Journal of Analytical Methods in Chemistry The latest articles from Hindawi Publishing Corporation © 2015 , Hindawi Publishing Corporation . All rights reserved. Solid-Phase Extraction Strategies to Surmount Body Fluid Sample Complexity in High-Throughput Mass Spectrometry-Based Proteomics Tue, 27 Jan 2015 09:51:03 +0000 For large-scale and standardized applications in mass spectrometry- (MS-) based proteomics automation of each step is essential. Here we present high-throughput sample preparation solutions for balancing the speed of current MS-acquisitions and the time needed for analytical workup of body fluids. The discussed workflows reduce body fluid sample complexity and apply for both bottom-up proteomics experiments and top-down protein characterization approaches. Various sample preparation methods that involve solid-phase extraction (SPE) including affinity enrichment strategies have been automated. Obtained peptide and protein fractions can be mass analyzed by direct infusion into an electrospray ionization (ESI) source or by means of matrix-assisted laser desorption ionization (MALDI) without further need of time-consuming liquid chromatography (LC) separations. Marco R. Bladergroen and Yuri E. M. van der Burgt Copyright © 2015 Marco R. Bladergroen and Yuri E. M. van der Burgt. All rights reserved. Regularities of Anthocyanins Retention in RP HPLC for “Water–Acetonitrile–Phosphoric Acid” Mobile Phases Mon, 26 Jan 2015 11:59:08 +0000 The influence of exchange of HCOOH (System 2) by phosphoric acid (System 1) for acidification of the “acetonitrile–water” mobile phases for reversed-phase HPLC of anthocyanins was investigated in the framework of relative retention analysis. The differences and similarities of anthocyanins separation were revealed. It has been shown that some common features of the quantitative relationships may be used for preliminary anthocyanins structure differentiation, according to the number of OH-groups in anthocyanidin backbone as well as to a number of saccharide molecules in glycoside radicals in position 3 of the anthocyanin without MS detection. V. I. Deineka, L. A. Deineka, and I. I. Saenko Copyright © 2015 V. I. Deineka et al. All rights reserved. General Analytical Procedure for Determination of Acidity Parameters of Weak Acids and Bases Mon, 26 Jan 2015 09:54:03 +0000 The paper presents a new convenient, inexpensive, and reagent-saving general methodology for the determination of values for components of the mixture of diverse chemical classes weak organic acids and bases in water solution, without the need to separate individual analytes. The data obtained from simple pH-metric microtitrations are numerically processed into reliable values for each component of the mixture. Excellent agreement has been obtained between the determined values and the reference literature data for compounds studied. Bogusław Pilarski, Roman Kaliszan, Dariusz Wyrzykowski, Janusz Młodzianowski, and Agata Balińska Copyright © 2015 Bogusław Pilarski et al. All rights reserved. Chitosan Hydrogels for Chondroitin Sulphate Controlled Release: An Analytical Characterization Wed, 31 Dec 2014 13:23:25 +0000 This paper provides an analytical characterization of chitosan scaffolds obtained by freeze-gelation toward the uptake and the controlled release of chondroitin sulphate (CS), as cartilage repair agent, under different pH conditions. Scanning electron microscopy (SEM), attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), and liquid chromatography-UV spectrophotometry (LC-UV) techniques were exploited to obtain qualitative and quantitative descriptions of polymer and drug behaviour in the biomaterial. As for morphology, SEM analysis allowed the evaluation of scaffold porosity in terms of pore size and distribution both at the surface (Feret diameter  μm) and on the cross section (Feret diameter  μm). LC and ATR-FTIR evidenced a pH-dependent CS loading and release behaviour, strongly highlighting the role of electrostatic forces on chitosan/chondroitin sulphate interactions. Annalisa Bianchera, Enrico Salomi, Matteo Pezzanera, Elisabeth Ruwet, Ruggero Bettini, and Lisa Elviri Copyright © 2014 Annalisa Bianchera et al. All rights reserved. Determination of β-Cyano-L-alanine, γ-Glutamyl-β-cyano-L-alanine, and Common Free Amino Acids in Vicia sativa (Fabaceae) Seeds by Reversed-Phase High-Performance Liquid Chromatography Mon, 22 Dec 2014 06:20:34 +0000 A method for determination of β-cyano-L-alanine, γ-glutamyl-β-cyano-L-alanine and other free amino acids in Vicia sativa is presented. Seed extracts were derivatized by reaction with diethyl ethoxymethylenemalonate and analyzed by reverse-phase high-performance liquid chromatography. Calibration curves showed very good linearity of the response. The limit of detection and quantification was 0.15 and 0.50 μM, respectively. The method has high intra- (RSD = 0.28–0.31%) and interrepeatability (RSD = 2.76–3.08%) and remarkable accuracy with a 99% recovery in spiked samples. The method is very easy to carry out and allows for ready analysis of large number of samples using very basic HPLC equipment because the derivatized samples are very stable and have very good chromatographic properties. The method has been applied to the determination of γ-glutamyl-β-cyano-L-alanine, β-cyano-L-alanine, and common free amino acids in eight wild populations of V. sativa from southwestern Spain. Cristina Megías, Isabel Cortés-Giraldo, Julio Girón-Calle, Javier Vioque, and Manuel Alaiz Copyright © 2014 Cristina Megías et al. All rights reserved. Quantification of Caffeoylquinic Acids in Coffee Brews by HPLC-DAD Sun, 21 Dec 2014 00:10:19 +0000 The influence of different brewing conditions on the concentration of the main caffeoylquinic acids (3-caffeoylquinic acid (3-CQA), 4-caffeoylquinic acid (4-CQA), and 5-caffeoylquinic acid (5-CQA)) was investigated. For this purpose, twenty-four coffee brews were extracted and analyzed using HPLC-DAD at 325 nm. Our findings demonstrate the great impact of brewing techniques on the caffeoylquinic acids (CQAs) content. The major isomer was 3-CQA, accounting for about 50% of the total CQAs, followed by 5-CQA and 4-CQA, accounting for about 24–36% for each one. The total content of CQAs was in the range of 45.79 to 1662.01 mg/L, found in iced cappuccino and pod espresso, respectively. In conclusion, this study demonstrates that coffee brews, in particular those prepared using pressurized methods, can be considered as the potential sources of antioxidants such as CQAs. Marzieh Moeenfard, Lígia Rocha, and Arminda Alves Copyright © 2014 Marzieh Moeenfard et al. All rights reserved. Research of Recognition Method of Discrete Wavelet Feature Extraction and PNN Classification of Rats FT-IR Pancreatic Cancer Data Thu, 04 Dec 2014 09:37:41 +0000 Sprague-Dawley (SD) rats’ normal and abnormal pancreatic tissues are determined directly by attenuated total reflectance Fourier transform infrared (ATR-FT-IR) spectroscopy method. In order to diagnose earlier stage of SD rats pancreatic cancer rate with FT-IR, a novel method of extraction of FT-IR feature using discrete wavelet transformation (DWT) analysis and classification with the probability neural network (PNN) was developed. The differences between normal pancreatic and abnormal samples were identified by PNN based on the indices of 4 feature variants. When error goal was 0.01, the total correct rates of pancreatic early carcinoma and advanced carcinoma were 98% and 100%, respectively. It was practical to apply PNN on the basis of ATR-FT-IR to identify abnormal tissues. The research result shows the feasibility of establishing the models with FT-IR-DWT-PNN method to identify normal pancreatic tissues, early carcinoma tissues, and advanced carcinoma tissues. Chayan Wan, Wenqing Cao, and Cungui Cheng Copyright © 2014 Chayan Wan et al. All rights reserved. Characteristic Fingerprint Based on Low Polar Constituents for Discrimination of Wolfiporia extensa according to Geographical Origin Using UV Spectroscopy and Chemometrics Methods Tue, 02 Dec 2014 13:01:19 +0000 The fungus species Wolfiporia extensa has a long history of medicinal usage and has also been commercially used to formulate nutraceuticals and functional foods in certain Asian countries. In the present study, a practical and promising method has been developed to discriminate the dried sclerotium of W. extensa collected from different geographical sites based on UV spectroscopy together with chemometrics methods. Characteristic fingerprint of low polar constituents of sample extracts that originated from chloroform has been obtained in the interval 250–400 nm. Chemometric pattern recognition methods such as partial least squares discriminant analysis (PLS-DA) and hierarchical cluster analysis (HCA) were applied to enhance the authenticity of discrimination of the specimens. The results showed that W. extensa samples were well classified according to their geographical origins. The proposed method can fully utilize diversified fingerprint characteristics of sclerotium of W. extensa and requires low-cost equipment and short-time analysis in comparison with other techniques. Meanwhile, this simple and efficient method may serve as a basis for the authentication of other medicinal fungi. Yan Li, Ji Zhang, Yanli Zhao, Zhimin Li, Tao Li, and Yuanzhong Wang Copyright © 2014 Yan Li et al. All rights reserved. Optimization of Ultrasonic Extraction of Phenolic Compounds from Epimedium brevicornum Maxim Using Response Surface Methodology and Evaluation of Its Antioxidant Activities In Vitro Wed, 12 Nov 2014 08:12:29 +0000 The ultrasound-assisted extraction of phenolic compounds from Epimedium brevicornu Maxim was modeled using response surface methodology. A Central Composite Design (CCD) was employed to optimize three extraction variables, including ethanol concentration (X1), extraction time (X2), and ratio of aqueous ethanol to raw material (X3), for the achievement of high extraction yield of the phenolic compounds. The optimized conditions are X1 of 50% (v/v), X2 of 27.5 min, and X3 of 250 mL/g. Under these conditions, the experimental yield is 4.29 ± 0.033% . The antioxidant activity was evaluated using the DPPH assay and ferric-reducing antioxidant power (FRAP). And it indicates that the phenolic compounds from Epimedium brevicornu Maxim possess significant antioxidant activity. HPLC analysis reveals that the main phenolic compound in the extract product was identified as gallic acid, catechin (Cianidanol), p-hydroxybenzoic acid, vanillic acid, caffeic acid, ferulaic acid, rutin, benzoic acid, and quercetin. Yan Zhao, Yingying Hou, Guosheng Tang, Enbo Cai, Shuangli Liu, He Yang, Lianxue Zhang, and Shijie Wang Copyright © 2014 Yan Zhao et al. All rights reserved. Simultaneous Determination of Uric Acid and Xanthine Using a Poly(Methylene Blue) and Electrochemically Reduced Graphene Oxide Composite Film Modified Electrode Tue, 11 Nov 2014 13:21:50 +0000 Poly(methylene blue) and electrochemically reduced graphene oxide composite film modified electrode (PMB-ERGO/GCE) was successfully fabricated by electropolymerization and was used for simultaneous determination of uric acid (UA) and xanthine (Xa). Based on the excellent electrocatalytic activity of PMB-ERGO/GCE, the electrochemical behaviors of UA and Xa were studied by cyclic voltammetry (CV) and square wave voltammetry (SWV). Two anodic sensitive peaks at 0.630 V (versus Ag/AgCl) for UA and 1.006 V (versus Ag/AgCl) for Xa were given by CV in pH 3.0 phosphate buffer. The calibration curves for UA and Xa were obtained in the range of 8.00 × 10−8~4.00 × 10−4 M and 1.00 × 10−7~4.00 × 10−4 M, respectively, by SWV. The detection limits for UA and Xa were  M and  M, respectively. Finally, the proposed method was applied to simultaneously determine UA and Xa in human urine with good selectivity and high sensitivity. Gen Liu, Wei Ma, Yan Luo, Deng-ming Sun, and Shuang Shao Copyright © 2014 Gen Liu et al. All rights reserved. A Modified o-Phthalaldehyde Fluorometric Analytical Method for Ultratrace Ammonium in Natural Waters Using EDTA-NaOH as Buffer Tue, 11 Nov 2014 12:48:36 +0000 In the existence of appropriate amount of disodium ethylenediaminetetraacetate (EDTA), precipitation would not occur in seawater and other natural waters even if the sample solution was adjusted to strong basicity, and the NH3-OPA-sulfite reaction at the optimal pH range could be used to determine ammonium in natural waters. Based on this, a modified o-phthalaldehyde fluorometric analytical method has been established to determine ultratrace ammonium in natural waters. Experimental parameters, including reagent concentration, pH, reaction time, and effect of EDTA, were optimized throughout the experiments based on univariate experimental design. The results showed that the optimal pH range was between 10.80 and 11.70. EDTA did not obviously affect the fluorometric intensity. The linearity range of the proposed method was 0.032–0.500 µmol/L, 0.250–3.00 µmol/L, and 1.00–20.0 µmol/L at the excitation/emission slit of 3 nm/5 nm, 3 nm/3 nm, and 1.5 nm/1.5 nm, respectively. The method detection limit was 0.0099 µmol/L. Compared to the classical OPA method, the proposed method had the advantage of being more sensitive and could quantify ultratrace ammonium without enrichment. Hongzhi Hu, Ying Liang, Shuo Li, Qing Guo, and Chancui Wu Copyright © 2014 Hongzhi Hu et al. All rights reserved. Determination of Sunset Yellow and Tartrazine in Food Samples by Combining Ionic Liquid-Based Aqueous Two-Phase System with High Performance Liquid Chromatography Wed, 05 Nov 2014 06:23:28 +0000 We proposed a simple and effective method, by coupling ionic liquid-based aqueous two-phase systems (IL-ATPSs) with high performance liquid chromatography (HPLC), for the analysis of determining tartrazine and sunset yellow in food samples. Under the optimized conditions, IL-ATPSs generated an extraction efficiency of 99% for both analytes, which could then be directly analyzed by HPLC without further treatment. Calibration plots were linear in the range of 0.01–50.0 μg/mL for both Ta and SY. The limits of detection were 5.2 ng/mL for Ta and 6.9 ng/mL for SY. This method proves successful for the separation/analysis of tartrazine and sunset yellow in soft drink sample, candy sample, and instant powder drink and leads to consistent results as obtained from the Chinese national standard method. Ou Sha, Xiashi Zhu, Yanli Feng, and Weixing Ma Copyright © 2014 Ou Sha et al. All rights reserved. Identification of the Related Substances in Ampicillin Capsule by Rapid Resolution Liquid Chromatography Coupled with Electrospray Ionization Tandem Mass Spectrometry Tue, 04 Nov 2014 07:27:41 +0000 Rapid Resolution Liquid Chromatography coupled with Electrospray Ionization Tandem Mass Spectrometry (RRLC-ESI-MSn) was used to separate and identify related substances in ampicillin capsule. The fragmentation behaviors of related substances were used to identify their chemical structures. Finally, a total of 13 related substances in ampicillin capsule were identified, including four identified components for the first time and three groups of isomers on the basis of the exact mass, fragmentation behaviors, retention time, and chemical structures in the literature. This study avoided time-consuming and complex chemosynthesis of related substances of ampicillin and the results could be useful for the quality control of ampicillin capsule to guarantee its safety in clinic. In the meantime, it provided a good example for the rapid identification of chemical structures of related substances of drugs. Lei Zhang, Xian Long Cheng, Yang Liu, Miao Liang, Honghuan Dong, Beiran Lv, Wenning Yang, Zhiqiang Luo, and Mingmin Tang Copyright © 2014 Lei Zhang et al. All rights reserved. Geographical and Seasonal Patterns of Geosmin and 2-Methylisoborneol in Environmental Water in Jiangsu Province of China Thu, 23 Oct 2014 10:00:02 +0000 This study was conducted to obtain the basic data of two common odorants—geosmin and 2-methylisoborneol (GSM and 2-MIB)—in environmental water. More specifically, the headspace solid-phase microextraction coupled to gas chromatography mass spectrometry (HS-SPME/GC-MS) was applied to determine the levels of GSM and 2-MIB in water samples, and the samples were collected depending on water sources, conventional treatment processes, and seasons. The significant difference was shown for the 2-MIB levels of source water , the concentrations of GSM and 2-MIB decreased significantly as treatment process of tap water moved forward , and the significant differences for the levels of GSM and 2-MIB were observed among three sampling periods . The levels of GSM and 2-MIB in all water samples were lower than 10 ng L−1, the odor threshold concentration (OTC), and the conventional treatment process plays a significant role in removing odorants in tap water. Zhen Ding, Shifu Peng, Yuqin Jin, Zhoubin Xuan, Xiaodong Chen, and Lihong Yin Copyright © 2014 Zhen Ding et al. All rights reserved. Isolation and Bioactivity Analysis of Ethyl Acetate Extract from Acer tegmentosum Using In Vitro Assay and On-Line Screening HPLC-ABTS+ System Wed, 15 Oct 2014 06:14:01 +0000 The Acer tegmentosum (3 kg) was extracted using hot water, and the freeze-dried extract powder was partitioned successively using dichloromethane (DCM), ethyl acetate (EA), butyl alcohol (n-BuOH), and water. From the EA extract fraction (1.24 g), five phenolic compounds were isolated by the silica gel, octadecyl silica gel, and Sephadex LH-20 column chromatography. Based on spectroscopic methods such as 1H-NMR, 13C-NMR, and LC/MS the chemical structures of the compounds were confirmed as feniculin (1), avicularin (2), (+)-catechin (3), (−)-epicatechin (4), and 6′-O-galloyl salidroside (5). Moreover, a rapid on-line screening HPLC-ABTS+ system for individual bioactivity of the EA-soluble fraction (five phenolic compounds) was developed. The results indicated that compounds 1 and 2 were first isolated from the A. tegmentosum. The anti-inflammatory activities and on-line screening HPLC-ABTS+ assay method of these compounds in LPS-stimulated murine macrophages were rapid and efficient for the investigation of bioactivity of A. tegmentosum. Kwang Jin Lee, Na-Young Song, You Chang Oh, Won-Kyung Cho, and Jin Yeul Ma Copyright © 2014 Kwang Jin Lee et al. All rights reserved. A Stability-Indicating HPLC-DAD Method for Determination of Stiripentol: Development, Validation, Kinetics, Structure Elucidation and Application to Commercial Dosage Form Tue, 14 Oct 2014 00:00:00 +0000 A rapid, simple, sensitive, and accurate isocratic reversed-phase stability-indicating high performance liquid chromatography method has been developed and validated for the determination of stiripentol and its degradation product in its bulk form and pharmaceutical dosage form. Chromatographic separation was achieved on a Symmetry C18 column and quantification was achieved using photodiode array detector (DAD). The method was validated in accordance with the ICH requirements showing specificity, linearity (, range of 1–25 μg/mL), precision (relative standard deviation lower than 2%), accuracy (mean recovery ), limits of detection and quantitation (LOD = 0.024 and LOQ = 0.081 μg/mL), and robustness. Stiripentol was subjected to various stress conditions and it has shown marked stability under alkaline hydrolytic stress conditions, thermal, oxidative, and photolytic conditions. Stiripentol degraded only under acidic conditions, forming a single degradation product which was well resolved from the pure drug with significantly different retention time values. This degradation product was characterized by 1H-NMR and 13C-NMR spectroscopy as well as ion trap mass spectrometry. The results demonstrated that the method would have a great value when applied in quality control and stability studies for stiripentol. Hany W. Darwish, Ali S. Abdelhameed, Mohamed I. Attia, Ahmed H. Bakheit, Nasr Y. Khalil, and Abdulrahman A. Al-Majed Copyright © 2014 Hany W. Darwish et al. All rights reserved. Content Determination of Active Component in Huangqi Yinyanghuo Group and Its Effects on hTERT and Bcl-2 Protein in Osteosarcoma Mon, 13 Oct 2014 08:25:03 +0000 To screen the optimal extraction process and content determination of active component of Huangqi Yinyanghuo group (HYG) and to study the effects of HYG on human telomerase reverse transcriptase (hTERT) and Bcl-2 protein in osteosarcoma (HOS) cells, providing the theoretical basis for clinical application of HYG in treatment of osteosarcoma, orthogonal design table L9(43) was used to design the extraction process of HYG, and icariin was taken as the investigation index to optimize the extraction process of HYG. 0.125, 0.25, 0.5, 1, 2, 4, and 8 μmol/L HYG were taken to act separately on logarithmic growth phase osteosarcoma HOS cells, CCK-8 assay was used to determine cell viability, and immunohistochemical SP assay was used to determine the expression of hTERT and Bcl-2 protein. Apoptosis rate was positively correlated with the dose of HYG, and the expressions of hTERT and Bcl-2 protein were significantly decreased with the prolonged duration of action. Under the effect of HYG, dose was negatively correlated with osteosarcoma cell survival fraction; osteosarcoma cell survival fraction was positively correlated with hTERT and Bcl-2 protein; duration of action was negatively correlated with hTERT and Bcl-2 protein; and hTERT and Bcl-2 protein were in a synchronous relationship. Ying Tan, Lei Tan, Shuai Huang, Junfan Lu, and Longtan Yu Copyright © 2014 Ying Tan et al. All rights reserved. Development of a Univariate Membrane-Based Mid-Infrared Method for Protein Quantitation and Total Lipid Content Analysis of Biological Samples Mon, 13 Oct 2014 08:11:56 +0000 Biological samples present a range of complexities from homogeneous purified protein to multicomponent mixtures. Accurate qualification of such samples is paramount to downstream applications. We describe the development of an MIR spectroscopy-based analytical method offering simultaneous protein quantitation (0.25–5 mg/mL) and analysis of total lipid or detergent species, as well as the identification of other biomolecules present in biological samples. The method utilizes a hydrophilic PTFE membrane engineered for presentation of aqueous samples in a dried format compatible with fast infrared analysis. Unlike classical quantification techniques, the reported method is amino acid sequence independent and thus applicable to complex samples of unknown composition. By comparison to existing platforms, this MIR-based method enables direct quantification using minimal sample volume (2 µL); it is well-suited where repeat access and limited sample size are critical parameters. Further, accurate results can be derived without specialized training or knowledge of IR spectroscopy. Overall, the simplified application and analysis system provides a more cost-effective alternative to high-throughput IR systems for research laboratories with minimal throughput demands. In summary, the MIR-based system provides a viable alternative to current protein quantitation methods; it also uniquely offers simultaneous qualification of other components, notably lipids and detergents. Ivona Strug, Christopher Utzat, Amedeo Cappione III, Sara Gutierrez, Ryan Amara, Joseph Lento, Florian Capito, Romas Skudas, Elena Chernokalskaya, and Timothy Nadler Copyright © 2014 Ivona Strug et al. All rights reserved. Development and Optimization of an UPLC-QTOF-MS/MS Method Based on an In-Source Collision Induced Dissociation Approach for Comprehensive Discrimination of Chlorogenic Acids Isomers from Momordica Plant Species Wed, 10 Sep 2014 11:27:07 +0000 Chlorogenic acids (CGA) have been profiled in the leaves of Momordica balsamina, Momordica charantia, and Momordica foetida. All three species were found to contain the trans and cis isomers of 4-acyl para-coumaroylquinic acid (pCoQA), caffeoylquinic acid (CQA), and feruloylquinic acid (FQA). To the best of our knowledge, this is the first report of pCoQA and FQA and their cis isomers in these Momordica species. These profiles were obtained by a newly developed UPLC-qTOF-MS method based on the in-source collision induced dissociation (ISCID) method optimized to mimic the MS2 and MS3 fragmentation of an ion trap-based MS. The presence of the cis isomers is believed to be due to high UV exposure of these plants. Furthermore, the absence of the 3-acyl and 5-acyl CGA molecules points to a metabolic mark that is unusual and represents a very interesting biochemical phenotype of these species. Our optimized ISCID method was also shown to be able to distinguish between the geometrical isomers of all three forms of CGA, a phenomenon previously deemed impossible with other common mass spectrometry systems used for CGA analyses. N. E. Madala, F. Tugizimana, and P. A. Steenkamp Copyright © 2014 N. E. Madala et al. All rights reserved. Validated Method for the Determination of Piroxicam by Capillary Zone Electrophoresis and Its Application to Tablets Tue, 09 Sep 2014 08:10:19 +0000 Simple and rapid capillary zone electrophoretic method was developed and validated in this study for the determination of piroxicam in tablets. The separation of piroxicam was conducted in a fused-silica capillary by using 10 mM borate buffer (pH 9.0) containing 10% (v/v) methanol as background electrolyte. The optimum conditions determined were 25 kV for separation voltage and 1 s for injection time. Analysis was carried out with UV detection at 204 nm. Naproxen sodium was used as an internal standard. The method was linear over the range of 0.23–28.79 µg/mL. The accuracy and precision were found to be satisfied within the acceptable limits (<2%). The LOD and LOQ were found to be 0.07 and 0.19 µg/mL, respectively. The method described here was applied to tablet dosage forms and the content of a tablet was found in the limits of USP-24 suggestions. To compare the results of capillary electrophoretic method, UV spectrophotometric method was developed and the difference between two methods was found to be insignificant. The capillary zone electrophoretic method developed in this study is rapid, simple, and suitable for routine analysis of piroxicam in pharmaceutical tablets. Arın Gül Dal, Zeynep Oktayer, and Dilek Doğrukol-Ak Copyright © 2014 Arın Gül Dal et al. All rights reserved. Quantification of Neurotransmitters in Mouse Brain Tissue by Using Liquid Chromatography Coupled Electrospray Tandem Mass Spectrometry Wed, 03 Sep 2014 00:00:00 +0000 A simple and rapid liquid chromatography tandem mass spectrometry method has been developed for the determination of BH4, DA, 5-HT, NE, EP, Glu, and GABA in mouse brain using epsilon-acetamidocaproic acid and isotopically labeled neurotransmitters as internal standards. Proteins in the samples were precipitated by adding acetonitrile, and then the supernatants were separated by a Sepax Polar-Imidazole (2.1 mm × 100 mm, i.d., 3 μm) column by adding a mixture of 10 mM ammonium formate in acetonitrile/water (75 : 25, v/v, 300 μl/min) for BH4 and DA. To assay 5-HT, NE, EP, Glu, and GABA; a Luna 3 μ C18 (3.0 mm × 150 mm, i.d., 3 μm) column was used by adding a mixture of 1% formic acid in acetonitrile/water (20 : 80, v/v, 350 μl/min). The total chromatographic run time was 5.5 min. The method was validated for the analysis of samples. The calibration curve was linear between 10 and 2000 ng/g for BH4 , 10 and 5000 ng/g for DA , 20 and 10000 ng/g for 5-HT , NE , and EP , and 0.2 and 200 μg/g for Glu and GABA in the mouse brain tissues. As stated above, LC-MS/MS results were obtained and established to be a useful tool for the quantitative analysis of BH4, DA, 5-HT, NE, EP, Glu, and GABA in the experimental rodent brain. Tae-Hyun Kim, Juhee Choi, Hyung-Gun Kim, and Hak Rim Kim Copyright © 2014 Tae-Hyun Kim et al. All rights reserved. A Comparative Study of Newly Developed HPLC-DAD and UHPLC-UV Assays for the Determination of Posaconazole in Bulk Powder and Suspension Dosage Form Wed, 03 Sep 2014 00:00:00 +0000 Objective. To develop and compare HPLC-DAD and UHPLC-UV assays for the quantitation of posaconazole in bulk powder and suspension dosage form. Methods. Posaconazole linearity range was 5–50 μg/mL for both assays. For HPLC-DAD assay, samples were injected through Zorbax SB-C18 (4.6 × 250 mm, 5 μm) column. The gradient elution composed of the mobile phase acetonitrile: 15 mM potassium dihydrogen orthophosphate (30 : 70 to 80 : 20, linear over 7 minutes) pumped at 1.5 mL/min. For UHPLC-UV assay, samples were injected through Kinetex-C18 (2.1 × 50 mm, 1.3 μm) column. The mobile phase composed of acetonitrile: 15 mM potassium dihydrogen orthophosphate (45 : 55) pumped isocratically at 0.4 mL/min. Detection wavelength was 262 nm in both methods. Results. The run time was 11 and 3 minutes for HPLC-DAD and UHPLC-UV assays, respectively. Both assays were linear () with CV% and % error of the mean <3%. Limits of detection and quantitation were 0.82 and 2.73 μg/mL for HPLC-DAD and 1.04 and 3.16 μg/mL for UHPLC-UV, respectively. The methods quantitated PSZ in suspension dosage form with no observable interferences. Conclusions. Both assays were proven sensitive and selective according to ICH guidelines. UHPLC-UV assay exhibited some economic and chromatographic separation superiority. Dalia A. Hamdy and Tarek S. Belal Copyright © 2014 Dalia A. Hamdy and Tarek S. Belal. All rights reserved. Extraction and Bioactivity Analysis of Major Flavones Compounds from Scutellaria baicalensis Using In Vitro Assay and Online Screening HPLC-ABTS System Mon, 01 Sep 2014 00:00:00 +0000 The extraction efficiency of a number of solvent compositions for the improvement of bioactive compounds yield from S. baicalensis has been investigated. Also, free radical scavengers in the glycoside baicalin (BG), wogonoside (WG), aglycon baicalein (B), and wogonin (W) compounds of S. baicalensis were screened, identified, and quantified using coupled offline ABTS and online screening HPLC-ABTS assay. Increasing ethanol content fractions resulted in decreased extract yield of bioactive compounds. In this case, the best yield of 37.01 mg/g in BG, WG, B, and W compounds was obtained by a dipping method with an extraction time of 4 h. In addition, the yield (43.05%) and IC50 (34.04 μg/mL) determined through ABTS assay of the 60% aqueous ethanol extract were the most satisfactory of all solvent solutions tested. This result shows that an online screening HPLC-ABTS assay can be a powerful technique for the rapid characterization of bioactivity compounds in plant extracts. Moreover, their anti-inflammatory activities were evaluated via analyzed inhibitory effect on NO and inflammatory cytokine production. Furthermore, WG and W exhibited the strong inhibitory effects on inflammatory mediator production including NO, IL-6, and IL-1β in LPS-stimulated RAW 264.7 macrophages. Kwang Jin Lee, Pil Mun Jung, You-Chang Oh, Na-Young Song, Taesoo Kim, and Jin Yeul Ma Copyright © 2014 Kwang Jin Lee et al. All rights reserved. Global Clustering Quality Coefficient Assessing the Efficiency of PCA Class Assignment Tue, 26 Aug 2014 00:00:00 +0000 An essential factor influencing the efficiency of the predictive models built with principal component analysis (PCA) is the quality of the data clustering revealed by the score plots. The sensitivity and selectivity of the class assignment are strongly influenced by the relative position of the clusters and by their dispersion. We are proposing a set of indicators inspired from analytical geometry that may be used for an objective quantitative assessment of the data clustering quality as well as a global clustering quality coefficient (GCQC) that is a measure of the overall predictive power of the PCA models. The use of these indicators for evaluating the efficiency of the PCA class assignment is illustrated by a comparative study performed for the identification of the preprocessing function that is generating the most efficient PCA system screening for amphetamines based on their GC-FTIR spectra. The GCQC ranking of the tested feature weights is explained based on estimated density distributions and validated by using quadratic discriminant analysis (QDA). Mirela Praisler and Stefanut Ciochina Copyright © 2014 Mirela Praisler and Stefanut Ciochina. All rights reserved. Electrochemical Study of Ni20Cr Coatings Applied by HVOF Process in ZnCl2-KCl at High Temperatures Mon, 25 Aug 2014 11:05:34 +0000 Corrosion behavior of Ni20Cr coatings deposited by HVOF (high velocity oxygen-fuel) process was evaluated in ZnCl2-KCl (1 : 1 mole ratio) molten salts. Electrochemical techniques employed were potentiodynamic polarization curves, open circuit potential, and linear polarization resistance (LPR) measurements. Experimental conditions included static air and temperatures of 350, 400, and 450°C. 304-type SS was evaluated in the same conditions as the Ni20Cr coatings and it was used as a reference material to assess the coatings corrosion resistance. Coatings were evaluated as-deposited and with a grinded surface finished condition. Results showed that Ni20Cr coatings have a better corrosion performance than 304-type SS. Analysis showed that Ni content of the coatings improved its corrosion resistance, and the low corrosion resistance of 304 stainless steel was attributed to the low stability of Fe and Cr and their oxides in the corrosive media used. J. Porcayo-Calderón, O. Sotelo-Mazón, M. Casales-Diaz, J. A. Ascencio-Gutierrez, V. M. Salinas-Bravo, and L. Martinez-Gomez Copyright © 2014 J. Porcayo-Calderón et al. All rights reserved. Alkaloids in Processed Rhizoma Corydalis and Crude Rhizoma Corydalis Analyzed by GC/MS Mon, 25 Aug 2014 07:39:54 +0000 The alkaloids in the processed Rhizoma Corydalis and the crude Rhizoma Corydalis were qualitatively and semiquantitatively analyzed using gas chromatography-mass spectrometry (GC/MS) method. The processing herb drug procedure was carried out according to the standard method of Chinese Pharmacopoeia. The samples were extracted using Soxhlet extractor with different solvents: methanol and acetone. The extraction effect on different solvents was investigated. The results showed that 11 kinds of alkaloids were identified from the crude Rhizoma Corydalis and only two were from the processed Rhizoma Corydalis. A total of 13 kinds of alkaloids were all based on two backbones. The alkaloids in the processed sample were less than those in the crude Rhizoma Corydalis significantly, while almost the corydaline has been changed in conformation after the sample had undergone processing, which provided support for the conclusion of reducing toxicity when the herbal medicine having been undergone a traditional drugs treatment process. Zhifeng Guo, Ru Cai, Huidan Su, and Yunlong Li Copyright © 2014 Zhifeng Guo et al. All rights reserved. Quality Control of Natural Product Medicine and Nutrient Supplements 2014 Thu, 21 Aug 2014 09:38:39 +0000 Ying-Yong Zhao, Shuang-Qing Zhang, Feng Wei, Yu-Ming Fan, Feng Sun, and Shuhua Bai Copyright © 2014 Ying-Yong Zhao et al. All rights reserved. Stacking and Analysis of Melamine in Milk Products with Acetonitrile-Salt Stacking Technique in Capillary Electrophoresis Mon, 18 Aug 2014 00:00:00 +0000 Melamine was measured in real milk products with capillary electrophoresis (CE) based on acetonitrile-salt stacking (ASS) method. Real milk samples were deproteinized with acetonitrile at a final concentration of 60% (v/v) and then injected hydrodynamically at 50 mBar for 40.0 s. The optimized buffer contains 80.0 mmol/L pH 2.8 phosphates. Melamine could be detected within 20.0 min at +10 kV with a low limit of detection (LOD) of 0.03 μmol/L. Satisfactory reproducibility (inter- and intraday RSD% both for migration time and peak area was lower than 5.0%) and a wide linearity range of 0.05 μmol/L ~ 10.0 μmol/L were achieved. The proposed method was suitable for routine assay of MEL in real milk samples that was subjected to a simple treatment step. Yu Kong, Chong Wei, Zhanwu Hou, Zilong Wang, Jiaqiang Yuan, Jiang Yu, Yongxi Zhao, Yuhai Tang, and Meili Gao Copyright © 2014 Yu Kong et al. All rights reserved. Determination of Flavanones in Orange Juices Obtained from Different Sources by HPLC/DAD Thu, 07 Aug 2014 10:56:09 +0000 Flavanones (hesperidin, naringenin, naringin, and poncirin) in industrial, hand-squeezed orange juices and from fresh-in-squeeze machines orange juices were determined by HPLC/DAD analysis using a previously described liquid-liquid extraction method. Method validation including the accuracy was performed by using recovery tests. Samples (36) collected from different Brazilian locations and brands were analyzed. Concentrations were determined using an external standard curve. The limits of detection (LOD) and the limits of quantification (LOQ) calculated were 0.0037, 1.87, 0.0147, and 0.0066 mg 100 g−1 and 0.0089, 7.84, 0.0302, and 0.0200 mg 100 g−1 for naringin, hesperidin, poncirin, and naringenin, respectively. The results demonstrated that hesperidin was present at the highest concentration levels, especially in the industrial orange juices. Its average content and concentration range were 69.85 and 18.80–139.00 mg 100 g−1. The other flavanones showed the lowest concentration levels. The average contents and concentration ranges found were 0.019, 0.01–0.30, and 0.12 and 0.1–0.17, 0.13, and 0.01–0.36 mg 100 g−1, respectively. The results were also evaluated using the principal component analysis (PCA) multivariate analysis technique which showed that poncirin, naringenin, and naringin were the principal elements that contributed to the variability in the sample concentrations. Lidércia C. R. Cerqueira e Silva, Jorge M. David, Rafael dos S. Q. Borges, Sérgio L. C. Ferreira, Juceni P. David, Pedro S. dos Reis, and Roy E. Bruns Copyright © 2014 Lidércia C. R. Cerqueira e Silva et al. All rights reserved. The Therapeutic Effect of Zuogui Wan in Gestational Diabetes Mellitus Rats Tue, 22 Jul 2014 08:59:48 +0000 In this experiment, we established an animal model of gestational diabetes mellitus rats using streptozotocin. Using the rat model of GDM, the pregnant rats in 1-19d were divided into three groups: (1) Zuogui Wan gestational diabetes mellitus group (group I, ). Compared with gestational diabetes mellitus rats as the control group, Zuogui Wan can change the indexes of fasting blood glucose, body weight, total cholesterol, insulin, and metabolism cage index significantly in Zuogui Wan gestational diabetes mellitus group. We can conclude that Zuogui Wan has the therapeutic effect on gestational diabetes mellitus. Yuwei Wang, Qianjin Feng, Xin Niu, Xinshe Liu, Kaixia Xu, Xiangzhu Yang, Huifeng Wang, and Qiuju Li Copyright © 2014 Yuwei Wang et al. All rights reserved.