ALBERT

Description:    An inexpensive, simple and environmentally friendly method based on dispersive liquid liquid microextraction (DLLME) for rapid determination of benzene derivatives in water samples was proposed. A significant improvement of DLLME procedure was achieved. Trace volume ethyl acetate (60 μL) was exploited as dispersion solvent instead of common ones such as methanol and acetone, the volume of which was more than 0.5 mL, and the organic solvent required in DLLME was reduced to a great extent. Only 83-μL organic solvent was consumed in the whole analytic process and the preconcentration procedure was less than 10 min. The advantageous approach coupled with gas chromatograph-flame ionization detector was proposed for the rapid determination of benzene, toluene, ethylbenzene and xylene isomers in water samples. Results showed that the proposed approach was an efficient method for rapid determination of benzene derivatives in aqueous samples. Content Type Journal Article Category Short Communication Pages 1-5 DOI 10.1007/s10337-012-2215-7 Authors Chun Peng Diao, College of Environmental Science and Engineering, South China University of Technology, Guangzhou Higher Education Mega Center, Guangzhou, 510006 China Chao Hai Wei, College of Environmental Science and Engineering, South China University of Technology, Guangzhou Higher Education Mega Center, Guangzhou, 510006 China Chun Hua Feng, College of Environmental Science and Engineering, South China University of Technology, Guangzhou Higher Education Mega Center, Guangzhou, 510006 China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Development of a reversed phase high performance liquid chromatographic method for determination of six related impurities in prilocaine substance is reported. The test of related impurities in European Pharmacopoeia (Ph. Eur.) cannot meet the demands with the chromatographic parameters given, therefore different types of chromatographic systems and eight columns have been evaluated in the present study. A new method with a Hypercarb column was developed and validated. This method fulfils the demands in the Ph. Eur., and the validation shows that the method is selective, reproducible, linear, accurate and robust with sufficient limits of detection (0.001–0.004% of 2.5 mg prilocaine mL −1 ) and quantification (0.002–0.009% of 2.5 mg prilocaine mL −1 ). Content Type Journal Article Category Original Pages 1-8 DOI 10.1007/s10337-012-2212-x Authors Janina Sroka-Markovic, Analytical Development, AstraZeneca, 15185 Södertälje, Sweden Linda Johansson, Analytical Development, AstraZeneca, 15185 Södertälje, Sweden Magnus B. O. Andersson, Analytical Development, AstraZeneca, 15185 Södertälje, Sweden Ingrid Granelli, Department of Analytical Chemistry, Stockholm University, 10691 Stockholm, Sweden Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    In the present study, baseline separation of the enantiomers of 16 β-carboline derivatives was successfully achieved using both capillary electrophoresis (CE) and high-performance liquid chromatography (HPLC) techniques in short run times (〈15 min) and thus permit the determination of enantiomeric excess. In HPLC methodology, cellulose chiral stationary phase (Chiralcel OD-H) was used with a binary mobile phase constituted of n -hexane/ethanol 85/15 leading to a resolution factor of 12.6 in 15 min. Preparative HPLC allowed to obtain pure enantiomers of two compounds. In CE, chiral selectivity was developed with an in-capillary stacking strategy using anionic (highly sulfated-γ) cyclodextrins 5% (w/v) as chiral selectors and a 60 mM phosphate buffer (pH 2.5) resulting in a resolution of 10.26 in 14 min of analysis. The analytical characteristics of the two developed methods were studied in terms of repeatability, limits of detection and limits of quantification showing their suitability to be extended to all the other molecules. Content Type Journal Article Category Original Pages 1-9 DOI 10.1007/s10337-012-2194-8 Authors Emmanuelle Lipka, Faculté des Sciences Pharmaceutiques et Biologiques, Univ Lille Nord de France, EA 4481, 3 rue du Pr. Laguesse, 59000 Lille, France Saïd Yous, Faculté des Sciences Pharmaceutiques et Biologiques, Univ Lille Nord de France, EA 4481, 3 rue du Pr. Laguesse, 59000 Lille, France Christophe Furman, Faculté des Sciences Pharmaceutiques et Biologiques, Univ Lille Nord de France, EA 4481, 3 rue du Pr. Laguesse, 59000 Lille, France Pascal Carato, Faculté des Sciences Pharmaceutiques et Biologiques, Univ Lille Nord de France, EA 4481, 3 rue du Pr. Laguesse, 59000 Lille, France Carole Deghaye, Faculté des Sciences Pharmaceutiques et Biologiques, Univ Lille Nord de France, EA 4481, 3 rue du Pr. Laguesse, 59000 Lille, France Jean-Paul Bonte, Faculté des Sciences Pharmaceutiques et Biologiques, Univ Lille Nord de France, EA 4481, 3 rue du Pr. Laguesse, 59000 Lille, France Claude Vaccher, Faculté des Sciences Pharmaceutiques et Biologiques, Univ Lille Nord de France, EA 4481, 3 rue du Pr. Laguesse, 59000 Lille, France Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    A capillary chromatography system was developed using an open capillary tube and a ternary solvents carrier solution of water-hydrophilic/hydrophobic organic solvent mixture. The chromatography is called a tube radial distribution chromatography (TRDC) system. The TRDC system works without applying high voltages or using specific columns, such as monolithic and packed columns. In this study, the effects of tube materials on separation performance were examined in the TRDC system, by using poly(tetrafluoroethylene) (PTFE; 100–400 μm inner diameter), polyethylene (PE; 200 μm inner diameter), and copolymer of (tetrafluoroethylene–perfluoroalcoxyethylene) (PTFE–PFAE; 100 μm inner diameter) capillary tubes. An analyte solution of 2,6-naphthalenedisulfonic acid and 1-naphthol as a model was subjected to the system with a water–acetonitrile–ethyl acetate carrier solution; 15:3:2 volume ratio (water-rich carrier) and 3:8:4 volume ratio (organic solvent-rich carrier). The flow rates were adjusted to be 0.5 μL min −1 for PTFE and PTFE–PFAE tubes as well as 2.0 μL min −1 for PE tube under laminar flow conditions. These analytes in the solution were separated in this order with the water-rich carrier solution with baseline separation in the three capillary tubes, while they were eluted in the reverse order or not separated with the organic solvent-rich carrier solution. The effects of tube temperature on separation were also examined with the water-rich carrier solution; the best resolutions were observed at 0 °C of the tube temperature. The obtained results were compared with those of fused-silica capillary tube and discussed. Content Type Journal Article Category Short Communication Pages 1-5 DOI 10.1007/s10337-012-2205-9 Authors Yudai Kudo, Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University, Kyotanabe, Kyoto 610-0321, Japan Naoya Jinno, Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University, Kyotanabe, Kyoto 610-0321, Japan Masahiko Hashimoto, Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University, Kyotanabe, Kyoto 610-0321, Japan Kazuhiko Tsukagoshi, Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University, Kyotanabe, Kyoto 610-0321, Japan Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description: R.E. Hester and R.M. Harrison (Eds.): Nuclear Power and the Environment Content Type Journal Article Category Book Review Pages 1-2 DOI 10.1007/s10337-012-2208-6 Authors Ken Jones, Knutsford, Cheshire, UK Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    In this study, improved homogeneous liquid–liquid extraction (HLLE), equipped with GC–ECD has been developed for the extraction and determination of organochlorinated pesticides (OCPs) in water. The phase separation phenomenon occurred by temperature in a ternary solvent (water/methanol/chloroform) system. Several factors influencing the extraction efficiency were investigated and optimized with orthogonal array design. Furthermore, in this study, for the first time, before immiscible organic phase formation, different volumes of deionized water were subjected to homogeneous solution to investigate the effect of this factor on the extraction performance of HLLE. Optimal results were as follows: volume of the extracting solvent (chloroform), 50 μL; volume of the consolute solvent (methanol), 1.2 mL; volume of the sample, 2.5 mL; volume of the deionized water, 0.5 mL; time of centrifuge, 7 min. Under the optimum conditions, repeatability was obtained by spiking OCPs at concentration level of 20 μg L −1 , the RSDs varied between 4.8 and 10.7% ( n  = 4). The limits of detection of 0.02–0.12 μg L −1 were obtained for the OCPs. Enrichment factors and the extraction percent of the studied compounds were in the range of 240–300 and 69.2–84.0%, respectively. Finally, the results of the proposed HLLE method were compared with the same HLLE method without addition of deionized water. The results indicated that the proposed method has higher enrichment factors and lower detection limits. Content Type Journal Article Category Original Pages 1-7 DOI 10.1007/s10337-012-2206-8 Authors Farahnaz Rezaei, Department of Analytical Chemistry, Faculty of Chemistry, Iran University of Science and Technology, Narmak, 16846 Tehran, Iran Mohammad-Reza Milani Hosseini, Department of Analytical Chemistry, Faculty of Chemistry, Iran University of Science and Technology, Narmak, 16846 Tehran, Iran Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    A sensitive and rapid liquid chromatography–mass spectrometry (LC–MS) assay was established for the quantitation of CYC-116, an antitumor drug, in rat plasma. The chromatographic separation was accomplished on a Kromasil C 18 column (150 mm × 4.6 mm i.d., 5 μm particle size) at an isocratic flow rate of 0.8 mL min −1 using acetonitrile–water–formic acid (23.5:76.5:0.1, v/v/v) as the mobile phase. The plasma extraction was performed by liquid–liquid extraction using ethyl acetate as the solvent, and the extracts were subjected to MS analysis using a quadrupole mass spectrometer. The calibration curve of CYC-116 was linear over the concentration range of 5–2,500 ng mL −1 ( r  = 0.9955). The mean recovery was 85.0 ± 8.0%, and the matrix effect ranged from 90.0 to 110.0%. The intra- and interday precisions were less than 11.8 and 6.6%, respectively, and the accuracy was within ±5.8%. The method was successfully applied to study the pharmacokinetics of CYC-116 in rats after oral administration. Content Type Journal Article Category Original Pages 1-6 DOI 10.1007/s10337-011-2175-3 Authors Jing Su, School of Pharmacy, Shenyang Pharmaceutical University, Wenhua Road 103, Shenyang, 110016 People’s Republic of China Xiaohui Chen, School of Pharmacy, Shenyang Pharmaceutical University, Wenhua Road 103, Shenyang, 110016 People’s Republic of China Qing Li, School of Pharmacy, Shenyang Pharmaceutical University, Wenhua Road 103, Shenyang, 110016 People’s Republic of China Zhiguo Yu, School of Pharmacy, Shenyang Pharmaceutical University, Wenhua Road 103, Shenyang, 110016 People’s Republic of China Xiaoduo Guan, School of Pharmacy, Shenyang Pharmaceutical University, Wenhua Road 103, Shenyang, 110016 People’s Republic of China Lulu Geng, School of Pharmacy, Shenyang Pharmaceutical University, Wenhua Road 103, Shenyang, 110016 People’s Republic of China Kaishun Bi, School of Pharmacy, Shenyang Pharmaceutical University, Wenhua Road 103, Shenyang, 110016 People’s Republic of China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Tube radical distribution chromatography (TRDC) uses an untreated open tubular capillary tube and a ternary mixture of solvents (water and hydrophilic/hydrophobic organic solvents) as a carrier solution. A model analyte mixture comprising 1-naphthol, 1-naphthoic acid, 1-naphthalenesulfonic acid, 2,6-naphthalenedisulfonic acid, and 1,3,6-naphthalenetrisulfonic acid was examined by the TRDC and capillary zone electrophoresis (CZE) systems that comprised mainly a capillary tube and a detector. In the TRDC system the elution order of analytes could be changed by altering the component ratios of the solvents, whereas in the CZE system the elution order was changed by altering the electroosmotic flow direction. The experimental data obtained provide clues about the features and utility of TRDC as a new separation method. Content Type Journal Article Category Short Communication Pages 1-6 DOI 10.1007/s10337-012-2203-y Authors Kisuke Tabata, Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University, Kyotanabe, Kyoto 610-0321, Japan Naoya Jinno, Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University, Kyotanabe, Kyoto 610-0321, Japan Keiichi Noda, Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University, Kyotanabe, Kyoto 610-0321, Japan Masahiko Hashimoto, Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University, Kyotanabe, Kyoto 610-0321, Japan Kazuhiko Tsukagoshi, Department of Chemical Engineering and Materials Science, Faculty of Science and Engineering, Doshisha University, Kyotanabe, Kyoto 610-0321, Japan Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description: K. Kamienska-Trela (Ed.): Nuclear Magnetic Resonance. Volume 40. Specialist Periodical Reports of the Royal Society of Chemistry Content Type Journal Article Category Book Review Pages 1-2 DOI 10.1007/s10337-012-2209-5 Authors John C. Lindon, Imperial College London, London, UK Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    A simple, selective and highly sensitive method was developed and optimized to determine the most commonly used UV filters with endocrine-disrupting potential in water, namely benzophenone-3 (BP-3), octocrylene (OC), ethylhexyl dimethyl p -aminobenzoate (OD-PABA), ethylhexyl methoxycinnamate, ethylhexyl salicylate (EHS) and homosalate (HMS). Samples were extracted by stir bar sorptive extraction followed by liquid desorption (SBSE-LD). The important factors influencing SBSE-LD were optimized. Under optimal conditions, assays were performed on 50 mL of water sample using stir bars (0.5 mm in film thickness, 10 mm in length) at room temperature. The analytes were determined by liquid chromatography–tandem mass spectrometry with triple quadrupole analyzer using atmospheric pressure chemical ionization. The main parameters in HPLC–APCI–MS/MS were also optimized to provide the best performances for all analytes. Moreover, matrix effect was investigated using two methods the post-column infusion system and the method of spiked matrices after extraction. As a result, no significant matrix effect on the analysis was observed. The method showed good linearity ( R 2 coefficients greater than 0.996 in different water samples after SBSE-LD). Recoveries of the analytes were close to 90%, except for BP-3 (64%) and OC (76%) with relative standard deviation lower than 11%. Detection limits were between 0.6 and 3.3 ng L −1 for all the analytes except for HMS (94 ng L −1 ) and EHS (114 ng L −1 ). This methodology was applied to measure UV filters in seawater, river water and wastewater in different sites of Liguria; BP-3 and OC were found in most of the considered samples at rather low concentration level. Content Type Journal Article Category Original Pages 1-10 DOI 10.1007/s10337-012-2202-z Authors Emanuele Magi, Department of Chemistry and Industrial Chemistry, University of Genoa, Via Dodecaneso 31, 16146 Genoa, Italy Marina Di Carro, Department of Chemistry and Industrial Chemistry, University of Genoa, Via Dodecaneso 31, 16146 Genoa, Italy Carlo Scapolla, Department of Chemistry and Industrial Chemistry, University of Genoa, Via Dodecaneso 31, 16146 Genoa, Italy Kieu T. N. Nguyen, Department of Chemistry and Industrial Chemistry, University of Genoa, Via Dodecaneso 31, 16146 Genoa, Italy Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Water-soluble sodium poly(aspartate- co -lactide) (PALNa) copolymers with a molar ratio of aspartate-to-lactide units equal to 1:0.6, 1:1.0 and 1:1.5 were studied using NMR spectroscopy to determine the composition as well as SEC-MALS and static light-scattering measurements to determine the molar-mass characteristics of the copolymers. In the copolymer aqueous solutions, high-molar-mass species were detected, most probably due to the incomplete dissolution of the samples. The molar-mass averages determined in water with added simple electrolyte, i.e., NaCl, were much lower than the values determined in pure water. The concentration of the salt, which allows dissolution on a molecular level, and the separation predominantly according to a size-exclusion mechanism depend on the chemical composition of the PALNa copolymers. The optimal mobile phase for the PALNa-1/0.6 and the PALNa-1/1.0 copolymers was 0.1 M NaCl at pH 9, and for the PALNa-1/1.5 copolymer with a higher content of lactide units it was 0.05 M NaCl at pH 9. The molar-mass averages of the PALNa-1/1.0 copolymer, determined by SEC-MALS and static light-scattering measurements, were comparable. Content Type Journal Article Category Original Pages 1-8 DOI 10.1007/s10337-012-2180-1 Authors Maja Gričar, Laboratory for Polymer Chemistry and Technology, National Institute of Chemistry, Hajdrihova 19, 1000 Ljubljana, Slovenia Majda Žigon, Laboratory for Polymer Chemistry and Technology, National Institute of Chemistry, Hajdrihova 19, 1000 Ljubljana, Slovenia Tina Šmigovec Ljubič, Laboratory for Polymer Chemistry and Technology, National Institute of Chemistry, Hajdrihova 19, 1000 Ljubljana, Slovenia Ema Žagar, Laboratory for Polymer Chemistry and Technology, National Institute of Chemistry, Hajdrihova 19, 1000 Ljubljana, Slovenia Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description: Luigi Mondello (Ed.): Comprehensive Chromatography in Combination with Mass Spectrometry Content Type Journal Article Category Book Review Pages 1-2 DOI 10.1007/s10337-012-2189-5 Authors Edward R. Adlard, Burton, South Wirral, UK Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Formaldehyde has been highlighted as potential genotoxic impurity (GTI). Trace-level quantification of GTIs in drug substances requires sensitive, precise and accurate analytical methodologies for their estimation in drug substances and control. Analysis and estimation of formaldehyde is very challenging due to its properties namely volatility, high polarity, low molecular weight and over and above the absence of chromophore. This article presents a validated HPLC–UV method which is sensitive to quantification of formaldehyde in active pharmaceutical ingredient. As formaldehyde does not possess chromophore, the developed HPLC method involves derivatization with 2,4-dinitrophenylhydrazine. Using this method, the detection and quantitation limits achieved are 0.5 and 1.5 ppm, respectively. The calibration curve of formaldehyde was linear over the concentration range of 1.5–20 ppm. The method was found to be sensitive, precise and accurate and the proposed method has been successfully applied to estimate formaldehyde content in scale-up batches of bulk drug. Content Type Journal Article Category Original Pages 1-6 DOI 10.1007/s10337-012-2186-8 Authors A. Nageswari, Center of Pharmaceutical Sciences, IST, Jawaharlal Nehru Technological University, Kukatpally, Hyderabad, 500072 India K. V. S. R. Krishna Reddy, Aptuit Laurus Private Limited, ICICI Knowledge Park, Turkapally, Hyderabad, 500078 India K. Mukkanti, Center of Pharmaceutical Sciences, IST, Jawaharlal Nehru Technological University, Kukatpally, Hyderabad, 500072 India Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    A method combining solid-phase extraction and high-performance liquid chromatography (SPE–HPLC) has been developed for analysis of ten rare ginsenosides including 20( R )-Rh 1 , 20( S )-Rh 1 , 20( R )-Rg 3 , 20( S )-Rg 3 , Rg 5 , Rk 1 , Rg 6 , F 4 , Rk 3 , and Rh 4 in three kinds of injection (Shenfu, Shenmai, and Shengmai). Waters Oasis HLB SPE columns were used to clean and enrich the sample. An Eclipse XDB-C 18 column was used to separate the analytes, with water and acetonitrile as mobile phase components under gradient elution conditions at 30 °C. There were good linear correlations between the signals measured and the concentrations of the ten analytes ( r 2  ≥ 0.9996). Intraday and interday precision were both better than 2.19% and average recovery ranged from 95.25 to 108.83%. The proposed method was successfully applied to determination of the ten analytes in different batches of the injections and the results indicated the method is simple, rapid, and accurate. The proposed method could be used for quality control during manufacture of the injections. Content Type Journal Article Category Original Pages 1-7 DOI 10.1007/s10337-012-2183-y Authors Rui-Jie Yang, College of Chemistry, Jilin University, ChangChun, 130012 Jilin Province, China Xu-Wen Li, College of Chemistry, Jilin University, ChangChun, 130012 Jilin Province, China Hua Yao, College of Chemistry, Jilin University, ChangChun, 130012 Jilin Province, China Mu-Chun Zhang, Department of Urology, China-Japan Union Hospital, Jilin University, ChangChun, 130033 Jilin Province, China Yong-Ri Jin, College of Chemistry, Jilin University, ChangChun, 130012 Jilin Province, China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    In this study, an end-point-based fluorescence assay for soluble epoxide hydrolase (sEH) was transformed into an on-line continuous-flow format. The on-line biochemical detection system (BCD) was coupled on-line to liquid chromatography (LC) to allow mixture analysis. The on-line BCD was based on a flow system wherein sEH activity was detected by competition of analytes with the substrate hydrolysis. The reaction product was measured by fluorescence detection. In parallel to the BCD data, UV and MS data were obtained through post-column splitting of the LC effluent. The buffer system and reagent concentrations were optimized resulting in a stable on-line BCD with a good assay window and good sensitivity (S/N 〉 60). The potency of known sEH inhibitors (sEHis) obtained by LC–BCD correlates well with published values. The LC–BCD system was applied to test how oxidative microsomal metabolism affects the potency of three sEHis. After incubation with pig liver microsomes, several metabolites of sEHis were characterized by MS, while their individual potencies were measured by BCD. For all compounds tested, active metabolites were observed. The developed method allows for the first time the detection of sEHis in mixtures providing new opportunities in the development of drug candidates. Content Type Journal Article Category Original Pages 1-9 DOI 10.1007/s10337-012-2343-0 Authors David Falck, Department of BioMolecular Analysis, VU University Amsterdam, Amsterdam, The Netherlands Nils Helge Schebb, Department of Entomology and UC Davis Comprehensive Cancer Center, University of California, Davis, CA, USA Setyo Prihatiningtyas, Department of BioMolecular Analysis, VU University Amsterdam, Amsterdam, The Netherlands Jiawen Zhang, Department of BioMolecular Analysis, VU University Amsterdam, Amsterdam, The Netherlands Ferry Heus, Department of BioMolecular Analysis, VU University Amsterdam, Amsterdam, The Netherlands Christophe Morisseau, Department of Entomology and UC Davis Comprehensive Cancer Center, University of California, Davis, CA, USA Jeroen Kool, Department of BioMolecular Analysis, VU University Amsterdam, Amsterdam, The Netherlands Bruce D. Hammock, Department of Entomology and UC Davis Comprehensive Cancer Center, University of California, Davis, CA, USA Wilfried M. A. Niessen, Department of BioMolecular Analysis, VU University Amsterdam, Amsterdam, The Netherlands Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    A quantitative structure–retention relationship study was performed for 286 flavor compounds with highly structural diversity on different stationary phases of different polarities. Firstly, the structure–retention relationship was statistically explored using constitutional, topological, molecular properties, charge descriptors and quantum chemical descriptors. Some recently developed chemometric methods, such as the ones for robust analysis and variable selection were firstly employed to predict accurately the gas chromatographic retention indices. The stability and validity of models have been tested by internal and external validation, and good robustness and predictive ability were obtained. The resulting QSRR models were well correlated, with the square of correlation coefficient for cross validation, Q 2 , values of 0.9847, 0.9838, 0.9745 and 0.9646 on stationary phase DB5, OV101, OV17 and C20M, respectively. The molecular properties known to be relevant for GC retention index, such as molecular size, branching, electron density distribution and hydrogen bond effect were well covered by generated descriptors. The descriptors used in models on four stationary phases were compared, and some reasonable explanations about gas chromatographic retention mechanism were obtained. The developed model may be useful for the prediction of flavor compounds while experimental data are unavailable. Content Type Journal Article Category Original Pages 1-13 DOI 10.1007/s10337-012-2349-7 Authors Xian Chen, Research Center of Modernization of Traditional Chinese Medicines, Central South University, Changsha, 410083 People’s Republic of China Hong-Dong Li, Research Center of Modernization of Traditional Chinese Medicines, Central South University, Changsha, 410083 People’s Republic of China Fang-Qiu Guo, Research Center of Modernization of Traditional Chinese Medicines, Central South University, Changsha, 410083 People’s Republic of China Jun Yan, Research Center of Modernization of Traditional Chinese Medicines, Central South University, Changsha, 410083 People’s Republic of China Dong-Sheng Cao, Research Center of Modernization of Traditional Chinese Medicines, Central South University, Changsha, 410083 People’s Republic of China Yi-Zeng Liang, Research Center of Modernization of Traditional Chinese Medicines, Central South University, Changsha, 410083 People’s Republic of China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    In this paper, the review on application of factorial-based designs in liquid chromatography (LC) is given. The most useful and applicable full factorial design and reduced forms of full factorial design (fractional factorial design and Plackett–Burman design) applied in LC are presented. Literature survey shows that experimental design presents very often used tool in screening, optimization and robustness testing of LC methods. Content Type Journal Article Category Original Pages 1-14 DOI 10.1007/s10337-012-2350-1 Authors Biljana Jančić Stojanović, Department of Drug Analysis, University of Belgrade, Vojvode Stepe 450, Belgrade, Serbia Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    A heart-cut two-dimensional high-performance liquid chromatography method for enantiomeric determination of salbutamol, salmeterol and atenolol in urine is presented. It involves the use of two separations in a liquid chromatography–liquid chromatography achiral–chiral coupling. Target compounds were previously separated in a primary column (Kinetex™ HILIC, 2.6 μm, 150 × 2.1 mm I.D.) with a mixture of MeOH:ACN:ammonium acetate buffer (5 mM, pH 6) 90:5:5 (v/v/v) as mobile phase at a flow rate of 0.40 mL min −1 . Enantiomeric separation was carried out by transferring peak of each compound through a switching valve to a vancomycin chiral column (Chirobiotic™ V, 2.6 μm, 150 × 2.1 mm I.D.) using MeOH:ammonium acetate buffer (2 mM, pH 4) 97:3 (v/v) as mobile phase at a flow rate of 0.50 mL min −1 . Ultraviolet detection was done at 227 nm. The method was applied to determine target analytes in urine samples after enzymatic hydrolysis with β-glucuronidase from Helix pomatia , followed by a solid-phase extraction procedure using Isolute ® HCX mixed-mode cartridges. Extraction recoveries ranged from 82 to 90 % in urine samples. Detection limits were 0.091–0.095 μg for each enantiomer of atenolol and between 0.058 and 0.076 and 0.18–0.14 μg for enantiomers of salbutamol and salmeterol, respectively (3 mL of urine). Linearity ranges were between 0.5 and 10 μg mL −1 . Intraday and interday reproducibilities of enantiomeric ratio and enantiomeric fraction, expressed as relative standard deviation, were between 1.9 and 9.0 %. The optimized method was successfully applied to the analysis of urine samples obtained from excretion studies in volunteers and in freeze-dried urine samples, containing urinary components with MW 〈 10,000 and components with MW 〉 10,000, spiked with different amounts of studied drugs. Content Type Journal Article Category Original Pages 1-11 DOI 10.1007/s10337-012-2353-y Authors Yung Yang, Department of Analytical Chemistry, Faculty of Chemistry, Complutense University of Madrid, Avda. Complutense s/n, 28040 Madrid, Spain Noelia Rosales-Conrado, Department of Analytical Chemistry, Faculty of Chemistry, Complutense University of Madrid, Avda. Complutense s/n, 28040 Madrid, Spain Vanesa Guillén-Casla, Department of Analytical Chemistry, Faculty of Chemistry, Complutense University of Madrid, Avda. Complutense s/n, 28040 Madrid, Spain María Eugenia León-González, Department of Analytical Chemistry, Faculty of Chemistry, Complutense University of Madrid, Avda. Complutense s/n, 28040 Madrid, Spain Luis Vicente Pérez-Arribas, Department of Analytical Chemistry, Faculty of Chemistry, Complutense University of Madrid, Avda. Complutense s/n, 28040 Madrid, Spain Luis María Polo-Díez, Department of Analytical Chemistry, Faculty of Chemistry, Complutense University of Madrid, Avda. Complutense s/n, 28040 Madrid, Spain Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description: Purpose   In this study, direct separation of ketoprofen enantiomers was performed on a Chirobiotic T column. Methods   The effects of the type and amount of the organic modifier, buffer concentration, pH value, temperature and flow rate on retention and selectivity were investigated. Experiments were carried out in the temperature range of 20–40 °C to study the effects of temperature. Thermodynamic parameters were calculated from plots of ln k or ln α versus 1/ T . Molecular dynamics simulation was done to investigate interactions between ketoprofen enantiomers and the chiral selector—teicoplanin. Results   It was observed that pH and flow rate had a large influence on resolution. Baseline separation of ketoprofen enantiomers could be achieved with low amounts of methanol, high temperature and high buffer concentrations. Conclusions   Results from a thermodynamic study and molecular dynamics simulation show that steric hindrance effect, π–π complexation, hydrogen bonding and electrostatic forces are the main driving forces which cause chiral recognition of ketoprofen enantiomers. Content Type Journal Article Category Original Pages 1-9 DOI 10.1007/s10337-012-2352-z Authors Xiaomei He, China Pharmaceutical University, 24 Tongjia Lane, Nanjing, 210009 Jiangsu province, China Rui Lin, Yancheng Health Vocational and Technical College, Yancheng, 224005 China Hua He, China Pharmaceutical University, 24 Tongjia Lane, Nanjing, 210009 Jiangsu province, China Meiling Sun, China Pharmaceutical University, 24 Tongjia Lane, Nanjing, 210009 Jiangsu province, China Deli Xiao, China Pharmaceutical University, 24 Tongjia Lane, Nanjing, 210009 Jiangsu province, China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    In this study, we focused on the studying of taurine complexes with phenol and sodium hypochlorite, and of taurine with sodium hypobromite by spectrometry, reverse phase chromatography and ion-exchange chromatography. The formed complexes were studied under various conditions such as temperature (10, 20, 30, 40, 50 and 60 °C), and/or time of interaction (0, 5, 10, 15, 20, 25 and 30 min). In addition, we optimized high performance liquid chromatography coupled with UV detector for detection of taurine and its complexes with the acids. Taurine–phenol–hypochlorite complex was effectively separated under isocratic elution, mobile phase water:methanol 30:70 %, v : v , flow rate 1 mL min −1 and 55 °C. Taurine-bromamine complex was isolated under the following optimized conditions as isocratic elution, mobile phase water:methanol 85:15 % v : v , flow rate 1 mL min −1 and 55 °C. The limits of detection (3 S/N) were estimated as 1 μM for both types of complexes, i.e. for taurine. Further, we estimated recovery in one sample of urine (male 25 years), commercially achieved energy drink and tea leaves and varied from 79 to 86 %. Further, we aimed our attention at investigating the ability of the above characterized taurine and taurine complexes to scavenge reactive oxygen species. For this purpose, an ion-exchange liquid chromatography with post-column derivatization with ninhydrin and VIS detector was used. It clearly follows from the results obtained that taurine itself reacts with peroxide more intensely than in a bound form, which can be associated with the highest signal decrease. Complexes stabilized structure taurine against peroxide radicals, resulting in slower decreasing of peak heights. The most stable was taurine complexes with phenol and hypobromite. Content Type Journal Article Category Original Pages 1-11 DOI 10.1007/s10337-012-2354-x Authors Lukas Nejdl, Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, 613 00 Brno, Czech Republic Jiri Sochor, Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, 613 00 Brno, Czech Republic Ondrej Zitka, Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, 613 00 Brno, Czech Republic Natalia Cernei, Central European Institute of Technology, Brno University of Technology, Technicka 3058/10, 616 00 Brno, Czech Republic Branislav Ruttkay-Nedecky, Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, 613 00 Brno, Czech Republic Pavel Kopel, Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, 613 00 Brno, Czech Republic Petr Babula, Central European Institute of Technology, Brno University of Technology, Technicka 3058/10, 616 00 Brno, Czech Republic Vojtech Adam, Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, 613 00 Brno, Czech Republic Jaromir Hubalek, Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, 613 00 Brno, Czech Republic Rene Kizek, Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, 613 00 Brno, Czech Republic Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    A capillary gas chromatography (GC) procedure has been developed for the determination of four pharmaceutical preparations (famotidine, ranitidine, cimetidine, and metformin) after precolumn derivatization with methylglyoxal (MGo). GC was carried out using an HP-5 column (30 m × 0.32 mm id) at an initial column temperature of 90 °C for 2 min, followed by heating rate of 25 °C min −1 up to 265 °C. Nitrogen flow rate was 2.5 mL min −1 with split ratio 10:1. A linear calibration curve was obtained within 50–1,000 ng mL −1 and the limit of detection (LOD) was within 17–25 ng mL −1 . The derivatization, GC elution, and separation were repeatable in terms of retention time and peak height/peak area with relative standard deviation within ±4.6 %. The procedure was applied to the determination of the drugs in pharmaceutical preparations and the sera of volunteers who were given oral doses of the drugs. The results of the analysis agreed with the labeled values of the pharmaceutical preparations and were 147–4,903 ng mL −1 in serum with an RSD within 1.0–4.2 %, after ingestion of a single dose of 40–500 mg of active ingredient in a tablet. Content Type Journal Article Category Original Pages 1-7 DOI 10.1007/s10337-012-2321-6 Authors Subhan Ali Majidano, Institute of Advanced Research Studies in Chemical Sciences (IARSCS), University of Sindh, Jamshoro, Pakistan Muhammad Yar Khuhawar, Institute of Advanced Research Studies in Chemical Sciences (IARSCS), University of Sindh, Jamshoro, Pakistan Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    High-mannose type N-linked glycan with 6 mannosyl residues, termed "M6Gn2", displayed clear binding to the same M6Gn2, conjugated with ceramide mimetic (cer-m) and incorporated in liposome, or coated on polystyrene plates. However, the conjugate of M6Gn2-cer-m did not interact with complex-type N-linked glycan with various structures having multiple GlcNAc termini, conjugated with cer-m. The following observations indicate that hamster embryonic fibroblast NIL-2 K cells display homotypic autoadhesion, mediated through the self-recognition capability of high-mannose type glycans expressed on these cells: (i) NIL-2 K cells display clear binding to lectins capable of binding to high-mannose type glycans ( e.g. , ConA), but not to other lectins capable of binding to other carbohydrates ( e.g. GS-II). (ii) NIL-2 K cells adhere strongly to plates coated with M6Gn2-cer-m, but not to plates coated with complex-type N-linked glycans having multiple GlcNAc termini, conjugated with cer-m; (iii) degree of NIL-2 K cell adhesion to plates coated with M6Gn2-cer-m showed a clear dose-dependence on the amount of M6Gn2-cer-m; and (iv) the degree of NIL-2 K adhesion to plates coated with M6Gn2-cer-m was inhibited in a dose-dependent manner by α1,4-L-mannonolactone, the specific inhibitor in high-mannose type glycans addition. These data indicate that adhesion of NIL-2 K is mediated by self-aggregation of high mannose type glycan. Further studies are to be addressed on auto-adhesion of other types of cells based on self interaction of high mannose type glycans. Content Type Journal Article Pages 1-12 DOI 10.1007/s10719-012-9449-3 Authors Seon-Joo Yoon, Division of Biomembrane Research, Pacific Northwest Research Institute, and Department of Global Health, University of Washington, Seattle, WA 98122, USA Natalia Utkina, Division of Biomembrane Research, Pacific Northwest Research Institute, and Department of Global Health, University of Washington, Seattle, WA 98122, USA Martin Sadilek, Depart of Chemistry, University of Washington, Seattle, WA 98195, USA Hirokazu Yagi, Graduate School of Pharmaceutical Sciences, Nagoya City University, Tanabe-dori 3-1, Mizuho-ku, Nagoya, 467-8603 Japan Koichi Kato, Graduate School of Pharmaceutical Sciences, Nagoya City University, Tanabe-dori 3-1, Mizuho-ku, Nagoya, 467-8603 Japan Sen-itiroh Hakomori, Division of Biomembrane Research, Pacific Northwest Research Institute, and Department of Global Health, University of Washington, Seattle, WA 98122, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    A liquid chromatography-mass spectrometry (LC-MS) assay was developed and validated for the quantification of lurasidone, an atypical antipsychotic drug, in rat plasma, bile, and urine. Rat plasma, bile, or urine samples were processed by liquid–liquid extraction and injected onto an LC-MS system for the quantification of lurasidone and ziprasidone (an internal standard). Lurasidone and ziprasidone were separated from endogenous substances using a Gemini C6-Phenyl column with mixture of acetonitrile and 0.1 % formic acid (80:20, v/v ) as the mobile phase. Quantification was performed using the selected ion monitoring mode at m/z 493 for lurasidone and m/z 413 for the IS. The detector response was specific and linear for lurasidone in the concentration range 5–5,000 ng mL −1 The intra- and inter-day accuracy and precision of the method were determined to be within the acceptable criteria for assay validation guidelines. In addition, lurasidone was stable under a variety of processing and handling conditions. Lurasidone concentrations could be readily measured in rat plasma, bile, and urine samples up to 24 h after an intravenous or oral administration, suggesting that the assay can be used in pharmacokinetic studies of lurasidone in rats. Content Type Journal Article Category Original Pages 1117-1128 DOI 10.1007/s10337-012-2294-5 Authors Yoon-Jee Chae, Department of Pharmaceutics, College of Pharmacy, Seoul National University, Gwanak 599, Gwanak-ro, Gwanak-gu, Seoul, 151-742 Republic of Korea Tae-Sung Koo, Graduate School of New Drug Discovery and Development, Chungnam National University, Yuseong, Daejeon, 305-764 Republic of Korea Kyeong-Ryoon Lee, Korea Research Institute of Chemical Technology, 141 Gajeongro, Yuseong, Daejeon, 305-343 Republic of Korea Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893 Journal Volume Volume 75 Journal Issue Volume 75, Numbers 19-20

Description:    Three alkoxyl-functionalized ionic liquids, 1-(3-triethoxysilyl propyl)-3-methyl imidazolium hexafluorophosphate (TESPMIM[PF 6 ]), 1-(3-triethoxysilyl propyl)-3-methyl imidazolium tetrafluoroborate (TESPMIM[BF 4 ]), and 1-(3-triethoxysilyl propyl)-3-methyl imidazolium bis(trifluoromethanesulfonyl)imide (TESPMIM[N(SO 2 CF 3 ) 2 ]), were synthesized and used as selective coating materials to prepare chemically bonded ionic-liquid-based organic–inorganic hybrid solid-phase microextraction (SPME) fibers by sol–gel technology. A possible mechanism of the sol–gel process is proposed, and the successful binding of ionic liquids to the formed silica substrate was confirmed by Fourier-transform infrared spectroscopy (FT-IR). These ionic-liquid-based sol–gel coatings have porous surface structure, high thermal stability, strong solvent resistance, wide pH application range, good coating preparation reproducibility, special selectivity, and extraction efficiency for both polar and nonpolar compounds, such as phenolic environmental estrogens, fatty acids, aromatic amines, alcohols, phthalate esters, and polycyclic aromatic hydrocarbons. The TESPMIM[PF 6 ]- and TESPMIM[BF 4 ]-coated fibers have much lower thermal stability (to 300 and 285 °C, respectively) than TESPMIM[N(SO 2 CF 3 ) 2 ]-coated fiber (to 454 °C). However, the selectivity of these two fibers is higher towards strong polar analytes, while lower towards medium polar or nonpolar analytes compared with TESPMIM[N(SO 2 CF 3 ) 2 ]-based fiber. This could be explained by the fact that different counteranions in ionic liquid structures have different steric hindrance, nucleophilicity, hydrophobicity, and ability to form hydrogen bonds, resulting in significant difference in the characteristics of the ionic-liquid-based SPME fibers, such as the surface morphology, thermal stability, selective extraction ability, etc. This work demonstrates that the performance of the ionic-liquid-based coatings can be simply tuned by changing the counteranions incorporated into the ionic liquid structures. Content Type Journal Article Category Original Pages 1-13 DOI 10.1007/s10337-012-2323-4 Authors Jianjun Shu, Key Laboratory of Arable Land Conservation (Middle and Lower Reaches of Yangtze River), Ministry of Agriculture, College of Resources and Environment, Huazhong Agricultural University, Wuhan, 430070 China Chan Li, Key Laboratory of Arable Land Conservation (Middle and Lower Reaches of Yangtze River), Ministry of Agriculture, College of Resources and Environment, Huazhong Agricultural University, Wuhan, 430070 China Mingming Liu, Key Laboratory of Arable Land Conservation (Middle and Lower Reaches of Yangtze River), Ministry of Agriculture, College of Resources and Environment, Huazhong Agricultural University, Wuhan, 430070 China Hanlan Liu, College of Science, Huazhong Agricultural University, Wuhan, 430070 China Xionghan Feng, Key Laboratory of Arable Land Conservation (Middle and Lower Reaches of Yangtze River), Ministry of Agriculture, College of Resources and Environment, Huazhong Agricultural University, Wuhan, 430070 China Wenfeng Tan, Key Laboratory of Arable Land Conservation (Middle and Lower Reaches of Yangtze River), Ministry of Agriculture, College of Resources and Environment, Huazhong Agricultural University, Wuhan, 430070 China Fan Liu, Key Laboratory of Arable Land Conservation (Middle and Lower Reaches of Yangtze River), Ministry of Agriculture, College of Resources and Environment, Huazhong Agricultural University, Wuhan, 430070 China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Human alpha-1-antitrypsin (α1AT) is a glycoprotein with protease inhibitor activity protecting tissues from degradation. Patients with inherited α1AT deficiency are treated with native α1AT (nAT) purified from human plasma. In the present study, recombinant α1AT (rAT) was produced in Chinese hamster ovary (CHO) cells and their glycosylation patterns, inhibitory activity and in vivo half-life were compared with those of nAT. A peptide mapping analysis employing a deglycosylation reaction confirmed full occupancy of all three glycosylation sites and the equivalency of rAT and nAT in terms of the protein level. N -glycan profiles revealed that rAT contained 10 glycan structures ranging from bi-antennary to tetra-antennary complex-type glycans while nAT displayed six peaks comprising majorly bi-antennary glycans and a small portion of tri-antennary glycans. In addition, most of the rAT glycans were shown to have only core α(1 - 6)-fucose without terminal fucosylation, whereas only minor portions of the nAT glycans contained core or Lewis X-type fucose. As expected, all sialylated glycans of rAT were found to have α(2 - 3)-linked sialic acids, which was in sharp contrast to those of nAT, which had mostly α(2 - 6)-linked sialic acids. However, the degree of sialylation of rAT was comparable to that of nAT, which was also supported by an isoelectric focusing gel analysis. Despite the differences in the glycosylation patterns, both α1ATs showed nearly equivalent inhibitory activity in enzyme assays and serum half-lives in a pharmacokinetic experiment. These results suggest that rAT produced in CHO cells would be a good alternative to nAT derived from human plasma. Content Type Journal Article Pages 1-11 DOI 10.1007/s10719-012-9453-7 Authors Kyung Jin Lee, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, 305-806 Korea Sang Mee Lee, Alteogen Inc., Bioventure town, Daejeon, 305-812 Korea Jin Young Gil, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, 305-806 Korea Ohsuk Kwon, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, 305-806 Korea Jin Young Kim, Korea Basic Science Institute, Ochang-eup, Cheongwon-gun, Chungbuk 363-883, Korea Soon Jae Park, Alteogen Inc., Bioventure town, Daejeon, 305-812 Korea Hye-Shin Chung, Alteogen Inc., Bioventure town, Daejeon, 305-812 Korea Doo-Byoung Oh, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, 305-806 Korea Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    As one of several biologically active compounds in milk, glycoproteins have been indicated to be involved in the protection of newborns from bacterial infection. As much of the physical and immune development of the tammar wallaby ( Macropus eugenii ) young occurs during the early phases of lactation and not in utero , the tammar is a model species for the characterization of potential developmental support agents provided by maternal milk. In the present study, the N - and O -linked glycans from tammar wallaby milk glycoproteins from six individuals at different lactation time points were subjected to glycomics analyses using porous graphitized carbon liquid chromatography electrospray ionization mass spectrometry. Structural characterization identified a diverse range of glycan structures on wallaby milk glycoproteins including sialylated, sulphated, core fucosylated and O -fucosylated structures. 30 % of N -linked structures contained a core (α1-6) fucose. Several of these structures may play roles in development, and exhibit statistically significant temporal changes over the lactation period. The N -glycome was found to contain structures with NeuGc residues, while in contrast the O -glycome did not. O -fucosylated structures were identified in the early stages of lactation indicating a potential role in the early stages of development of the pouch young. Overall the results suggest that wallaby milk contains structures known to have developmental and immunological significance in human milk and reproduction in other animals, highlighting the importance of glycoproteins in milk. Content Type Journal Article Pages 1-14 DOI 10.1007/s10719-012-9452-8 Authors Katherine Wongtrakul-Kish, Biomolecular Frontiers Research Centre, Department of Chemistry and Biomolecular Sciences, Macquarie University, Building E8C Room 307, North Ryde, NSW 2109, Australia Daniel Kolarich, Biomolecular Frontiers Research Centre, Department of Chemistry and Biomolecular Sciences, Macquarie University, Building E8C Room 307, North Ryde, NSW 2109, Australia Dana Pascovici, Australian Proteome Analysis Facility, Macquarie University, North Ryde, NSW 2109, Australia Janice L. Joss, Department of Biological Sciences, Macquarie University, North Ryde, NSW 2109, Australia Elizabeth Deane, Department of Biological Sciences, Macquarie University, North Ryde, NSW 2109, Australia Nicolle H. Packer, Biomolecular Frontiers Research Centre, Department of Chemistry and Biomolecular Sciences, Macquarie University, Building E8C Room 307, North Ryde, NSW 2109, Australia Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description: Glycosylation effects on cancer development Content Type Journal Article Pages 1-2 DOI 10.1007/s10719-012-9448-4 Authors Sen-itiroh Hakomori, Division of Biomembrane Research, Pacific Northwest Research Institute, 720 Broadway, Seattle, WA 98122, USA Richard D. Cummings, Department of Biochemistry, Emory University School of Medicine, Atlanta, GA 30322, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    In clinical medicine, urine creatinine concentration is an important marker in the evaluation of renal function and muscular dysfunctions. Herein, we reported a novel method for rapid determination of creatinine in urine by microchip electrophoresis with light-emitting diode induced fluorescence detection. Creatinine was derivatized by fluorescein isothiocyanate, and then quantitatively detected by the developed microchip LED induced fluorescence detection system. The excitation and emission wavelengths were 490 and 523 nm, respectively. The urine samples were analyzed after centrifuge and filtration. A baseline separation was obtained in 〈30 s using 10 mM borate buffer (pH 9.0, containing 45 mM sodium dodecylsulfate), with separation voltage of 1.5 kV. Good linearity was obtained ( r 2  = 0.9978) in the concentration range of 10.0–2.00 × 10 3  μM, and the limit of detection was 2.87 μM ( S/N  = 3). The recovery was 96.0–107 %, and the interday precision was 〈4.5 % ( n  = 6). To validate assay results, we compared the present method with the Jaffe’s colorimetric assay by measuring real urine samples. The method was reliable, sensitive, high-speed, low-cost and suitable for the routine analysis of creatinine in biofluids. Content Type Journal Article Category Original Pages 1-7 DOI 10.1007/s10337-012-2324-3 Authors Shuping Wang, School of Pharmaceutical Sciences, Sun Yat-sen University, 132 Waihuan East Road of Higher Education Mega Centre, Guangzhou, 510006 People’s Republic of China Xinchun Li, School of Pharmaceutical Sciences, Sun Yat-sen University, 132 Waihuan East Road of Higher Education Mega Centre, Guangzhou, 510006 People’s Republic of China Jianping Yang, School of Pharmaceutical Sciences, Sun Yat-sen University, 132 Waihuan East Road of Higher Education Mega Centre, Guangzhou, 510006 People’s Republic of China Xiujuan Yang, School of Pharmaceutical Sciences, Sun Yat-sen University, 132 Waihuan East Road of Higher Education Mega Centre, Guangzhou, 510006 People’s Republic of China Fenghua Hou, School of Pharmaceutical Sciences, Sun Yat-sen University, 132 Waihuan East Road of Higher Education Mega Centre, Guangzhou, 510006 People’s Republic of China Zuanguang Chen, School of Pharmaceutical Sciences, Sun Yat-sen University, 132 Waihuan East Road of Higher Education Mega Centre, Guangzhou, 510006 People’s Republic of China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description: Mike S. Lee: Mass Spectrometry Handbook Content Type Journal Article Category Book Review Pages 1-2 DOI 10.1007/s10337-012-2325-2 Authors Colin F. Poole, Wayne State University, Detroit, MI, USA Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    CE offers the possibility of fast, cheap and reproducible separations for active compounds of Chinese herbs. Lotusine ( 1 ), liensinine ( 2 ), isoliensinine ( 3 ) and neferine ( 4 ) were the major bioactive alkaloids in lotus plumule, which was used as an important Chinese herb. In this paper, simultaneous separation of 1 – 4 in lotus plumule by nonaqueous CZE has been achieved within 11 min by use of buffer consisting of 80 mM sodium acetate and 40 mM ammonium acetate methanol solution of pH 5.4. Analysis of the four alkaloids in ten plumule samples of Nelumbo nucifera was conducted. Limits of detection (LOD) of 1 – 4 by UV absorbance at 203 nm were achieved in the range of 1.5–2.8 μg mL −1 . Reproducibility (percent RSD) of the analysis by CZE were 2.33, 2.31, 0.78 and 2.58 %, respectively ( n  = 5). Content Type Journal Article Category Original Pages 1-6 DOI 10.1007/s10337-012-2312-7 Authors Guangming Yang, National First-Class Key Discipline for Traditional Chinese Medicine of Nanjing University of Chinese Medicine, Nanjing, 210046 China Weidong Li, National First-Class Key Discipline for Traditional Chinese Medicine of Nanjing University of Chinese Medicine, Nanjing, 210046 China Yang Pan, National First-Class Key Discipline for Traditional Chinese Medicine of Nanjing University of Chinese Medicine, Nanjing, 210046 China Xia Tu, Department of Bio-Pharmacy and Laboratory of Medical Fungi and Phyto-Biotech, Nanjing University of Chinese Medicine, Nanjing, 210029 China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Phospholipids have been shown to cause matrix effects particularly in liquid chromatography–mass spectrometry (LC–MS) analysis of small molecules. This results in suppression of the analyte signal. This study provides a versatile validated method for the analysis of serotonin in serum along with dopamine and melatonin using LC–MS/MS. It utilises HybridSPE-Precipitation cartridges for the clean-up of serum samples. This technology involves a simple protein precipitation step together with a fast and robust SPE method that is designed to remove phospholipids. Serotonin and dopamine are major neurotransmitters in the brain which affect various functions both in the brain and in the rest of the body. Melatonin plays an important role in the regulation of circadian sleep–wake cycle. Good linear calibrations were obtained for the multiplex assay of analytes in serum samples (0.021–3.268 μmol L −1 ; R 2  = 0.9983–0.9993). Acceptable intra- and inter-day repeatability was achieved for all analytes in serum. Excellent limits of detection (LOD) and limits of quantitation (LOQ) were achieved with LODs of 3.2–23.5 nmol L −1 and the LOQs of 15.4–70.5 nmol L −1 for these analytes in serum. The sample clean-up procedure that was developed provided efficient recovery and reproducibility while also decreasing preparation time and solvent use. A sample storage protocol was established, this was achieved by investigation of sample stability under different storage conditions. Evaluation of matrix effects was also carried out and the influence of ion suppression on analytical results reported. This clean-up protocol was then applied to the analysis of clinical serum samples. Content Type Journal Article Category Original Pages 1-13 DOI 10.1007/s10337-012-2330-5 Authors Merisa Moriarty, Department of Chemistry, PROTEOBIO (Mass Spectrometry Centre for Proteomic and Biotoxin Research), Cork Institute of Technology, Cork, Ireland Aoife Lee, Department of Biological Sciences, Cork Institute of Technology, Bishopstown, Cork, Ireland Brendan O’Connell, Department of Biological Sciences, Cork Institute of Technology, Bishopstown, Cork, Ireland Mary Lehane, Department of Chemistry, PROTEOBIO (Mass Spectrometry Centre for Proteomic and Biotoxin Research), Cork Institute of Technology, Cork, Ireland Helen Keeley, Child and Adolescent Mental Health Services, Health Service Executive, South, North Cork Area and the National Suicide Research Foundation, Cork, Ireland Ambrose Furey, Team Elucidate and PROTEOBIO (Mass Spectrometry Centre for Proteomic and Biotoxin Research) research groups, Cork Institute of Technology, Cork, Ireland Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    We have analyzed the structures of glycosphingolipids and intracellular free glycans in human cancers. In our previous study, trace amounts of free N -acetylneuraminic acid (Neu5Ac)-containing complex-type N -glycans with a single GlcNAc at each reducing terminus (Gn1 type) was found to accumulate intracellularly in colorectal cancers, but were undetectable in most normal colorectal epithelial cells. Here, we used cancer glycomic analyses to reveal that substantial amounts of free Neu5Ac-containing complex-type N -glycans, almost all of which were α2,6-Neu5Ac-linked, accumulated in the pancreatic cancer cells from three out of five patients, but were undetectable in normal pancreatic cells from all five cases. These molecular species were mostly composed of five kinds of glycans having a sequence Neu5Ac-Gal-GlcNAc-Man-Man-GlcNAc and one with the following sequence Neu5Ac-Gal-GlcNAc-Man-(Man-)Man-GlcNAc. The most abundant glycan was Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3Manβ1-4GlcNAc, followed by Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-6Manβ1-4GlcNAc. This is the first study to show unequivocal evidence for the occurrence of free Neu5Ac-linked N -glycans in human cancer tissues. Our findings suggest that free Neu5Ac-linked glycans may serve as a useful tumor marker. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9435-9 Authors Masahiko Yabu, Department of Immunology, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari-ku, Osaka, 537-8511 Japan Hiroaki Korekane, Systems Glycobiology Research Group, Chemical Biology Department, RIKEN Advanced Science Institute, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan Hidenori Takahashi, Department of Surgery, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari-ku, Osaka, 537-8511 Japan Hiroaki Ohigashi, Department of Surgery, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari-ku, Osaka, 537-8511 Japan Osamu Ishikawa, Department of Surgery, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari-ku, Osaka, 537-8511 Japan Yasuhide Miyamoto, Department of Immunology, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari-ku, Osaka, 537-8511 Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Graphene, a novel class of carbon nanostructure, possesses an ultra-high specific surface area (theoretical value 2,630 m 2  g −1 ), and both sides of the planar sheets of graphene are available for molecule adsorption. Graphene has already been used for preconcentration, extraction, and electrochemical selective determination. In this study, we used graphene to clean up pigments in cucumber for analysis, and measured eight pyrethroid model analytes using GC with electron capture detection (ECD). The recoveries of the 8 pyrethroids were 75–116 % with RSDs below 10 %, and LOQs ranged from 2.5 to 10 μg kg −1 . Comparative studies showed that graphene was superior to graphitized carbon black for the purification of pigments. We also investigated the ability of graphene to clean up spinach. A promising new adsorbent for pesticide residue analysis was developed. Graphene has significant potential as an effective adsorbent of pigments. Content Type Journal Article Category Original Pages 1-7 DOI 10.1007/s10337-012-2299-0 Authors Xiaoli Wu, College of Science, China Agricultural University, Beijing, 100193 People’s Republic of China Hongyan Zhang, College of Science, China Agricultural University, Beijing, 100193 People’s Republic of China Lixuan Meng, College of Science, China Agricultural University, Beijing, 100193 People’s Republic of China Xiaotong Liu, College of Science, China Agricultural University, Beijing, 100193 People’s Republic of China Yongqiang Ma, College of Science, China Agricultural University, Beijing, 100193 People’s Republic of China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    A highly sensitive and rapid method was developed that involves capillary electrophoresis for separation and determination of the stereoisomeric impurity of folinic acid diastereomers. In this method, vancomycin was used as the chiral selector, and a solution of poly(dimethylacrylamide) (PDMA) was prepared for dynamic coating of the capillary wall to minimize the adsorption of vancomycin. This method was optimized for six factors including concentrations of the organic modifier and vancomycin, pH and concentration of the background electrolyte, column temperature, and separation voltage. The following conditions were established: 100 mM Tris-phosphate buffer (pH 6.0) containing 1.0 mM vancomycin and 5 % acetonitrile at 30 °C, and −15 kV applied voltage on the PDMA dynamically coated capillary. Preliminary validation was performed with the determination of limit of quantification and detection, accuracy, precision, and linearity. Under our optimized method, the folinic acid diastereomers were baseline-separated within 7.5 min, and a (6 S ,2′ S )-calcium folinate sample with 0.08 % stereoisomeric impurity was determined. Content Type Journal Article Category Short Communication Pages 1-5 DOI 10.1007/s10337-012-2304-7 Authors Zhaoyan Wang, School of Pharmacy, Lanzhou University, Lanzhou, 730000 China Changjun Mu, School of Life Science, Lanzhou University, Lanzhou, 730000 China Jingwu Kang, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, 200032 China Zhide Hu, School of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou, 730000 China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    A simple and sensitive method for the determination of free aliphatic amines using 10-phenyl-acridone-2-sulfonyl chloride (PASC) as a labeling reagent by high-performance liquid chromatography with fluorescence detection and online mass spectrometry identification (HPLC-FLD-MS) has been developed. Derivatization conditions including reagent concentration, buffer pH, reaction time and temperature were optimized. PASC reacted with aliphatic amines at 50 °C for 4 min in aqueous acetonitrile (ACN) in the presence of sodiumtetraborate–NaOH buffer (0.10 mol L −1 , pH 9.0) to give high yields of PASC-amine derivatives. Derivatives exhibited intense fluorescence with an excitation maximum at λ ex 265 nm and an emission maximum at λ em 418 nm. The separation of derivatives was performed by a reversed-phase Hypersil BDS C8 column in combination with a gradient elution. The identification of derivatives was carried out by online post-column mass spectrometry with atmospheric pressure chemical ionization (APCI) source in positive-ion detection mode. Excellent linear responses were observed with the correlation coefficients of larger than 0.9997, and detection limits (at a signal-to-noise of 3:1) were from 3.0 to 24.3 fmol. Comparing with 10-ethyl-acridine-2-sulfonyl chloride (EASC), PASC exhibited more intense fluorescence and ultraviolet absorbance. The proposed method is sensitive and reproducible for the determination of aliphatic amines from water and soil samples. Content Type Journal Article Category Original Pages 1-10 DOI 10.1007/s10337-012-2298-1 Authors Shujing Ning, Key Laboratory of Life-Organic Analysis of Shandong Province, Qufu Normal University, Qufu, 273156 China Jinmao You, Key Laboratory of Life-Organic Analysis of Shandong Province, Qufu Normal University, Qufu, 273156 China Zhiwei Sun, Key Laboratory of Life-Organic Analysis of Shandong Province, Qufu Normal University, Qufu, 273156 China Shijuan Zhang, Key Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining, 810008 China Zhongyin Ji, Key Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining, 810008 China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Sulfatides, 3- O -sulfogalactosylceramides, are known to have multifunctional properties. These molecules are distributed in various tissues of mammals, where they are synthesized from galactosylceramides by sulfation at C3 of the galactosyl residue. Although this reaction is specifically catalyzed by cerebroside sulfotransferase (CST), the mechanisms underlying the transcriptional regulation of this enzyme are not understood. With respect to this issue, we previously found potential sequences of peroxisome proliferator-activated receptor (PPAR) response element on upstream regions of the mouse CST gene and presumed the possible regulation by the nuclear receptor PPARα. To confirm this hypothesis, we treated wild-type and Ppara -null mice with the specific PPARα agonist fenofibrate and examined the amounts of sulfatides and CST gene expression in various tissues. Fenofibrate treatment increased sulfatides and CST mRNA levels in the kidney, heart, liver, and small intestine in a PPARα-dependent manner. However, these effects of fenofibrate were absent in the brain or colon. Fenofibrate treatment did not affect the mRNA level of arylsulfatase A, which is the key enzyme for catalyzing desulfation of sulfatides, in any of these six tissues. Analyses of the DNA-binding activity and conventional gene expression targets of PPARα has demonstrated that fenofibrate treatment activated PPARα in the kidney, heart, liver, and small intestine but did not affect the brain or colon. These findings suggest that PPARα activation induces CST gene expression and enhances sulfatide synthesis in mice, which suggests that PPARα is a possible transcriptional regulator for the mouse CST gene. Content Type Journal Article Pages 1-8 DOI 10.1007/s10719-012-9454-6 Authors Takero Nakajima, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Yuji Kamijo, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Huang Yuzhe, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Takefumi Kimura, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Naoki Tanaka, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Eiko Sugiyama, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Kozo Nakamura, Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, Minami-Minowa, Kami-Ina, Nagano, Japan Mamoru Kyogashima, Division of Microbiology and Molecular Cell Biology, Nihon Pharmaceutical University, Ina, Kita-Adachi, Saitama, Japan Atsushi Hara, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Toshifumi Aoyama, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    A simple, rapid and sensitive LC–MS/MS method in positive ion mode was developed and validated to determine CKD-501, lobeglitazone, in human plasma and urine using glipizide as an internal standard (IS). Lobeglitazone is a novel thiazolidinedione (TZDs)-based peroxisome proliferator-activated receptor (PPAR) agonist, used for the management of type-2 diabetes. After mixing the IS, dissolved in acetonitrile, with a plasma or urine sample containing lobeglitazone, 10 μL of supernatant was injected into the LC–MS/MS system. Quantification was performed in the multiple reaction monitoring (MRM) mode using transition of 481.5 → 152.2 ( m / z ) for lobeglitazone and 446.1 → 321.2 ( m / z ) for the IS. The method showed good linearity over concentration ranges of 0.5–1,000 ng mL −1 for plasma and 0.2–250 ng mL −1 for urine ( r 2  ≥ 0.9996). The mean percent extraction recovery of lobeglitazone was 90.8 % for plasma and 87.3 % for urine, while the recoveries of the IS were greater than 86.4 % for both. The intra-day and inter-day precision of plasma ranged from 1.1 to 3.7 and 2.5 to 3.3 % (RSD), respectively, and the intra- and inter-day precision of urine ranged from 1.5 to 2.7 and 3.2 to 3.5 %, respectively. This method is simple, sensitive, and applicable for the pharmacokinetic study of lobeglitazone in human plasma. Most of the urine concentrations of lobeglitazone were below the LLOQ because the lobeglitazone is extensively metabolized. Content Type Journal Article Category Short Communication Pages 1-7 DOI 10.1007/s10337-012-2238-0 Authors Bora Kim, Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine and Hospital, 101 Daehangno, Jongno-gu, Seoul, 110-744 Republic of Korea Hyun-Suk Shin, Analytical Chemistry Laboratory, Clinical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea Jung-Ryul Kim, Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine and Hospital, 101 Daehangno, Jongno-gu, Seoul, 110-744 Republic of Korea Kyung-Soo Lim, Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine and Hospital, 101 Daehangno, Jongno-gu, Seoul, 110-744 Republic of Korea Seo Hyun Yoon, Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine and Hospital, 101 Daehangno, Jongno-gu, Seoul, 110-744 Republic of Korea Kyung-Sang Yu, Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine and Hospital, 101 Daehangno, Jongno-gu, Seoul, 110-744 Republic of Korea Sang-Goo Shin, Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine and Hospital, 101 Daehangno, Jongno-gu, Seoul, 110-744 Republic of Korea In-Jin Jang, Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine and Hospital, 101 Daehangno, Jongno-gu, Seoul, 110-744 Republic of Korea Joo-Youn Cho, Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine and Hospital, 101 Daehangno, Jongno-gu, Seoul, 110-744 Republic of Korea Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Expanded bed adsorption (EBA) is a practical method for the separation of nanoparticulates. In order to analysis the local hydrodynamic and adsorption behavior of nanoparticle (NP)-based biological feedstock, a modified Nano Biotechnology Group EBA column with a 26-mm inner diameter was used to withdraw liquid from different axial positions of the column. Fabricated egg albumin (EA) NPs with an average size of 70 nm were employed as a model system and viral size/charge mimic to assess the relationship between hydrodynamic and adsorption performance of NPs at the different column regions. The effects of influential factors, including flow velocity and initial concentration of NPs, on NP hydrodynamic behavior and adsorption kinetics along the bed height were investigated. NP hydrodynamic studies confirmed that non-uniform behavior dominated the system and a decreasing trend of liquid mixing/dispersion with increase of bed height was observed in this column. The results demonstrated an increase in the mixing/dispersion at certain bed heights with the increase in both the velocity and feed initial concentration. Breakthrough curves were measured at various column points to determine the adsorption performance [dynamic binding capacity (DBC) and yield] in different bed positions/zones. Yield and DBC of NPs were improved along the bed height, whereas liquid velocity had the opposite effect. Increasing the initial concentration of NPs enhanced only the DBC. Separation of EA NPs under optimal conditions was 87 %, which is an excellent result for a one-pass frontal chromatography method. Content Type Journal Article Category Original Pages 1-12 DOI 10.1007/s10337-012-2235-3 Authors Mohsen Jahanshahi, Nanotechnology Research Institute, Faculty of Chemical Engineering, Babol University of Technology, P.O. Box 484, Babol, Iran Mohammad Taghi Hamed Mosavian, Faculty of Chemical Engineering, Ferdowsi University of Mashhad, Mashhad, Iran Elham Sadat Taheri Otaghsara, Nanotechnology Research Institute, Faculty of Chemical Engineering, Babol University of Technology, P.O. Box 484, Babol, Iran Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    A liquid chromatographic tandem mass spectrometric (LC–MS–MS) method for the determination of five chemical coccidiostats (decoquinate, diclazuril, halofuginone, nicarbazin, and robenidine) and five ionophore coccidiostats (maduramicin, monensin, narasin, salinomycin, and semduramicin) in yoghurt, kefir, and sour cream is presented. Lasalocid, the sixth ionophore listed in 124/2009/EC was not included because of its extremely dissimilar behavior during sample preparation. Main steps of the method include extraction with acetonitrile, centrifugation, clean-up on Oasis HLB solid phase extraction cartridge, evaporation under nitrogen stream, and LC–MS–MS determination. Selectivity, linearity, sensitivity, accuracy, repeatability, within-laboratory reproducibility, limit of determination, and limit of quantitation were determined during the validation procedure. The method proved to be applicable for both qualitative and quantitative determination of the ten above-mentioned target compounds. In our in-house fermentation experiments, milk fortified with coccidiostats was fermented to get yoghurt, kefir, and sour cream. Our results show that the coccidiostat content did not change significantly during fermentation for any of the target compounds. Content Type Journal Article Category Original Pages 1-9 DOI 10.1007/s10337-012-2236-2 Authors Szilárd Nász, Joint Research and Training Laboratory on Separation Techniques, Eötvös Loránd University, Pázmány Péter sétány 1/A, Budapest, 1117 Hungary Etelka M. Károlyné, Alföldi Tej Kft, Seregélyesi út 127, Székesfehérvár, 8000 Hungary Tamás Rikker, Wessling International Research and Educational Center, Fóti út 56, Budapest, 1047 Hungary Zsuzsanna Eke, Joint Research and Training Laboratory on Separation Techniques, Eötvös Loránd University, Pázmány Péter sétány 1/A, Budapest, 1117 Hungary Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    The use of the large retention index database for identification and filtering of false positive hits in GC–MS analysis of the ylang-ylang essential oil is illustrated. Differences between experimental retention indices and database values of retention indices of candidate compounds provide additional constraints on the list of candidates for a target compound. Over 100 components of ylang-ylang essential oil (total grade) were identified. The main components, with concentrations more than 4 %, are β-caryophyllene, germacrene D, benzyl benzoate, linalool, geranyl acetate, α- (E,E) -farnesene and isobornyl acetate. Content Type Journal Article Category Short Communication Pages 1-8 DOI 10.1007/s10337-012-2231-7 Authors V. I. Babushok, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA N. R. Andriamaharavo, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    The objective of current investigation was to study the degradation behavior of l -DOPA under different conditions by high performance liquid chromatography (HPLC), and to develop and validate a stability-indicating HPLC method. The developed RP-HPLC method was validated with respect to linearity, accuracy, precision and specificity. Oxidation was found to occur in alkaline and to some extent in thermal conditions, while the drug was stable when incubated at acidic conditions and under photolytic stress. The oxidation of l -DOPA was observed to follow first-order kinetics. The degradation rate constants and half-life were calculated. The cytotoxicity and enzymatic degradation of l -DOPA was examined using the human intestinal epithelial Caco-2 cells. The drug was rapidly decarboxylated by aromatic amino acid decarboxylase to dopamine. The conversion of l -DOPA to dopamine was dose- and time-dependent. Content Type Journal Article Category Original Pages 1-10 DOI 10.1007/s10337-012-2229-1 Authors Yong Zhi Zhou, Drug Delivery Research Unit, School of Pharmacy, Faculty of Medical and Health Sciences, The University of Auckland, Auckland, New Zealand Raid G. Alany, Drug Delivery Research Unit, School of Pharmacy, Faculty of Medical and Health Sciences, The University of Auckland, Auckland, New Zealand Victor Chuang, School of Pharmacy, Faculty of Health Sciences, Curtin Health Innovation Research Institute, Curtin University, GPO Box U1987, Perth, WA 6845, Australia Jingyuan Wen, Drug Delivery Research Unit, School of Pharmacy, Faculty of Medical and Health Sciences, The University of Auckland, Auckland, New Zealand Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Aerial parts of Epimedium koreanum Nakai have been used as Herba Epimedii in China. Its anti-osteoporosis effect has attracted much attention in recent years. In this study, a method involving osteoblastic cell (MC3T3-E1 cell line) extraction and high-performance liquid chromatography–electrospray ionisation–mass spectrometry (HPLC–ESI-MS n ) was developed for screening of potential anti-osteoporosis agents in E. koreanum . Four compounds identified as epimedin A, epimedin B, epimedin C and icariin were found to interact with MC3T3-E1 cells and possessed potent osteoblast-stimulating activity as evaluated by cell proliferation [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2 H -tetrazolium bromide (MTT) assay] and differentiation [alkaline phosphatase (ALP) activity and Ca content] in vitro. The results suggest that these four flavonoids are the anti-osteoporosis constituents of Herba Epimedii and that the method combining MC3T3-E1 cell extraction with HPLC–ESI-MS n is rapid and effective in screening anti-osteoporosis agents from traditional Chinese medicines. Content Type Journal Article Category Original Pages 1-9 DOI 10.1007/s10337-012-2232-6 Authors Rui Wang, State Key Laboratory of Natural Medicines, Department of Natural Medicinal Chemistry, China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing, 210009 People’s Republic of China Jian-Guang Luo, State Key Laboratory of Natural Medicines, Department of Natural Medicinal Chemistry, China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing, 210009 People’s Republic of China Ling-Yi Kong, State Key Laboratory of Natural Medicines, Department of Natural Medicinal Chemistry, China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing, 210009 People’s Republic of China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    A simple method is described for efficient isolation of compounds having an antibacterial effect. Two thyme ( Thymus vulgaris ) essential oils, obtained from the market, were chosen as prospective materials likely to feature several bioactive components when examined by thin layer chromatography coupled with direct bioautography as a screening method. The newly developed infusion overpressured layer chromatographic separation method coupled with direct bioautography assured that only the active components were isolated by means of overrun overpressured layer chromatography (OPLC) with on-line detection and fractionation. Each of the 5 collected fractions represented one of the five antimicrobial essential oil components designated at the screening. The purity and the activity of the fractions were confirmed with chromatography coupled with various detection methods (UV, vanillin–sulphuric acid reagent, direct bioautography). The antibacterial components were identified with GC–MS as thymol, carvacrol, (−)-linalool, diethyl-phthalate, and α-terpineol. The oil component diethyl-phthalate is an artificial compound, used as a plasticizer or detergent base in the industry. Our results support that exploiting its flexibility and the possible hyphenations, OPLC is especially attractive for isolation of antimicrobial components from various matrixes. Content Type Journal Article Category Original Pages 1-9 DOI 10.1007/s10337-012-2233-5 Authors Ágnes M. Móricz, Centre for Agricultural Research, Plant Protection Institute, Hungarian Academy of Sciences, Herman O. Str. 15, Budapest, 1022 Hungary Péter G. Ott, Centre for Agricultural Research, Plant Protection Institute, Hungarian Academy of Sciences, Herman O. Str. 15, Budapest, 1022 Hungary Andrea Böszörményi, Institute of Pharmacognosy, Faculty of Pharmacy, Semmelweis University, Üllői út 26, Budapest, 1085 Hungary Éva Lemberkovics, Institute of Pharmacognosy, Faculty of Pharmacy, Semmelweis University, Üllői út 26, Budapest, 1085 Hungary Emil Mincsovics, OPLC-NIT Engineering Co., Ltd, Budapest Andor Str. 60, Budapest, 1119 Hungary Ernő Tyihák, Centre for Agricultural Research, Plant Protection Institute, Hungarian Academy of Sciences, Herman O. Str. 15, Budapest, 1022 Hungary Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description: Response to Letter to the Editor Regarding: Comparison of Several Methods of Chromatographic Baseline Removal with a New Approach Based on Quantile Regression Content Type Journal Article Category Letter to the Editor Pages 315-316 DOI 10.1007/s10337-012-2191-y Authors Ł. Komsta, Department of Medicinal Chemistry, Medical University of Lublin, Jaczewskiego 4, 20-090 Lublin, Poland Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893 Journal Volume Volume 75 Journal Issue Volume 75, Numbers 5-6

Description:    The bisecting GlcNAc is transferred to the core mannose residue of complex or hybrid N-glycans on glycoproteins by the β1,4- N -acetylglucosaminyltransferase III (GlcNAcT-III) or MGAT3. The addition of the bisecting GlcNAc confers unique lectin recognition properties to N-glycans. Thus, LEC10 gain-of-function Chinese hamster ovary (CHO) cells selected for the acquisition of ricin resistance, carry N-glycans with a bisecting GlcNAc, which enhances the binding of the erythroagglutinin E-PHA, but reduces the binding of ricin and galectins-1, -3 and -8. The altered interaction with galactose-binding lectins suggests that the bisecting GlcNAc affects N-glycan conformation. LEC10 mutants expressing polyoma middle T antigen (PyMT) exhibit reduced growth factor signaling. Furthermore, PyMT-induced mammary tumors lacking MGAT3, progress more rapidly than tumors with the bisecting GlcNAc on N-glycans of cell surface glycoproteins. In recent years, evidence for a new paradigm of cell growth control has emerged involving regulation of cell surface residency of growth factor and cytokine receptors via interactions and cross-linking of their branched N-glycans with a lattice of galectin(s). Specific cross-linking of glycoprotein receptors in the lattice regulates their endocytosis, leading to effects on growth factor-induced signaling. This review will describe evidence that the bisecting GlcNAc of N-glycans regulates cellular signaling and tumor progression, apparently through modulating N-glycan/galectin interactions. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9373-6 Authors Hazuki E. Miwa, Department of Cell Biology, Albert Einstein College of Medicine, New York, NY 10461, USA Yinghui Song, Department of Cell Biology, Albert Einstein College of Medicine, New York, NY 10461, USA Richard Alvarez, Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA Richard D. Cummings, Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA Pamela Stanley, Department of Cell Biology, Albert Einstein College of Medicine, New York, NY 10461, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    The retention behavior of melamine (MEL), ammeline (AMN), ammelide, and cyanuric acid on various hydrophilic interaction liquid chromatography (HILIC) stationary phases was studied. Using fully optimized electrospray conditions and ammonium formate buffer pH 4/acetonitrile (20:80 v/v) as a mobile phase, precursor and product ions for each compound were identified. For the LC separation study, the effect of the following parameters was investigated: the type of buffer, the effect of the pH of ammonium formate buffer, the dilution percentage of the standard solutions in the vial, the stability of the standard solutions, and the column equilibration time. The retention and separation of melamine and its hydrolysis products on several HILIC columns was also investigated and retention models were proposed. Retention mechanisms were discussed for all the compounds. When the percentage of acetonitrile in the mobile phase was increased, the retention times of ammelide, ammeline and melamine were shifted to higher values, while the retention time of cyanuric acid did not change significantly. The separation of compounds with isobaric transitions (MEL, AMN) was achieved on four HILIC columns, namely TSKgel Amide-80, Luna HILIC, XBridge HILIC, and ZIC-HILIC (at either 10/90 or 15/85 ammonium formate buffer pH 4/ACN). Content Type Journal Article Category Original Pages 1-11 DOI 10.1007/s10337-012-2228-2 Authors Maroula G. Kokotou, Laboratory of Analytical Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, 15771 Athens, Greece Nikolaos S. Thomaidis, Laboratory of Analytical Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, 15771 Athens, Greece Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Malondialdehyde (MDA) is an end-product of lipid peroxidation and a side product of thromboxane A 2 synthesis. Moreover, it is not only a frequently measured biomarker of oxidative stress, but its high reactivity and toxicity underline the fact that this molecule is more than “just” a biomarker. Additionally, MDA was proven to be a mutagenic substance. Having said this, it is evident that there is a major interest in the highly selective and sensitive analysis of this molecule in various matrices. In this review, we will provide a brief overview of the most recent developments and techniques for the liquid chromatography (LC) and gas chromatography (GC)-based analysis of MDA in different matrices. While the 2-thiobarbituric acid assay still is the most prominent methodology for determining MDA, several advanced techniques have evolved, including GC–MS(MS), LC–MS(MS) as well as several derivatization-based strategies. Content Type Journal Article Category Review Pages 1-8 DOI 10.1007/s10337-012-2237-1 Authors Martin Giera, Biomolecular Mass Spectrometry Unit, Leiden University Medical Center (LUMC), Albinusdreef 2, 2300 RC Leiden, The Netherlands Henk Lingeman, BioMolecular Analysis, VU University Amsterdam, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands Wilfried M. A. Niessen, BioMolecular Analysis, VU University Amsterdam, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Acepromazine maleate (Sedalin ® ) was administered orally to six thoroughbred horses at a dose of 0.15 mg kg −1 . Urine and blood samples were collected up to 412 h post-administration. Plasma and urine were hydrolysed; plasma samples were then processed using liquid–liquid extraction and urine samples using solid-phase extraction. A sensitive tandem mass spectrometric method was developed in this study, achieving a lower limit of quantification for acepromazine of 10 pg mL −1 in plasma and 100 pg mL −1 in urine. Acepromazine, hydroxyethylpromazine, hydroxyacepromazine, hydroxyethylpromazine sulphoxide, hydroxyethylhydroxypromazine, dihydroxyacepromazine and dihydroxyhydroxyethylpromazine were detected in the post-administration samples. The parent drug and its metabolites were identified using a combination of UPLC–MS/MS and accurate mass measurement. Separation of the structural isomers hydroxyethylpromazine sulphoxide and hydroxyethylhydroxypromazine was another significant outcome of this work and demonstrated the advantages to be gained from investing in chromatographic method development. Content Type Journal Article Category Original Pages 1-9 DOI 10.1007/s10337-012-2234-4 Authors M. E. Wieder, HFL Sport Science, Cambridgeshire, UK B. P. Gray, HFL Sport Science, Cambridgeshire, UK P. R. Brown, HFL Sport Science, Cambridgeshire, UK S. Hudson, HFL Sport Science, Cambridgeshire, UK C. M. Pearce, HFL Sport Science, Cambridgeshire, UK S. W. Paine, British Horseracing Authority, London, UK L. Hillyer, British Horseracing Authority, London, UK Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    High-performance liquid chromatography (HPLC), over-pressured-layer chromatography (OPLC) and thin-layer chromatography (TLC) techniques with micellar mobile phases were proposed to evaluate the lipophilicity of 21 newly synthesized 1,2,4-triazoles, compounds of potential importance in medicine or agriculture as fungicides. Micellar parameters log  k m were compared with extrapolated R M 0 values determined from reversed-phase (RP) TLC experimental data obtained on RP-8 stationary phases as well as with log  P values (Alog  Ps , AClog  P , Alog  P , Mlog  P , KowWin, xlog  P 2 and xlog  P 3) calculated from molecular structures of solutes tested. The results obtained by applying principal component analysis (PCA) and linear regression showed considerable similarity between partition and retention parameters as alternative lipophilicity descriptors, and indicated micellar chromatography as a suitable technique to study lipophilic properties of organic substances. In micellar HPLC, RP-8e column (Purospher) was applied, whereas in OPLC and TLC, RP-CN plates were applied, which was the novelty of this study and allowed the use of micellar effluents in planar chromatography measurements. Content Type Journal Article Category Original Pages 1-8 DOI 10.1007/s10337-012-2227-3 Authors Małgorzata Janicka, Department of Physical Chemistry, Faculty of Chemistry, Maria Curie-Skłodowska University, Maria Curie-Skłodowska Sq. 3, 20-031 Lublin, Poland Katarzyna Stępnik, Department of Physical Chemistry, Faculty of Chemistry, Maria Curie-Skłodowska University, Maria Curie-Skłodowska Sq. 3, 20-031 Lublin, Poland Anna Pachuta-Stec, Department of Organic Chemistry, Faculty of Pharmacy, Medical University, 6 Staszica St, 20-081 Lublin, Poland Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Several months ago, a paper by Komsta describing a method of baseline removal based on quantile regression for chromatographic datasets was published in this journal entitled “Comparison of Several Methods of Chromatographic Baseline Removal with a New Approach Based on Quantile Regression”. By comparing baseline removal methods based on polynomial, spline, loess and Whittaker smoother with the newly introduced method, Komsta concluded that the main advantage of this baseline removal method is visible better performance and short computational time. Unfortunately several wrong conclusions about the baseline removal methods based on Whittaker smoother contained in this paper. We feel that wrong conclusions about the baseline removal methods based on Whittaker smoother needs to be corrected so that it will not mislead the reader. Content Type Journal Article Category Letter to the Editor Pages 313-314 DOI 10.1007/s10337-012-2192-x Authors Zhi-Min Zhang, College of Chemistry and Chemical Engineering, Research Center of Modernization of Chinese Medicines, Central South University, Changsha, 410083 People’s Republic of China Yi-Zeng Liang, College of Chemistry and Chemical Engineering, Research Center of Modernization of Chinese Medicines, Central South University, Changsha, 410083 People’s Republic of China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893 Journal Volume Volume 75 Journal Issue Volume 75, Numbers 5-6

Description:    The past 25 years have seen significant advances in understanding the diversity and functions of glycoprotein glycans in Drosophila melanogaster . Genetic screens have captured mutations that reveal important biological activities modulated by glycans, including protein folding and trafficking, as well as cell signaling, tissue morphogenesis, fertility, and viability. Many of these glycan functions have parallels in vertebrate development and disease, providing increasing opportunities to dissect pathologic mechanisms using Drosophila genetics. Advances in the sensitivity of structural analytic techniques have allowed the glycan profiles of wild-type and mutant tissues to be assessed, revealing novel glycan structures that may be functionally analogous to vertebrate glycans. This review describes a selected set of recent advances in understanding the functions of N-linked and O-linked (non-glycosaminoglycan) glycoprotein glycans in Drosophila with emphasis on their relatedness to vertebrate organisms. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9442-x Authors Toshihiko Katoh, The Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA Michael Tiemeyer, The Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    In this study, we purified and characterized the β-xylosidase involved in the turnover of plant complex type N -glycans to homogeneity from mature red tomatoes. Purified β-xylosidase (β-Xyl’ase Le-1) gave a single band with molecular masses of 67 kDa on SDS-PAGE under a reducing condition and 60 kDa on gelfiltration, indicating that β-Xyl’ase Le-1 has a monomeric structure in plant cells. The N- terminal amino acid could not be identified owing to a chemical modification. When pyridylaminated (PA-) N- glycans were used as substrates, β-Xyl’ase Le-1 showed optimum activity at about pH 5 at 40 °C, suggesting that the enzyme functions in a rather acidic circumstance such as in the vacuole or cell wall. β-Xyl’ase Le-1 hydrolyzed the β1-2 xylosyl residue from Man 1 Xyl 1 GlcNAc 2 -PA, Man 1 Xyl 1 Fuc 1 GlcNAc 2 -PA, and Man 2 Xyl 1 Fuc 1 GlcNAc 2 -PA, but not that from Man 3 Xyl 1 GlcNAc 2 -PA or Man 3 Xyl 1 Fuc 1 GlcNAc 2 -PA, indicating that the α1-3 arm mannosyl residue exerts significant steric hindrance for the access of β-Xyl’ase Le-1 to the xylosyl residue, whereas the α1-3 fucosyl residue exerts little effect. These results suggest that the release of the β1-2 xylosyl residue by β-Xyl’ase Le-1 occurs at least after the removal the α-1,3-mannosyl residue in the core trimannosyl unit. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9441-y Authors Daisuke Yokouchi, Department of Biofunctional Chemistry, Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka 1-1-1, Okayama, 700-8530 Japan Natsuko Ono, Department of Biofunctional Chemistry, Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka 1-1-1, Okayama, 700-8530 Japan Kosuke Nakamura, Kagome Research Institute, Kagome Co., Ltd., 17 Nishitomiyama, Nasushiobara, Tochigi 329-2762, Japan Megumi Maeda, Department of Biofunctional Chemistry, Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka 1-1-1, Okayama, 700-8530 Japan Yoshinobu Kimura, Department of Biofunctional Chemistry, Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka 1-1-1, Okayama, 700-8530 Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Many post-translational modifications, including glycosylation, are pivotal for the structural integrity, location and functional activity of glycoproteins. Sub-populations of proteins that are relocated or functionally changed by such modifications can change resting proteins into active ones, mediating specific effector functions, as in the case of monoclonal antibodies. To ensure safe and efficacious drugs it is essential to employ appropriate robust, quantitative analytical strategies that can (i) perform detailed glycan structural analysis, (ii) characterise specific subsets of glycans to assess known critical features of therapeutic activities (iii) rapidly profile glycan pools for at-line monitoring or high level batch to batch screening. Here we focus on these aspects of glycan analysis, showing how state-of-the-art technologies are required at all stages during the production of recombinant glycotherapeutics. These data can provide insights into processing pathways and suggest markers for intervention at critical control points in bioprocessing and also critical decision points in disease and drug monitoring in patients. Importantly, these tools are now enabling the first glycome/genome studies in large populations, allowing the integration of glycomics into other ‘omics platforms in a systems biology context. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9443-9 Authors Tharmala Tharmalingam, NIBRT Glycobiology Laboratory, NIBRT - The National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland Barbara Adamczyk, NIBRT Glycobiology Laboratory, NIBRT - The National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland Margaret A. Doherty, NIBRT Glycobiology Laboratory, NIBRT - The National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland Louise Royle, Ludger Ltd., Culham Science Centre, Oxfordshire, OX14 3EB UK Pauline M. Rudd, NIBRT Glycobiology Laboratory, NIBRT - The National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    In this paper, we present an optimized procedure for metabolomic analysis of endogenous metabolites in mouse fibroblast (L929) cell line using gas chromatography/time-of-flight mass spectrometry with multivariate statistics. The optimization of metabolite extraction was performed using three solvents: methanol, water, and chloroform, and then followed by methoxymation and silylation. This method was subsequently validated using 29 reference standards and cell line samples. The intra- and inter-day relative standard deviations (RSDs) of the standard compounds were lower than 15.0 and 25.0 %, respectively. As for most of the tested metabolites in cell line samples, RSDs were below 20.0 % for reproducibility and stability, respectively. We applied this approach in metabolomic study of L929 cells obtained from TiO 2 nanoparticle-induced cytotoxicity model samples ( n  = 5) and control samples ( n  = 5). Metabolite markers associated with TiO 2 nanoparticle-induced cytotoxicity were identified and validated by statistical methods and reference standards. Our work highlights the potential of this method for cell metabolomic study. Content Type Journal Article Category Original Pages 1-10 DOI 10.1007/s10337-012-2315-4 Authors Yumin Liu, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, 200240 China Yu Cheng, Department of Chemistry, East China Normal University, Shanghai, 200240 China Tianlu Chen, Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, 200240 China Yinan Zhang, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, 200240 China Xiaoyan Wang, Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, 200240 China Aihua Zhao, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, 200240 China Wei Jia, Department of Nutrition, University of North Carolina at Greensboro, North Carolina Research Campus, Kannapolis, NC 28081, USA Yang Bo, Center for Instrumental Analysis, Shanghai Jiao Tong University, Shanghai, 200240 China Chengyu Jin, Center for Instrumental Analysis, Shanghai Jiao Tong University, Shanghai, 200240 China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    The direct enantiomeric resolution of racemic 2-(1 H -imidazole-1-yl)-1-naphthalene-2-yl)ethanol esters, 1-(naphthalene-2-yl)ethanol esters, and 1-(1-hydroxynaphthalene-2-yl)-2-(1 H -imidazole-1-yl)ethanol on silica-based cellulose tris(3,5-dimethylphenylcarbamate) (Chiralcel OD) column is described. The separations were performed using mobile phases which consist of alcohol (methanol, ethanol or 2-propanol)/ n -hexane in various proportions. The effect of structural features of the solutes along with the nature and concentration of alcohol in the mobile phase on the discrimination between the enantiomers was examined for different mobile phase compositions. The results suggest that not only the structure and concentration of alcohol in the mobile phase, but also the subtle structural differences in racemates can have a pronounced effect on enantiomeric separation and retention. Baseline separations were obtained for 2-(1 H -imidazole-1-yl)-1-naphthalene-2-yl)ethanol esters carrying imidazole ring in addition to ester functional group in their structures. The α values of the resolved enantiomers of 2-(1 H -imidazole-1-yl)-1-naphthalene-2-yl)ethanol esters were in the range of 1.49–1.62 while the R s values varied from 4.20 to 6.75 when methanol/ n -hexane (70:30 v/v) was used as mobile phase. Content Type Journal Article Category Short Communication Pages 1-7 DOI 10.1007/s10337-012-2303-8 Authors A. Karakurt, Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Hacettepe University, Sihhiye, 06100 Ankara, Turkey S. Saraç, Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Hacettepe University, Sihhiye, 06100 Ankara, Turkey S. Dalkara, Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Hacettepe University, Sihhiye, 06100 Ankara, Turkey Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    To investigate the effects of QuTan HuaYu TongMai granule (QHTG) on the perturbed metabolism of atherosclerosis mini-pig model, a serum metabonomics method based on Liquid chromatography/mass spectrometry was developed. Principal component analysis and partial least squares-discriminate analysis models were established for the metabonomics analysis. The effect of high dose QHTG group is equivalent with those of positive drugs group in PCA score plots. Some significantly changed metabolites, including lyso-PCs, metabolites of vitamin D 3 and fatty acids were found to be reasonable in explaining the anti-atherosclerosis mechanism of QHTG. Metabonomic approach is helpful to further understand the pathophysiology of atherosclerosis in mini-pigs and the therapeutic mechanism of QHTG. Our work also indicated that the LC–MS based metabonomics method is a potential tool for performing intervention and mechanism research of traditional Chinese medicine (TCM). Content Type Journal Article Category Original Pages 1-8 DOI 10.1007/s10337-012-2316-3 Authors Sun Ming-Qian, Research Center, Xiyuan Hospital, China Academy of Chinese Medical Sciences, 100091 Beijing, People’s Republic of China Liu Jian-Xun, Research Center, Xiyuan Hospital, China Academy of Chinese Medical Sciences, 100091 Beijing, People’s Republic of China Lin Cheng-Ren, Research Center, Xiyuan Hospital, China Academy of Chinese Medical Sciences, 100091 Beijing, People’s Republic of China Li Lei, Research Center, Xiyuan Hospital, China Academy of Chinese Medical Sciences, 100091 Beijing, People’s Republic of China Ren Jian-Xun, Research Center, Xiyuan Hospital, China Academy of Chinese Medical Sciences, 100091 Beijing, People’s Republic of China Miao Lan, Research Center, Xiyuan Hospital, China Academy of Chinese Medical Sciences, 100091 Beijing, People’s Republic of China Cao Jin, Research Center, Xiyuan Hospital, China Academy of Chinese Medical Sciences, 100091 Beijing, People’s Republic of China Lin Li, Research Center, Xiyuan Hospital, China Academy of Chinese Medical Sciences, 100091 Beijing, People’s Republic of China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    The modernization and marketing of traditional Chinese medicine is difficult primarily because its pharmacodynamic material basis has not been elucidated. Moreover, a central problem is the lack of effective research methods and strategies. After 〉20 years of study, here, we propose the theory of pharmacodynamic differential serum chromatography. The method described here can be used to identify (and possibly develop) effective pharmacodynamic materials. In light of the theory, we identified hydroxysafflor yellow A (HSYA) to be the effective pharmacodynamic material of Carthamus tinctorius L. (safflower). With the help of the methodology described here, the modernization and understanding of traditional Chinese medicine may be accelerated. Content Type Journal Article Category Original Pages 1-6 DOI 10.1007/s10337-012-2319-0 Authors Chao Wang, Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, 17 Chang Le Xi Road Xi’an, Shaanxi, People’s Republic of China Qian Xiao, Department of Dermatology, Xijing Hospital, Fourth Military Medical University, 17 Chang Le Xi Road, Xi’an, Shaanxi, People’s Republic of China Yuwen Li, Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, 17 Chang Le Xi Road Xi’an, Shaanxi, People’s Republic of China Zhifu Yang, Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, 17 Chang Le Xi Road Xi’an, Shaanxi, People’s Republic of China Yin Wu, Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, 17 Chang Le Xi Road Xi’an, Shaanxi, People’s Republic of China Yanyan Jia, Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, 17 Chang Le Xi Road Xi’an, Shaanxi, People’s Republic of China Yi Qiao, Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, 17 Chang Le Xi Road Xi’an, Shaanxi, People’s Republic of China Miaomiao Xi, Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, 17 Chang Le Xi Road Xi’an, Shaanxi, People’s Republic of China Aidong Wen, Department of Pharmacy, Xijing Hospital, Fourth Military Medical University, 17 Chang Le Xi Road Xi’an, Shaanxi, People’s Republic of China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Catalytic reactions involved in the synthesis of the promising kinds of novel fuel and products formed in these reactions were systematized according to the resulting fuel type. Generalization of the retention of the substances comprising these products is presented. Chromatograms exhibiting their separation on chromatographic materials with the surface of different chemical properties are summarized. We propose procedures for gas-chromatographic analysis of the catalytic reactions products formed in the synthesis of hydrogen, methanol, dimethyl ether and hydrocarbons as a new generation of fuel alternative to petroleum and coal. For partial oxidation of methane into synthesis gas, on-line determination of the components obtained in the reaction was carried out by gas chromatography and gas analyzer based on different physicochemical methods (IR spectroscopy and electrochemical methods). Similarity of the results obtained using these methods is demonstrated. Content Type Journal Article Category Original Pages 1057-1068 DOI 10.1007/s10337-012-2281-x Authors V. Zheivot, Boreskov Institute of Catalysis, Siberian Branch, Russian Academy of Sciences, Lavrentyev Ave., 5, Novosibirsk, 630090 Russian Federation N. Sazonova, Boreskov Institute of Catalysis, Siberian Branch, Russian Academy of Sciences, Lavrentyev Ave., 5, Novosibirsk, 630090 Russian Federation Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893 Journal Volume Volume 75 Journal Issue Volume 75, Numbers 17-18

Description:    Recently, we demonstrated that the human xylosyltransferase II (XT-II) has enzymatic activity and is able to catalyze the initial and rate-limiting step in the biosynthesis of glycosaminoglycans (GAGs) like chondroitin and dermatan sulfate, as well as heparan sulfate and heparin. Therefore, this enzyme also very likely assumes a crucial regulatory role in the biosynthesis of proteoglycans (PGs). In this study, we identified and characterized for the first time the XYLT2 gene promoter region and transcription factors involved in its regulation. Several binding sites for members of the Sp1 family of transcription factors were identified as being necessary for transcriptional regulation of the XYLT2 gene. This was determined by mithramycin A treatment, electrophoretic mobility shift and supershift assays, as well as numerous site-directed mutagenesis experiments. Different 5′ and 3′ deletion constructs of the predicted GC rich promoter region, which lacks a canonical TATA and CAAT box, revealed that a 177 nts proximal promoter element is sufficient and indispensable to drive the constitutive transcription in full strength in HepG2 hepatoma cells. In addition, we also detected the transcriptional start site using 5′-RACE (rapid amplification of cDNA ends). Our results provide an insight into transcriptional regulation of the XYLT2 gene and may contribute to understanding the manifold GAG-involving processes in health and disease. Content Type Journal Article Pages 1-9 DOI 10.1007/s10719-012-9439-5 Authors Benjamin Müller, Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum NRW, Universitätsklinik der Ruhr-Universität Bochum, 32545 Bad Oeynhausen, Germany Christian Prante, Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum NRW, Universitätsklinik der Ruhr-Universität Bochum, 32545 Bad Oeynhausen, Germany Cornelius Knabbe, Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum NRW, Universitätsklinik der Ruhr-Universität Bochum, 32545 Bad Oeynhausen, Germany Knut Kleesiek, Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum NRW, Universitätsklinik der Ruhr-Universität Bochum, 32545 Bad Oeynhausen, Germany Christian Götting, Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum NRW, Universitätsklinik der Ruhr-Universität Bochum, 32545 Bad Oeynhausen, Germany Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Natural killer gene complex (NKC) encodes a group of proteins with a single C-type lectin-like domain, (CTLD) which can be subdivided several subfamilies according to their structures and expression patterns. The receptors containing the conserved calcium binding sites in the CTLD fold belong to group II of C-type lectin superfamily and are expressed on myeloid cells and non- myeloid cells. The receptors lacking conserved calcium binding sites in the CTLD fold have evolved to bind ligands other than carbohydrates independently on calcium and thereby are named as C-type lectin-like receptors. The C-type lectin-like receptors are previously thought to be exclusively expressed on natural killer (NK) cells and enable NK cells to discriminate self, missing self or altered self. However, some C-type lectin-like receptors are identified in myeloid cells and are intensely investigated, recently. These myeloid C-type lectin-like receptors, especially Dectin-1 cluster, have a wide variety of ligands, including those of exogenous origin, and play important roles in the physiological functions and pathological processes including immune homeostasis, immune defenses, and immune surveillance. In this review, we summarize each member of the Dectin-1 cluster, including their structural profiles, expression patterns, signaling properties as well as known physiological functions. Content Type Journal Article Pages 1-12 DOI 10.1007/s10719-012-9419-9 Authors Jianhui Xie, Key Laboratory of Glycoconjugate Research, Ministry of Health, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, Shanghai, 200032 China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    A high ionization efficiency derivatization reagent with a diazonium moiety, 4-diazomethylpyridine, has been designed for prostaglandins labeling. The labeled prostaglandins are then determined by LC–MS/MS. Prostaglandin E 2 (PGE 2 ) in rat brain homogenate was used for evaluation of the analytical potential of this reagent. 4-Diazomethylpyridine was synthesized in three steps. Analytes were purified in one step via C 18 solid-phase extraction, derivatized with 4-diazomethylpyridine, separated by reverse-phase HPLC and quantified on a triple-quadrupole mass spectrometer using selected reaction monitoring in positive ionization. The derivatization reaction was completed at 40 °C within 10 min without catalyst. Chromatographic separation of PGE 2 derivative was achieved on a Waters Symmetry C 18 column (100 × 2.1 mm, 3.5 μm) using a mobile phase consisting of 10-mM ammonium acetate in deionized water and methanol (25:75, v/v ). The flow rate was 0.2 mL min −1 and the total run time was 5 min. The mass transition was m/z 444 → 110. The calibration curve was linear in the range from 0.65 to 7.10 pmol per injection volume of 5 μL and the detection limit was about 0.21 pmol on column. Recoveries of spiked PGE 2 were above 70 % with intra-day relative standard deviation 〈6.8 % and inter-day relative standard deviation 〈9.8 %. This method has been successfully applied to the determination of PGE 2 in rat brain tissue. Content Type Journal Article Category Original Pages 1-7 DOI 10.1007/s10337-012-2271-z Authors Ying Zeng, Bioanalytical Laboratory, Shantou University Medical College, Guangdong, 515041 Shantou, China Hui Li, Bioanalytical Laboratory, Shantou University Medical College, Guangdong, 515041 Shantou, China Zhexuan Lin, Bioanalytical Laboratory, Shantou University Medical College, Guangdong, 515041 Shantou, China Hongjun Luo, Bioanalytical Laboratory, Shantou University Medical College, Guangdong, 515041 Shantou, China Jinhong Zheng, Department of Chemistry, Shantou University Medical College, Guangdong, 515041 Shantou, China Wenhong Luo, Bioanalytical Laboratory, Shantou University Medical College, Guangdong, 515041 Shantou, China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Polysaccharide-based chiral columns were evaluated for the separation of enantiomers of four chiral epoxides under normal-phase and polar organic mode. Since all four studied compounds contain two centers of chirality and, thus, theoretically, the coexistence of four stereoisomers is possible, the columns were evaluated from the viewpoints of both, chemoselectivity and enantioselectivity. For two of the epoxides, enantioseparation was achieved in the normal-phase mode as well as in the polar organic mode on several polysaccharide columns. The polar organic mode is especially attractive for the preparative separation of the enantiomers due to the enhanced solubility of the compounds in these solvents, as well as due to a shorter run time. A new and unusual (stereoselective) peak focusing phenomenon was observed in some experiments. Content Type Journal Article Category Original Pages 1-7 DOI 10.1007/s10337-012-2289-2 Authors Ketevan Lomsadze, Institute of Physical and Analytical Chemistry, School of Exact and Natural Sciences, Tbilisi State University, Chavchavadze Ave 3, 0179 Tbilisi, Georgia Maia Merlani, Laboratory of Plant Biopolymers, Kutateladze Institute of Pharmacochemistry, Tbilisi State Medical University, 0159 Tbilisi, Georgia Vakhtang Barbakadze, Laboratory of Plant Biopolymers, Kutateladze Institute of Pharmacochemistry, Tbilisi State Medical University, 0159 Tbilisi, Georgia Tivadar Farkas, Phenomenex Inc, 411 Madrid Ave., Torrance, 90501 CA, USA Bezhan Chankvetadze, Institute of Physical and Analytical Chemistry, School of Exact and Natural Sciences, Tbilisi State University, Chavchavadze Ave 3, 0179 Tbilisi, Georgia Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Human LOX-1/OLR 1 plays a key role in atherogenesis and endothelial dysfunction. The N-glycosylation of LOX-1 has been shown to affect its biological functions in vivo and modulate the pathogenesis of atherosclerosis. However, the N-glycosylation pattern of LOX-1 has not been described yet. The present study was aimed at elucidating the N-glycosylation of recombinant human LOX-1 with regard to N-glycan profile and N-glycosylation sites. Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS 3 ) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn 139 is readily identified and found to carry heterogeneous complex type N-glycans. In addition, manual annotation of multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans was addressed for proper spectra interpretation. The findings described herein will shed new light on further research of the structure-function relationships of LOX-1 N-glycan. Content Type Journal Article Pages 1-11 DOI 10.1007/s10719-012-9408-z Authors Yifan Qian, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Xingwang Zhang, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Lei Zhou, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Xiaojing Yun, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Jianhui Xie, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Jiejie Xu, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Yuanyuan Ruan, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Shifang Ren, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Rheumatoid arthritis (RA) is an inflammatory disorder that is characterized by persistent recurrence of joint inflammation leading to cartilage and bone destruction. The present anti-arthritis therapies failed to achieve satisfactory remission in all patients; therefore, it is still necessary to develop novel approaches to fulfill the demand in clinic. Here, we reported the therapeutic effects of lactosyl derivative Gu-4, a synthetic compound that was previously identified as a selective inhibitor against leukocyte integrin CD11b, in a bovine type II collagen induced arthritis (CIA) rat model. First, prophylactic administration of Gu-4 (1.2728 mg/kg) to rats by intraperitoneal injection every 2 days from the first day of collagen immunization significantly decreased the incidence of CIA, diminished the mean paw volume increase, and reduced the number of swollen paws. Second, administration of Gu-4 (1.2728 mg/kg) to rats at early-onset stage of CIA prevented the progression of the pathological process of RA, accelerated the remission of paw edema, and declined the arthritis score; after 5 weeks treatment, X-ray and histological examinations were carried out, the ankle joint of hind limb of Gu-4 treated CIA rats exhibited slighter bone erosion and much less inflammatory cell infiltration compared to those of saline treated animals; furthermore, Gu-4 remarkably attenuated the production of rheumatoid factor (RF) in the serum of CIA rats as determined by ELISA. Moreover, we performed in vitro lymphocyte proliferation assay and found that Gu-4 significantly inhibited the proliferation of splenic lymphocytes isolated from CIA rats in a dose-dependent manner. Our results suggest that Gu-4 can effectively ameliorate CIA and might be an alternative option for the treatment of RA. Content Type Journal Article Pages 1-9 DOI 10.1007/s10719-012-9407-0 Authors Jie Fan, Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046 China Huiting Zhou, Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046 China Shihui Wang, Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046 China Hailian Wang, Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046 China Yushun Zhang, Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046 China Yingtao Guo, Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046 China Qing Li, State Key Laboratory of Natural and Biomimetic Drugs, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Beijing, 100083 China Zhongjun Li, State Key Laboratory of Natural and Biomimetic Drugs, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Beijing, 100083 China Zhihui Zhao, Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046 China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Despite recent technical advances in glycan analysis, the rapidly growing field of glycomics still lacks methods that are high throughput and robust, and yet allow detailed and reliable identification of different glycans. LC-MS-MS 2 methods have a large potential for glycan analysis as they enable separation and identification of different glycans, including structural isomers. The major drawback is the complexity of the data with different charge states and adduct combinations. In practice, manual data analysis, still largely used for MALDI-TOF data, is no more achievable for LC-MS-MS 2 data. To solve the problem, we developed a glycan analysis software GlycanID for the analysis of LC-MS-MS 2 data to identify and profile glycan compositions in combination with existing proteomic software. IgG was used as an example of an individual glycoprotein and extracted cell surface proteins of human fibroblasts as a more complex sample to demonstrate the power of the novel data analysis approach. N-glycans were isolated from the samples and analyzed as permethylated sugar alditols by LC-MS-MS 2 , permitting semiquantitative glycan profiling. The data analysis consisted of five steps: 1) extraction of LC-MS features and MS 2 spectra, 2) mapping potential glycans based on feature distribution, 3) matching the feature masses with a glycan composition database and de novo generated compositions, 4) scoring MS 2 spectra with theoretical glycan fragments, and 5) composing the glycan profile for the identified glycan compositions. The resulting N-glycan profile of IgG revealed 28 glycan compositions and was in good correlation with the published IgG profile. More than 50 glycan compositions were reliably identified from the cell surface N-glycan profile of human fibroblasts. Use of the GlycanID software made relatively rapid analysis of complex glycan LC-MS-MS 2 data feasible. The results demonstrate that the complexity of glycan LC-MS-MS 2 data can be used as an asset to increase the reliability of the identifications. Content Type Journal Article Pages 1-12 DOI 10.1007/s10719-012-9412-3 Authors Hannu Peltoniemi, Applied Numerics Ltd, Nuottapolku 10 A 8, 00330 Helsinki, Finland Suvi Natunen, Finnish Red Cross Blood Service, Helsinki, Finland Ilja Ritamo, Finnish Red Cross Blood Service, Helsinki, Finland Leena Valmu, Finnish Red Cross Blood Service, Helsinki, Finland Jarkko Räbinä, Finnish Red Cross Blood Service, Helsinki, Finland Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Proteoglycans have been studied to a limited extent in lymphoid cells. In this study we have investigated the expression of proteoglycans in B-cells, CD4+ T-cells, CD8+ T-cells, natural killer cells, as well as in nine different cell lines established from patients with lymphoid malignancies. Serglycin was the major proteoglycan expressed at mRNA level by the primary lymphocytes. None of the syndecans or glycpicans was detected at mRNA level in the primary lymphocytes, except for syndecan-4 in CD4+ T-cells and CD8+ T-cells. All lymphoid cell lines expressed serglycin mRNA, as well as one or several members of the syndecan and glypican families. Further, increased synthesis of proteoglycans was found in the cell lines compared to the primary lymphocytes, as well as the presence of heparan sulfate on the cell surface of five of the cells lines. Western blot analysis showed a close correlation between serglycin mRNA level and expression of serglycin core protein. Our results show that serglycin is a major proteoglycan in all the normal lymphoid cells and that these cells carry little, or none, proteoglycans on the cell surface. Serglycin was also a major proteoglycan in the malignant lymphoid cells, but these also expressed one or more types of cell surface proteoglycans. Thus, malignant transformation of lymphoid cells may be followed by increased synthesis of proteoglycans and expression of cell surface proteoglycans. Content Type Journal Article Pages 1-11 DOI 10.1007/s10719-012-9427-9 Authors Bodil Fadnes, Institute of Medical Biology, Faculty of Health Sciences, University of Tromsø, Tromsø, Norway Anne Husebekk, Institute of Medical Biology, Faculty of Health Sciences, University of Tromsø, Tromsø, Norway Gunbjørg Svineng, Institute of Medical Biology, Faculty of Health Sciences, University of Tromsø, Tromsø, Norway Øystein Rekdal, Institute of Medical Biology, Faculty of Health Sciences, University of Tromsø, Tromsø, Norway Masaki Yanagishita, Tokyo Medical and Dental University, Tokyo, Japan Svein O. Kolset, Department of Nutrition, University of Oslo, Oslo, Norway Lars Uhlin-Hansen, Institute of Medical Biology, Faculty of Health Sciences, University of Tromsø, Tromsø, Norway Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    A new preparative column for the vortex counter-current chromatograph was fabricated by making many (966) cylindrical separation units to a high-density polyethylene disk and then threading them with 6–40 taps. The resulting column had a total capacity of 364 mL. The performance of this vortex column was examined with three different two-phase solvent systems each using a set of suitable test samples: hexane–ethyl acetate–methanol–0.1 M hydrochloric acid (1:1:1:1, v/v) for the separation of DNP-amino acids; 1-butanol–acetic acid–water (4:1:5, v/v) for the separation of dipeptides; and hexane–acetonitrile–water (20:15:2, v/v) for the separation of Sudan dyes. Most of the separations show high partition efficiency of over a thousand theoretical plates, as expected based on the results previously obtained in preliminary separations with a small column. Overall, the results of the present study suggest that further improvement of the partition efficiency can be obtained by the modifying column configuration. Content Type Journal Article Category Original Pages 1-7 DOI 10.1007/s10337-012-2285-6 Authors Yoichiro Ito, Bioseparation Technology Laboratory, Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bldg. 10, Room 8N230, 10 Center Drive, Bethesda, MD 20892-1762, USA Robert Clary, Machine Instrumentation Design and Fabrication, National Institutes of Health, Bethesda, MD 20892, USA Jacob J. Witten, Bioseparation Technology Laboratory, Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bldg. 10, Room 8N230, 10 Center Drive, Bethesda, MD 20892-1762, USA Yun Zeng, Bioseparation Technology Laboratory, Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bldg. 10, Room 8N230, 10 Center Drive, Bethesda, MD 20892-1762, USA Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    A binary supercritical fluid chromatography (SFC) system was further modified by the addition of a quaternary HPLC pump. The flow outlet of this pump was connected to the flow inlet of the existing modifier pump (“pump B”) by way of a custom-designed reservoir in such a way that blends of methanol with up to three additional solvents and/or additives could be delivered to pump B on-the-fly. Small amounts of modifiers (0.1–5 %) are typically mixed with mobile phase B in SFC to improve either the peak shape or to optimize the selectivity of the separation. As the critical influence of these modifiers on the separation of complex mixtures cannot a priori be predicted, incremental changes in mobile phase B compositions typically have to be evaluated on a trial-and-error basis. The instrumental modifications described in this paper eliminate the need to prepare these combinations off-line and the concomitant time requirement to re-purge supply and de-gasser solvent lines with each change in composition. By eliminating these steps, a significantly decrease in the overall SFC method development time can be achieved. Furthermore, such an on-line arrangement allows for greater flexibility in method development, that is, in the “fine-tuning” of the method once the stationary phase and gradient conditions have been established. Content Type Journal Article Category Short Communication Pages 1-6 DOI 10.1007/s10337-012-2291-8 Authors Anthony J. Alexander, Analytical and Bioanalytical Development, Bristol Myers Squibb Company, 1 Squibb Drive, New Brunswick, NJ 08903, USA Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    An efficient and reliable technique to evaluate the degree of coverage of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) was described. The method was based on a quantitative analysis of RDX leakage carried out with a high performance of liquid chromatography (HPLC). For this study, replicated analyses were performed on coated samples prepared by different kinds of coating materials and methods. The efficiency of characterization by using both HPLC and scanning electron microscope was compared. To note, the evaluation through former technique is more on macroscopic perspective rather than morphological observation of sample. Meanwhile, the HPLC method also provided characterization results that were in good agreement with morphology observation. A noteworthy advantage of this original technique is that the evaluation of coating quality of melt-cast explosives can be carried out under similar conditions. The experimental data were provided for deep understanding of the soluble behavior of coated RDX and its possible applications in practical problems. Content Type Journal Article Category Short Communication Pages 1-6 DOI 10.1007/s10337-012-2290-9 Authors Shuai Zhang, Institute of Chemical Materials, China Academy of Engineering Physics, Mianyang, 621900 People’s Republic of China Tianbo Zhao, Department of Chemistry, Key Laboratory of Cluster Science, Ministry of Education of China, Beijing Institute of Technology, Beijing, 100081 People’s Republic of China Guan Luo, Institute of Chemical Materials, China Academy of Engineering Physics, Mianyang, 621900 People’s Republic of China Hui Huang, Institute of Chemical Materials, China Academy of Engineering Physics, Mianyang, 621900 People’s Republic of China Zhongzhan Cai, Institute of Chemical Materials, China Academy of Engineering Physics, Mianyang, 621900 People’s Republic of China Pingsheng Wang, Institute of Chemical Materials, China Academy of Engineering Physics, Mianyang, 621900 People’s Republic of China Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Mucin-type O -linked glycoproteins are known for regulating many aspects of cell activity but remains a challenge to detect under physiological conditions which is due to the diversity of O -glycosylation and the lack of universal method. Here a direct labeling strategy for in situ visualizing of mucin-type O -linked glycoproteins on living cells has been developed. The strategy utilizes the combination of metabolic engineering and chemical probing technologies. Treating cells with an unnatural sugar, 2-keto Ac 4 GalNAc analogue (2-keto isostere of GalNAc) to generate keto groups upon cells, followed by chemoselective ligation of keto groups on cells with a fluorescent tag, fluorescein-5-thiosemicarbazide (FTSC), provides a promising platform to probing mucin-type O -glycosylation on living cells. The FTSC conjugates illustrated very similar fluorescent spectra as FITC, a fluorescent tag widely used in proteomics, indicating good compatibility with commonly used fluorescent equipments. The established method eliminated the need of an additional fluorescent amplification step. Cells after being treated with the method maintained a rather high level of viability of 84.3 %. Finally, the assay has been successfully applied to image the expression of mucin-type O -linked glycoproteins within CHO and HeLa cells. Content Type Journal Article Pages 1-8 DOI 10.1007/s10719-012-9425-y Authors Ying Zhang, Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, Xi’an, 710069 People’s Republic of China Yujiao Sun, Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, Xi’an, 710069 People’s Republic of China Zhongfu Wang, Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, Xi’an, 710069 People’s Republic of China Linjuan Huang, Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, Xi’an, 710069 People’s Republic of China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    This paper proposes an innovative concept for robustness evaluation guided by two crucial aims: indubitable identification of the factors that significantly affect the LC method and avoidance of unnecessary time and money wasting. The first phase of the proposed strategy includes robustness screening during the method optimization. Initial assumptions of the method robustness can be tracked as the rate of the response change while the factors deviate within the expected range. Therefore, partial and total robustness criteria are calculated. If the results obtained are not satisfactory, re-optimization of the method should be considered. Otherwise, extensive robustness testing defined by experimental design and multi-level factors estimation should be performed to confirm the method robustness. Firstly, the important factors are investigated by the standard graphical (normal probability plots) and statistical (algorithm of Dong and error estimation based on a priori declared negligible effects) procedures. Since these approaches have several drawbacks, they can result in the appearance of false negative or false positive results. Thus, the modification of the statistical tests is advised in order to make the final conclusions. Special attention was dedicated to the advantages of the adapted algorithm of Dong (so-called 75 % approach) in the absence of the effect sparsity. The new approach is presented on the optimization and robustness testing of LC method for determination of ramipril and its five impurities. It is proved that the proposed strategy can perform an overall robustness estimation and successfully reveal all important factors. Content Type Journal Article Category Original Pages 1-11 DOI 10.1007/s10337-012-2317-2 Authors Tijana Rakić, Department of Drug Analysis, University of Belgrade-Faculty of Pharmacy, Vojvode Stepe 450, 11000, Belgrade, Serbia Sava Vemić, Department of Drug Analysis, University of Belgrade-Faculty of Pharmacy, Vojvode Stepe 450, 11000, Belgrade, Serbia Biljana Jančić-Stojanović, Department of Drug Analysis, University of Belgrade-Faculty of Pharmacy, Vojvode Stepe 450, 11000, Belgrade, Serbia Mirjana Medenica, Department of Physical Chemistry and Instrumental Methods, University of Belgrade-Faculty of Pharmacy, Vojvode Stepe 450, 11000, Belgrade, Serbia Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    The retention of fifty structurally different compounds has been studied using linear solvation energy relationships. Investigations were performed with the use of six various stationary phases with two mobile phases (50/50 % v/v methanol/water and 50/50 % v/v acetonitrile/water). Packing materials were home-made and functionalized with octadecyl, alkylamide, cholesterol, alkyl-phosphate and phenyl molecules. This is the first attempt to compare all of these stationary phases synthesized on the same silica gel batch. Therefore, all of them may be compared in more complex and believable way, than it was performed earlier in former investigations. The phase properties (based on Abraham model) were used to the classification of stationary phases according to their interaction properties. The hydrophilic system properties s , a , b indicate stronger interactions between solute and mobile phase for most of the columns. Both e and v cause greater retention as a consequence of preferable interactions with stationary phase by electron pairs and cavity formation as well as hydrophobic bonds. However, alkyl-phosphate phase has different retention properties, as it was expressed by positive sign of s coefficient. It may be concluded that most important parameters influencing the retention of compounds are volume and hydrogen bond acceptor basicity. The LSER coefficients showed also the dependency on the type of organic modifier used as a mobile phase component. Content Type Journal Article Category Original Pages 1-12 DOI 10.1007/s10337-012-2310-9 Authors S. Studzińska, Chair of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University, 7 Gagarin St., 87-100 Toruń, Poland B. Buszewski, Chair of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University, 7 Gagarin St., 87-100 Toruń, Poland Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    A gas chromatography–mass spectrometry (GC–MS) method was investigated for the simultaneous analysis of two types of endocrine disrupting compounds (EDCs), i.e., alkylphenol ethoxylates and brominated flame retardants (BFRs), by extraction and derivatization followed by GC–MS. Different solid phase extraction (SPE) cartridges (Cleanert PestiCarb, C 18 , Cleanert-SAX and Florosil), solvents (toluene, tetrahydrofuran, acetone, acetonitrile and ethyl acetate) and bases (NaHCO 3 , triethylamine and pyridine) were tested and the best chromatographic analysis was achieved by extraction with Strata-X (33 μm, Reverse Phase) cartridge and derivatization with heptafluorobutyric anhydride at 55 °C under Na 2 CO 3 base in hexane. It was observed that APE together with lower substituted PBBs (PBB1, PBB10, PBB18 and PBB49), HBCD and TBBPA can be determined simultaneously under the same GC conditions. This simple and reliable analytical method was applied to determining trace amounts of these compounds from wastewater treatment plant samples. The recoveries of the target compounds from simulated water were above 60 %. The limit of detection ranged from 0.01 to 0.15 μg L −1 and the limit of quantification ranged from 0.05 to 0.66 μg L −1 . There were no appreciable differences between filtered and unfiltered wastewater samples from Leeuwkil treatment plant although concentration of target analytes in filtered influent was slightly lower than the concentration of target analytes in unfiltered influent water. The concentrations of the target compounds from the wastewater treatment were determined from LOQ upwards. Content Type Journal Article Category Original Pages 1-12 DOI 10.1007/s10337-012-2293-6 Authors Tlou B. Chokwe, Scientific Services, Rand Water, Vereeniging, 1930 South Africa Jonathan O. Okonkwo, Department of Environmental, Water and Earth Sciences, Tshwane University of Technology, Pretoria, 0001 South Africa Linda L. Sibali, Directorate of Research and Innovation, Tshwane University of Technology, Pretoria, 0001 South Africa Esper J. Ncube, Scientific Services, Rand Water, Vereeniging, 1930 South Africa Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Inverse gas chromatography (IGC) was used to analyze the secondary transition temperatures and the miscibility of binary mixtures of poly (ether imide) (Ultem™) and a copolyester of bisphenol-A with terephthalic and isophthalic acids (50/50) (Ardel™) in three compositions (25/50, 50/50 and 75/25). Retention diagrams of the mixtures of Ultem™ and Ardel™ for n -nonane, n -decane, n -butyl acetate and isoamyl acetate were obtained at temperatures between 60 and 285 °C. Second-order transition temperatures of the mixtures were determined according to the slope change in retention diagrams of the solvents. The glass transition temperatures of the mixtures suggested the miscibility of the polymers. Polymer–polymer interaction parameters of binary mixtures of the polymers were determined at temperatures between 260 and 285 °C by Flory–Huggins theory. The polymer–polymer interaction parameters were dependent on the solvent used. The small values of polymer–polymer interaction parameters close to zero suggest some weak interactions between the polymers in the mixture. It was concluded that it was possible to obtain more meaningful information related to the interactions of polymers in a mixture from IGC measurements, if binary polymer–solvent interaction parameters of the used solvent probes were around 0.5. Content Type Journal Article Category Original Pages 1-8 DOI 10.1007/s10337-012-2302-9 Authors Fatih Cakar, Department of Chemistry, Yildiz Technical University, 34220 Istanbul, Esenler, Turkey Ozlem Cankurtaran, Department of Chemistry, Yildiz Technical University, 34220 Istanbul, Esenler, Turkey Ferdane Karaman, Department of Chemistry, Yildiz Technical University, 34220 Istanbul, Esenler, Turkey Journal Chromatographia Online ISSN 1612-1112 Print ISSN 0009-5893

Description:    Glycoconjugates (GCs) are recognized as stimulation and signaling agents, affecting cell adhesion, activation, and growth of living organisms. Among GC targets, macrophages are considered ideal since they play a central role in inflammation and immune responses against foreign agents. In this context, we studied the effects of highly selective GCs in neutralizing toxin factors produced by B. anthracis during phagocytosis using murine macrophages. The effects of GCs were studied under three conditions: A) prior to , B) during , and C) following exposure of macrophages to B. anthracis individual toxin (protective antigen [PA], edema factor [EF], lethal factor [LF] or toxin complexes (PA-EF-LF, PA-EF, and PA-LF). We employed ex vivo phagocytosis and post-phagocytosis analysis including direct microscopic observation of macrophage viability, and macrophage activation. Our results demonstrated that macrophages are more prone to adhere to GC-altered PA-EF-LF, PA-EF, and PA-LF toxin complexes. This adhesion results in a higher phagocytosis rate and toxin complex neutralization during phagocytosis. In addition, GCs enhance macrophage viability, activate macrophages, and stimulate nitric oxide (NO) production. The present study may be helpful in identifying GC ligands with toxin-neutralizing and/or immunomodulating properties. In addition, our study could suggest GCs as new targets for existing vaccines and the prospective development of vaccines and immunomodulators used to combat the effects of B. anthracis . Content Type Journal Article Pages 1-12 DOI 10.1007/s10719-012-9446-6 Authors Olga Tarasenko, Department of Biology, University of Arkansas at Little Rock, 2801 South University Ave., Little Rock, AR 72204, USA Ashley Scott, Department of Biology, University of Arkansas at Little Rock, 2801 South University Ave., Little Rock, AR 72204, USA April Jones, Department of Biology, University of Arkansas at Little Rock, 2801 South University Ave., Little Rock, AR 72204, USA Lee Soderberg, Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, AR, USA Pierre Alusta, Department of Biology, University of Arkansas at Little Rock, 2801 South University Ave., Little Rock, AR 72204, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    In the past decade, the identification of most genes involved in Congenital Disorders of Glycosylation (CDG) (type I) was achieved by a combination of biochemical, cell biological and glycobiological investigations. This has been truly successful for CDG-I, because the candidate genes could be selected on the basis of the homology of the synthetic pathway of the dolichol linked oligosaccharide in human and yeast. On the contrary, only a few CDG-II defects were elucidated, be it that some of the discoveries represent wonderful breakthroughs, like e.g , the identification of the COG defects. In general, many rare genetic defects have been identified by positional cloning. However, only a few types of CDG have effectively been elucidated by linkage analysis and so-called reverse genetics. The reason is that the families were relatively small and could—except for CDG-PMM2—not be pooled for analysis. Hence, a large number of CDG cases has long remained unsolved because the search for the culprit gene was very laborious, due to the heterogeneous phenotype and the myriad of candidate defects. This has changed when homozygosity mapping came of age, because it could be applied to small (consanguineous) families. Many novel CDG genes have been discovered in this way. But the best has yet to come: what we are currently witnessing, is an explosion of novel CDG defects, thanks to exome sequencing: seven novel types were published over a period of only two years. It is expected that exome sequencing will soon become a diagnostic tool, that will continuously uncover new facets of this fascinating group of diseases. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9445-7 Authors Gert Matthijs, Center for Human Genetics, University of Leuven, Herestraat 49, 3000 Leuven, Belgium Daisy Rymen, Center for Human Genetics, University of Leuven, Herestraat 49, 3000 Leuven, Belgium María Beatriz Bistué Millón, Center for Human Genetics, University of Leuven, Herestraat 49, 3000 Leuven, Belgium Erika Souche, Center for Human Genetics, University of Leuven, Herestraat 49, 3000 Leuven, Belgium Valérie Race, Center for Human Genetics, University of Leuven, Herestraat 49, 3000 Leuven, Belgium Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    This review summarizes the analytical advances made during the last several years in the structural and quantitative determinations of glycoproteins in complex biological mixtures. The main analytical techniques used in the fields of glycomics and glycoproteomics involve different modes of mass spectrometry and their combinations with capillary separation methods such as microcolumn liquid chromatography and capillary electrophoresis. The need for high-sensitivity measurements have been emphasized in the oligosaccharide profiling used in the field of biomarker discovery through MALDI mass spectrometry. High-sensitivity profiling of both glycans and glycopeptides from biological fluids and tissue extracts has been aided significantly through lectin preconcentration and the uses of affinity chromatography. Content Type Journal Article Pages 1-29 DOI 10.1007/s10719-012-9444-8 Authors Milos V. Novotny, Department of Chemistry, Indiana University, Bloomington, IN, USA William R. Alley Jr., Department of Chemistry, Indiana University, Bloomington, IN, USA Benjamin F. Mann, Department of Chemistry, Indiana University, Bloomington, IN, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080