pharmacokinetics of triclosan in rat plasma by using UPLC-triple quadrupole MS WU Jianlin, YUE Hao, CAI Zongwei 2009, 27 (5):  724-730.  Abstract ( 2458 )   [Full Text(HTML)] () PDF (274KB) ( 692 )   Triclosan has been widely used as a disinfectant in human health care products. Although this particular chemical is less toxic, its biotransformation products might have toxicity to human. Therefore, understanding the pharmacokinetics and metabolism of triclosan in animal and human body is important. Plasma samples from SD rats collected after the oral administration of 5 mg/kg triclosan were analyzed by ultra performance liquid chromatography (UPLC)-triple quadrupole mass spectrometry (TQMS) to support the pharmacokinetic study of triclosan. The method development was conducted with selected-ion-recording (SIR, also called SIM for selected-ion-monitoring) mode in ESI-MS and multiple-ion-monitoring (MRM) mode in MS/MS and the obtained results were compared. While MRM provided lower detection limits, its other method validation parameters were worse than those of SIR due to the poor fragmentation of triclosan. The developed SIR method provided limit of quantification of 10.8 ng/mL in plasma. The recovery, accuracy, precision and repeatability were satisfactory. The pharmacokinetic data of triclosan in the rats were presented including the half time of elimination that was (48.5±10.5) h, indicating that the elimination of triclosan in the rat was slow. Two hydroxylated and sulfonated triclosan, one glucuronidated triclosan, and one sulfonated triclosan were identified in the rat plasma samples

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Preparation of a strong cation-exchange polymer monolith and its application in determination of melamine in milk products MA Qiao, HU Xizhou, HUANG Jincui, FENG Yuqi 2009, 27 (5):  731-736.  Abstract ( 2750 )   [Full Text(HTML)] () PDF (351KB) ( 1134 )   A poly(2-acrylamido-2-methyl-1-propanesulfonic acid-co-ethylene dimethacrylate) (AMPS-co-EDMA) monolith was prepared in a fused-silica capillary (530 μm i.d.) and applied for polymer monolith microextraction (PMME). With the optimal ratio of N,N-dimethylformamide (DMF, porogen) and polyethylene glycol (PEG, co-porogen), the resulting monolith exhibited satisfactory permeability, high mechanical strength and good stability in aqueous buffer. The effects of several parameters to PMME were investigated, such as pH value, inorganic salt and organic phase concentration of the sample matrix. It demonstrated that the melamine was captured on the poly(AMPS-co-EDMA) monolith mainly through strong cation-exchange and hydrophobic interactions. A novel approach is presented for the determination of melamine in milk products by coupling PMME to high performance liquid chromatography with ultraviolet detection. Because of the high extraction capacity of the monolith towards melamine, low detection limits (S/N=3, 0.09 mg/kg) and quantification limits (S/N=10, 0.3 mg/kg) were obtained. The method showed good linearity ranging from 0.5 to 80 mg/kg. Excellent reproducibility of the method was exhibited by intraday and interday precisions, yielding the relative standard deviations not larger than 7.5%. The proposed method is simple, rapid, sensitive, and low cost.