Simultaneous monitoring of inorganic cations, amines and amino acids in human sweat by capillary electrophoresis

The determination of cationic constituents of sweat is widely recognized as a difficult analytical task due to its complex composition and minute sample volumes available for the individual analysis. Capillary electrophoresis (CE) has been evaluated as a simple routine method to measure sweat metal cations, biogenic amines, and amino acids using a sampling procedure previously developed in one of collaborative teams. The carrier electrolyte, which consisted of 10 mM 4-methylbenzylamine, 6.5 mM α-hydroxyisobutyric acid, and 2 mM 18-crown-6 at pH 4.25, allowed the separation of five cations (NH₄⁺, K⁺, Ca²⁺, Na⁺, Mg²⁺) and four amino acids (ornithine, histidine, lysine, arginine) to be completed in about 13 min with a positive polarity of the applied voltage (30 kV). By increasing the sample volume (due to employing hydrodynamic instead of hydrostatic injection mode), it was also possible to detect indirect UV signals of Zn²⁺, diethanolamine, and trithanolamine. Sweat samples were collected from the fingers and forearms of three healthy male volunteers and analyzed by CE. A good repeatability and reproducibility of peak area responses based on five intraday and three inter-day assays (average %R.S.D. less than 3.5 and 2.5, respectively) were obtained. The limits of detection were in the range of 3.2-5.8 μM for alkali and alkaline-earth cations (hydrostatic injection) and 0.27-0.79 μM for other target analytes (hydrodynamic injection). The analytical results for particular analytes were found to vary, depending on the sampling spot and individual, but in general correspond well to clinical concentration ranges.     

Potent platelet-derived growth factor-beta receptor (PDGF-betaR) inhibitors: Synthesis and structure-activity relationships of 7-[3-(cyclohexylmethyl)ureido]-3-{1-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl}quinoxalin-2(1H)-one derivatives

We found previously that 7-[3-(cyclohexylmethyl)ureido]-3-{1-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl}quinoxalin-2(1H)-one (7d-6) has considerable potency as a PDGF inhibitor. This compound showed potent inhibitory activity in a PDGF-induced CPA (Cell Proliferation Assay) and APA (Auto-Phosphorylation Assay) (IC50 = 0.05 micromol/l in CPA, 0.03 micromol/l in APA). Therefore, we tried to develop a novel and effective PDGF-betaR inhibitor by optimizing a series of its derivatives. We found that trifluoroacetic acid (TFA)-catalyzed coupling of pyrrolo[2,3-b]pyridines with quinoxalin-2-ones proceeded efficiently under mild oxidation condition with manganese(IV) oxide (MnO2) in situ, so this method was applied to prepare a series of derivatives. Results of in vitro screening of newly synthesized derivatives identified compound 7d-9 as having potent (IC50 = 0.014 micromol/l in CPA, 0.007 micromol/l in APA) and selective [IC50 values against vascular endothelial growth factor receptor 2 (VEGFR2, kinase domain region, KDR), epidermal growth factor receptor (EGFR), c-Met (hepatocyte growth factor receptor) and insulin growth factor I receptor (IGF-IR)/IC50 against PDGFR were each >1000] inhibitory activity. Moreover, in this series of derivatives, 7b-2 showed potent inhibitory activity toward both PDGF- and VEGF-induced signaling (PDGFR: IC50 = 0.004 micromol/l in CPA, 0.0008 micromol/l in APA, KDR: IC50 = 0.008 micromol/l in APA). Herein we report a new and convenient synthetic method for this series of derivatives and its SAR study.     

A novel hybrid mode of sample injection to enhance CZE sensitivity for simultaneous determination of a pyridine-triphenylborane anti-fouling agent and its degradation products

We developed a novel hybrid sample injection mode (HSIM) that presents the combination of electrokinetic injection and vacuum injection to enhance detection sensitivity in CZE. Samples were introduced using both vacuum and electrokinetic injections simultaneously, with a water plug injected into the capillary prior to sample introduction (i.e. similarly to field-amplified sample injection, FASI). Using a sample mixture containing an anti-fouling agent applied to ship hulls, pyridine-triphenylborane and its degradation products (diphenylborinic acid, phenylboronic acid, and phenol) dissolved in ACN, the length of water plug, time, and voltage for sample introduction were optimized. The signal intensity (peak height) was found to be up to a 30-fold increased using HSIM by applying 4 kV for 4 s at the inlet end of the capillary as the cathode with supplementary vacuum in comparison with only vacuum injection for 4 s. The LODs (at a S/N of 3) for pyridine-triphenylborane, diphenylborinic acid, phenylboronic acid, and phenol were 0.88, 1.0, 21, and 23 μg/L, respectively. At the level of 0.04 mg/L, the RSDs (n=4, intra-day) for the above analytes were in the ranges of 1.9-11, 4.3-9.2, and 0.34-0.66% for peak area, peak height, and migration time, respectively. The HSIM is a simple and promising procedure useful for enhancing the sensitivity for both low-and high-mobility ions in CZE.     

Diffusion-weighted echo-planar imaging of the head and neck using 3-T MRI: Investigation into the usefulness of liquid perfluorocarbon pads and choice of optimal fat suppression method

Purpose

To investigate whether image quality can be improved using liquid perfluorocarbon pads (Sat Pad) and clarify the optimal fat-suppression method among chemical shift selective (CHESS), water excitation (WEX), and short TI inversion recovery (STIR) methods in diffusion-weighted imaging (DWI) of the head and neck using 3-T magnetic resonance imaging. Correlations between results of visual inspection and quantitative analysis were also examined.

Material and Methods

This study was approved by our Institutional Review Board and informed consent was waived. DWI was performed on 25 subjects with/without Sat Pad and using three fat-suppression methods (6 patterns). Image quality was evaluated visually (4-point scales and lesion-depiction capability) and by quantitative analysis (signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR)). Two-way repeated-measures analysis of variance (ANOVA) was used to detect significant differences in scores of visual evaluation, SNR, and CNR.

Results

Mean visual evaluation scores were significantly higher with Sat Pad using STIR than without Sat Pad for all fat-suppression methods (P < 0.05). DWI with Sat Pad using STIR tended to be useful for depicting lesions. DWI using STIR showed reduced W-SNR (W: whole area of depicted structure) and CNR (between semispinalis capitis muscle and subcutaneous fat) due to fewer artifacts and uniform fat suppression.

Conclusion

Combining Sat Pad with STIR provides good image quality for visual inspections. When numerous artifacts are present and fat suppression is insufficient, higher SNR and CNR do not always provide good diagnostic image quality.     

离子排斥色谱用于厌氧消化处理水中脂肪族羧酸的分析（英文）

The analysis of seven aliphatic carboxylic acids(formic,acetic,propionic,iso-butyric,n-butyric,iso-valeric and n-valeric acid) in anaerobic digestion process waters for biogas production was examined by ion-exclusion chromatography with dilute acidic eluents(benzoic acid,perfluorobutyric acid(PFBA) and sulfuric acid) and non-suppressed conductivity/ultraviolet(UV) detection.The columns used were a styrene/divinylbenzene-based strongly acidic cation-exchange resin column(TSKgel SCX) and a polymethacrylate-based weakly acidic cation-exchange resin column(TSKgel Super IC-A/C).Good separation was performed on the TSKgel SCX in shorter retention times.For the TSKgel Super IC-A/C,peak shape of the acids was sharp and symmetrical in spite of longer retention times.In addition,the mutual separation of the acids was good except for iso-and n-butyric acids.The better separation and good detection was achieved by using the two columns(TSKgel SCX and TSKgel Super IC-A/C connected in series),lower concentrations of PFBA and sulfuric acid as eluents,non-suppressed conductivity detection and UV detection at 210 nm.This analysis was applied to anaerobic digestion process waters.The chromatograms with conductivity detection were relatively simpler compared with those of UV detection.The use of two columns with different selectivities for the aliphatic carboxylic acids and the two detection modes was effective for the determination and identification of the analytes in anaerobic digestion process waters containing complex matrices.     

Investigation of the pH gradient formation and cathodic drift in microchip isoelectric focusing with imaged UV detection

This paper reports the protein analysis by using microchip IEF carried on an automated chip system. We herein focused on two important topics of microchip IEF, the pH gradient and cathodic drift. The computer simulation clarified that the EOF could delay the establishment of pH gradient and move the carrier ampholytes (CAs) to cathode, which probably caused a cathodic drift to happen. After focusing, the peak positions of components in a calibration kit with broad pI were plotted against their pI values to know the actual pH gradient in a microchannel varying time. It was found that the formed pH gradient was stable, not decayed after readily steady state, and migrated to cathode at a rate of 10.0 μm/s that determined by the experimental conditions such as chip material, internal surface coating and field strength. The theoretical pH gradient was parallel with the actual pH gradient, which was demonstrated in two types of microchip with different channel lengths. No compression of pH gradient was observed when 2% w/v hydroxypropyl methyl cellulose was added in sample and electrolytes. The effect of CAs concentration on current and cathodic drift was also explored. With the current automatic chip system, the calculated peak capacity was 23–48, and the minimal pI difference was 0.20–0.42 for the used single channel microchip with the effective length of 40.5 mm. The LOD for the analysis of CA‐I and CA‐II was around 0.32 μg/mL by using normal imaged UV detection, the detected amount is ca. 0.07 ng.