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    Chinese Journal of Chromatography
    2014, Vol. 32, No. 4
    Online: 08 April 2014

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    Recent advances in sample preparation methods of plant hormones
    WU Qian, WANG Lu, WU Dapeng, DUAN Chunfeng, GUAN Yafeng
    2014, 32 (4):  319-329.  DOI: 10.3724/SP.J.1123.2014.01039
    Abstract ( 943 )   [Full Text(HTML)] () PDF (4625KB) ( 578 )  

    Plant hormones are a group of naturally occurring trace substances which play a crucial role in controlling the plant development, growth and environment response. With the development of the chromatography and mass spectroscopy technique, chromatographic analytical method has become a widely used way for plant hormone analysis. Among the steps of chromatographic analysis, sample preparation is undoubtedly the most vital one. Thus, a highly selective and efficient sample preparation method is critical for accurate identification and quantification of phytohormones. For the three major kinds of plant hormones including acidic plant hormones & basic plant hormones, brassinosteroids and plant polypeptides, the sample preparation methods are reviewed in sequence especially the recently developed methods. The review includes novel methods, devices, extractive materials and derivative reagents for sample preparation of phytohormones analysis. Especially, some related works of our group are included. At last, the future developments in this field are also prospected.

    Advances and applications of selective reaction monitoring technology in proteomics study
    SHAN Yichu, ZHANG Lihua, ZHANG Yukui
    2014, 32 (4):  330-335.  DOI: 10.3724/SP.J.1123.2013.12019
    Abstract ( 776 )   [Full Text(HTML)] () PDF (1107KB) ( 258 )  

    As an important technology for targeted protein analysis, selective reaction monitoring technology (SRM) improves the detection sensitivity and quantification accuracy by eliminating the interference of impurities and co-eluting peptides by selective detection of specific mother ions and daughter ions. It has been widely applied to the quantitative proteomics study due to the advantages of high selectivity, excellent reproducibility, high sensitivity and wide dynamic range and plays an important role in the area of life science. For the quantitative analysis of the complex samples with wide dynamic range, the throughput of analysis and detection sensitivity still need to be improved. Moreover, various quantification strategies have been proposed to improve the accuracy and precision of quantification. Furthermore, data processing becomes more and more important with the application of SRM technology to the analysis of complex samples. In this work, the recent development of SRM technology is reviewed from the above mentioned aspects. Since SRM technology gains wider applications along with the technological development, its applications in the area of proteomics quantitative study including biomarker validation, post-translational proteomics study (phosphorylation, glycosation, acetylation and so on), biotechnology and signaling pathway analysis are briefly described. Finally, the future developments, applications and outlook of SRM technology are described.

    Recent advancement of photonic-crystal-based analytical chemistry
    CHEN Yun, GUO Zhenpeng, WANG Jinyi, CHEN Yi
    2014, 32 (4):  336-342.  DOI: 10.3724/SP.J.1123.2014.01011
    Abstract ( 899 )   [Full Text(HTML)] () PDF (3173KB) ( 385 )  

    Photonic crystals are a type of novel materials with ordered structure, nanopores/channels and optical band gap. They have hence important applications in physics, chemistry, biological science and engineering fields. This review summarizes the recent advancement of photonic crystals in analytical chemistry applications, with focus on sensing and separating fields happening in the nearest 5 years.

    Development of online conventional array-based two-dimensional liquid chromatographic system for proteins separation in human plasma
    HUANG Zhi, HONG Guangfeng, GAO Mingxia, ZHANG Xiangmin
    2014, 32 (4):  343-348.  DOI: 10.3724/SP.J.1123.2013.10020
    Abstract ( 755 )   [Full Text(HTML)] () PDF (2291KB) ( 195 )  

    Human plasma is one of the proteins-containing samples most difficult to characterize on account of the wide dynamic concentration range of its intact proteins. Herein, we developed a high-throughput conventional array-based two-dimensional liquid chromatographic system for proteins separation in human plasma in online mode. In the system, a conventional strong-anion exchange chromatographic column was used as the first separation dimension and eight parallel conventional reversed-phase liquid chromatographic columns were integrated as the second separation dimension. The fractions from the first dimension were sequentially transferred into the corresponding reversed-phase liquid chromatographic precolumns for retention and enrichment using a 10-port electrically actuated multi-position valve. The second dimensional solvent flow was directly and identically split into 8 channels. The fractions were concurrently back-flushed from the precolumns into the 8 conventional RP columns and were separated simultaneously. An 8-channel fraction collector was refitted to collect the reversed-phase liquid chromatographic fractions for further investigation. Bicinchoninic acid (BCA) dyeing solution was conveniently used for high-abundance protein location. Two separation dimensions were relatively independent parts, as well as each channel of the second dimensional array separation. Therefore, the new system could improve the separation throughput and total peak capacity. The system was successfully applied for the separation of human plasma intact proteins. The results indicated the established system is an effective method for removing high abundance proteins in plasma and in-depth research in plasma proteomics.

    A method using spectrum alignment to improve analysis efficiency of liquid chromatography combined with mass spectrometry
    LU Wenyuan, ZHANG Yang, SHEN Chengpin, YIN Xuefei, LIU Xiaohui, YANG Pengyuan
    2014, 32 (4):  349-354.  DOI: 10.3724/SP.J.1123.2013.10030
    Abstract ( 720 )   [Full Text(HTML)] () PDF (2056KB) ( 203 )  

    In the analysis of proteins in human umbilical vein endothelial cells (HUVEC) treated with dimethyl sulfoxide (DMSO) and NEDD8-activating enzyme inhibitor (MLN4924,MLN), the Progenesis LC-MS software (Nonlinear Dynamics Ltd) was applied to liquid chromatography spectrum alignment, while spectrum similarities were figured out among several experiments of the same sample, and also among different samples. After double enzymolysis, the sample was added with digested QconCAT standard proteins. They were separated by HPLC-MS/MS, followed by spectrum alignment and data analysis. This established experiment flow offered a better identification result of more than 8000 proteins, while the original result was about 7000 proteins, ensuring a relatively high identification efficiency. On the basis of relative quantification with spectrum count, the described procedure can analyze the differential expression of proteins induced by DMSO and MLN. The similarities of total ion chromatograms after alignment were also compared. This method was proved to be quick and easy, with the advantages of high throughput and high sensitivity.

    Establishment and application of integrated platform for proteome quantification
    ZHOU Yuan, ZHANG Shen, YUAN Huiming, ZHANG Lihua, ZHANG Yukui
    2014, 32 (4):  355-360.  DOI: 10.3724/SP.J.1123.2014.01002
    Abstract ( 599 )   [Full Text(HTML)] () PDF (1117KB) ( 182 )  

    To improve the accuracy, throughput and automation of proteome quantification analysis, an integrated platform including a microflow mixed-bed ion exchange column, a hydrophilic immobilized enzymatic reactor (hIMER) and nanoRPLC-electrospray ionization (ESI)-MS/MS system was established. Online separation and digestion of dimethylated proteins, followed by peptide separation, identification and quantification can be realized automatically by this platform. High and light dimethyl-labeled (H/L) proteins with the mass ratio of 1:1 were used to evaluate the quantification performance of the platform. The results showed that the dimethyl labeling efficiency at protein level was 90%. The incomplete digestion resulting from 10 min online digestion by the hIMER column and the non-specific adsorption of protein digests on the column had little adverse effect on the accuracy of protein quantification results. The mean value of H/L (mass ratio) of all the quantified proteins was 1.01. This platform was finally applied to analyze the different protein expression levels of two mice hepatocarcinoma ascites cell lines with high and low lymph node metastasis rates (Hca-F and Hca-P cell lines). Finally 15 up-regulated and 12 down-regulated proteins (Hca-F/Hca-P) were successfully obtained. All these results demonstrated that the integrated platform can be used for proteome quantification with the advantages of high accuracy and high throughput.

    Articles
    Metal-tag labeling coupled with high performance liquid chromatography-selected ion monitoring mass spectrometry for absolute quantitation of proteins
    LI Jiabin, ZHOU Lianqi, YAN Hui, LI Nannan, HAO Feiran, TIAN Fang, ZHANG Yangjun
    2014, 32 (4):  361-368.  DOI: 10.3724/SP.J.1123.2013.11042
    Abstract ( 753 )   [Full Text(HTML)] () PDF (1129KB) ( 248 )  

    A novel method has been established based on metal element chelated tags coupled with high performance liquid chromatography-selected ion monitoring mass spectrometry (HPLC-SIM/MS). The labeling efficiency and stability of metal element chelated tags, the chromatographic retention behavior and MS behavior of the labeled peptides, the linear range and accuracy of this method were examined. The results showed that the metal element chelated tag method has high labeling efficiency and high labeling stability, and the labeled peptides with different kinds of metal tags have consistent chromatographic retention behavior. The method of metal tags coupled with HPLC-SIM/MS has high sensitivity with the limit of quantification (LOQ) up to 1 fmol. The linear range for the method was between 1 fmol to 500 fmol with R2>0.99, which means the method has a good linearity. Moreover, this method had an average recovery of 117.01%. The method was used in the absolute quantitation of a protein enolase in Thermoanaerobacter tengcongensis (TTE) with a relative standard deviation of 5.74%, which means high precision. All the results showed that this method is accurate and reliable for the absolute quantitation of proteins. This gives us an alternative for the quantitative determination of proteins in relatively simple biological samples.

    Enrichment of glycoproteins in human serum using concanavalin A-functionalized magnetic nanoparticles and identification by mass spectrometry
    LI Feng, KANG Jingwu
    2014, 32 (4):  369-375.  DOI: 10.3724/SP.J.1123.2013.12018
    Abstract ( 820 )   [Full Text(HTML)] () PDF (4277KB) ( 464 )  

    Biomedical sciences, and in particular biomarker research, demand efficient glycoprotein enrichment platforms. Herein novel magnetic nanoparticles with an average size around 135 nm in diameter were prepared for the enrichment of glycoproteins in human serum. The prepared magnetic nanoparticles possessed uniform core/shell/shell structure which was composed of 8 nm magnetite internal core and double layers consisting of silica and poly glycidyl methacrylate (GMA). The latter was constructed by seed polymerization. Modified by a polyethylene hydrophilic linker, it made the surfaces of the magnetic nanoparticles highly hydrophilic so as to reduce the nonspecific adsorption of proteins. We examined affinity purification of glycoprotein in diluted human serum using our prepared magnetic nanoparticles with immobilization of concanavalin A (MNP@ConA). The enriched proteins were reduced, alkylated and digested with trypsin. These peptides then were separated by offline two-dimensional chromatography. Protein identification was realized with nano-high performance liquid chromatography-orbitrap mass spectrometry. A total of 80 proteins were identified, among them 76 proteins were found to be glycoproteins by use of bioinformatic tools. β-2-Glycoprotein 1 present in serum at low mass concentration around 0.00001 g/L was also identified. This demonstrates the capability of magnetic nanoparticle for recovering minute amounts of glycoproteins from a fluid exhibiting a dynamic concentration range more than 12 orders of magnitude. Overall, MNP@ConA has been proven to be an efficient alternative to currently available immobilization supports.

    Effect of the lysine guanidination on proteomic analysis
    ZHENG Hao, MAO Jiawei, PAN Yanbo, LIU Zhongshan, LIU Zheyi, YE Mingliang, ZOU Hanfa
    2014, 32 (4):  376-380.  DOI: 10.3724/SP.J.1123.2014.01003
    Abstract ( 888 )   [Full Text(HTML)] () PDF (1425KB) ( 280 )  

    The guanidination of lysine side chain was paid great attention in recent years. It plays an important role in qualitative and quantitative proteomics. In this study, based on the results of separated peptides extracted from HeLa cells before and after the guanidination by liquid chromatography-tandem mass spectrometry (LC-MS/MS), the effect of the guanidination of three different kinds of peptides was systematically analyzed. It was found that the selectivity of the guanidination of the lysine side chain was as high as 96.8%. The ratio of identified peptides with lysine at C-term to all peptides increased from 51.7% to 57.3% and more new peptides were identified, while the ratio of peptides with lysine in the middle or without lysine changed little. Further study on the ratio of b and y ions indicated that there were more y ions of peptides with lysine at C-term after the guanidination. The results proved that the selective conversion of lysine to homoarginine by the guanidination could increase the sensitivity and selectivity of mass spectrum. The increased basicity and ability to sequester proton of lysine produced more y ions fragmentation information, which contributed to more identified peptides. It concluded that the lysine guanidination can improve the coverage of proteomic analysis.

    Preparation and retention mechanism of a mixed-mode reversed-phase/weak-cationic-exchange chromatographic packing
    PENG Xitian, FENG Yuqi
    2014, 32 (4):  381-387.  DOI: 10.3724/SP.J.1123.2013.10038
    Abstract ( 811 )   [Full Text(HTML)] () PDF (1119KB) ( 428 )  

    A "mixed ligand" octyl-carboxylic co-bonded silica (OCS) packing was prepared by the method of "click chemistry". The resulting OCS packing was characterized by elemental analysis (EI) and Fourier transform infrared spectroscopy (FT-IR) to prove the successful immobilization of octyl and carboxylic groups on the surface of silica gel. Then the mixed-mode reversed-phase/weak-cationic-exchange (RP/WCX) retention mechanism of the OCS packing was quantitatively probed by studying the retention factors of a homologous series of three cationic surfactants on the mixed-mode stationary phase column as a function of the ammonium concentration in the eluent or the number of methylene groups in the solute. The one-site and two-site mixed-mode retention models of the three cationic surfactants on the OCS phases were studied by investigating the logarithm and reciprocal relationships of retention factors and salt concentrations, demonstrating that the two-site model was more suitable for the description of the retention mechanism of the three cationic surfactants on the OCS phases. Furthermore, the individual RP or WCX contribution to total retention was obtained according to the mathematical equations of two-site retention mechanism, which can provide some valuable guidance for the separation of real samples. This study developed the qualitative model of retention mechanism of the mixed-mode OCS packing, and a series of standard basic mixtures were well separated on the OCS packing, demonstrating the great application potential of OCS packing for the separation of various basic compounds.

    Synthesis and chromatographic evaluation of sulfobetaine-based capillary zwitterionic hydrophilic monolithic column using a binary porogenic agent of polyethylene glycol/methanol
    KUANG Yuanyuan, WANG Yan, GU Xue, LI Jing, YAN Chao
    2014, 32 (4):  388-394.  DOI: 10.3724/SP.J.1123.2013.11003
    Abstract ( 908 )   [Full Text(HTML)] () PDF (3100KB) ( 256 )  

    Sulfobetaine-based capillary zwitterionic hydrophilic monolithic columns were synthesized with a novel binary porogenic agent of polyethylene glycol (PEG)/methanol. The polymer was prepared with (3-(metharyloylamino) propyl) dimethyl (3-sulfopropyl) ammonium hydroxide inner salt (SPP) as monomer, pentaerythritol triacrylate (PETA) as crosslinker, and azobisisobutyronitrile (AIBN) as initiator. In order to optimize the properties, the contents of the polymerization mixture were investigated. The optimum preparation conditions were as follows: the mass ratio of monomer and porogenator=1:2.5; the mass ratio of SPP and PETA=1:1 in the monomer; the mass ratio of PEG and methanol=2:1 in the binary porogenic agent; the content of the initiator (AIBN) =0.1% (m/m). With the binary porogenic agent of PEG/methanol addition, good mechanical stability, homogeneous column bed, good permeability and narrow pore size distribution were obtained. In the capillary liquid chromatography mode, the hydrophilic monolith provided column efficiency up to 2.4×105 plates/m which was much higher than that fabricated by traditional method without PEG/methanol. The columns were used in capillary liquid chromatography and pressurized capillary electrochromatography for the separation of a mixture of phenols, nucleosides and so on.

    Preliminary exploration of energy transfer about sample ionization process in electrospray ionization source
    ZHANG Weibing, GAO Fangyuan, GUAN Yafeng, ZHANG Yukui
    2014, 32 (4):  395-401.  DOI: 10.3724/SP.J.1123.2013.11001
    Abstract ( 752 )   [Full Text(HTML)] () PDF (2288KB) ( 218 )  

    Electrospray ionization source (ESI) can be used not only for the detection of small molecules, but also for large molecules such as proteins and peptides. This article proposes energy transfer theory based on the lowest energy principle by systematically analyzing ionization processes. The transference of the analyte from liquid phase to gas phase can be influenced by multiple forces during the ionization and vaporization processes, such as electrostatics force and van der Waals' force. The ionization of samples is the result of the interactions among multiple forces. During different stages of the ionization process, different forces lead to different effects. There are competition between evaporation and formation of multi-charge ions for charges. For molecules with different structures, Gibbs free energy between two phases from the changes of molecule shape or conformation may lead to ion evaporation, multiply charged macromolecule and chain ejection, etc. The energy transfer theory can simplify the three existing theories, as well as explain the solvent effect and electrolyte ion effect during ionization process. The proposed theory provides foundation to optimize the detection condition for different samples and to understand the real process of ionization.

    Determination of nitrofuran residues in feed and water samples by monolith-based stir bar sorptive extraction and high performance liquid chromatography
    ZHANG Yong, MEI Meng, HUANG Xiaojia, YUAN Dongxing
    2014, 32 (4):  402-406.  DOI: 10.3724/SP.J.1123.2013.10013
    Abstract ( 765 )   [Full Text(HTML)] () PDF (1382KB) ( 320 )  

    A simple, efficient and sensitive method for the simultaneous determination of nitrofurantoin (NFT), furazolidone (FZD) and nitrofurazone (NFZ) in feed and water samples was developed by stir bar sorptive extraction (SBSE) coupled to high performance liquid chromatography with diode array detection. The SBSE based on poly (vinylimidazole-divinylbenzene) (VIDB) monolithic material as coating was used to concentrate the three target analytes. To obtain the optimum extraction performance, several VIDB-SBSE parameters were investigated and studied, including pH value, ionic strength of sample matrix, extraction and desorption time. Under the optimized experimental conditions, the linear ranges were 0.5-200 μg/L for FZD, 0.25-200 μg/L for NFT and NFZ. The limits of detection (S/N=3) were in the range of 0.068-0.11 μg/L for the three analytes. The precision of the proposed method was evaluated in terms of intra- and inter-day repeatability calculated as RSD, and it was found that the RSDs were all below 6%. The developed method was successfully applied to the determination of nitrofuran residues in animal feed and water samples. The satisfactory recoveries of the spiked target compounds were in the range of 80.6%-108%.

    Monolith column solid-phase extraction coupled with high performance liquid chromatography in online mode for the determination of forchlorfenuron in fruits
    LUO Weiqiang, XIAO Xiaohua, DU Zhuo, LI Gongke
    2014, 32 (4):  407-412.  DOI: 10.3724/SP.J.1123.2013.11011
    Abstract ( 721 )   [Full Text(HTML)] () PDF (3952KB) ( 369 )  

    A novel method based on the monolithic column solid-phase extraction (SPE) coupled with high performance liquid chromatography in online mode was developed for the determination of trace forchlorfenuron in fruits. The enrichment factor of SPE for forchlorfenuron was 214. The method was fully validated. It showed a linear range of 0.01-50.00 μg/L and a limit of detection of 25 ng/L, with the relative standard deviation (RSD) of 3.9%. The proposed method was successfully applied for the determination of forchlorfenuron in grape, kiwifruit and watermelon samples, with the recoveries of 87.0%-120.7% and the RSDs of 0.6%-9.2%. This online method was proved to be selective, sensitive and convenient for the determination of trace forchlorfenuron in complex samples.

    Dispersive solid phase microextraction of vanillins in milk using magnetic nanoparticles of ferroferric oxide/carbon nanotubes combining with high performance liquid chromatography analysis
    LI Haifang, YANG Hongyun, ZHANG Ying, WANG Peilong, LIN Jin-Ming
    2014, 32 (4):  413-418.  DOI: 10.3724/SP.J.1123.2013.11015
    Abstract ( 2212 )   [Full Text(HTML)] () PDF (2563KB) ( 453 )  

    Magnetic nanoparticles of ferroferric oxide/carbon nanotubes (Fe3O4/CNTs) were synthesized by chemically bonding single-walled carbon nanotubes (SWCNTs) to the Fe3O4 nanoparticles. The pretreated carboxyl SWCNTs were modified on the surface of amino-functioned Fe3O4 nanoparticles through the cross-linking reaction between 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS). The synthesized Fe3O4/CNTs nanoparticles presented high super paramagnetic property and good dispersion ability, and were considered as good candidate adsorbents for dispersive solid phase microextraction. In this work, the synthesized Fe3O4/CNTs nanoparticles were applied to the extraction of vanillin additives, which was combined with high performance liquid chromatography (HPLC) analysis. The rapid and efficient preconcentration of vanillin and ethyl vanillin from different milk samples were successfully obtained with the detection limits of 10 μg/L and satisfactory recoveries of more than 92%. The results indicated that Fe3O4/CNTs magnetic nanoparticles were good pretreatment alternatives for the concentration of vanillin additives in milk products.

    Fast analysis of malachite green, leucomalachite green, crystal violet and leucocrystal violet in fish tissue based on a modified QuEChERS procedure
    ZHU Chengyun, WEI Jie, DONG Xuefang, GUO Zhimou, LIU Mingyang, LIANG Xinmiao
    2014, 32 (4):  419-425.  DOI: 10.3724/SP.J.1123.2014.01016
    Abstract ( 1105 )   [Full Text(HTML)] () PDF (1799KB) ( 617 )  

    Triphenylmethane dyes malachite green (MG) and crystal violet (CV) have been used as antimicrobial, antiparasitic and antiseptic agents in aquaculture. However, MG and CV, as well as their metabolites leucomalachite green (LMG) and leucocrystal violet (LCV) are potential mutagens and carcinogens. Thus, the efficient determination of dye residues is of great concern. Considering the complexity of the aquatic products, the sample pretreatment is significant for decreasing matrix interference and improving detection sensitivity. In this study, a simple and rapid QuEChERS procedure was developed and combined with HPLC analysis for the simultaneous determination of the four dyes in fish tissue. An XCharge C18 column was applied in HPLC analysis to achieve good peak shape and selectivity. The pretreatment method involved the extraction of dyes from fish tissue and further clean-up with dispersive solid phase extraction (d-SPE) material. The extraction volume, extraction time as well as d-SPE materials were systematically optimized. The results indicated that reversed-phase/strong anion exchange (C18SAX) adsorbent in the d-SPE procedure could effectively improve the recovery compared with conventional C18 or C18 incorporated with primary secondary amine (PSA) material. Under optimized conditions, good linearity was achieved in the concentration range of 0.5-100 mg/L with R2 greater than 0.998. The recoveries were 73%-91% and the precisions were 0.66%-5.41%. The results demonstrated the feasibility and efficiency of QuEChERS procedure incorporated with HPLC for dye monitoring.

    Determination of n-octanol/water partition coefficients for persistent organic pollutants by reversed-phase high performance liquid chromatography with dual-point retention time correction
    LIANG Chao, QIAO Junqin, GE Xin, LIAN Hongzhen
    2014, 32 (4):  426-432.  DOI: 10.3724/SP.J.1123.2013.10034
    Abstract ( 1019 )   [Full Text(HTML)] () PDF (917KB) ( 408 )  

    n-Octanol/water partition coefficients (logKow) for persistent organic pollutants (POPs) including polycyclic aromatic hydrocarbons (PAHs), polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and decabromodiphenylethane (DBDPE) have been determined by a modified method of reversed-phase high performance liquid chromatography (RP-HPLC). A dual-point retention time correction (DP-RTC) was used to rectify chromatographic retention time (tR) shift resulted from stationary phase aging and so on. Based on this correction, the relationship model between logKow and logkw, the logarithm of the retention factor extrapolated to pure water, was trained by a set of model compounds (a total of 37) with reliable experimental logKow as training set, including benzene homologues, PAHs and PCDD/Fs-related compounds. A linear regression equation of logKow=(1.18±0.02) logkw+(0.36±0.11) was established with correlation coefficient (R2) of 0.985, cross-validated correlation coefficient (Rcv2) of 0.983 and standard deviation (SD) of 0.16. This quantitative structure retention relationship (QSRR) model was further validated using four verification compounds, biphenyl, fluorene, PCDD 1 and PCDF 114, with reliable experimental logKow values. The RP-HPLC-determined Kow values showed good consistency with shake-flask (SFM) or slow-stirring (SSM) results, especially for highly hydrophobic compounds. Then, the logKow values for 29 POPs of wide interest were evaluated by the improved RP-HPLC method for the first time. The DP-RTC-HPLC method is recommended for the determination of the logKow values of POPs with strong hydrophobicity.

    Determination of N-nitrosodimethylamine in beer by frozen zone melting liquid-liquid extraction/gas chromatography
    PENG Qiaorong, TANG Tao, YU Shuxin, SUN Yuanshe, LEI Wu, WANG Fengyun, ZHANG Weibing, LI Tong
    2014, 32 (4):  433-437.  DOI: 10.3724/SP.J.1123.2013.12030
    Abstract ( 725 )   [Full Text(HTML)] () PDF (907KB) ( 246 )  

    A simple and effective sample enrichment method of frozen zone melting liquid-liquid extraction was optimized and validated for the analysis of trace N-nitrosodimethylamine (NDMA) in beer samples. The method was based on high pressure liquid-liquid extraction with a low temperature frozen step. The 90 mL beer was placed in a container with 10 mL dichloromethane. After agitation, the sample was kept in a freezer for 16 h at -19 ℃. The organic extract was analyzed by gas chromatography with a flame ionization detector (GC-FID). The accuracy, precision, detection and quantification limits and linearity of the method were evaluated. The results showed that the calibration curve of NDMA was linear in the range of 5-200 mg/L with a good correlation coefficient (r2) of 0.9996. The recoveries at the spiked levels of 5, 10 and 20 mg/L were 84.94%, 83.24%, 85.14% with the relative standard deviations (n=7)of 3.06%, 3.19%, 2.63%, respectively. The ordinary extraction method of N-nitrosodimethylamine in beer includes the four steps of low-temperature distillation, liquid-liquid extraction, rotary evaporation and nitrogen blowing concentration. With the extremely low volume of solvent used, the proposed extraction method proved to be easy and simple, and adequate for high-throughput analysis at low cost.

    Analysis of tartrazine aluminum lake and sunset yellow aluminum lake in foods by capillary zone electrophoresis
    ZHANG Yiding, CHANG Cuilan, GUO Qilei, CAO Hong, BAI Yu, LIU Huwei
    2014, 32 (4):  438-442.  DOI: 10.3724/SP.J.1123.2013.11020
    Abstract ( 1253 )   [Full Text(HTML)] () PDF (1510KB) ( 394 )  

    A novel analytical method for tartrazine aluminum lake and sunset yellow aluminum lake using capillary zone electrophoresis (CZE) was studied. The pigments contained in the color lakes were successfully separated from the aluminum matrix in the pre-treatment process, which included the following steps: dissolve the color lakes in 0.1 mol/L H2SO4, adjust the pH of the solution to 5.0, then mix it with the solution of EDTA·52Na and heat it in a water bath, then use polyamide powder as the stationary phase of solid phase extraction to separate the pigments from the solution, and finally elute the pigments with 0.1 mol/L NaOH. The CZE conditions systematically optimized for tartrazine aluminum lake were: 48.50 cm of a fused silica capillary with 40.00 cm effective length and 50 μm i. d., the temperature controlled at 20.0 ℃, 29.0 kV applied, HPO42--PO43- (0.015 mol/L, pH 11.45) solution as running buffer, detection at 263 nm. The conditions for sunset yellow aluminum lake were: the same capillary and temperature, 25.0 kV applied, HPO42--PO43- (0.025 mol/L, pH 11.45) solution as running buffer, detection at 240 nm. The limits of detection were 0.26 mg/L and 0.27 mg/L, and the linear ranges were 0.53-1.3×102 mg/L and 0.54-1.4×102 mg/L for tartrazine aluminum lake and sunset yellow aluminum lake, respectively. The RSDs were 4.3% and 5.7% (run to run, n=6), 5.6% and 6.0% (day to day, n=6) for tartrazine aluminum lake and sunset yellow aluminum lake, respectively. Further developments for this method could make it a routinely used method analyzing color lakes in foods.