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Journal of Chromatography B (v.899, #C)
Editorial Board (i).
Simultaneous determination of lercanidipine, benazepril and benazeprilat in plasma by LC–MS/MS and its application to a toxicokinetics study by Keguang Chen; Jing Zhang; Sha Liu; Dujuan Zhang; Yanni Teng; Chunmin Wei; Benjie Wang; Xiaoyan Liu; Guiyan Yuan; Rui Zhang; Wenjing Zhao; Ruichen Guo (1-7).
► Simultaneous determination of LER, BEN and benazeprilat in plasma with single IS. ► The method was fully validated to meet the requirements of FDA and GCP guidelines. ► It can meet the requirement in analyzing large number of samples from clinical trials.We aim to develop a rapid, simple, sensitive and specific LC–MS/MS method for the simultaneous quantification of lercanidipine, benazepril and benazeprilat in plasma. It is performed on the Agilent 6410 LC–MS/MS under the multiple-reaction monitoring (MRM) mode with electrospray ionization. Gliclazide was used as the internal standard (IS). Analytes and IS were extracted from plasma by solid-phase extraction. The reconstituted samples were chromatographed on a Diamond C18 (150 mm × 4.6 mm, 5 μm) column. The mobile phase was composed of 0.1% acetic acid–acetonitrile (50:50, v/v), with gradient flow rates: 0.6 mL/min (0–4.55 min); 4.55–4.65 min, 1 mL/min; 1 mL/min (4.65–9.5 min); 9.5–9.6 min, 0.6 mL/min; 0.6 mL/min (9.6–10 min). Method validation demonstrated that the method was of satisfactory specificity, sensitivity, precision and accuracy in linear ranges of 1–2000 ng/mL for lercanidipine, 1–2000 ng/mL for benazepril and 1–1600 ng/mL for benazeprilat, respectively. The precision (RSD%) was better than 15, and the lower limit of quantitation was identifiable and reproducible at 1 ng/mL for the three analytes. The plasma samples were stable after being stored for more than 60 days and after two freeze–thaw cycles (−20 to −25 °C). It is demonstrated that this method was successfully applied to samples from a toxicokinetics study of a compound of lercanidipine and benazepril in beagle dogs.
Keywords: Lercanidipine; Benazepril; Benazeprilat; Determination; LC–MS/MS;
Interference by metabolites and the corresponding troubleshooting during the LC–MS/MS bioanalysis of G004, a bromine-containing hypoglycemic agent by Linlin Hu; Li Ding; Xiaobing Li; Na Zhou; Shuisheng Zhong; Guanzhong Wu; Huibin Zhang (8-13).
► G004, a novel bromine-containing hypoglycemic agent with antithrombosis effect. ► The detection of an interfering metabolite in the LC–MS/MS bioanalysis of G004. ► MS conditions were optimized to eliminate the interference from the metabolite. ► Metabolite-related matrix effect was evaluated by the standard addition method. ► Pharmacokinetics of G004 in beagle dogs was reported for the first time.The quantitative determination of drugs in bio-samples may be interfered by the drug-related metabolites during the high-throughput LC–MS/MS analysis. When quantifying bromine or chlorine containing compounds, the 81Br/37Cl isotopic forms of their mono-hydroxylated metabolites after in-source dehydration could produce ions which are isobaric with the precursor ions of the parent compounds at the 79Br/35Cl isotopic form. In this report, we described the identification of an interfering hydroxylated metabolite of G004, a novel bromine-containing hypoglycemic agent, during LC–MS/MS analysis of plasma samples. Several different MRM transitions were tested and evaluated to minimize the metabolite influence on the quantification of G004. Furthermore, the standard addition method using incurred samples was used to evaluate the matrix effect caused by the interfering metabolite. The lower limit of quantitation of the established method was 0.2 ng/ml, which was 10 times lower than the existing one. The method was successfully applied to investigate the single-dosing pharmacokinetic profile of G004 in beagle dogs. The above results indicated that when quantifying chlorine or bromine containing compounds, evaluation of the interference from mono-hydroxylation or dehydrogenation metabolites should be undertaken, and if such metabolites existed, their impact on quantification of the parent compounds could be eliminated by the proper selection of the MRM transitions.
Keywords: G004; Liquid chromatography–tandem mass spectrometry; Bromine or chlorine containing compounds; Metabolite interference; Metabolite-related matrix effect; Pharmacokinetics;
Analysis of colistin A and B in fishery products by ultra performance liquid chromatography with positive electrospray ionization tandem mass spectrometry by Yingjiang Xu; Xiuhui Tian; Chuanbo Ren; Hui Huang; Xiuzhen Zhang; Xianghong Gong; Huihui Liu; Zhaoqiang Yu; Limin Zhang (14-20).
A rapid and simple method for the determination of colistin A and B in fishery products by reversed phase ultra performance liquid chromatography with positive electrospray ionization tandem spectrometry (UPLC–ESI-MS/MS) method was described. The samples were extracted with 1.0 mol/L of hydrochloric acid in methanol–water and then purified on the PLS solid phase extraction columns. Then the eluate was evaporated to less than 1 mL under a gentle stream of nitrogen at 40 °C and formic acid–acetonitrile–water (0.2/10/90, v/v/v) was added to adjust volume to 1 mL final volume. An aliquot (10 μL) was injected onto the LC column for analysis with the mobile phase of 0.2% formic acid in acetonitrile and 0.2% formic acid in water at 0.20 mL min−1. Multiple reaction monitoring was performed using precursor-product ion combinations. Calibration curves were linear from 200 ng/mL to 2000 ng/mL for colistin A and B. Mean recoveries were between 72.9% and 82.9%. The LOD was 10.0 μg/kg and LOQ was 40.0 μg/kg. The intra-day assay precision values for QC samples were between 2.17% and 9.00%, and inter-day values were between 2.80% and 6.97%. The method has merits of simplicity, sensitivity and rapidity, and it can be used for the determination of colistin A and B in fishery products.
Keywords: Colistin A; Colistin B; UPLC–ESI-MS/MS; Fishery products;
An LC–MS/MS method for determination of jujuboside A in rat plasma and its application to pharmacokinetic studies by Changhui Liu; Yingyi Li; Yanhua Zhong; Xiaotao Huang; Xia Zheng; Neng Li; Shaoling He; Suiqing Mi; Ningsheng Wang (21-26).
► It firstly developed a LC–MS/MS method for measurement of jujuboside A in rat plasma. ► It firstly reported the pharmacokinetic profile of jujuboside A in rats. ► The lower limit of quantification (LLOQ) of developed LC–MS/MS method was lower than the previous report.A novel, simple, and sensitive method for the determination of jujuboside A in rat plasma using liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS) was developed. Following solid-phase extraction, measurement of jujuboside A was performed by negative ion electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode. The limit of detection was 1.25 ng/mL, and the lower limit of quantification was 5 ng/mL in rat plasma. Good linearity was obtained over the range of 6.25–500 ng/mL, and the correlation coefficient was better than 0.998. The intra- and inter-day precisions ranged 4.4–7.5% and 2.9–10.7%, respectively. The accuracy derived from QC samples ranged 3.2–7.8% and 2.2–3.5%, respectively. The recovery ranged from 72.9 to 75.1% and the matrix effect from 96.7 to 105.3%. The analyte was stable under various conditions (at room temperature, during freeze–thaw, in the autosampler and under deep-freeze conditions). The developed method was successfully applied to the pharmacokinetic study in rats.
Keywords: Jujuboside A; LC–MS/MS; Rat plasma; Pharmacokinetics;
Simultaneous quantification of metronidazole, tinidazole, ornidazole and morinidazole in human saliva by Yongqing Wang; Peipei Zhang; Ningling Jiang; Xiaojian Gong; Ling Meng; Dewang Wang; Ning Ou; Haibo Zhang (27-30).
► Simultaneous quantification of metronidazole, tinidazole, ornidazole and morinidazole. ► Ornidazole and morinidazole (500 mg) reach more than 4100 ng/ml after i.v. infusion. ► The method is simple, sensitive and specific, with a short analysis time (6 min). ► It was used to find out who had taken other 5-nitroimidazole derivatives before trial.The aim of this study was to develop a rapid and sensitive method for the simultaneous quantification of metronidazole (MEZ), tinidazole (TNZ), ornidazole (ONZ) and morinidazole (MNZ) in human saliva. A reversed-phase high-performance liquid chromatography (HPLC) method with ultraviolet (UV) detection at 318 nm was carried out on a C18 column, using a mixture of potassium dihydrogen phosphate buffer, acetonitrile, and methanol (55:15:30, v/v/v) as a mobile phase with a flow rate of 1.0 ml/min. The saliva samples (100 μl) were firstly deproteinized by precipitation with methanol (400 μl), after which they were centrifuged and the supernatants were directly injected into the HPLC system. This method produced linear responses in the concentration ranges of 25.2–5040.0, 23.9–4790.0, 25.4–5080.0, 25.0–5000.0 ng/ml with detection limits of 6.0, 17.6, 10.0 and 11.3 ng/ml for MEZ, TNZ, ONZ and MNZ (S/N = 3), respectively. The methods were validated in terms of intra- and inter-batch precision (within 7.3% and 9.1%, respectively), accuracy, linearity, recovery and stability. The study proved that HPLC is both sensitive and selective for the simultaneous quantification of MEZ, TNZ, ONZ and MNZ in human saliva using a single mobile phase.
Keywords: Metronidazole; Tinidazole; Ornidazole; Morinidazole; Saliva; Periodontitis;
Determination of the active metabolite of moguisteine in human plasma and urine by LC–ESI-MS method and its application in pharmacokinetic study by Yanni Teng; Haibo Song; Fanlong Bu; Chunmin Wei; Wenjing Zhao; Rui Zhang; Guiyan Yuan; Xiaoyan Liu; Benjie Wang; Ruichen Guo (31-35).
► It is the first LC–MS method to determine main active metabolite of moguisteine. ► This method is quicker, more accurate and specific than the previous literatures. ► It is the first pharmacokinetic study of moguisteine in healthy Chinese volunteers.In this study, a sensitive and reproducible electro-spray ionization liquid chromatography–mass spectrometry (LC–ESI-MS) method was established to determine the concentration of M1, the main active metabolite of moguisteine in human plasma and urine. The analysis was performed on a Diamonsil® C18(2) column (150 mm × 4.6 mm, 5 μm) with the mobile phase consisting of 0.1% formic acid–acetonitrile (57:43, v/v, pH = 3.0) at a flow rate of 0.8 mL min−1. The pseudo-molecular ions [M+H]+ (m/z 312.2 for M1 and 446.3 for glipizide) were selected as the target ions for quantification in the selected ion monitoring (SIM) mode. Plasma samples were analyzed after being processed by acidification with formic acid and protein precipitation with acetonitrile. Urine samples were appropriately diluted with blank urine for analysis. Calibration curve was ranged from 0.02 to 8 μg mL−1. The extraction recovery in plasma was over 90%. Both the inter- and intra-day precision values were less than 7.5%, and the accuracy was in the range from −6.0% to 6.0%. This is the first reported LC–ESI-MS method for analyzing M1 in human plasma and urine. The method was successfully applied to the pharmacokinetic study after oral administration of single-dose and multiple-dose of moguisteine tablets in healthy Chinese subjects.
Keywords: Moguisteine; LC–ESI-MS; Pharmacokinetics; Plasma; Urine;
An improved high performance liquid chromatography–diode array detection–mass spectrometry method for determination of carotenoids and their precursors phytoene and phytofluene in human serum by B.Y. Hsu; Y.S. Pu; B. Stephen Inbaraj; B.H. Chen (36-45).
► We develop HPLC–DAD–MS method for analysis of carotenoids in human serum. ► 30 carotenoids are separated within 45 min, identified and quantified. ► Sensitivity, recovery and reproducibility of the developed method are good. ► Method applied to samples from subjects fed with carotenoid-rich capsules.An improved high performance liquid chromatography–diode array detection–mass spectrometry method was developed for determination of various carotenoids and their precursors phytoene and phytofluene in human serum. A polymeric C30 column and mobile phase of (A) methanol/acetonitrile/water (84:14:4, v/v/v) and (B) dichloromethane (100%) were employed with the gradient condition of 100% A and 0% B initially, raised to 10% B at 4 min, 18% B at 12 min, 21% B at 17 min, 30% B at 20 min and maintained until 25 min and increased further to 39% B at 28 min, 60% B at 40 min and returned to 100% A and 0% B at 45 min. A total of 30 carotenoids, including 6 all-trans forms, 20 cis-isomers, 2 β-carotene epoxides, phytoene and phytofluene, were resolved within 45 min at a flow-rate of 1 mL/min, column temperature 25 °C and detection wavelengths 450, 348 and 286 nm. Identification of carotenoids was carried out by comparing retention behavior, absorption and mass spectral data with those of reference standards, isomerized standards and reported values. An internal standard parared was found appropriate for quantitation of all the carotenoids. The developed method provided high sensitivity with low detection and quantitation limits (2–14 and 6–43 ng/mL), high recovery (91–99%), and small intra-day and inter-day variations (0.14–6.01% and 0.31–7.28%). Application of the developed method to Taiwan subjects supplemented with carotenoid-rich capsules revealed β-carotene plus its cis isomers as well as epoxide derivatives to be present in largest amount (1069.8–2783.1 ng/mL) in serum, followed by lutein plus its cis isomers (511.6–2009.5 ng/mL), phytofluene plus its cis isomer (515.0–1765.0 ng/mL), lycopene plus its cis isomers (551.1–1455.1 ng/mL), β-cryptoxanthin plus its cis isomers (458.0–965.0 ng/mL), all-trans-zeaxanthin (110.0–177.0 ng/mL), phytoene (41.8–165.0 ng/mL) and all-trans-α-carotene (37.5–95.9 ng/mL).
Keywords: Carotenoid; Phytoene; Phytofluene; Human serum; HPLC–DAD–MS;
Simultaneous determination of dextromethorphan, dextrorphan and doxylamine in human plasma by HPLC coupled to electrospray ionization tandem mass spectrometry: Application to a pharmacokinetic study by J.L. Donato; F. Koizumi; A.S. Pereira; G.D. Mendes; G. De Nucci (46-56).
► HPLC–ESI-MS method to quantify dextromethorphan, dextrorphan and doxylamine. ► Method was applied in pharmacokinetic study in human plasma. ► Extraction performed with a simple liquid–liquid extraction. ► High sensitivity, specificity and high throughput.In the present study, a fast, sensitive and robust method to quantify dextromethorphan, dextrorphan and doxylamine in human plasma using deuterated internal standards (IS) is described. The analytes and the IS were extracted from plasma by a liquid–liquid extraction (LLE) using diethyl-ether/hexane (80/20, v/v). Extracted samples were analyzed by high performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry (HPLC–ESI-MS/MS). Chromatographic separation was performed by pumping the mobile phase (acetonitrile/water/formic acid (90/9/1, v/v/v) during 4.0 min at a flow-rate of 1.5 mL min−1 into a Phenomenex Gemini® C18, 5 μm analytical column (150 × 4.6 mm i.d.). The calibration curve was linear over the range from 0.2 to 200 ng mL−1 for dextromethorphan and doxylamine and 0.05 to 10 ng mL−1 for dextrorphan. The intra-batch precision and accuracy (%CV) of the method ranged from 2.5 to 9.5%, and 88.9 to 105.1%, respectively. Method inter-batch precision (%CV) and accuracy ranged from 6.7 to 10.3%, and 92.2 to 107.1%, respectively. The run-time was for 4 min. The analytical procedure herein described was used to assess the pharmacokinetics of dextromethorphan, dextrorphan and doxylamine in healthy volunteers after a single oral dose of a formulation containing 30 mg of dextromethorphan hydrobromide and 12.5 mg of doxylamine succinate. The method has high sensitivity, specificity and allows high throughput analysis required for a pharmacokinetic study.
Keywords: Quantification; Volunteer; LC–MS/MS;
Quechers methodologies as an alternative to solid phase extraction (SPE) for the determination and characterization of residues of cephalosporins in beef muscle using LC–MS/MS by R. Pérez-Burgos; E.M. Grzelak; G. Gokce; J. Saurina; J. Barbosa; D. Barrón (57-65).
► We compared SPE and QuEChERS methods to determine seven cephalosporins in cow muscle. ► An experimental design was used to optimize the two clean-up methods. ► The validation of the two methods was done according to the Directive 2002/657/EC. ► The separation and detection of the cephalosporins was carried out by LC–MS/MS. ► Comparable results were found for two methods in terms of quality parameters.This work was focused on the comparison of two clean-up methods to be used for the simultaneous determination of seven cephalosporins in cow muscle. In particular, the performance of novel dispersive solid phase extraction (d-SPE) procedures based on QuEChERS methodologies was assessed and compared with conventional SPE. The separation and detection of the analytes using both methods was carried out by LC–MS/MS to reach enough sensitivity to be compatible with the detection of the maximum residue limits (MRL) of cephalosporins as regulated by EU directives. The optimization of the clean-up step relied on experimental design in order to find the most suitable conditions with a reduced number of assays. Besides, multi-objective responses were used to reach an overall compromise in the recovery of all analytes simultaneously. The validation of the two methods was done according to the Directive 2002/657/EC. Linearity, decision limit, detection capability, detection and quantification limits (4–50 μg kg−1), precision (RSD less than 15% except for PIR) and recoveries were determined and adequate results with comparable values using QuEChERS and SPE methodologies. LOQ were better for SPE method (0.1–10 μg kg−1) but both methods show LOQ below MRL values. Precision was slightly better for the QuEChERS method, that also presents better recoveries, higher than 85% except for cephalexin.
Keywords: Cephalosporins; Beef tissues; QuEChERS; SPE; Experimental design; LC–MS/MS; Validation according 2002/657/EC;
Low-density solvent-based dispersive liquid–liquid microextraction followed by high performance liquid chromatography for determination of warfarin in human plasma by Hoda Ghambari; Mohammadreza Hadjmohammadi (66-71).
► LDS-DLLME presented a high recovery for the extraction of warfarin in human plasma. ► LDS-DLLME–HPLC-UV yielded good sensitivity for warfarin determination. ► Wide linear range of this method can be applied in therapeutic drug monitoring.Extraction and determination of warfarin, a widely used anticoagulant drug, in human plasma were performed using a new generation of dispersive liquid–liquid microextraction (DLLME) and high performance liquid chromatography (HPLC). The extraction procedure is based on extraction solvents lighter than water and performing of extraction in a specially designed extraction cell. Some important parameters, including kind and volume of extraction and disperser solvents, pH of the sample solution, salt concentration in the sample solution and extraction time were investigated and optimized. Under the optimized conditions (150 μL 1-octanol as extraction solvent, 150 μL methanol as disperser solvent, pHsample = 2.3, extraction time of 2 min, without salt addition), limit of detection (LOD) of 5 ng mL−1 and extraction recovery of 91.0% were obtained. The calibration curve was linear within the range of 15–3000 ng mL−1 with the square of correlation coefficient (R 2) of 0.998. Repeatability and reproducibility of method based on five replicate extraction and determination were 2.8% and 6.5%, respectively. The proposed method was applied successfully for the determination of warfarin in plasma sample from a patient under treatment with this drug, and was demonstrated to be sensitive, efficient, and convenient.
Keywords: Low-density solvent; Dispersive liquid–liquid microextraction; Warfarin; HPLC; Sample preparation; Plasma sample analysis;
Optimization and correlation of HPLC-ELSD and HPLC–MS/MS methods for identification and characterization of sophorolipids by Isabel A. Ribeiro; M. Rosário Bronze; Matilde F. Castro; Maria H.L. Ribeiro (72-80).
► TLC, HPLC-ELSD and HPLC–ESI-MS/MS were optimized and correlated for SLs analysis. ► HPLC–ESI-MS/MS analysis allowed identification of SLs from different sources. ► A [M+Na]+ fragmentation pattern of acidic and lactonic SLs was established. ► For the first time C24:0 monoacetylated and diacetylated SLs were identified.High-performance liquid chromatography (HPLC) with evaporative light scattering detection (ELSD) and HPLC with electrospray ionization tandem mass spectrometry (HPLC–ESI-MS/MS) methods were implemented and optimized to separate and identify sophorolipids (SLs) produced by Rhodotorula bogoriensis and Starmerella bombicola. SLs are carbohydrate-based amphiphilic biosurfactants with increased interest in pharmaceutical and environmental areas. Rhodotorula bogoriensis and Starmerella bombicola are mainly producers of respectively C22, and C16 and C18 SLs. Mass fragmentation patterns of SLs produced by both yeasts were investigated by HPLC–ESI-MS/MS in the positive mode for [M+Na]+. Based on the established fragmentation pattern, SLs produced by both yeasts were identified and characterized. A correlation between HPLC-ELSD and HPLC- ESI-MS/MS methods was established and made possible the identification of SLs by the HPLC-ELSD technique. TLC is a common tool for the analysis of SLs mixtures. In this work, TLC scrapped bands were analysed by HPLC-ELSD and HPLC–MS allowing the correlation between R F values and the identification of sophorolipids by this technique. Identification of monoacetylated and diacetylated C24:0 hydroxy fatty acids sophorolipids produced by Rhodotorula bogoriensis was for the first time accomplished with this study. Although present in lower quantity these longer chain SLs can assume special importance regarding their biological activity and surface active properties.
Keywords: Sophorolipid; TLC; HPLC-ELSD; HPLC–ESI-MS/MS; Rhodotorula bogoriensis; Starmerella bombicola;
Development and validation of a liquid chromatography–tandem mass spectrometry method for quantification of decitabine in rat plasma by Haiyan Xu; Shaoqiong lv; Mingxi Qiao; Yao Fu; Xue Jiang; Yi Jin; Chibing Li; Bo Yuan (81-85).
Decitabine is chemically unstable at physiological temperature and pH. In addition, the bioanalysis of decitabine is easily interfered by endogenous 2-deoxycytidine. A simple, sensitive and specific LC–MS/MS method was developed for the analysis of decitabine in rat plasma. No exogenous stabilizers were used to prevent the degradation of decitabine in rat plasma. After deproteinized with acetonitrile at room temperature, rat plasma samples were analyzed on a Hypersil APS-2 NH2 column interfaced with a triple quadrupole tandem mass spectrometer in positive electrospray ionization mode. Decitabine was completely separated from 2-deoxycytidine using gradient elution of water (solvent A) and acetonitrile (solvent B) at a flow rate of 1 mL/min. To quantify decitabine and daidzin (internal standard), respectively, multiple reaction monitoring (MRM) transitions of m/z 251.1 → 134.7 and m/z 417.3 → 255.3 was performed. The assay was linear over the concentration range of 5.0–2000 ng/mL. The intra- and inter-day precision was within 12.0% in terms of relative standard deviation (RSD%) and the accuracy within 5.9% in terms of relative error. The LC–MS/MS method was fully validated for its sensitivity, selectivity, stability study, matrix effect and recovery. The data indicate that this LC–MS/MS method is a specific and effective method for the pharmacokinetic study of decitabine in rat plasma. Compared with the previously reported analytical methods, this method showed easy and economic sample preparation, good specificity and high sensitivity with less plasma (50 μL).
Keywords: Decitabine; LC–MS/MS; Rat plasma; Pharmacokinetics;
Application of nanoLC–MS/MS to the shotgun proteomic analysis of the nematocyst proteins from jellyfish Stomolophus meleagris by Rongfeng Li; Huahua Yu; Ronge Xing; Song Liu; Yukun Qing; Kecheng Li; Bing Li; Xiangtao Meng; Jinhui Cui; Pengcheng Li (86-95).
► We analyzed the nematocyst proteins of jellyfish Stomolophus meleagris by using the nanoLC–MS/MS with the shotgun proteomic method. ► A total of 181 proteins had been identified with the molecular weight ranging from 5268.06 to 843,487.57 and the pI from 4.49 to 11.39. ► The identified nematocyst proteins were analyzed by bioinformatic method including gene ontology (GO) annotation, pathways and gene network analysis.The nematocyst proteins of jellyfish Stomolophus meleagris, a complicated mixture, contain many important bioactive molecules. In present study, to gain comprehensive insight into the protein component and search some novel bioactive molecules in the nematocyst proteins, shotgun proteomic analysis of the nematocyst proteins was carried out by nano liquid chromatography tandem mass spectrometry (nanoLC–MS/MS) for the first time. Digested peptides of the nematocyst proteins were analyzed by nanoLC–MS/MS and all MS/MS spectra were then automatically searched by the SEQUEST program. A total of 181 proteins had been identified, with the molecular weight ranging from 5268.06 to 843,487.57 and the pI from 4.49 to 11.39. Bioinformatic analysis was also applied to better understand the identified proteins. In the gene ontology (GO) annotation, all the identified proteins were classified into 13, 9 and 7 groups according to biological process, cellular component and molecular function, respectively. Pathways analysis of the identified proteins was conducted with 33 corresponding pathways found. On the basis of pathways analysis, we also constructed the gene network to analyze the relationship of those genes each other, which contained enzyme–enzyme relation, protein–protein interaction and gene expression interaction.
Keywords: Jellyfish; Stomolophus meleagris; Nematocyst proteins; Shotgun; NanoLC–MS/MS; Proteomic analysis;
Click chemistry: A route to designing and preparing pseudo-biospecific immunoadsorbent for IgG adsorption by Xiaoyan Hu; Guangji Li; Ende Huang (96-102).
► Exploring a novel route to a pseudo-biospecific immunoadsorbent – click chemistry. ► Preparing "clickable" reaction modules using l-histidine and original sepharose. ► Click reaction can greatly increase the immobilization efficiency of ligands. ► The 1,2,3-triazole in spacer-arm helps IgG binding without non-specific adsorption. ► The prepared immunoadsorbent exhibits excellent adsorption performance for IgG. l-histidine is a promising alternative to expensive protein ligands for the adsorption of IgG due to its high selectivity, no toxicity and low cost; while click chemistry can improve the reaction selectivity between the ligands and the support matrix under mild reaction conditions. Thus, using l-histidine as a ligand and original sepharose gel as a support, a novel immunoadsorbent possessing pseudo-biospecific affinity for IgG from human plasma, Sep-triazole-His, was designed and prepared according to the principle of Click-reaction between alkyne and azide functional groups; while both sepharose-based control samples Sep-His and Sep-PA were prepared by a conventional method using l-histidine and protein A as a ligand, respectively. The ligand density and IgG adsorption performance of Sep-triazole-His from human plasma were measured and evaluated. The influences of click chemistry on the preparation, structure and performance of sepharose-based immunoadsorbent were also investigated. The results indicate that the ligand density immobilized on Sep-triazole-His is 319.1 μmol/g sepharose gel, almost 4-fold as high as that on Sep-His; the IgG adsorption capacity of Sep-triazole-His from human plasma reaches 16.49 mg/g at pH 7.0, or increases 5.72-fold with respect to Sep-His, and does not decrease noticeably after being repeatedly used for 10 times; and Sep-triazole-His can exhibit high adsorption selectivity for IgG comparable to Sep-PA. The further studies prove that the 1,2,3-triazole ring in the spacer-arm of Sep-triazole-His, can facilitate the binding of IgG without non-specific adsorption.
Keywords: Pseudo-biospecific immunoadsorbent; Click chemistry; Histidine ligand; IgG; Adsorption;
Analysis of N′-nitrosonornicotine and its metabolites in rabbit blood with liquid chromatography/tandem mass spectrometric method by Beibei Zhao; Sheng Wang; Juan Wang; Hailei Lang; Shihao Sun; Jian Mao; Jianxun Zhang (103-108).
► A method to analyze NNN and its metabolites with HPLC–MS/MS was established. ► Hydroxy acid was the major metabolite of NNN in rabbit blood. ► 5′-Hydroxylation of NNN might be the main metabolic pathway of NNN in rabbit blood. ► Metabolic curves and pharmacokinetic parameters were obtained for NNN and its metabolites.N′-nitrosonornicotine (NNN) is a strong carcinogen. The metabolic study of NNN in vivo will help us to further understand it, however, trace detection in complex matrices requires highly sensitive detection methods. After the chromatographic conditions and mass spectrometric conditions had been optimized and confirmed, a method for determining NNN and its metabolites in rabbit blood by high-performance liquid chromatography/tandem mass spectrometry (HPLC–MS/MS) was established. The results showed that precisions (R.S.Ds) were between 0.5% and 8.62%, the recoveries ranged from 80% to 111%. Linearity was observed for all compounds with detection limits ranging from 0.039 ng mL−1 to 0.217 ng mL−1. Metabolic curves and pharmacokinetic parameters were obtained for NNN and its metabolites. The elimination half-life of NNN was 30 min and the main metabolite of NNN was 4-hydroxy-4-(3-pyridyl)-butyric acid (hydroxy acid) and the major metabolic pathway was 5′-hydroxylation and subsequent secondary metabolite formation.
Keywords: N′-nitrosonornicotine (NNN); Metabolites; Pharmacokinetic; HPLC–MS/MS;
Biotransformation of flavonols and taxifolin in hepatocyte in vitro systems as determined by liquid chromatography with various stationary phases and electrospray ionization-quadrupole time-of-flight mass spectrometry by Jan Vacek; Barbora Papoušková; Pavel Kosina; Jiří Vrba; Vladimír Křen; Jitka Ulrichová (109-115).
► A robust LC/MS metabolic study focused on flavonols and flavanonols is presented. ► For analysis of the metabolites C18, C8, CNP and phenyl-based LC stationary phases was used. ► The study describes retention behaviour and MS data for individual metabolites. ► The metabolites were obtained using an in vitro human hepatocyte model system.Liquid chromatography (LC) on various stationary phases was used for the metabolite profile analysis of quercetin, rutin, isoquercitrin and taxifolin. The metabolites were obtained using an in vitro model system of human and rat hepatocytes in the form of cell suspensions and the primary cultures. For separations of the parent compounds and their metabolites, stationary phases based on C18, C8, cyanopropyl (CNP) or phenyl (PHE) modifications of silica were tested. CNP and PHE stationary phases operating in reversed-phase mode have been shown to be efficient for separation of parent flavonoids and their polar metabolites. Individual metabolites were identified on the basis of an elemental composition determination using electrospray ionization-quadrupole time-of-flight mass spectrometry (ESI-QqTOF MS) on-line connected with an LC system. Detailed analytical parameters such as retention times, selectivity, resolution of chromatographic peaks, MS fragmentation and UV–vis absorption maxima were determined for individual metabolites, namely for phase II biotransformation products. The predominant metabolites were methylated flavonols and flavonol glucuronides. The highest biotransformation rate was found with taxifolin, which was mainly converted to sulfates. The HPLC/ESI-QqTOF MS analyses revealed that quercetin and taxifolin were metabolized more extensively than the studied glycosides, rutin and isoquercitrin.
Keywords: Flavonols; Flavanonol; Taxifolin; Metabolism; Liquid chromatography; Retention behaviour; Mass spectrometry; Hepatocytes;
Design of high productivity antibody capture by protein A chromatography using an integrated experimental and modeling approach by Candy K.S. Ng; Hector Osuna-Sanchez; Eric Valéry; Eva Sørensen; Daniel G. Bracewell (116-126).
► A lumped parameter transport-dispersive model is used. ► An experimental strategy for critical parameter estimation is designed. ► The impact of process variables on productivity is studied via use of model.An integrated experimental and modeling approach for the design of high productivity protein A chromatography is presented to maximize productivity in bioproduct manufacture. The approach consists of four steps: (1) small-scale experimentation, (2) model parameter estimation, (3) productivity optimization and (4) model validation with process verification. The integrated use of process experimentation and modeling enables fewer experiments to be performed, and thus minimizes the time and materials required in order to gain process understanding, which is of key importance during process development. The application of the approach is demonstrated for the capture of antibody by a novel silica-based high performance protein A adsorbent named AbSolute. In the example, a series of pulse injections and breakthrough experiments were performed to develop a lumped parameter model, which was then used to find the best design that optimizes the productivity of a batch protein A chromatographic process for human IgG capture. An optimum productivity of 2.9 kg L−1 day−1 for a column of 5 mm diameter and 8.5 cm length was predicted, and subsequently verified experimentally, completing the whole process design approach in only 75 person-hours (or approximately 2 weeks).
Keywords: Protein A chromatography; Batch chromatography; Antibody capture; Productivity; Integrated process experimentation and modeling;
Direct pharmacokinetic analysis of puqietinone by in vivo microdialysis sampling and turbulent-flow chromatography coupled with liquid chromatography–mass spectrometry by Gui-Zhong Xin; Liu Cao; Zi-Qi Shi; Hui-Jun Li; Xiao-Dong Wen; Jun Chen; Lian-Wen Qi; Ping Li (127-134).
► Microdialysis sampling and turbulent-flow chromatography were coupled to LC/MS. ► The system is able to achieve online desalting and concentrating within 1 min. ► Influences of matrix effect and carry-over were eliminated effectively. ► Lifetime of extraction column is much extended. ► The temporal resolution of sampling points is improved by using MD technology.Sample pretreatment is a key step in bioanalytical process because of possible interference and matrix effects in mass spectrometry analysis. In this work, a novel strategy towards high speed and sensitivity was developed combining in vivo microdialysis (MD) sampling, turbulent-flow chromatography (TFC), and liquid chromatography–mass spectrometry (LC–MS). The procedures of cleanup, preconcentration, and separation were completed on-line in one step within 10 min. During the MD optimization procedure, 1% hydroxypropyl-β-cyclodextrin (HP-β-CD) was used to improve the relative recovery of the analyte. Untreated MD samples were directly injected, and a TFC precolumn was flushed for 1 min with aqueous phase of 4 mL/min flow rate to desalt and concentrate biosamples. The retained analytes were then back-flushed by a switching valve onto a fast LC column (4.6 mm × 50 mm, 1.8 μm) for separation. Another diverter valve was employed to prevent the HP-β-CD that interferes with the ESI process from entering the MS. Puqietinone, a lipophilic alkaloid from Fritillaria puqiensis, was used as a case for validation. Results showed that the limit of quantification for puqietinone was 0.10 ng/mL, and good linearity (R 2 = 0.9993) was maintained over the range of 1.02–200.02 ng/mL. Accuracy and precision were satisfactory within the range of the standard curve. This approach was able to effectively eliminate the influences of matrix effect and carry-over as the injection volume increased up to 20 μL. The developed method was successfully applied to pharmacokinetic study of puqietinone after intravenous administration to rat. Results demonstrate the potential of using MD with TFC-LC/MS for in vivo monitoring experiments.
Keywords: Microdialysis; Turbulent-flow chromatography; Mass spectrometry; Desalting; Pharmacokinetic; Puqietinone;
Bile acid profiling in human biological samples: Comparison of extraction procedures and application to normal and cholestatic patients by Lydie Humbert; Marie Anne Maubert; Claude Wolf; Henri Duboc; Myriam Mahé; Dominique Farabos; Philippe Seksik; Jean Maurice Mallet; Germain Trugnan; Joëlle Masliah; Dominique Rainteau (135-145).
► Four extraction protocols were compared to optimize bile acid quantification by HPLC MS/MS. ► The best protocols were the Acetonitrile one for liquid (serum, urine, bile) and the NaOH one for stool samples. ► Allowed to detect 14 different species in serum, 11 in urine and 25 in stool. ► Applied to control and cholestatic patients and useful for quantification in routine analysis.The role of bile acids in cell metabolism, membrane biology and cell signaling is increasingly recognized, thus making necessary a robust and versatile technique to extract, separate and quantify a large concentration range of these numerous molecular species. HPLC–MS/MS analysis provides the highest sensitivity to detect and identify bile acids. However, due to their large chemical diversity, extraction methods are critical and quite difficult to optimize, as shown by a survey of the literature. This paper compares the performances of four bile acid extraction protocols applied to either liquid (serum, urine, bile) or solid (stool) samples. Acetonitrile was found to be the best solvent for deproteinizing liquid samples and NaOH the best one for stool extraction. These optimized extraction procedures allowed us to quantitate as much as 27 distinct bile acids including sulfated species in a unique 30 min HPLC run, including both hydrophilic and hydrophobic species with a high efficiency. Tandem MS provided a non ambiguous identification of each metabolite with a good sensitivity (LOQ below 20 nmol/l except for THDCA and TLCA). After validation, these methods, successfully applied to a group of 39 control patients, detected 14 different species in serum in the range of 30–800 nmol/l, 11 species in urine in the range of 20–200 nmol/l and 25 species in stool in the range of 0.4–2000 nmol/g. The clinical interest of this method has been then validated on cholestatic patients. The proposed protocols seem suitable for profiling bile acids in routine analysis.
Keywords: Bile acids; Extraction; Liquid chromatography–mass spectrometry; Profiling; Routine analysis;
Liquid chromatography coupled to ion trap-tandem mass spectrometry to evaluate juvenile hormone III levels in bee hemolymph from Nosema spp. infected colonies by A.M. Ares; M.J. Nozal; J.L. Bernal; R. Martín-Hernández; M.Higes; J. Bernal (146-153).
► JH III was determined in bee hemolymph by LC–MS/MS for the first time. ► The proposed sample treatment was fast and simple. ► A mixture of methanol and phenylthiourea was chosen to dilute bee hemolymph. ► Matrix (bee hemolymph) did not have any influence onto the analyte signal. ► The highest concentration of JH III was found in bees infected with Nosema ceranae. It has been described a fast, simple and sensitive liquid chromatography–tandem mass spectrometry (LC–MS/MS) method to measure juvenile hormone III (JH III), which was used to study of the effects of Nosema spp. infection on JH III levels in bee hemolymph. Honey bee hemolymph was extracted by centrifugation and mixed with a solution of phenylthiourea in methanol. This mixture was then centrifuged and the supernatant removed and evaporated to dryness. The residue was reconstituted in methanol containing the internal standard (methoprene) and injected onto an LC–MS/MS (ion-trap) system coupled to electrospray ionization (ESI) in positive mode. Chromatography was performed on a Synergi Hydro-RP column (4 μm, 30 mm × 4.60 mm i.d.) using a mobile phase of 20 mM ammonium formate and methanol in binary gradient elution mode. The method was fully validated and it was found to be selective, linear from 15 to 14,562 pg/μL, precise and accurate, with %RSD values below 5%. The limits of detection and quantification were: LOD, 6 pg/μL; LOQ, 15 pg/μL. Finally, the proposed LC–MS/MS method was used to analyze JH III levels in the hemolymph of worker honey bees (Apis mellifera iberiensis) experimentally infected with different Nosema spp. (Nosema apis, Spanish and Dutch Nosema ceranae strains). The highest concentrations of JH III were detected in hemolymph from bees infected with Spanish N. ceranae.
Keywords: Juvenile hormone III; Honey bee hemolymph; LC–MS/MS; Nosema ceranae; Nosema apis;
Simultaneous determination in hair of multiclass drugs of abuse (including THC) by ultra-high performance liquid chromatography–tandem mass spectrometry by D. Di Corcia; F. D'Urso; E. Gerace; A. Salomone; M. Vincenti (154-159).
► Hair analysis is a tool to evaluate drug exposure in several application fields. ► We present a simple UHPLC–MS/MS method to detect 13 common drugs of abuse in hair. ► The method proved excellent analytical performances and drastic reduction of time. ► High sample throughput is essential for laboratories. ► The increased global productivity makes the workplace testing of hair affordable.A simple procedure for the quantitative determination in hair samples of 13 common drugs of abuse or metabolites (morphine, 6-acetylmorphine, codeine, amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine, 3,4-methylenedioxymethamphetamine, 3,4-methylenedioxyethylamphetamine, benzoylecgonine, cocaine, buprenorphine, methadone and Δ9-tetrahydrocannabinol) has been developed and fully validated. The analytes were extracted from the matrix by a simple overnight incubation with methanol at 55 °C. An aliquot of the extract was directly injected into an ultra-high performance liquid chromatography system equipped with Waters Acquity UHPLC BEH C18 column (100 mm × 2.1 mm, 1.7 μm). The mobile phase eluted with a linear gradient (water/formic acid 5 mM:acetonitrile; v:v) from 98:2 to 0:100 in 4.5 min, followed by isocratic elution at 100% B for 1.0 min. The flow rate was 0.6 mL/min and the total run time was 8.0 min including re-equilibration at the initial conditions. The compounds were revealed by a triple quadrupole mass spectrometer operating in the selected reaction monitoring mode. The absence of matrix interferents, together with excellent repeatability of both retention times and relative abundances of diagnostic transitions, allowed the correct identification of all analytes tested. The method proved linear in the interval from the limit of quantification to 5.0 ng/mg (1.0 ng/mg for Δ9-tetrahydrocannabinol) with correlation coefficient values ranging from 0.9970 to 0.9997. Quantitation limits were below the cut-off values recommended by the Society of Hair Testing and ranged from 0.02 to 0.08 ng/mg. Application of the present UHPLC–MS/MS procedure and instrumentation to hair analysis allows high sample-throughput, together with excellent sensitivity and selectivity, in workplace drug-screening controls and forensic investigations. These qualities, combined with minimal sample workup, make the cost of this screening affordable for most private and public administrations.
Keywords: Hair; Ultra-high performance liquid chromatography; Multidrugs; Validation;
Simple, cheap and effective high-performance liquid chromatographic method for determination of praziquantel in bovine muscle by Yu Sun; Shi-Jin Bu (160-162).
► This article describes the development of a more sensitive HPLC method for determination of PZQ in bovine muscle for the first time. ► The purpose of this work was to develop a detection method for PZQ residues in bovine muscle. ► This method was validated and used to detect praziquantel residues in a plan survey. ► The work presented here used HPLC with UV detector for the determination of praziquantel following a solid-phase (SPE) extraction. ► The method is specific and sensitive, with a quantification limit of 0.02 mg/kg at and a detection limit of 0.01 mg/kg.A simple high-performance liquid chromatographic method using ultraviolet detection was developed for the determination of praziquantel in bovine muscle. The sample was extracted with ethyl acetate, cleaned up by alumin B cartridge. Analyses were run at a flow-rate of 0.8 ml/min with the detector operating at a detection wavelength of 220 nm. The method is specific and sensitive, with a quantification limit of 0.02 mg/kg and a detection limit of 0.01 mg/kg. The standard calibration curve of drugs solution was linear in the range of 0.02–2.0 μg/ml (r 2 = 0.9999). At the fortified levels of 0.02, 0.05, 0.2 mg/kg, the mean recoveries of praziquantel ranged from 75% to 85%, while the intra-day and inter-day coefficient of variation (CV) of the assay were all less than 9%.
Keywords: Praziquantel; HPLC; Determination; Bovine muscle;
Determination of celecoxib in human plasma by liquid chromatography–tandem mass spectrometry by Pavel Ptáček; Josef Klíma; Jan Macek (163-166).
► The sample preparation step was simplified using protein precipitation. ► The run time was shortened with a more selective LC–MS/MS procedure. ► Isotopically labeled internal standard was used to improve precision and accuracy. ► A rapid method for determination of celecoxib in human plasma has been developed. ► The method was successfully applied to a pharmacokinetic study.A liquid chromatography–electrospray tandem mass spectrometry method was developed and validated to quantitate celecoxib in human plasma. The assay was based on protein precipitation with methanol and liquid chromatography on a C18 column (55 mm × 2 mm, 3 μm), the mobile phase consisted of methanol – 10 mM ammonium acetate (75:25, v/v). Quantification was performed by mass spectrometry in the multiple reaction monitoring mode with negative electrospray ionization at m/z 380 → 316 and 384 → 320 for celecoxib and the internal standard celecoxib-D4, respectively. The lower limit of quantitation was 7.0 ng/ml using 0.1 ml of plasma and linearity was demonstrated up to 1800 ng/ml. Intra-assay and inter-assay precision expressed by relative standard deviation was less than 4% and inaccuracy did not exceed 6% at all levels. The assay was applied to the analysis of samples from a pharmacokinetic study.
Keywords: Celecoxib; Plasma; LC–MS/MS; Protein precipitation; Pharmacokinetics;
Corrigendum to "Bioanalysis in clinical development of tasquinimod using liquid chromatography/tandem mass spectrometry" [J. Chromatogr. B: Anal. Technol. Biomed. Life Sci. 879 (30) (2011) 3401–3406] by G.P. Hansson; M. Olin; C. Svanström; L.D. Svensson; C.J. Sennbro (167).


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Like most different issues, choosing the proper website hosting platform can also be a problem, and it turns into extra tedious if you're new to the web occupation. There are principally three sorts of internet hosting platforms - Shared, VPS and Devoted Server. As well as, you may additionally have free internet hosting, however I can't suggest you to make use of it, as a result of it has vital limitations and issues, and it's possible you'll encounter some technical inconveniences later.
You, being a brand new blogger, I assume that you simply don't have sufficient cash to take a position big funds in putting your blogs on WordPress. As well as, your necessities are usually not such that you simply select an costly choice - devoted or VPS, I strongly suggest that you simply swap to digital internet hosting.
On this article, I'll cowl numerous facets that specify why shared WordPress internet hosting is the most suitable choice for brand new blogs.
The perfect website hosting platform for brand new blogs
Technically, website hosting platforms may be divided into a number of totally different classes, resembling Shared, VPS, Devoted, Reseller and Cloud. It may also be categorised in Home windows and Linux internet hosting. Linux is very really useful for any WordPress internet hosting.
Sharing a number of internet hosting platforms, shared internet hosting is the primary selection for any new blogger who plans to create an internet site. The reason being that it doesn't beat the value, and the sources supplied are greater than sufficient to fulfill your whole fundamental necessities. Contemplating that devoted internet hosting and VPS internet hosting are costly, which may be an extreme quantity for freshmen such as you. Subsequently, it's extremely really useful to make use of a shared internet hosting platform once you begin running a blog. Sooner or later, when you want extra server sources with elevated web page site visitors, you may improve to VPS or devoted internet hosting. There are a variety of bloggers who began their on-line profession with shared internet hosting, however now they're on a VPS or devoted server. After some time, once you really feel that your WordPress website is consuming extreme server sources, initially, earlier than you alter the host, optimize your website. To hurry up your web site, you may optimize your database, clear up unused tables and eliminate plugins. If issues exist even in any case these actions have been accomplished, it's time to consider altering the host.
Beneath are the explanation why I like to recommend digital internet hosting for brand new blogs:
· At first of your website might not have heavy site visitors.
Shared internet hosting prices lower than devoted and VPS internet hosting.
· Most startups are simply dealt with by shared internet hosting.
Many individuals don't select digital internet hosting, as a result of they imagine that digital internet hosting impacts the rating of the positioning in serps. However this can be a fable. Search engine rankings don't have anything to do with the internet hosting platform.
Probably the most dependable internet hosting for WordPress
There are a number of website hosting firms, every of which affords wonderful sources at a low price, however I counsel you to be clever sufficient when selecting a internet hosting to your WordPress website. There are a number of elements to contemplate earlier than getting into right into a website hosting contract. For me, efficiency and round the clock technical help are two of an important elements.

subset: commoncrawl


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Let me explain. I teach only Science and STEM.
- NO background work.
- No lesson.
- NO research.
- Just- here are your supplies.
- Get busy!
Then I started thinking of all the things we could do. We now have about 25 of these Quick Challenges now- in many variations.
So, how does it work?
The Pom-Pom Blaster
- Think about safety first. I do not allow students to make slingshots. My experience with models made in that style is that they come apart too easily and parts go flying.
- My rule about the slingshot is simple: Your device must rest on the tabletop or the floor. It cannot be handheld.
- Another rule: Never aim at anyone. Always look before you let go of the pom-pom. (Yes, I know it's a pom-pom, but sometimes the devices come apart and fly.)
- And another safety rule: Your device must remain together while being fired. (Safety first!)


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Intikana Tantra
Return To Ecstasy
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In our Edgecliff training we deliver detailed guidance for shiva worship massage by employing sensual massage procedures.
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By advanced appointment only
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Copyright ©2016 IntikanaTantra.com all rights reserved.


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Kinvey AI/ML
Differentiate with data science capabilities engineered for modern apps.
Data Science Remains Elusive for Business Apps
Organizations often have limited data science practices that revolve around business intelligence and analytics and are far removed from software development teams. As a result, data science is often an afterthought for modern business apps, which focus on delivering operational efficiency and engaging customers across new channels. To tackle complex business problems with the latest tech, enterprises need better collaboration between software development and data science.
Kinvey AI/ML
Kinvey AI/ML is a data science capability that analyzes enterprise data to uncover the previously inaccessible insights required to address complex business problems. Develop and operationalize data science solutions to deliver differentiated app experiences.


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Digested: The Secrets of Docker Secrets
digested.blogspot.com/2019/11/the-secrets-of-docker-secrets.html, posted 14 Nov by peter in automation continuousdelivery deployment docker howto toread
Most web apps need login information of some kind, and it is a bad idea to put them in your source code where it gets saved to a git repository that everyone can see. Usually these are handled by environment variables, but Docker has come up with what they call Docker secrets. The idea is deceptively simple in retrospect. While you figure it out it is arcane and difficult to parse what is going on.
Frida • Inject JavaScript to explore native apps on Windows, macOS, GNU/Linux, iOS, Android, and QNX
https://frida.re/, posted 31 Jul by peter in automation development free hack javascript opensource software testing
Inject your own scripts into black box processes. Hook any function, spy on crypto APIs or trace private application code, no source code needed. Edit, hit save, and instantly see the results. All without compilation steps or program restarts.
Geb - Very Groovy Browser Automation
gebish.org/, posted Apr '18 by peter in automation free software testing
Geb is a browser automation solution.
It brings together the power of WebDriver, the elegance of jQuery content selection, the robustness of Page Object modelling and the expressiveness of the Groovy language.
It can be used for scripting, scraping and general automation — or equally as a functional/web/acceptance testing solution via integration with testing frameworks such as Spock, JUnit & TestNG.
A Tutorial on Portable Makefiles « null program
nullprogram.com/blog/2017/08/20/, posted 2017 by peter in automation development howto reference
In my first decade writing Makefiles, I developed the bad habit of liberally using GNU Make's extensions. I didn't know the line between GNU Make and the portable features guaranteed by POSIX. Usually it didn't matter much, but it would become an annoyance when building on non-Linux systems, such as on the various BSDs. I'd have to specifically install GNU Make, then remember to invoke it (i.e. as
gmake) instead of the system's
I've since become familiar and comfortable with make's official specification, and I've spend the last year writing strictly portable Makefiles. Not only has are my builds now portable across all unix-like systems, my Makefiles are cleaner and more robust. Many of the common make extensions — conditionals in particular — lead to fragile, complicated Makefiles and are best avoided anyway. It's important to be able to trust your build system to do its job correctly.
This tutorial should be suitable for make beginners who have never written their own Makefiles before, as well as experienced developers who want to learn how to write portable Makefiles.
In the age of robots, our schools are teaching children to be redundant | George Monbiot
https://www.theguardian.com/commentisfree/2017/feb/15/robots-schools-teaching-children-redundant-testing-learn-future, posted 2017 by peter in automation education opinion parenting toread
In the future, if you want a job, you must be as unlike a machine as possible: creative, critical and socially skilled. So why are children being taught to behave like machines?
A Perl toolchain for building micro-services at scale
https://engineering.semantics3.com/2016/06/15/a-perl-toolchain-for-building-micro-services-at-scale/, posted 2016 by peter in automation development perl toread
Our extensive use of Perl to build many of our internal services often comes as a surprise to many and we can understand why. Perl is a dinosaur among mainstream programming languages. It lacks the glamour that other, relatively younger languages have. There is also a common misconception in the programming world that modern software engineering practices cannot be followed with a language like Perl. In this post, we hope to debunk that myth. We want to give you a glimpse of the developer experience (DX) here at Semantics3 where we write a lot of Perl code but still manage to employ the latest engineering best-practices. We would like to highlight that we are able to do so with the help of a tool-chain written entirely in Perl.
How to Deploy Software
https://zachholman.com/posts/deploying-software, posted 2016 by peter in automation continuousdelivery deployment opinion
This is a long — sorry not sorry! — written piece specifically about the high-level aspects of deployment: collaboration, safety, and pace. There's plenty to be said for the low-level aspects as well, but those are harder to generalize across languages and, to be honest, a lot closer to being solved than the high-level process aspects. I love talking about how teams work together, and deployment is one of the most critical parts of working with other people. I think it's worth your time to evaluate how your team is faring, from time to time.
Nest Reminds Customers That Ownership Isn't What It Used to Be | Electronic Frontier Foundation
https://www.eff.org/deeplinks/2016/04/nest-reminds-customers-ownership-isnt-what-it-used-be, posted 2016 by peter in automation crapification opensource opinion
Nest Labs, a home automation company acquired by Google in 2014, will disable some of its customers' home automation control devices in May. This move is causing quite a stir among people who purchased the $300 Revolv Hub devices—customers who reasonably expected that the promised "lifetime" of updates would enable the hardware they paid for to actually work, only to discover the manufacturer can turn their device into a useless brick when it so chooses.
CI as Code Part I: Stateless Jenkins Deployments Using Docker | zwischenzugs
https://zwischenzugs.wordpress.com/2016/01/24/ci-as-code-stateless-jenkins-deployments-using-docker/, posted 2016 by peter in automation continuousdelivery howto toread virtualization
I don't know about you, but I've always been uncomfortable with Jenkins' apparent statefulness. You set up your Jenkins server, configure it exactly as you want it, then DON'T TOUCH IT.
Fortunately I now have a solution. With a combination of Docker, Python's Jenkins API modules, the Jenkins job builder Python module, and some orchestration using docker-compose, I can reproduce my Jenkins state at will from code and run it in isolated environments, improving in iterable, track-able steps.
https://github.com/a-r-d/Bellman-Form-BTCe-Arbitrager, posted 2015 by peter in automation cryptocurrency opensource trading
This is a simple Bellman-Ford bot that uses the negative cycle detection feature of the algorithm to find favorable currency trades to make in forex markets (in this case we are targeting bitcoin/fiat/scryptcoin markets on btc-e and other exchanges).
Typically the trades the bot finds are less than 0.5% profit, will take 3 steps, and must be filled quickly to be profitable.


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But there is an argument to be made for shortening the. The key word in Torre's statement, though, is "time." Baseball games are too damn long. That's the thesis, and there's evidence to support it.
Persuasive meaning you can make a case for whatever your thesis is. And your thesis has to be buttressed with all manner of things. It can be documents; it can be interviews; it can be the person's.
Video game movies have never been good — one could make an argument for Silent Hill. while Donkey Kong was featured in this summer's Pixels (which is basically a thesis statement on why we.
In an argumentative essay, your goal is to convince the reader to align with your viewpoint on an issue. You may also be using your words to compel the reader.
Math taught well by our definition (that is, math not focused on memorizing answers) offers a guide for teaching writing because a word problem, like an essay, is an argument. we ask students to.
Image Comparison Research Paper Dr. Nada Jabado, Paediatric Hemato-Oncologist and Senior Scientist in the Child Health and Human Development Program at the. Synonyms for comparison at Thesaurus.com with free online thesaurus, antonyms, and definitions. Find descriptive alternatives for comparison. Image · PPPs used in tracking SDG progress. January 15, 2019 — Blog looks at use. Overview; History; Governance; Regions; Data; Methodology; Research.
A thesis statement expresses the main idea of essay. The best way to look at it is this: if you could boil your entire essay down to one sentence and still get your message across, you've got a.
The last sentence of the first paragraph is essential: the thesis statement contains your main argument in one sentence, providing a summary of your supporting evidence. The structure of the thesis.
Mar 15, 2017 · For example, if your thesis statement subjectively states that there were not that many soldiers killed during the WWII, this fact can be easily refuted by the official statistical data. And if you say that the WWII was the most disastrous war of all, it is a debatable topic.
In a statement following the announcement. of financial experience." Hobson's argument was that imperialism is caused by.
The right thesis position. A statement always belongs to the beginning of your persuasive speech because it's a strong sentence that tells the audience what you will discuss. Most instructors have their unique preferences for a specific position of a thesis, but the rule of thumb is the end of an opening.
If you want a fully laid out argument for why oil prices might head down, read and watch Tony Seba's thesis. I don't agree with Seba and. President Trump's next statement makes it clear to me what.
Examples of Argumentative Language. Below are examples of signposts that are used in argumentative essays. Signposts enable the. Thesis statement/claim.
Argumentative Essay Examples. In an argumentative essay, the writer is taking a position, or a "side," related to a topic about which opinions could vary.The position of the writer is established in the thesis statement.Then the essay presents reasons, evidence, and examples that support the writer's position.
Composition classes stress the role of the thesis statement because it is the. There are two major types of thesis statements: explanatory and argumentative.
Speech that damages democratic processes will find itself unprotected by the self-government thesis. And if the autonomy argument is compelling we will not want to protect speech that undermines this.
What I believe to be the most important result is that the derivation of the equation contains a purely mathematical proof of the existence. Church-Turing thesis in that Descartes must be able to.
Argumentative thesis statement. Examples of thesis statement for an Argumentative essay.
Again, you must decide which side of the argument you are for, public or private. Yet, there are many pros and cons to this debate. Some pros for public.
Law school applicants using a thesis for a personal statement are required to present coherent evidence to support the ideas being explored. (Getty Images) The goal of the personal statement in a law.
Step 4: Summarizing with Conclusion The purpose of a concluding part is twofold — it serves as a summary of your main argument and a reaffirmation of its relevance. Start by restating your thesis.
According to The Holt Handbook (HH), an argumentative essay takes a. If you can create an antithesis, your thesis statement takes a stand, if not, then your.
Jul 21, 2017. Analytical and argumentative essays require different approaches. When you're asked to analyze something, for example a piece of literature,
How to Identify Nonargumentative Uses of Language. This lesson reviews more about: The differences in informational versus argumentative The type of text when you can see all perspectives, not just.
This is a broad topic, which you can take in many directions. So, you need to be selective. In other words, you need a good thesis statement that will drive your.
Argumentative Thesis Statements In an argumentative paper, you are making a claim about a topic and justifying this claim with reasons and evidence. This claim could be an opinion, a policy proposal, an evaluation, a cause-and-effect statement, or an interpretation.
Although the commentary promises to use "biblical teaching" to prove its thesis, his cited texts demonstrate the. So what is his biblical argument? "At no time did Jesus use deadly force," writes.
State Your Thesis. The thesis is the very important essence of an argumentative essay. In a clear and single sentence it sums up what point you are trying to make. The thesis statement should be.
Suggestions for Developing Argumentative Essays 1. Select an arguable topic, Take a position on your topic, and form a thesis statement. Your thesis must be.
After reading your thesis statement, the reader should think, "This essay is going to try to convince me of something. I'm not convinced yet, but I'm interested to.
Purdue Owl Lecture Citation Scholarly Articles On Immunizations The article is titled "Weaponized Health Communications: Twitter Bots and Russian Trolls Amplify the Vaccine Debate. Dr. Amesh Adalja, an infectious diseases physician and senior scholar at the. In fact, he had published an article titled, "James Comey Did the Right Thing. In our chats, he made false claims about. For example, the perception
Apr 10, 2019. Step-by-step instructions on how to write an argumentative essay, including how to craft an enticing introduction, how to write a thesis statement.
Oct 17, 2013 · What Is a Thesis Statement. A thesis is a specific statement that is usually placed in the introductory part of an essay, research paper, dissertation etc., and which is explained in subsequent body paragraphs. A thesis of an article, report or other scientific work is a set of certain statements.
argument | thesis |. is that argument is a fact or statement used to support a proposition; a reason while thesis is a statement supported by arguments.
Thesis statements, introductions, conclusions, and quotations. While you might use the same sources in writing an argumentative essay as your classmate.
A good argumentative essay starts with a line to catch the reader's attention. The thesis statement should assert a position on a particular issue — one that a.
Most students are capable of solid expository writing. It's their skill with persuasive writing that's the problem. Specifically, they're weak at writing a thesis statement that can be argued. I spend.
Writing Thesis Statements for Argumentative Essays Introduction University writing often requires students to use persuasion: they need to convince readers of a.
A thesis statement expresses the main idea of essay. The best way to look at it is this: if you could boil your entire essay down to one sentence and still get your message across, you've got a.
Tips of writing the best euthanasia argumentative essay and guidelines of. The thesis statement of every essay is a summary of the main points of the essay.
What is an Argumentative Essay? Argumentative essays are a common assignment given by educators and are an exciting opportunity for you to practice the valuable skill of persuasion. Argumentative essays are, at their core, pieces of writing that aim to convince the reader of the writer's own opinion.
. a thesis statement. Once you have an idea of what you want to say, and have some grasp of what others have said, you need to make your ideas more concrete by coming up with a thesis sentence(s). A.
After much soul searching and research on the issue, I got the idea that writing a business intelligence report can, and even should, be viewed in the context of writing a thesis. Consider the.
Noun (theses) A statement supported by arguments. A written essay, especially one submitted for a university degree. * Goldsmith ; I told them of the grave, becoming, and sublime deportment they should assume upon this mystical occasion, and read them two homilies and a thesis of my own composing,
What is an Argumentative or Position Paper? In this type of assignment, (See the Writing Center's handout on thesis statements for more help with this stage!)
The rest of the paper is made of the supporting points. "What is a thesis statement?" is the initial question to answer. A thesis statement is the main argument or point that is set out to be proven using tools such as logical/emotional reasoning. It is the root from where everything grows.
Most of us have experience with arguing, but an argumentative essay is quite. An effective thesis statement is an important foundational element of any essay,
A thesis statement expresses the main idea of essay. The best way to look at it is this: if you could boil your entire essay down to one sentence and still get your message across, you've got a.
Jan 09, 2017 · The thesis statement tells your view of the presented problem, directly and explicitly, in a sentence or two. Your paper then builds an academic argument to support the thesis. Thesis statements examples based on the type of the paper you want to write. When you're developing your thesis statement…
Thesis statement generator for argumentative essay sample. Writing an expository essay thesis statement amount of homework for students conway complex analysis solutions homework i need to write an essay faster, dissertation structure uk template jane eyre research essay topics. Plan small.
Noun (theses) A statement supported by arguments. A written essay, especially one submitted for a university degree. * Goldsmith ; I told them of the grave, becoming, and sublime deportment they should assume upon this mystical occasion, and read them two homilies and a thesis of my own composing,
Have a well defined thesis. Start with a thesis statement/research question/ statement of intent. Make sure you answer your question or do what you say you set.
Apr 10, 2019 · Step-by-step instructions on how to write an argumentative essay, including how to craft an enticing introduction, how to write a thesis statement, and how to outline your essay. Classical, Rogerian, and Toulmin argument strategies will also be discussed.
His central thesis is that by allowing ourselves to get swept up in. tech narrative that civic data can provide a useful.
I'll be curating submissions for the best videos as measured by a compelling macro thesis, a bold mission and a persuasive founder. Your answer should be in the form of a statement and be no longer.
Argumentative Thesis Overview. Are you ready to review what you have learned about argumentative thesis statements? The video below will help with that. Be sure the volume on your computer is turned up as you review information on building a strong thesis statement for your argumentative essay. <<
Korean S And Sh Sounds Linguistics If you were looking for a story about this week's Netflix releases being bland. that a dope movie title or what? It sounds. The government's policy to ban English education. policy at all," Robert Kluender, a linguistics professor at the University of California San Diego, told The Korea Times. "Is it based on any. About phonemes. an extract
Apr 10, 2011 · Autism Thesis Statement Examples Autism is a neural development disorder which inhibits the normal growth and development of a child within a social environment. An autistic person finds it difficult to socially communicate with people and can't relate to social norms in a way a normal.


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