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    Chinese Journal of Chromatography
    2016, Vol. 34, No. 11
    Online: 08 November 2016

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    Mini-reviews
    Recent advances of nanomaterials assisted negative ion laser desorption/ionization-time-of-flight mass spectrometry in the analysis of small molecules
    ZHANG Xiaona, NIU Jiahua, LU Minghua, CAI Zongwei
    2016, 34 (11):  1017-1021.  DOI: 10.3724/SP.J.1123.2016.09014
    Abstract ( 440 )   [Full Text(HTML)] () PDF (1178KB) ( 172 )  

    Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) is a soft ionization technique that has been widely used in the analysis of macromolecules, such as proteins, peptides, nucleic acids and polymers. However, it has difficulty in the analysis of small molecules with relative molecular masses below 700 Da because of the interferences from the matrix. To overcome these drawbacks, interference-free nanomaterials assisted laser desorption/ionization (LDI) has been paid more and more attention. Compared with the multiple peaks of one compound obtained in the positive ion mode, nanomaterials assisted negative ion LDI provides only deprotonated ion peak. In this article, recent advances of nanomaterials assisted negative ion LDI in the analysis of small molecules are reviewed.

    Reviews
    Applications of polydopamine surface modification technique in separation science
    CHAI Weibo, WANG Huijuan, AN Xuehan, DING Guosheng
    2016, 34 (11):  1022-1030.  DOI: 10.3724/SP.J.1123.2016.07018
    Abstract ( 489 )   [Full Text(HTML)] () PDF (1969KB) ( 223 )  

    As a novel bio-inspired material, polydopamine (PDA) coatings are easily formed in environmentally friendly conditions on virtually all kinds of inorganic and organic substrates. PDA has been applied in many areas such as chemical, biomedical, pharmaceutical, sensor and battery manufacturing, etc. In the field of separation science, PDA can be used to prepare new enrichment materials, as well as stationary phases for chromatographic separation. In this review, the research status of the formation mechanism of PDA is introduced firstly; emphases are put on the recent applications of PDA in the areas of separation and enrichment, such as capillary electrophoresis, liquid chromatography, molecularly imprinted solid-phase extraction, dispersive solid-phase microextraction and solid phase microextraction, etc.

    Advances of Caenorhabditis elegans research on microfluidic device
    YIN Fangchao, WEN Hui, ZHU Guoli, QIN Jianhua
    2016, 34 (11):  1031-1042.  DOI: 10.3724/SP.J.1123.2016.07017
    Abstract ( 487 )   [Full Text(HTML)] () PDF (8196KB) ( 110 )  

    The nematode Caenorhabditis elegans (C. elegans) has been widely used as a multicellular organism for biological research due to its simplicity, short life cycle, and relevance to human genetics and biology. In the past decade, microfluidics technology was emerging as a promising platform for the study of C. elegans both in continuous flow-based or droplet-based format. Worm long-term culture, immobilization, sorting, stimuli exposure and individual worm encapsulation have been realized on microfluidic platform. By providing unique capabilities with precise manipulations and controlled microenvironment, this platform has been used for the study of neurobiology, behaviors, aging and development, and drug screening in C. elegans. In this review, we highlight the recent advances of microfluidic technology in worm manipulations and its biomedical applications. The prospects and challenges of microfluidic technology for future studies are discussed as well.

    Articles
    Detection of microenvironment acidification in breast cancer cells by microfluidic chips
    YAN Wei, XU Deshun, ZHA Yunfeng, WU Xiaofang
    2016, 34 (11):  1043-1047.  DOI: 10.3724/SP.J.1123.2016.07011
    Abstract ( 394 )   [Full Text(HTML)] () PDF (1572KB) ( 115 )  

    An analytical method was established for dynamic detection of microenvironment acidification in a breast cancer cell model based on microfluidic chips. A multilayer hybrid microfluidic chip was designed. When the suspension of the breast cancer cells entered the chips the hydrogel grids would become skeleton for 3D cultures. Continuous perfusion in microfluidic chip channels was used to mimic blood flow in vivo. And the non-electrochemical pH sensor was introduced into the chips to give the real-time changes of pH by image analysis. Cell viabilities, cell proliferations, lactic acid levels and pH values of cancer cells were detected to analyze microenvironment acidification, while comparing with the normal cells. Continuous perfusion for 7 d, the cell viability remained above 90%. With the increase of culturing days, the pH of the tumor microenvironment decreased gradually. The lower perfusion rates, the lower pH values, but the pH of the normal cells microenvironment had no obvious change. The microenvironment acidification detection platform of the microfluidic chip could detect microenvironment pH in real time, and it is a useful tool for cancer research.

    Determination of vitamin C in fruits and vegetables by ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry
    LIU Zhenzhen, QI Peipei, WANG Xinquan, CHEN Wenxue, WU Liyu, WANG Qiang
    2016, 34 (11):  1048-1054.  DOI: 10.3724/SP.J.1123.2016.07015
    Abstract ( 537 )   [Full Text(HTML)] () PDF (902KB) ( 211 )  

    A method was developed for the determination of vitamin C in fruits and vegetables by ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry. The main factors which affect the sensitivity and stability of vitamin C in the sample preparation and instrument analysis were investigated. Fruit and vegetable samples were prepared by high speed homogenate under the protection of dry CO2, and then extracted by 0.1% (v/v) formic acid solution. The analysis was performed using an Acquity-UPLC T3 column (100 mm×2.1 mm, 1.8 μm) with the column temperature of 30℃. The injection volume was 5 μL. The mobile phase consisted of 0.05% (v/v) formic acid solution and methanol with a volume ratio of 8:2. The flow rate was set at 0.25 mL/min. The extracted ion [M-H]-(m/z 175.0238) was selected for quantification, while extracted ion [M-C3H4O3]-(m/z 87.0077) was selected for qualification. The mass error was 5×10-6 (5 ppm). The results displayed that the linearity of vitamin C ranged from 0.02 to 5.0 mg/L with the correlation coefficient (R2) of 0.9992. The limit of detection (LOD) and the limit of quantification (LOQ) were 0.008 mg/L (S/N=3) and 0.02 mg/L (S/N=10), respectively. The recoveries of the vitamin C in the spiked samples ranged from 89.3% to 113% with the RSD less than 5.40%. The developed method was then utilized to analyze the real kiwifruit samples. The results showed that the contents of the vitamin C ranged from 498 to 1625 mg/kg for the domestic kiwifruit samples. Regarding the New Zealand kiwifruit samples, the contents of the vitamin C ranged from 247 to 449 mg/kg. The present method demonstrated the advantages of easy operation, good precision and repeatability, making it feasible for accurate determination of vitamin C in fruits and vegetables.

    Rapid screening of 27 pesticide residues in chili powder by ultra performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry
    LIU Yongqiang, LI Kai, XU Wenjuan, GUO Liqiang, SUN Jun
    2016, 34 (11):  1055-1062.  DOI: 10.3724/SP.J.1123.2016.07007
    Abstract ( 493 )   [Full Text(HTML)] () PDF (2897KB) ( 133 )  

    An analytical method was established for the simultaneous determination of 27 pesticide residues in chili powder by ultra performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry (UPLC-Q-TOF/MS). The samples were extracted with acetonitrile, and cleaned-up by solid phase extraction (SPE) with Carb/NH2 cartridges, and eluted with acetonitrile-ethyl acetate (3: 1, v/v). The eluent was determined by UPLC-Q-TOF/MS with electrospray ionization in positive mode. The confirmatory analysis was carried out by comparing the retention times, accurate masses, isotope abundance and isotope spacing of all the compounds and fragment ions with Targeted MS/MS. The correlation coefficients (r) were greater than 0.997 in the linear ranges of the 27 pesticides. The limits of quantification (LOQs) of the 27 pesticides were 2.5-5.0 μg/kg. At the three spiked levels, the recoveries were between 72.3% and 118.9%, with the relative standard deviations (RSDs, n=6) ranging from 0.21% to 12.7%. The method is efficient, sensitive and accurate, and can meet the requirement of the determination of the 27 pesticides in chili powder.

    Determination of nine fluoroquinolone residues in chicken by ultra performance liquid chromatography-tandem mass spectrometry coupled with molecularly imprinted solid phase extraction
    YANG Yanfei, CAO Xumin, LI Xuelian, SONG Jiande, SUN Xiaoliang, WANG Shuting, WANG Xiaoyin, LI Lin, ZHAO Sijun
    2016, 34 (11):  1063-1069.  DOI: 10.3724/SP.J.1123.2016.07024
    Abstract ( 510 )   [Full Text(HTML)] () PDF (1757KB) ( 118 )  

    An ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method coupled with molecularly imprinted solid phase extraction (MISPE) for the determination of nine fluoroquinolone residues in chicken was developed. The samples were extracted with phosphate buffer solution, and cleaned up with MISPE columns. Then the drugs were analyzed on a BEH C18 column by gradient elution with acetonitrile and 0.1% (v/v) formic acid solution as mobile phase. The samples were analyzed in electrospray positive ion mode and multiple reaction monitoring (MRM) mode. The analytes were quantified by external standard method. The specific adsorption of MISPE for the nine fluoroquinolones in chicken samples was investigated. The linear ranges were 0.25-100 μg/L with correlation coefficients (r) greater than 0.9965. The limits of detection (LODs, S/N=3) and the limits of quantification (LOQs, S/N=10) were 0.08 μg/kg and 0.25 μg/kg, respectively. At the spiked levels of 0.25, 2.5, 5.0 μg/kg, the recoveries were 65.8%-112.2% with the intra-RSDs of 0.6%-13.5% and inter-RSDs of 0.5%-14.9%, respectively. The maximum capacities of the MISPE column were 464.7-932.4 μg/L. The method is sensitive, simple and rapid for the simultaneous determination of the nine fluoroquinolone residues.

    Determination of eight novel succinate dehydrogenase inhibitor fungicides in vegetables and fruits by modified QuEChERS
    YANG Huan, SUN Weihua, CAO Zhaoyun, MA Youning, CHAI Shuangshuang, QIN Meiling, CHEN Mingxue
    2016, 34 (11):  1070-1076.  DOI: 10.3724/SP.J.1123.2016.07005
    Abstract ( 582 )   [Full Text(HTML)] () PDF (2098KB) ( 157 )  

    A method was developed for the simultaneous determination of eight novel succinate dehydrogenase inhibitor fungicides (boscalid, penflufen, bixafen, sedaxane, fluopyram, penthiopyrad, isopyrazam, fluxapyroxad) residues in vegetables and fruits by liquid chromatography-tandem mass spectrometry (LC-MS/MS) coupled with modified QuEChERS. The amount of the two solid phase extraction agents (octadecyl bonded silica (C18) and primary secondary amine (PSA)) in QuEChERS method was optimized according to their adsorption and purification capacities. The eight compounds were extracted from samples by acetonitrile and purified by solid phase extraction agents. The eight compounds were separated on a Poroshell 120 EC-C18 column by the gradient elution using acetonitrile-0.1% (v/v) formic acid aqueous solution as mobile phases, detected by MS/MS with positive/negative electrospray ionization (ESI+/ESI-) in multiple reaction monitoring (MRM) mode, and finally quantified by external standard method. The correlation coeffcients (r) of the eight compounds were all above 0.998 in the mass concentration ranging from 0.5 to 500.0 μg/L. The limits of detection and the limits of quantification of the method were 0.2-1.7 μg/kg and 0.5-5.0 μg/kg, respectively. The recoveries of the eight analytes spiked at three levels (5.0, 25.0 and 125.0 μg/kg) in eight matrices ranged from 71.4% to 121.3% with the relative standard deviations (RSDs, n=6) from 0.8% to 17.2%. The method is easy to operate and with good purification effect. It can be applied to the rapid determination of the novel succinate dehydrogenase inhibitor fungicides in vegetables and fruits.

    Influence of column selection in assay of emodin and chrysophanol in Liuwei Anxiao capsules
    WANG Mingjuan, LIU Yadan, HU Xiaoru, HE Fengyan, XIANG Xinhua, DAI Zhong, YU Jiandong, MA Shuangcheng
    2016, 34 (11):  1077-1083.  DOI: 10.3724/SP.J.1123.2016.08008
    Abstract ( 538 )   [Full Text(HTML)] () PDF (895KB) ( 83 )  

    Column selection is a factor of critical risk that may influence the accuracy of high performance liquid chromatography (HPLC) method. Assay of emodin and chrysophanol in Liuwei Anxiao capsules described in Chinese Pharmacopeia was taken as a typical example. Though 155 laboratories in the proficiency testing program all used C18 columns, the 20 ones obtained incorrect results due to the co-elution of emodin and acid hydrolysis product of Liuwei Anxiao capsules in the selected C18 columns. Statistical results showed that A-type (early developed columns with higher silicon hydroxyls residues) and E-type (with embedded or end-capping polar group) C18 columns were more suitable for the assay of emodin and chrysophanol in Liuwei Anxiao capsules. It might attribute to the presence of phenolic hydroxyl groups in emodin and chrysophanol and interact with residual silicon hydroxyls in A-type columns or polar groups in E-type ones. To reduce the risk of unsuitable column selection, related laboratories should pay more attention to the robustness of columns when HPLC methods are established. The column selection for robustness could be based on peer-reviewed column classification databases, and the resolution of critical pair should be recommended to use as system suitability parameter.

    Simultaneous determination of 26 organic acids and anions in fruit juice by ion chromatography with suppressed conductivity detection using gradient elution
    SONG Weide, YUAN Xiaoying, LV Ning, CHEN Taifa, HUI Xidong, SU Zheng, JIN Wei, LIU Bing
    2016, 34 (11):  1084-1090.  DOI: 10.3724/SP.J.1123.2016.06057
    Abstract ( 535 )   [Full Text(HTML)] () PDF (888KB) ( 160 )  

    A method for simultaneous determination of 26 organic acids and anions in fruit juice by ion chromatography (IC) with suppressed conductivity detection using gradient elution was established. The effects of selection of columns, flow rates, column temperatures, pH values of solution, and eluent concentrations were studied, and then the suitable gradient elution conditions were found for the determination. The testing results of the 26 components in fruit juice were accurate under the conditions that the flow rate was 1.00 mL/min, column temperature was 30℃ and pH values of the solution were in the range of 5.5-6.8. As a result, the linear relationships of the 26 components were good in the range of 0.02-10.0 mg/L and the correlation coefficients (r) were greater than 0.995, the limits of detection (S/N=3) of the 26 components were in the range of 0.17-52.0 μg/L. In the added levels of 0.20-2.00 mg/L, the recoveries of the 26 components in fruit juice ranged from 85.58% to 108.86%, and the relative standard deviations (RSDs) were in the range of 0.15%-7.65% (n=6). The method is simple, quick, sensitive and accurate, and it is absolutely suitable for the trace analysis and determination of the 26 organic acids and anions in fruit juice.

    Optimization of rapid separation conditions of Ralstonia solanacearum using high performance ion-exchange chromatography
    ZHENG Xuefang, LIU Bo, ZHU Yujing, CHEN Deju
    2016, 34 (11):  1091-1096.  DOI: 10.3724/SP.J.1123.2016.06044
    Abstract ( 496 )   [Full Text(HTML)] () PDF (1517KB) ( 89 )  

    Rapid separation of Ralstonia solanacearum with intact cell using high performance ion-exchange chromatography equipped with a UV detector was described. The bacterial growth abilities and cell morphologies in different suspension buffers were compared. The results showed that the bacterial numbers in the balance buffer (0.02 mol/L piperazine-hydrochloric acid), elution buffer (0.02 mol/L piperazine-hydrochloric acid+1 mol/L NaCl) and double distilled water were 6.467×109, 6.267×109 and 6.233×109 cfu/mL, respectively, exhibiting no significant difference. Moreover, the cell morphologies were observed under a transmission electron microscope which showed that the bacterial cells could remain intact. Effects of buffer pH, flow velocity and bacterial cell concentration on the separation efficiency were analyzed. The results showed that the optimum chromatographic separation parameters were as follows: the buffer pH was 8, the flow velocity was 2 mL/min and the bacterial cell concentration was from 1.0×108 cfu/mL to 1.0×1010 cfu/mL. Under the chromatographic separation conditions, the separation time was shortened and the separation efficiency was improved. Therefore, the chromatographic method developed here may be a promising tool for the rapid separation of Ralstonia solanacearum and other bacteria.

    Rapid screening of 182 pesticide residues in foods by gas chromatography coupled with quadrupole-time of flight mass spectrometry
    YI Xionghai, SHI Yiyin, ZHAO Shanzhen, MENG Linghua, PAN Xiaobo, SHENG Yonggang, HAN Li, ZHU Jian, DENG Xiaojun, GUO Dehua
    2016, 34 (11):  1097-1105.  DOI: 10.3724/SP.J.1123.2016.06056
    Abstract ( 509 )   [Full Text(HTML)] () PDF (1701KB) ( 144 )  

    A method for rapid screening of 182 pesticide residues in foods according to the Hongkong regulation was developed by gas chromatography coupled with quadrupole-time of flight mass spectrometry (GC-Q-TOF/MS). The target analytes were extracted by acetonitrile with 0.1% (v/v) formic acid, then purified by a modified QuEChERS method. The separation was performed on an Agilent HP-5MS column (30 m×0.25 μm×0.25 mm). The compounds were ionized under electron impact ion source. Qualitative and quantitative detection of compounds were completed by searching accurate mass database with full scan mode. Suspected compounds can be confirmed by the spectrum database of the secondary fragment ions. The matrix effects in 10 kinds of typical foods (rice, mushrooms, soybeans, spinaches, tomatoes, broccoli, grapefruits, carrots, lettuces, cucumbers) were evaluated. The results demonstrated that the linear ranges were from 10 to 500 μg/kg with good correlation coefficients (r>0.99). The limits of quantitation were in the range of 10-100 μg/kg (S/N≥10). The average recoveries at three spiked levels were in the range of 66.1%-121.5%, 75.4%-125.8%, 77.2%-128.9% with relative standard deviations (RSDs) of 0.8%-17.6% (n=6). The method is simple, sensitive and robust, as well as having practical application for its low costs.

    Application of Fisher discrimination in gas chromatography-mass spectrometry analysis on combustion residues of typical combustion improvers
    CHEN Zhenbang, JIN Jing
    2016, 34 (11):  1106-1112.  DOI: 10.3724/SP.J.1123.2016.06017
    Abstract ( 419 )   [Full Text(HTML)] () PDF (983KB) ( 89 )  

    To find out a more accurate and effective pattern recognition method for the identification of combustion residues of typical combustion improvers, gas chromatography-mass spectrometry (GC-MS) was applied to analyze the combustion residues of seven combustion improvers loaded on different carriers and one submitted sample, and gasoline was identified directly by the GC-MS analysis from the unknown specimen. Two discrimination methods, including Fisher discrimination and PCA (principal component analysis) combined with Fisher discrimination, were used for the data analysis. The results showed that nitro paint thinner was discriminated from the submitted sample by PCA combined with Fisher discrimination, while gasoline was discriminated from unidentified sample by Fisher discrimination, which was in accordance with the direct GC-MS identification result. By comparing the results obtained by these two discrimination methods, Fisher discrimination is believed to be more efficient in classification of combustion residues of the seven combustion improvers and identification of the submitted sample. The results of this study provide a new analytical method for the combustion improvers identification.

    Comparison of the methylation methods for the determination of fatty acids in Yak bones by gas chromatography
    LIU Wenyuan, JIA Wei, WU Ting, ZHANG Chunhui, LI Xia, CHEN Xuefeng
    2016, 34 (11):  1113-1119.  DOI: 10.3724/SP.J.1123.2016.07009
    Abstract ( 460 )   [Full Text(HTML)] () PDF (924KB) ( 140 )  

    The acid hydrolysis-esterification system with six methylation methods (acetyl chloride-methanol method, H2SO4-methanol method, HCl-methanol method, KOH-methanol method, KOH-methanol+H2SO4-methanol method and KOH-methanol+HCl-methanol method) were compared for the fatty acids (FAs) profiling of Yak bones by gas chromatography (GC) in this study. Good linearities were obtained in the range of 0.28-250.00 mg/L with correlation coefficient R2>0.99, except for C4:0 (R2=0.9878). FAs can hardly be detected by the base-catalytic method and the combined acid-base method (the content of total FAs was less than 0.20 g/100 g) in the present study. The FAs determined by the acetyl chloride-methanol method (content of total FAs was 13.61 g/100 g) was superior to the H2SO4-methanol method (content of total FAs was 11.68 g/100 g) and the HCl-methanol method (content of total FAs was 3.18 g/100 g). The precisions for intra-day and inter-day of acetyl chloride-methanol method and H2SO4-methanol method were 0.27%-8.60% and 0.34%-2.64%, respectively. And the recoveries for the FAs of these two methods were in the range of 83.06%-105.54%. This study suggested that the acid hydrolysis-acetyl chloride-methanol method was the best method in the six methods. It was convenient for the accurate determination of FAs in Yak bones. C18: 1n9c, C16: 0, C18: 0 and conjugated linoleic acid (CLA) were the mainly FAs in Yak bones, and the sum of the four FAs was more than 85% of the total FAs. The ratio of the contents of the saturated and unsaturated FAs in Yak bones was about 1: 2. The study of fatty acids in Yak bones provides an important basis for the effective utilization of bone resources.

    Solidification of floating organic drop dispersive liquid-liquid microextraction combined with capillary electrophoresis for the determination of sulfonylurea herbicides in water
    ZHU Jing, LI Mingxin, WANG Qian, REN Lin, Zheng Bo, YONG Li, ZOU Xiaoli
    2016, 34 (11):  1120-1125.  DOI: 10.3724/SP.J.1123.2016.06012
    Abstract ( 428 )   [Full Text(HTML)] () PDF (949KB) ( 113 )  

    A novel method of solidification of floating organic drop dispersive liquid-liquid microextraction for the simultaneous determination of sulfonylurea herbicides in water samples by capillary electrophoresis was established. Sulfonylurea herbicides in water samples were extracted by solidification of floating organic drop dispersive liquid-liquid microextraction using dodecanol as extractant and methanol as dispersive solvent, and were determined with capillary electrophoresis. Nine kinds of sulfonylurea herbicides such as tribenuron-methyl, pyrazosulfuron-methyl, bensulfuron-methyl and so on were detected by this method. The good linearities were obtained in the range of 10.0-1000 μg/L. The correlation coefficiencies were not less than 0.992. The method detection limits (MDLs) ranged from 2.40 to 7.50 μg/L. The relative standard derivations (RSDs) of the method were 6.55%-13.9%. The method was applied for the determination of water samples and showed satisfactory results. The spiked recoveries ranged from 82.0% to 104%. The method is quick, sensitive and suitable for the batch analysis of sulfonylurea herbicides in water samples.