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    Chinese Journal of Chromatography
    2008, Vol. 26, No. 5
    Online: 30 September 2008

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    Articles
    Preparation of magnesia-zirconia based mimetic biomembrane stationary phase and its applications in evaluating drug-membrane interactions
    HU Zhixiong, ZHANG Weinong, HE Haibo, FENG Yuqi, DA Shilu
    2008, 26 (5):  529-533. 
    Abstract ( 2418 )   [Full Text(HTML)] () PDF (353KB) ( 547 )  
    A novel mimetic biomembrane chromatographic stationary phase of magnesia-zirconia composite matrix was prepared based on the Lewis acid-base interaction between the phosphonate group of phosphatidylcholine residue and the Lewis acid sites of magnesia-zirconia composite. The infrared absorption spectrum and X-ray photoelectron spectrum of the stationary phase illustrated that the magnesia-zirconia composite was successfully modified with phosphatidylcholine. The interactions between the membrane and the drugs were evaluated. It is observed that the log Kmbm values have good relationships with the log Papp, and the linear slope is 1.049, which is near unity. Moreover, on the basis of the thermodynamics derivation, the difference in standard free energies (Δ(ΔG°)) is introduced to describe the drug-membrane interaction. The results show that the log Kmbm and Δ (ΔG°) value provide key information on the transport properties of the drugs. The establishment of this chromatographic model may be a new way for the evaluation of the drug-membrane interactions.
    Identification of Mycobacterium species using reversed-phase high performance liquid chromatographic analysis of mycolic acid
    DU Rong, CHEN Baowen, GUO Lei, LI Yang, XIE Jianwei, WANG Guozhi, ZHOU Hongbing
    2008, 26 (5):  534-539. 
    Abstract ( 3210 )   [Full Text(HTML)] () PDF (747KB) ( 587 )  
    A method for the identification of Mycobacterium species using reversed-phase high performance liquid chromatography (RP-HPLC) was developed. The fingerprints library was constructed on the basis of RP-HPLC chromatograms of the mycolic acids derivatives from 49 Mycobacterium species cultures. Two inoculation loops of Mycobacterium, for fast growth with one week incubation or for slow growth with three weeks incubation, were saponified for 1 h and stocked at 4 ℃. The mycolic acids from each culture of Mycobacterium species were acidified, extracted, derivatized, and analyzed by the RP-HPLC method. On the basis of the HPLC patterns of relative retention time and relative peak height, the identifications of Mycobacterium species were performed. This established method has a good precision of retention times with the relative standard deviations (RSDs) ranging from 0.13% to 1.07%. The mycolic acids fingerprints library of HPLC patterns was set up, including 49 species that were recorded in “Bergey's Manual of Systematic Bacteriology”. Three patterns were observed from the chromatographic behaviors of mycolic acids derivatives, including single peak-cluster, double peak-clusters, triple and multiple peak-clusters. Forty-one species were successfully identified according to the different relative retention times of the peaks and the relative peak heights. The established method can identify Mycobacterium species with rapidity and high reliability.
    Analysis of the related substances in phosphorothioate antisense oligonucleotide FT19
    ZHANG Hongyan, LU Dandan, WU Lixia, ZHOU Zhe, WANG Shengqi
    2008, 26 (5):  540-543. 
    Abstract ( 2771 )   [Full Text(HTML)] () PDF (431KB) ( 652 )  
    A method using anion exchange high performance liquid chromatography (AX-HPLC) and capillary gel electrophoresis (CGE) was established for the analysis of the related substances in phosphorothioate antisense oligonucleotide FT19 for quality control. The FT19 and its analogs of partial phosphodiester compounds (P=O)1 as well as deletion sequences (n-1) were analyzed. AX-HPLC with the DNA Pac PA-100 column (4 mm×250 mm) was used for the separation of phosphorothioate modifications. The size of the capillary column used in the CGE was 31 cm × 100 μm with an effective length of 20 cm. It was found that full phosphorothioate and partial phosphodiester oligonucleotides of the same length were successfully separated, and the deletion sequences (n-1) and the full-length sequence n were successfully separated in CGE. The results demonstrated that the related substances of phosphorothioate oligonucleotides can be well analyzed by AX-HPLC and CGE.
    Simultaneous determination of olmesartan medoxomil and irbesartan and hydrochlorothiazide in pharmaceutical formulations and human serum using high performance liquid chromatography
    SULTANA Najma, ARAYNE M Saeed, ALI S Shahid, SAJID Shahnawaz
    2008, 26 (5):  544-549. 
    Abstract ( 2491 )   [Full Text(HTML)] () PDF (280KB) ( 462 )  
    A simple, selective, sensitive, precise, simultaneous high performance liquid chromatographic analysis of serum samples and commercial tablet formulation containing hydrochlorothiazide, olmesartan medoxomil and irbesartan are reported. Good chromatographic separation was achieved using a μ-Bondapak, C18 column (15 cm×4.6 mm, 5 μm), and a mobile phase consisting of acetonitrile-0.2% acetic acid aqueous solution (50∶50, v/v) at a flow rate of 1.0 mL/min. The ultraviolet detector was set at a wavelength of 260 nm. Hydrochlorothiazide, olmesartan medoxomil, and irbesartan were eluted at 1.2, 3.8, and 4.4 min, respectively. No extraneous materials were found to interfere. The method uses protein precipitation with acetonitrile for the preparation of serum sample. The linear ranges for hydrochlorothiazide, olmesartan medoxomil, and irbesartan were 6.25-18.75, 20-60, and 75-225 ng/mL, respectively. The recoveries of hydrochlorothiazide, olmesartan medoxomil, and irbesartan in spiked samples were all greater than 98%, and their relative standard deviations were less than 2.0%. The limits of detection were 1, 2, and 2 ng/mL for hydrochlorothiazide, olmesartan medoxomil, and irbesartan, respectively, and the limits of quantification were 3 ng/mL, which allow their determination at the expected serum concentration levels.
    Determination of kinetic transformation of two geometrical isomers of the [Fe(PDT)3]2+ by high performance liquid chromatography
    ZHU Weihuang, WU Fengchang, HUANG Tinglin
    2008, 26 (5):  550-553. 
    Abstract ( 2498 )   [Full Text(HTML)] () PDF (393KB) ( 522 )  
    The kinetic transformation of the two isomers of the [Fe(PDT)3]2+ (PDT: 3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine) was studied by high performance liquid chromatography. The transformation between two isomers was proved to be treated kinetically as the first-order reaction. At different reaction temperatures, the linear regression equations between xeln[(xe-x0)/(xe-x)] and t(min) were as follows: xeln[(xe-x0)/(xe-x)]=0.082t+0.729 (r2=0.9911, T=45 ℃), xeln[(xe-x0)/(xe-x)]=0.049t+0.598 (r2=0.9987, T=40 ℃), xeln[(xe-x0)/(xe-x)]=0.022t+0.586 (r2=0.9987, T=35 ℃), xeln[(xe-x0)/(xe-x)]=0.012t+0.591(r2=0.9988, T=30 ℃). The activation enthalpy (ΔH), activation entropy (ΔS), and activation energy (ΔEa) characterizing the kinetic transformation were as follows: ΔH=103.84 kJ·mol-1, ΔS=271.93 J·mol-1·K-1, ΔEa=86.74 kJ·mol-1 (fac-isomer→mer-isomer); ΔH=106.47 kJ·mol-1, ΔS=257.65 J·mol-1·K-1, ΔEa=94.43 kJ·mol-1 (mer-isomer→fac-isomer).
    Determination of 22 components in hair dyes by high performance liquid chromatography
    ZHU Huijuan, YANG Yanwei, ZHANG Weiqiang, ZHU Ying
    2008, 26 (5):  554-558. 
    Abstract ( 2735 )   [Full Text(HTML)] () PDF (297KB) ( 1037 )  
    A high performance liquid chromatographic (HPLC) method was developed for the determination of 22 components in hair dyes. The target analytes were separated on an amide bonded C16 silica column (250 mm×4.6 mm, 5 μm), employing acetonitrile-0.025 mol/L phosphate buffer (pH 6.0) containing 0.1% 1-heptanesulfonic acid sodium salt as the mobile phase and detected by a photodiode array detector (DAD) with the detection wavelengths of 260 nm and 280 nm. The flow rate was 1.0 mL/min and the column temperature was 25 ℃. The linear range was from 10 mg/L to 500 mg/L with good relationship. The relative standard deviations were less than 10%(except toluene-2,5-diamine sulfate, 2-methylresorcinol, N,N-diethyltoluene-2,5-diamine HCl at low concentrations) and the recoveries were from 77.6% to 122.8%. The method is simple, rapid and accurate, and is suitable for the analysis of various hair dyes.
    Resolution of thyroxine hormone enantiomers by precolumn derivatization with a fluorescent chiral reagent using reversed-phase high performance liquid chromatography
    JIA Shaodong, ZHANG Meina, JIN Dongri
    2008, 26 (5):  559-562. 
    Abstract ( 2542 )   [Full Text(HTML)] () PDF (290KB) ( 560 )  
    A highly fluorescent chiral tagging reagent, R(-)-4-(N,N-dimethylaminosulfonyl)-7-(3-isothiocy-anatopyrrolidino)-2,1,3-benzoxadiazole (R(-)-DBD-PyNCS), was employed for the enantiomer separation of thyroxine hormone, D,L-3,5,3′,5′-tetraiodothyronine (T4) and L-3,5,3′-triiodothyronine (T3). The reaction of R(-)-DBD-PyNCS with the thyroxine enantiomers was carried out at 40 ℃ for 20 min under a basic medium surrounding to yield the corresponding pair of diastereomers. No racemization occurs during the tagging reaction under the optimized conditions. Various experimental parameters for the derivatization reaction including the concentration of tagging reagent, reaction temperature and reaction time have been studied in order to get the highest yield of T4/T3 derivatives. The structures of T4/T3 derivatives were identified based on high performance liquid chromatography-mass spectrometry (HPLC-MS) measurement in negative mode. The efficient separation of derivatives have been achieved by isocratic elution with a water-acetonitrile mobile phase containing 1% acetic acid in a reversed-phase column utilizing a conventional fluorescence detector. The calibration curves of L-T3, D,L-T4 were linear over the concentration ranges of 0.0067-0.22 μg/μL and 0.016-0.30 μg/μL, respectively. The limits of detection (S/N=3) for L-T3 and D,L-T4 were 0.85 μg/mL and 0.02 μg/mL, respectively. The proposed method has been successfully applied to the determination of T3 and T4 in clinical pharmaceutics.
    Determination of benomyl, carbendazim and thiabendazole in apple juice concentrate using solid-phase extraction coupled with ion exchange chromatography
    HE Qiang, KONG Xianghong, ZHAO Jie, LI Jianhua, YUE Aishan, ZHANG Ying
    2008, 26 (5):  563-567. 
    Abstract ( 2900 )   [Full Text(HTML)] () PDF (375KB) ( 789 )  
    A method was developed for the determination of benomyl, carbendazim and thiabendazole in apple juice concentrate by solid-phase extraction coupled with ion exchange chromatography (IEC). The sample was diluted with water, and then benomyl was degradated completely to carbendazim at 80 ℃, and purified by an SCX solid-phase extraction column. Liquid chromatographic analysis was performed on the instrument of Agilent 1200 series equipped with a diode-array detector and autosampler. The column was LC-SCX (25 cm×4.6 mm, 5 μm). The mobile phase was 0.1 mol/L KH2PO4 (pH 2.5)-acetonitrile (70∶30, v/v) with a flow rate of 1.0 mL/min. The presented method showed good linear relationship with good precision and accuracy at the range of 0.02-2.0 mg/L for carbendazim and hiabendazole. The detection limits were 0.004 mg/kg for carbendazim and thiabendazole. The method was characterized with cceptable sensitivity to meet the requirements for monitoring these pesticides in apple juice concentrate.
    Determination of 92 pesticide residues in tea by gas chromatography with solid-phase extraction
    LOU Zhengyun, CHEN Zongmao, LUO Fengjian, TANG Fubin, LIU Guangming
    2008, 26 (5):  568-576. 
    Abstract ( 2870 )   [Full Text(HTML)] () PDF (535KB) ( 1091 )  
    A multiresidue analytical method was developed for the determination of ninety-two pesticides in tea using solid-phase extraction (SPE) and gas chromatographic determination. Tea samples were extracted with acetonitrile, organophosphorus pesticides were clean-up by an Envi-Carb SPE cartridge, eluted with 10 mL acetonitrile-toluene (3∶1, v/v) and determined by gas chromatography-flame photometric detection (GC-FPD); organochlorine pesticides and pyrethroids pesticides were cleaned-up by Envi-Carb+NH2 SPE cartridges, eluted with 5 mL acetonitrile-toluene (3∶1, v/v) and determined by gas chromatography-electron capture detection (GC-ECD). The recoveries of those pesticides were ranged from 80.3% to 117.1% by fortification test with the relative standard deviations being from 1.5% to 9.8%. The limits of detection were 0.0025-0.10 mg/kg. The proposed method is characterized by simplicity, higher sensitivity and accuracy.
    A rapid multi-residual analysis for organophosphorus pesticides in the products of animal origin using gas chromatography coupled with accelerated solvent extraction and gel permeation chromatographic purification
    WU Gang, BAO Xiaoxia, WANG Huaxiong, YU Chunyan, WU Huiming, YE Qingfu
    2008, 26 (5):  577-582. 
    Abstract ( 2661 )   [Full Text(HTML)] () PDF (391KB) ( 1080 )  
    A rapid method has been developed to determine the multi-residues of 36 organophosphorus pesticides in the products of animal origin using capillary gas chromatography with flame photometric detector (GC-FPD (P)). The organophosphorus pesticides were extracted with acetonitrile by accelerated solvent extraction, and cleaned up by auto gel permeation chromatography and primary secondary amine (PSA) packing material. The collected solution was analyzed by the GC-FPD (P) and quantified by internal standard method. The 36 organophosphorus pesticides were separated efficiently from impurity in high sensitivity and reproducibility by GC-FPD (P). The limits of detection (LODs) ranged from 0.0012 mg/kg (phorate) to 0.014 mg/kg (pyraclofos), and the limits of quantitation (LOQs) ranged from 0.004 mg/kg (phorate) to 0.047 mg/kg (pyraclofos). The recoveries ranged from 58.2% to 106.3% in blank samples spiked with 0.05, 0.1 and 0.2 mg/kg of 36 organophosphorus pesticides. The LODs, LOQs and the recoveries of the method all satisfy the requirement of pesticide residue analysis.
    Determination of 28 organochlorine and synthetic pyrethroid pesticides in greasy wool using accelerated solvent extraction technique and gas chromatography
    FAN Yuanmu, HUANG Shaotang, YU Xuejun, GU Xiaojun, QIU Yajun, ZHAN Jia, CHEN Shubing, HE Xiaoyu, CHEN Jun, WANG Shijie, LI Yaping, LI Zhongbang
    2008, 26 (5):  583-589. 
    Abstract ( 2741 )   [Full Text(HTML)] () PDF (497KB) ( 651 )  
    A method for the simultaneous determination of 28 organochlorine and synthetic pyrethroid pesticides in greasy wool was developed by accelerated solvent extraction (ASE) coupled with gas chromatography-electron capture detection (GC-ECD). The organochlorine and synthetic pyrethroid pesticides in greasy wool were extracted with acetonitrile saturated with n-hexane at 80 ℃, 10.34 MPa by ASE. The extract was pretreated by a series of procedures such as freezing-lipid filtration, concentration and purification by solid-phase extraction prior to the determination with GC-ECD. The linear ranges were 0.005-1.0 mg/L for 16 organochlorines, 0.02-4.0 mg/L for flumethrin and 0.01-2.0 mg/L for the others. There were good linear relationships between the peak area and concentration in the linear ranges. The average recoveries of 28 organochlorine and synthetic pyrethroid pesticides were 67.2%-107.7%, and the relative standard deviations were 2.6%-29.0%. The method is simple, sensitive and suitable for preliminary screening of organochlorine and synthetic pyrethroid pesticides in greasy wool.
    Analysis of phenolic compounds in aqueous samples by gas chromatography coupled with headspace solid-phase microextraction using poly(phthalazine ether sulfone ketone) coated fiber
    YAO Guiyan, GUAN Wenna, XU Feng, WANG Hua, GUAN Yafeng
    2008, 26 (5):  590-594. 
    Abstract ( 2697 )   [Full Text(HTML)] () PDF (482KB) ( 648 )  
    The direct trace analysis of phenolic compounds in aqueous samples was performed by headspace solid-phase microextraction/gas chromatography (HS-SPME/GC). A laboratory made poly(phthalazine ether sulfone ketone) (PPESK, 30 μm) coated fiber was used to extract the phenols from aqueous samples. The parameters affecting the extraction efficiency, such as extraction temperature and time, pH value, and salt concentration, were optimized. The low pH value and high salt concentration can increase the extraction efficiency of phenols. The limits of detection (LODs) were from 0.003 to 0.041 μg/L, which were within the range of EPA Method 604. The relative standard deviations (RSDs) were less than 16%. Compared with commercial polyacrylate (PA) fiber (85 μm), the PPESK fiber shows high affinity toward phenolic compounds, and therefore, high absolute recoveries. The phenols were detected with the recoveries of 100.5%-111.8% for a tap water sample and 94.8%-117.3% for a seawater sample at the spiked level of 20 μg/L.
    Determination of hydroxyproline in collagenous proteins by high performance liquid chromatography-mass spectrometry
    XIA Jingen, CHEN Bo, YAO Shouzhuo
    2008, 26 (5):  595-598. 
    Abstract ( 2760 )   [Full Text(HTML)] () PDF (385KB) ( 1068 )  
    A simple, rapid and accurate method for the determination of hydroxyproline in collagenous proteins by high performance liquid chromatography-electrospray mass spectrometry (HPLC-ESI/MS) was developed. After hydrolysis of collagenous proteins with hydrochloric acid, the hydroxyproline was separated on a Spherigel C8 column using acetonitrile-0.05% trifluoroacetic acid aqueous solution (5∶95, v/v) at a flow rate of 1.0 mL/min. Theanine was used as the internal standard for the quantification. Hydroxyproline was identified and quantified by electrospray ionization mass spectrometry (ESI-MS), which was operated in positive ion mode while the m/z 132 and m/z 175 ions were recorded in the selective ionization monitoring (SIM) mode. The peak area ratio of hydroxyproline to theanine versus the hydroxyproline concentration was linear over a concentration range of 11.7-117 mg/L for hydroxyproline. The correlation coefficient was 0.9993. The reproducibility and average recovery of the method were 1.87% and 98.85% respectively. The method has the advantages of easy operation, without derivatization, less analysis time, good precision and accuracy.
    Simultaneous determination of five effective components in Qingrejiedu oral liquid using high performance liquid chromatography-mass spectrometry
    XU Haitang, HUANG Lihan, XU Yuanjin,
    2008, 26 (5):  599-602. 
    Abstract ( 3172 )   [Full Text(HTML)] () PDF (392KB) ( 702 )  
    A high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI/MS) method for the simultaneous determination of five effective components in Qingrejiedu oral liquid was developed. The HPLC separation was performed on a Zorbax SB C18 column (250 mm×3.0 mm, 5 μm) using 0.4 mmol/L sodium acetate solution containing 0.2% formic acid (A) and acetonitrile (B) as the mobile phase with gradient elution (0 min, 10% B; 06 min, 10%B30%B; 615 min, 30%B39%B; 1516 min, 39%B80%B) at a flow rate of 0.35 mL/min. The analytes were detected by ESI(+)-MS under selected ion monitoring mode (09.7 min, m/z 377; 9.712 min, m/z 411; 1214.7 min, m/z 447; 14.718 min, m/z 557; 1825 min, m/z 263). The linear ranges were 0.050-50 mg/L, 0.020-20 mg/L, 0.005-30 mg/L, 0.010-15 mg/L and 0.010-10 mg/L with detection limits of 0.010, 0.005, 0.001, 0.002 and 0.003 mg/L for chlorogenic acid, geniposide, baicalin, forsythin and indirubin, respectively. The average recoveries ranged from 97.0% to 101.7%. The relative standard deviations were less than 2.2%. This method is rapid, accurate, and suitable for the quality control of the five effective components in Qingrejiedu oral liquid.
    Fast separation and identification of nine carcinogenic dyes in textiles using liquid chromatography-electrospray tandem mass spectrometry
    DING Youchao, CAO Xizhong, WU Lina, ZHANG Qian
    2008, 26 (5):  603-607. 
    Abstract ( 3332 )   [Full Text(HTML)] () PDF (531KB) ( 896 )  
    A qualitative method of the identification of nine carcinogenic dyes prohibited in textiles was developed using high performance liquid chromatography-tandem mass spectrometry interfaced with electro-spray ionization (HPLC-ESI-MS/MS) in the selective reaction monitoring (SRM) mode. The dyes were extracted from textiles composed of natural or chemical fibers by methanol under ultrasounication, and then eluted with gradient by acetonitrile and 5 mmol/L ammonium acetate from an RP-C18 column with two segments in effluents. The first effluents accommodated Acid Red 26, Direct Blue 6, Direct Black 38 and Direct Red 28 with negative ionization mode, while the second accommodated Basic Red 9, Basic Violet 14, Disperse Blue 1, Disperse Orange 11 and Disperse Yellow 3 with positive ionization mode. Thus the investigated compounds could be identified simultaneously with single-run analysis no matter which type of the fibre the sample was and no matter which category of the dye the analyte was. The established method was successfully applied to identify the carcinogenic dyes in textile samples through comparing the chromatographic retention time and the relative abundance of characteristic product ions with the standards.
    Determination of 14 pesticide residues in sulfur-containing vegetables by gas chromatography-negative chemical ionization mass spectrometry
    HU Beizhen, SONG Weihua, XIE Liping, SHAO Tiefeng
    2008, 26 (5):  608-612. 
    Abstract ( 2716 )   [Full Text(HTML)] () PDF (442KB) ( 784 )  
    A gas chromatography-negative chemical ionization mass spectrometric (GC-NCI/MS) method has been developed for analyzing 14 pesticide residues in sulfur-containing vegetables (scallion, garlic, garlic bolt, leek, etc.). The samples were first heated in a microwave oven to eliminate most of the sulfur-containing interfering impurities and then extracted with acetonitrile. The extracts were further cleaned-up by gel permeation chromatography (GPC) and a primary-secondary amine (PSA) cartridge. The target analytes were determined using GC-NCI/MS in the selected ion monitoring (SIM) mode. The recoveries of all the pesticides (at spiked level of 50 μg/kg) were from 49.2% to 113.1% with the relative standard deviations between 1.42% and 8.70%. The detection limits (S/N=3) were in the range of 0.5-10.0 μg/kg. The method is selective without interference and suitable for the determination and confirmation of pesticides in the sulfur-containing vegetables.
    Pyrolysis-gas chromatographic fingerprints with hierarchical cluster analysis for Dendrobium candidum Wall. ex Lindl.
    WANG Lili, WANG Cong, PAN Zaifa, SUN Fa
    2008, 26 (5):  613-617. 
    Abstract ( 2511 )   [Full Text(HTML)] () PDF (329KB) ( 729 )  
    The pyrogram fingerprints of Dendrobium candidum Wall. ex Lindl. from different regions were studied by pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) and compared with hierarchical cluster analysis. The effect of pyrolysis temperature on the fingerprint was examined by evolved gas analysis, and then 450 ℃ was selected as the optimized pyrolysis temperature. An amount of 0.4 mg of raw drug powder was pyrolysed in a vertical microfurnace pyrolyzer, and the products were directly introduced into a gas chromatograph equipped with a flame ionization detector (FID) and a fused-silica capillary column (30 m×0.25 mm×0.25 μm). The pyrogram fingerprints of 10 samples from different regions showed a high similarity and a good reproducibility with the relative standard deviations (RSDs) of the retention times less than 0.33% and the RSDs of the relative peak areas less than 4.8%. Therefore, each sample was characterized by the peak area of 31 peaks in each pyrogram and these peaks were employed for hierarchical cluster analysis. Furthermore, the discrimination of the sample from different regions was achieved by hierarchical cluster analysis via recognizing the 10×31 data matrix. Thus, the results proved it is a simple, rapid and accurate method suitable for the quality control of the traditional Chinese medicines.
    Articles
    Determination of environmental estrogens in pharmacy wastewater using solid-phase extraction-gas chromatography/mass spectrometry with derivatization
    HUANG Cheng, JIANG Liying, CHEN Jianmeng, CHEN Xiao
    2008, 26 (5):  618-621. 
    Abstract ( 2547 )   [Full Text(HTML)] () PDF (198KB) ( 970 )  
    A method for the determination of estrone (E1), estradiol (E2), ethinylestradiol (EE2) and estriol (E3) in pharmacy wastewater was developed using solid-phase extraction-gas chromatography/mass spectrometry (SPE-GC/MS) with derivatization. The sample was extracted by an SPE column, derivatized by bis(trimethylsilyl) rifluoroacetamide (BSTFA), and analyzed by GC/MS. The detection limits were 1.8-4.7 ng/L, and the relative standard deviations were 2.3%-9.1% (n=8). The recoveries of above four environmental estrogen compounds were (94.0±2.9)% to (101±3.8)%. The method can be applied in the determination of the estrogenic compounds in wastewater samples successfully. The concentrations of EE2 and E1 in wastewater were 396.6 ng/L and 39.9 ng/L, respectively, and the removal rates of EE2 and E1 were 35%-40% after traditional biological treatment. The results demonstrated that the removal efficiency was not satisfactory and the traditional treatment process of wastewater containing estrogen compounds from pharmaceaticals factory should be improved.
    Urine profiling by capillary electrophoresis with online stacking of moving reaction boundary
    WU Jin, FAN Liuyin, ZHANG Wei, WANG Qiuling, CAO Chengxi
    2008, 26 (5):  622-625. 
    Abstract ( 2792 )   [Full Text(HTML)] () PDF (266KB) ( 528 )  
    Rapid and sensitive profiling of urine holds evident significance for the discovery of new biomarkers and clinical diagnosis. In this study, a simple, rapid and sensitive moving reaction boundary (MRB)-induced stacking technique is described for the analysis of urine profiling by capillary zone electrophoresis (CZE). The MRB was formed with acidic Gly-HCl as the sample buffer and alkaline Gly-NaOH as the running buffer. The optimum conditions of stacking and separation were: 25 mmol/L Gly-HCl (pH 2.5) as the sample buffer and 50 mmol/L Gly-NaOH (pH 12.3) as the running buffer, 15 kV of applied voltage, 4.82 kPa (0.7 psi)×20 s of pressure injection, 60.2 cm×75 μm i.d. (50 cm effective length) of capillary, 214 nm of detection wavelength, 24 ℃of separation temperature. Under the optimum conditions, the MRB can significantly improve the sensitivity of urine profiling. More than 80 stacking peaks have been observed and more than 10 fold sensitivity enhancement was achieved in contrast with the conventional CZE, which with only ten observable peaks and poor sensitivity. The results indicate that the MRB-induced stacking is a useful and potential tool for urine profiling. Furthermore, the present method is easy to perform, and the preparation of sample is quite simple.
    Analyse of sugars in beer and wort using high performance anion-exchange chromatography coupled with pulsed amperometric detection
    PAN Yuanyuan, LIANG Lina, CAI Yaqi, MOU Shifen
    2008, 26 (5):  626-630. 
    Abstract ( 3124 )   [Full Text(HTML)] () PDF (273KB) ( 702 )  
    The method of the simultaneous and direct analysis of eleven sugars, including monosaccharides, disaccharides and oligosaccharides, was established using high performance anion-exchange chromatography (HPAEC) coupled with pulsed amperometric detection (PAD) using an Au working electrode and a pH-Ag/AgCl reference electrode. The separation was accomplished on a CarboPac PA-100 column by using gradient elution consisting of water, 0.25 mol/L NaOH and 1 mol/L NaAc as mobile phase at a flow rate of 0.25 mL/min. Under these conditions, glucose, fructose, sucrose, maltose, isomaltose, maltotriose, isomaltotriose, maltotetraose, maltopentaose, maltohexaose and maltoheptaose were separated in 40 min. Then sugars were detected directly with PAD method without any derivatization reaction, and there was no need to employ complicated pretreatment procedures to the samples before analysis. The detection limits (S/N=3) for the sugars were 13-88 pg. The proposed method was applied for the determination of the 11 sugars in beer and wort samples satisfactorily, with the spiked recoveries of most sugars ranging from 81% to 107%. In addition, some potential transition mechanism of the sugars during the brewage procedure can be educed from the comparison of sugar contents between beer and wort.
    Technical Notes
    Chiral separation of racemic epoxiconazole on cellulose tris(3,5-dimethylphenylcarbamate) chiral stationary phase using high performance liquid chromatography
    HAN Xiaoqian, WEI Yan, LIU Yanhua, CHANG Jing, QIU Wei, CHEN Feng
    2008, 26 (5):  631-633. 
    Abstract ( 2695 )   [Full Text(HTML)] () PDF (183KB) ( 687 )  
    The two pairs of enantiomers of epoxiconazole were resolved using normal phase, reversed-phase and polar organic solvent high performance liquid chromatography on cellulose tris(3,5-dimethylphenylcarbamate) (CDMPC) chiral stationary phase, separately. The effects of different mobile phase compositions on the retention factor, separation factor and resolution in the chiral separation of epoxiconazole were investigated. It can be baseline separated on a Chiralcel OD-H column packed with CDMPC chiral stationary phase when methanol-water (80∶20, v/v) was used as the mobile phase, the resolutions of the two pairs of enantiomers were 1.64 and 6.50, respectively.
    Determination of 1-deoxynojirimycin in Morus alba L. leaves using reversed-phase high performance liquid chromatography-fluorescence detection with pre-column derivatization
    XIE Huiming, WU Fangrui, YANG Yi, LIU Jie
    2008, 26 (5):  634-636. 
    Abstract ( 2437 )   [Full Text(HTML)] () PDF (233KB) ( 639 )  
    A rapid, reliable and suitable method for the determination of 1-deoxynojirimycin (DNJ) in Morus alba L. leaves has been developed. The DNJ in 100 mg dried leaves was extracted twice with 10 mL aqueous 0.05 mol/L HCl, then derivatized by 6-aminoquinoiyi-N-hydroxysuccinimidyl carbamate (AQC) in K3BO3 buffer (pH 8.5), and analyzed using a high performance liquid chromatograph equipped with a fluorescence detector. The analyte was eluted with a mobile phase of 0.02 mol/L potassium dihydrogen phosphate buffer-acetonitrile (85∶15, v/v) at the rate of 1.0 mL/min. The derivatized DNJ was well dissolved from the hydrolysis products of AQC. The linearity ranged from 0.5 to 25 mg/L, and the detection limit was 0.02 mg/L (S/N=3). The content of DNJ in Morus alba L. leaves was 0.12%, the recovery was 96.1%-98.6%.
    Simultaneous determination of atractylenolide Ⅲ, atractylenolideⅠ and atractylon in Artactylodis
    SHOU Dan, DAI Shiwen, ZHANG Jianmin, LI Hongyu, YU Zhongming
    2008, 26 (5):  637-639. 
    Abstract ( 2041 )   [Full Text(HTML)] () PDF (201KB) ( 550 )  
    A microbore liquid chromatography (microbore LC) has been developed for the simultaneous determination of atractylenolide Ⅲ, atractylenolideⅠ and atractylon in Artactylodis macrocephala. In the method, a Microsil C18 column (150 mm×1.0 mm) was used with the mobile phase containing methanol-acetonitrile-water for gradient elution, the flow rate was 50 μL/min, and the detection wavelength was set at 220 nm. Under the optimized separation conditions, every component was separated thoroughly. The relationships between the concentrations and the peak areas of the three components were all linear. The recoveries were 96.86% for atractylenolide Ⅲ, 97.13% for atractylenolideⅠand 98.06% for atractylon, the relative standard deviations (RSD) were 1.63%, 1.31% and 0.39%, respectively. The method is simple, convenient, and can be used for the quality control of Artactylodis macrocephala.
    Simultaneous determination of adapalene, 2-phenoxyethanol and methyl-4-hydroxybenzoate in adapalene gels using high performance liquid chromatography
    ZHANG Chunhong, ZHAO Yingchun, HAN Chunhui, GUO Xingjie
    2008, 26 (5):  640-642. 
    Abstract ( 2630 )   [Full Text(HTML)] () PDF (189KB) ( 723 )  
    A reversed-phase high performance liquid chromatographic (RP-HPLC) method was developed for the simultaneous determination of the contents of adapalene and the preservatives (2-phenoxyethanol and methyl-4-hydroxybenzoate) in adapalene gels. The chromatographic analysis was carried out on a Tigerkin C18 column (150 mm×4.6 mm, 5 μm). The mobile phase was 0.02 mol/L ammonium acetate buffer (pH 3.0) and tetrahydrofuran-acetonitrile with gradient elution, and the detection wavelength was set at 270 nm. The calibration curves were linear over the ranges of 10-100 mg/L (r=0.9999), 4-40 mg/L (r=0.9999) and 4-40 mg/L (r=0.9999) for 2-phenoxyethanol, methyl-4-hydroxybenzoate and adapalene, respectively. The average recoveries of the three substances were within 98.0%-98.6%. The method is simple, reliable and suitable for the simultaneous determination of adapalene, 2-phenoxyethanol and methyl-4-hydroxybenzoate in adapalene gels.
    Enantiomeric separation of liarozole on amylose chiral stationary phase
    CHEN Yuqing, MA Zheng, AN Fang, GUO Xingjie
    2008, 26 (5):  643-645. 
    Abstract ( 2408 )   [Full Text(HTML)] () PDF (142KB) ( 588 )  
    A high performance liquid chromatographic method was established for the enantiomeric separation of liarozole. Baseline chiral separation of liarozole was achieved under normal-phase chromatographic mode by the Chiralpak AD-H chiral stationary phase. The influences of the concentration and kind of organic solvent, the proportion of acid and base, column temperature and flow rate on the enantiomeric separation were studied. The optimized chromatographic conditions were hexane-ethanol (containing 0.3% diethylamine and 0.1% glacial acetic acid) (80∶20, v/v) as the mobile phase with a flow rate of 0.6 mL/min and detection at the wavelength of 254 nm. The column temperature was set at 20 ℃. The resolution of 3.4 for liarozole was achieved under the above chromatographic conditions. The method is simple, rapid and with good reproducibility.
    Determination of decabromodiphenyl ethane using high performance liquid chromatography
    YANG Yang, CHEN Jianhai, CHANG Liping, HUANG Qi
    2008, 26 (5):  646-648. 
    Abstract ( 2559 )   [Full Text(HTML)] () PDF (227KB) ( 745 )  
    A method was developed for the quantitative determination of decabromodiphenyl ethane (DBDPE) using high performance liquid chromatography (HPLC). The sample was separated on a Zorbax C18 column (5 μm, 150 mm×4.6 mm) at the temperature of 40 ℃ with the elution of methanol-tetrahydrofuran (70∶30, v/v) as mobile phase at a flow rate of 0.8 mL/min. The detection wavelength was 230 nm. The assay exhibited a good linearity in decabromodiphenyl ethane concentration range from 0.001 to 0.100 g/L with a correlation coefficient of 0.9991. The limit of detection was 0.2 mg/L. The recovery was more than 96%(n=4), and the relative standard deviation was 4.0%(n=6). This method can be applied in the industrialized production for replacing the thermal analysis.