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    Chinese Journal of Chromatography
    2019, Vol. 37, No. 7
    Online: 08 July 2019

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    Contents
    Volume 37, Number 7 Content
    2019, 37 (7):  0-0. 
    Abstract ( 66 )   PDF (4690KB) ( 36 )  
    Articles
    Synthesis and applications of epoxy-functionalized bi-functional magnetic molecularly imprinted polymers for polysaccharide adsorption
    HUANG Weiwei, ZHAO Qianyu, YANG Xin, YAO Lei, ZHAO Haitian
    2019, 37 (7):  673-682.  DOI: 10.3724/SP.J.1123.2019.01018
    Abstract ( 201 )   [Full Text(HTML)] () PDF (7350KB) ( 169 )  
    A polysaccharide-imprinted nanoparticle composite material with bi-functional monomers molecularly imprinted polymers (Bi-MMIPs) was successfully synthesized in aqueous solution by using starch as the template, 3-aminobenzeneboronic acid (APBA) as the functional monomer, 2-acrylamide-2-methylpropanesulfonic acid (AMPS) as the co-functional monomer, and ammonium persulfate (APS) as the initiator. Transmission electron microscopy (TEM), scanning electron microscopy (SEM), and Fourier-transform infrared (FT-IR) spectroscopy were used to characterized Bi-MMIPs. The adsorption and recognition characteristics of Bi-MMIPs for starch were investigated in detail by using the static method, dynamic method, and competitive adsorption experiments. The TEM, SEM and FT-IR results showed that the two functional monomers were loaded on the surface of Bi-MMIPs. Bi-MMIPs possessed very strong adsorption affinity and specific recognition for starch. The saturated adsorption capacity reached 13.88 mg/g, and the selectivity coefficients relative to glucosans Mr 5000 Da and 70000 Da were 2.67 and 3.77, respectively. Despite the imprinting factor of Bi-MMIPs (α=3.04), Bi-MMIPs were regenerated easily and exhibited excellent recognition, selectivity, and reusability for adsorbing starch. To elucidate the mechanism, APBA and AMPS showed synergy effect in synthesizing bi-functional monomers by providing reversible covalent bonds and hydrogen bonds, respectively, which could effectively enhance the spatial arrangement of combining sites for template molecular.
    Preparation and chromatographic evaluation of zwitterionic stationary phases with controllable ratios of positively and negatively charged groups
    CHENG Xiaodong, LI Yunping, HE Yinju
    2019, 37 (7):  683-691.  DOI: 10.3724/SP.J.1123.2019.01038
    Abstract ( 140 )   [Full Text(HTML)] () PDF (983KB) ( 84 )  
    The present study focuses on the preparation and application of zwitterionic stationary phases with controllable ratios of positively charged amino groups and negatively charged phosphonic acid groups. Successful grafting was confirmed by elemental analysis and evaluation of pore structure characteristics. Additionally, the proportion of amino and phosphonic acid groups could be easily determined by the mass fractions of N and P. In this study, three types of amino phosphate-based zwitterionic surface-bonded stationary phases (APS) were prepared with different ratios of amino and phosphonic acid groups. The resulting APS silica materials were slurry-packed into stainless-steel columns (150 mm×4.6 mm). Various parameters such as column temperature, water content, pH, and ionic strength of the mobile phase were investigated to evaluate the chromatographic characteristics of the APS columns. A mixed-mode retention mechanism involving surface adsorption, partitioning, and electrostatic interactions was revealed. Finally, a set of test mixtures (nucleosides, water-soluble vitamins, benzoic acids, and basic compounds) were employed to evaluate the separation selectivity and retention characteristics of the three prepared APS stationary phases. The APS phases demonstrated entirely different selectivity and retention behavior for various polar analytes under the same conditions, thus demonstrating excellent application potential for the analysis of polar compounds in hydrophilic interaction chromatography (HILIC).
    Synthesis and application of a surface molecularly imprinted adsorbent for di(2-ethylhexyl)phthalate base on graphite oxide
    QIU Haotian, ZHENG Ningning, FANG Quanhui, LIN Meiying, ZHOU Sunying
    2019, 37 (7):  692-700.  DOI: 10.3724/SP.J.1123.2019.01049
    Abstract ( 117 )   [Full Text(HTML)] () PDF (4203KB) ( 68 )  
    A molecularly imprinted polymer (MIP) base on the surface of graphite oxide (GO) has been developed for the selective recognition of di(2-ethylhexyl)phthalate (DEHP), and applied for the extraction of DEHP in a milk bag sample, with detection by high-performance liquid chromatography. The surface-molecularly imprinted material was prepared by precipitation polymerization in N,N-dimethylformamide (DMF) solvent, using GO as the supporting material, DEHP as the template molecule, and methacrylic acid (MAA) as the functional monomer; the synthesis conditions were also optimized. The obtained GO-MAA-MIP was characterized by Fourier transform infrared (FTIR) spectroscopy and transmission electron microscopy (TEM). The adsorption properties of the product were evaluated by adsorption experiments, including adsorption isotherms, kinetics, selectivity, saturated adsorption capacity, adsorption equilibrium time, and reusability. Under optimized conditions, DEHP was effectively extracted in the real sample (milk bag) and detected by HPLC. Linearity was obtained with a correlation coefficient (R2) of 0.9979 in a linear range of 0.5-50 mg/L. The limits of detection and quantitation were 0.03 mg/L and 0.1 mg/L, respectively. The average recoveries of the spiked samples at three concentration levels of DEHP ranged from 81.6% to 92.4% with relative standard deviations (RSDs) less than 7%. The results indicated that the proposed GO-MAA-MIP-SPE (solid phase extraction) protocol with HPLC-UV detection could be applied for the selective analysis of DEHP in real samples.
    Determination of 50 non-edible additives in health food using the QuEChERS method and liquid chromatography-tandem mass spectrometry
    HU Shengjie, LI You, ZHOU Ying, YI Xionghai, DENG Xiaojun, CHEN Qin, XU Dunming, NIU Bing
    2019, 37 (7):  701-711.  DOI: 10.3724/SP.J.1123.2019.01046
    Abstract ( 103 )   [Full Text(HTML)] () PDF (4760KB) ( 86 )  
    A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established for the determination of 50 non-food additives in health food. The samples were extracted with methanol and purified using the QuEChERS method. The separation was performed on an Agilent Phoroshell SB C18 column (150 mm×3.0 mm, 2.7 μm) with a mobile phase of 0.1% (v/v) formic acid aqueous solution and acetonitrile. The determination was performed by MS/MS in positive or negative electrospray ionization and multiple reaction monitoring mode. The results demonstrated that all 50 compounds could be well separated with a good linear relationship (r2>0.99). The limits of detection (LODs, S/N ≥ 3) and limits of quantification (LOQs, S/N ≥ 10) were in the range of 0.010-1 mg/kg and 0.020-2 mg/kg, respectively. The matrix effects in five kinds of typical health foods (oral liquid, tablets, ointments, pills, and capsules) were evaluated and reduced by means of matrix matching. At the spiked levels of 1 LOQ, 2 LOQ, and 10 LOQ, the recoveries of all drugs varied from 63.1% to 115.7% with relative standard deviations (RSDs) of no more than 8.9% (n=6). The method is simple, sensitive, practical, and suitable for monitoring 50 non-edible additives in health food.
    Detection of seven kinds of aquatic product allergens in meat products and seasoning by liquid chromatography-tandem mass spectrometry
    MENG Jia, GU Shuqing, FANG Zhen, NIU Bing, DENG Xiaojun, GUO Dehua, ZHU Jian, HAN Fang
    2019, 37 (7):  712-722.  DOI: 10.3724/SP.J.1123.2019.01042
    Abstract ( 173 )   [Full Text(HTML)] () PDF (2147KB) ( 255 )  
    A liquid chromatography-tandem mass spectrometry method for the identification of marker peptides of aquatic product allergens and quantitative detection of multiple allergens in meat products and seasonings was developed. The samples were prepared by protein extraction, protein purification, and trypsin hydrolysis. The proteins and peptides were identified using ProteinPilot by the data analysis of the ion spectrum of polypeptide fragments using ultra-performance liquid chromatography-quadrupole/electrostatic orbitrap high-resolution mass spectrometry (UPLC-Q/Exactive-HRMS). The identification of 30 species-specific marker peptides in Penaeus vannamei, Eriocheir, Scylla serrata, Thunnus thynnus, and Atlantic salmon by comparison of the basic local alignment search tool (BLAST) with the UniProt database was achieved. The verification and multiple reaction monitoring (MRM) quantitative studies of these marker peptides were performed using a triple quadrupole mass spectrometry (UPLC-QqQ-MS) system. The proposed method showed a good linear relationship in the range of 5-250 mg/kg. The limits of quantitation and observed recoveries were in the range of 2-3.5 mg/kg and 88.7%-110.2%, respectively. This method presents various advantages such as good repeatability and high throughput, suitability for rapid screening, and quantitative analysis of seven aquatic allergens in meat products and seasonings.
    Determination of cylindrospermopsin, nodularin and microcystins in freshwater fish by dispersive solid phase extraction-liquid chromatography-tandem mass spectrometry
    CHEN Lihui, JIA Yuzhu, ZHANG Bin, PAN Qiuren, CHEN Qinqin, CAI Weipeng
    2019, 37 (7):  723-728.  DOI: 10.3724/SP.J.1123.2019.01022
    Abstract ( 206 )   [Full Text(HTML)] () PDF (1170KB) ( 137 )  
    A method was developed for the determination of cylindrospermopsin (CYN), nodularin (NOD), microcystin-RR (MC-RR), microcystin-YR (MC-YR) and microcystin-LR (MC-LR) in freshwater fish by dispersive solid phase extraction-liquid chromatography-tandem mass spectrometry (DSPE-LC-MS/MS). The analytes were extracted from fish tissues with acetonitrile-water-formic acid (89:10:1, v/v/v), and purified by DSPE using C18 as the adsorbent. The separation of analytes was performed on an Agilent ZORBAX Eclipse XDB C18 column with the gradient elution of acetonitrile and water as mobile phases. Qualitative analysis was performed using the multiple reaction monitoring (MRM) mode. The analytes were quantified by matrix-matched external standard curves. The chromatographic and MS parameters were optimized. Major factors affecting the extraction and cleanup efficiencies including the type of extraction solvent and cleanup sorbent were investigated. The linear correlation coefficients (R2) of the five target compounds were no less than 0.9954. The limits of detection (LODs, S/N=3) and limits of quantification (LOQs, S/N=10) of the five target compounds were 5-10 μg/kg and 15-40 μg/kg, respectively. The spiked recoveries of the five target compounds ranged from 62.3% to 101.2%. The developed method is simple, rapid, accurate, sensitive, and is suitable for the determination of cylindrospermopsin, nodularin and microcystins in freshwater fish.
    Determination of chlorpyrifos residues in the paddy field aquatic products by ultra performance liquid chromatography-tandem mass spectrometry
    PENG Jie, GAN Jinhua, JU Xiaoqian, CHEN Jianwu, HE Li
    2019, 37 (7):  729-734.  DOI: 10.3724/SP.J.1123.2019.01026
    Abstract ( 182 )   [Full Text(HTML)] () PDF (1100KB) ( 77 )  
    A method was developed for the determination of chlorpyrifos residues in paddy field aquatic products by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Samples were extracted with acetonitrile and purified by using 0.2 g primary secondary amine (PSA) sorbent and 1.2 g MgSO4. The target compound was separated on a Hypersil GOLD C18 column (100 mm×2.1 mm, 5 μm), and was determined using a heated electrospray ionization (HESI) source in the positive ion selective reaction monitoring (SRM) mode. The analyte was quantified with external standard using the matrix-matched standard calibration curve method. The results showed that good linearities were obtained in the range of 0.5-100.0 μg/L, and the correlation coefficients were greater than 0.999. The spiked recoveries of chlorpyrifos ranged from 86.2% to 103.6% with RSDs of 3.5%-7.6% (n=6). The limits of detection and quantification were 0.25 μg/kg and 0.5 μg/kg, respectively. This method is simple, quick, sensitive, and suitable for the rapid determination and analysis of chlorpyrifos residues in paddy field aquatic products.
    Determination of streptomycin and dihydrostreptomycin residues in honey by hydrophilic interaction liquid chromatography-tandem mass spectrometry
    WEI Lili, XUE Xia, LIU Yanming, SUN Lizhen, CHENG Zhi, SU Shufang, ZHAO Yin
    2019, 37 (7):  735-741.  DOI: 10.3724/SP.J.1123.2019.01029
    Abstract ( 192 )   [Full Text(HTML)] () PDF (1196KB) ( 128 )  
    An analytical method was developed for the determination of streptomycin and dihydrostreptomycin in honey using hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS). The streptomycin and dihydrostreptomycin residues in the test samples were extracted with 20 g/L trichloroacetic acid aqueous solution (including 50 mmol/L phosphate, pH 6.8) and cleaned on an Oasis HLB solid phase extraction column. The products were separated on a SIELC Obelisc R column with gradient elution using 0.5% (v/v) formic acid aqueous solution and acetonitrile as mobile phases. Streptomycin and dihydrostreptomycin were detected by liquid chromatography-tandem mass spectrometry in the positive ion mode using the external standard method. Under the optimal conditions, streptomycin and dihydrostreptomycin showed good linearity (r>0.99) in the range of 2.5-100 μg/L. The LOD and LOQ of the method was 2.0 μg/kg and 5.0 μg/kg, respectively. The spiked recoveries of the analytes from blank honey samples at the three levels of 5.0, 20.0 and 100.0 μg/kg were in the range of 86.9%-113.2% with the relative standard deviations less than 10%. With the advantages of convenience, rapidity, sensitivity and good repeatability, the method is suitable for the detection of streptomycin and dihydrostreptomycin in honey.
    Determination of seven indicative ingredients in oral liquids of Chinese medicine by liquid chromatography-electrospray ionization-ion mobility spectrometry
    FANG Kang, GUO Xiangyu, WANG Hongwei, XIONG Xingchuang, BAI Hua, LEI Haimin, MA Qiang
    2019, 37 (7):  742-749.  DOI: 10.3724/SP.J.1123.2019.01013
    Abstract ( 132 )   [Full Text(HTML)] () PDF (3152KB) ( 142 )  
    A two-dimensional separation and analysis method, based on liquid chromatography-electrospray ionization-ion mobility spectrometry (LC-ESI-IMS), is developed for the determination of seven indicative ingredients (danshensu, glycyrrhizic acid, gastrodin, chlorogenic acid, puerarin, baicalin, and rutin) in oral liquids of Chinese medicine. The sample was first separated on an ACQUITY UPLC BEH C18 column (50 mm×1 mm, 1.7 μm). The post-column effluent was directed to an adjustable flow splitter with a split ratio of 50:1. The low-flow and high-flow outlets were connected to an ion mobility spectrometer and a triple quadrupole mass spectrometer, respectively. The experimental conditions for LC, spray voltage, drift tube temperature, gas pre-heating temperature, and drift gas velocity were systematically optimized. The limits of detection (LODs) and quantitation (LOQs) for the seven analytes were 2-10 μg/mL and 5-25 μg/mL, respectively. The proposed method was applied for the analysis of real oral liquids of Chinese medicine samples. By coupling LC and IMS, two-dimensional separation could be achieved based on hydrophobicity difference and ionic mobility disparity, thus providing more comprehensive measurement information than LC or IMS used alone.
    Development and application of two-dimensional liquid chromatography for the analysis of polycyclic aromatic sulfur heterocycles in heavy oil
    SONG Chunxia, WANG Wei, LIU Zelong, LIU Yingrong, CAI Xinheng
    2019, 37 (7):  750-758.  DOI: 10.3724/SP.J.1123.2019.01014
    Abstract ( 141 )   [Full Text(HTML)] () PDF (1781KB) ( 72 )  
    A two-dimensional liquid chromatographic (2D LC) system was developed for the analysis of polycyclic aromatic sulfur heterocycles (PASHs) in heavy oil. The first dimension is a ligand-exchange chromatographic column packed with a palladium chloride/silica (PdCl2-SiO2) stationary phase, while the second dimension is a Spherisorb-NH2 column. The PASHs in vacuum gas oil were enriched online, and polycyclic aromatics hydrocarbons (PAHs) were separated according to their aromatic ring numbers. These separated fractions were analyzed by Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS), and detailed molecular characterization of PASHs and PAHs was achieved. Typical molecular structures of PASHs and PAHs in these fractions could be provided, and the side chain as well as the core structures were predicted. It is significant for the feedstocks selection and optimization of the hydrodesulfurization process by using these molecular characterization information.
    Simultaneous determination of 33 hair dyes in oxidative hair dye products by high performance liquid chromatography
    ZUO Xue, DI Zheng, ZHANG Rong, WU Guoqing
    2019, 37 (7):  759-765.  DOI: 10.3724/SP.J.1123.2019.01043
    Abstract ( 192 )   [Full Text(HTML)] () PDF (856KB) ( 129 )  
    A method was developed for the simultaneous determination of 33 hair dyes in oxidative hair dye products by high-performance liquid chromatography (HPLC). The analytes were separated on a Waters Atlantis® T3 MV Kit column (250 mm×4.6 mm, 5 μm) by gradient elution using phosphate buffer-acetonitrile as mobile phases. The components were detected at two different wavelengths of 235 and 280 nm using a diode array detector. The results showed that the method had good linearity with correlation coefficients exceeding 0.999. Precision was good for the 33 analytes, with the relative standard deviations (RSDs) less than 2%. Stabilities of tetraaminopyrimidine sulfate and 2,4-diaminophenoxyethanol HCl in 12 h, as well as other analytes in 24 h, were good, with RSDs less than 5%. Recoveries of the method ranged from 77.6% to 116.3% at three different concentration levels. The method proved to be simple, rapid, and accurate, suitable for the determination of various hair dyes in oxidative hair dye products
    Rapid determination of six banned fragrance components in tea by QuEChERS-gas chromatography-tandem mass spectrometry
    WANG Yujiao, LIU Tong, DENG Yamei, WANG Jiaqi, CHEN Hong, HUA Erbing, ZHANG Feng
    2019, 37 (7):  766-772.  DOI: 10.3724/SP.J.1123.2019.01032
    Abstract ( 173 )   [Full Text(HTML)] () PDF (1703KB) ( 112 )  
    QuEChERS purification was established as a sample pretreatment method and combined with gas chromatography-tandem mass spectrometry (GC-MS/MS) for the simultaneous detection of six banned fragrance components in tea (coumarin, thujone, 2-exylthiophene, trans-beta-acacene, p-mentha-1,8(10)diene-9-ethyl acetate, and butyl cinnamate). Tea samples were extracted with ethyl acetate and subjected to whirlpool oscillation, following which anhydrous magnesium sulfate, primary secondary amine (PSA), and octadecylsilyl silica gel (C18) were added to the supernatant. After centrifugation, the supernatant was filtered over a membrane and separated on a DB-5MS column. The six compounds were determined by GC-MS/MS in MRM mode, and quantified by a matrix-matched external standard method. The six banned flavor compounds exhibited good linearity over the range of 1-200 μg/L, and the linear correlation coefficients (R2) were greater than 0.999. The limits of detection were 0.005-1 μg/kg and the limits of quantification were 0.02-2 μg/kg, the average recoveries were 82.3%-93.1%, and the intra-day and inter-day RSDs were each less than 10%. The method is simple, rapid, accurate, and sensitive, and can be used for the simultaneous detection of six banned fragrance components in tea.
    Simultaneous analysis of five organic acids in aqueous, lotion, and cream cosmetics by capillary electrophoresis
    WANG Huimin, SUN Miao, QU Feng
    2019, 37 (7):  773-777.  DOI: 10.3724/SP.J.1123.2019.01021
    Abstract ( 169 )   [Full Text(HTML)] () PDF (811KB) ( 73 )  
    An indirect ultraviolet detection method for simultaneous analysis of five organic acids by capillary electrophoresis was established. The concentration of the tetradecyltrimethylammonium bromide (TTAB) surfactant in the background electrolyte solution and the pH of the solution were optimized. Five organic acids formic acid, malic acid, citric acid, glycolic acid, lactic acid were effectively separated from each other within 6 min with an electrolyte solution (15 mmol/L potassium hydrogen phthalate and 0.4 mmol/L TTAB, pH 5.6) under conditions of -15 kV, 25℃, and 254 nm, while the injection pressure was 5 kPa and the injection time was 5 s. The linear ranges were 15-600 mg/L, 30-800 mg/L, 20-700 mg/L, 40-500 mg/L, and 30-5000 mg/Lfor formic acid, malic acid, citric acid, glycolic acid, and lactic acid, respectively, with correlation coefficients of 0.9983-0.9998. The detection limits were between 0.1 μg/g and 2.0 μg/g. The method can be used for the detection of five organic acids in three types of cosmetics including aqueous, lotion and cream. At three spiked levels, the recoveries of organic acids were between 95.0% and 101.6%, and relative standard deviations (RSDs) were within 2.0%. The proposed method is easy, rapid, environment friendly, and sensitive with good reproducibility. It can be used for the monitoring of five organic acids in the production and storage of cosmetics including aqueous, lotion and cream.
    Technical Notes
    Simultaneous determination of 21 illegally added compounds in health foods by ultra high performance liquid chromatography-tandem mass spectrometry with solid phase extraction
    XU Dunming, LAI Guoyin, CHEN Yan, LUO Chao, YI Xionghai, DENG Xiaojun, FENG Feng, ZHANG Feng
    2019, 37 (7):  778-785.  DOI: 10.3724/SP.J.1123.2019.01016
    Abstract ( 160 )   [Full Text(HTML)] () PDF (1395KB) ( 96 )  
    Supporting Information
    A rapid method for the simultaneous determination of 21 illegally added compounds in health foods was developed based on solid phase extraction (SPE) combined with ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). After ultrasonic extraction with acetonitrile, the samples were purified on an HLB SPE column, separated on a Waters BEH C18 column (100 mm×2.1 mm, 1.7 μm) using 10 mmol/L ammonium acetate aqueous solution and methanol as the mobile phases. The compounds were detected by an electrospray ionization (ESI) source in positive ion mode with multi-reaction monitoring (MRM). The 21 illegal additives showed good linear relationships with correlation coefficients no less than 0.995. The limits of detection (LODs) were 3-160 μg/kg for different matrixes. The recoveries ranged from 61.8% to 109.3%, and the relative standard deviations ranged from 1.6% to 14.7%. The method can be used for the simultaneous determination of the illegally added drugs in the losing weight, lipid-lowering, anti-diabetic and anti-hypertension health foods.