Determination of estrone and 17 beta-estradiol in human hair by gas chromatography-mass spectrometry

An efficient procedure is described for the determination of estrone and 17 beta-estradiol in hair by gas chromatography-mass spectrometry (GC-MS). The method involves alkyloxycarbonylation with isobutyl chloroformate (isoBCF) of phenolic hydroxy groups after alkaline digestion of hair samples. The resulting isobutyloxycarbonyl derivatives of estrone and 17 beta-estradiol are extracted with hexane and subjected to chlorodifluoroacetyl derivatization in order to protect the remaining alcoholic hydroxy groups. When GC-MS with selected ion monitoring (SIM) was used, the quantitative ions were at m/z 270 and 384 in the electron ionization mass spectra for estrone and 17 beta-estradiol, respectively. The detection limits for SIM of the steroids were 1 and 2 pg, respectively, and the SIM responses were linear with correlation coefficients varying from 0.991 to 0.994 in the concentration range 0.2-4.0 ng g-1 for the estrogens studied. The detection of estrone and 17 beta-estradiol in hair samples was possible in the concentration range of 0.24-1.30 ng g-1. The concentrations of the two estrogens detected were different in male and female hair samples.     

Complexometric determination of magnesium oxide in flyash blended cement

A complexometric titration method is proposed to determine magnesium oxide in flyash blended cement. A 0.50 g of sample was heated with hydrochloric acid for 10 min. The solution was diluted to 500 ml, and 50 ml was pipetted and heated to boiling with 2.5 ml of 5% ammonium oxalate solution. The solution was then made alkaline by ammonium hydroxide. The suspension was cooled and filtered. The filtrate was titrated by standard 0.002M EDTA solution. The concentration of MgO in sample was calculated. The flyash content of the sample was determined by British Standard method and the recovery factor (f) was calculated by the equation of f = 100/(99 - 0.315 x %FA). Concentration of MgO in sample was corrected by multiplying the recovery factor with concentration initially found by EDTA titration. The precision of the method is better with more time saving than the official methods.     

A novel oxygen and/or carbon dioxide-sensitive optical transducer

A novel oxygen (O(2)) and/or carbon dioxide (CO(2))-sensitive transducer for the measurement of both gaseous O(2) and CO(2) over the concentration ranges of O(2), 0-100% and CO(2), 0-10% has been described employing a solution of 10.6 muM fluorescein (FL) and 190 muM potassium hydroxide in a solvent mixtures of 1:1 (v/v) N,N'-diethylaniline (DEA) and N,N-dimethylformamide. Increasing O(2) concentrations cause the absorbance of the solution at a wavelength of 400 nm to increase owing to a contact charge transfer reaction existing between O(2) and DEA molecules, and increasing CO(2) concentrations produce a non-linear fall in absorbance at 520 nm as the colour of FL changes from its orange dianion form to the colourless neutral, lactonic form. Both processes are independent of each other and reversible. The response to changes in O(2) concentrations is in good agreement with Beer-Lambert's law and the response to changes in CO(2) concentrations in non-linear. A fibre optic sensing system based on this solvent-dye solution has been set up for continuous and reversible determination of both gaseous O(2) and CO(2). Possible applications include environmental and physiological monitoring of O(2) over the ranges of 0-100% and CO(2), 0-10%.     

用手性高效液相色谱对β—羟基酯化合物进行对映体分离

本文利用手性ＯＤ柱在高效液相色谱上对光活性β－羟基酯类化合物进行了对映体分离，达到了较好的分离效果，并通过讨论手性ＯＤ涂附物的结构与该类化合物的作用方式，解释了不同的该类化合物的色谱分离效果。     

Dual effect of oxidative stress on NF-kappakB activation in HeLa cells

Reactive oxygen species (ROS) has been implicated as an inducer of NF-kappaB activity in numbers of cell types where exposure of cells to ROS such as H(2)O(2) leads to NF-kappaB activation. In contrast, exposure to oxidative stress in certain cell types induced reduction of tumor necrosis factor (TNF)- induced NF-kappaB activation. And various thiol-modifying agents including gold compounds and cyclopentenone prostaglandins inhibit NF-kappaB activation by blocking IkappaB kinase (IKK). To understand such conflicting effect of oxidative stress on NF- kappakB activation, HeLa cells were incubated with H(2)O(2) or diamide and TNF-induced expression of NF-kappaB reporter gene was measured. NF-kappaB activation was significantly blocked by these oxidizing agents, and the inhibition was accompanied with reduced nuclear NF-kappaB and inappropriate cytosolic IkappaB degradation. H(2)O(2) and diamide also inhibited IKK activation in HeLa and RAW 264.7 cells stimulated with TNF and lipopolysaccharide, respectively, and directly blocked IKK activity in vitro. In cells treated with H(2)O(2) alone, nuclear NF-kappaB was induced after 2 h without detectable degradation of cytosolic IkappaBalphaa or activation of IKK. Our results suggest that ROS has a dual effect on NF-kappaB activation in the same HeLa cells: it inhibits acute IKK-mediated NF-kappakB activation induced by inflammatory signals, while longer-term exposure to ROS induces NF-kappaB activity through an IKK-independent pathway.     

An organic-phase optical phenol biosensor coupling enzymatic oxidation with chemical reduction

This paper presents a recycle amplification optical biosensor to monitor phenol in hydrophobic organic solvents. Tyrosinase was first immobilised by entrapping it in a copolymer membrane of poly(vinyl alcohol)-hydroxyethyl carboxymethylcellulose doped with octadecylsilica particles. The biosensor was then constructed by co-mixing small particles of the immobilised tyrosinase with the adduct of L-ascorbic acid-poly(vinyl alcohol) (AsA-PVA) in conjunction with an optical oxygen transducer. The biosensor was characterised by its amplifying response to phenol, stable biocatalytic activity of entrapped-tyrosinase, free of interference from o-quinone polymerisation, and large water buffer capacity in hydrophobic organic solvents. The working range of the biosensor to phenol was 0.08-40 mmol dm(-3) in the flow mode. The response times (95%) of the biosensor were 4-7 min for phenol. The operational lifetime was more than 40 assays and the shelf lifetime of the biosensor was longer than 3 months. The biosensor has been successfully applied to quantify the phenol contents in some commercial ointment samples.     

Development of an indirect competitive ELISA for the determination of papaverine

Papaverine (1-(3,4-dimethoxybenzyl)-6,7-dimethoxyisoquinoline, PAP) is a member of the benzylisoquinoline sub-group of the opium alkaloids. It has been widely used for treating diseases like pulmonary arterial embolism and renal or biliary colic. In this paper, a specific conjugate of mono-demethylated papaverine-O-carboxylmethyl ether (MDMPAP-O-CME) and bovine serum albumin (BSA) was synthesized and used as the complete antigen (PAP-BSA), with which we successfully obtained a high-titer anti-PAP polyclonal antibody (pAb) by immunization of rabbits. The anti-PAP pAb showed high affinity to papaverine with an affinity constant (K_aff) of 7.3 × 10⁷ L/mol. With this antibody, we established a sensitive immunochemical method for the determination of papaverine based on indirect competitive enzyme-linked immunosorbent assay (ELISA). The optimal concentrations of the coated antigen (PAP-OVA) and purified pAb used in the ELISA were 5 and 1.2 μg/mL, respectively. The cross reactivity of other benzylisoquinoline derived substances, including 1-(3,4-dihydroxybenzyl)-7-hydroxy-6-methoxy-isoquinoline (6-methoxy-papaveroline, MPAPO), morphine (MP) and codeine (CD) were all lower than 1%. The linear range of the calibration curve was 0.1-1000 ng/mL. Normal human serum samples were spiked with known amount of papaverine and measured by the ELISA. Recoveries were between 102% and 105%. Papaverine content in a commercial papaverine hydrochloride injection sample was also determined using the established ELISA. Compared with the results given by the control experiment of HPLC, the recoveries of ELISA to detect injection samples were 102-110%. The limits of detection for synthetic serum samples and injection samples of papaverine hydrochloride were 0.25 and 0.06 ng/mL, respectively.     

Study on the oligomerization of cyclopentadiene and dicyclopentadiene to tricyclopentadiene through diels-alder reaction

Summary The oligomerization of cyclopentadiene (CPD) and its dimer, dicyclopentadiene (DCPD), to tricyclopentadiene (TCPD) through Diels-Alder reaction at temperatures between 120 and 150°C was investigated. The results show that reaction temperature, pressure and solvent influence the product yield; at 150°C up to 50% yield of TCPD was obtained in the absence of solvents. The ratios of isomers A to B in the product can be adjusted by using different solvents. The kinetics indicate that the rate is more sensitive to the concentration of CPD than to that of DCPD.     

Modeling of the Permeate Flux during Microfiltration of BSA-Adsorbed Microspheres in a Stirred Cell

A study on the variation of the permeate flux was performed in a stirred cell charged with microspheres, to investigate the effects of the stirrer speeds (300, 400, and 600 rpm) and the BSA concentration (0.1, 0.2, 0.4, and 0.8 g/L) under constant pressure. The permeate flux increased over time before the saturation point, but it began to decrease after that point. An increase of the BSA concentration and the stirrer speed resulted in the rapid increase of the permeate flux. This is contrary to the observation of the conventional filtration experiments using a stirred cell. A resistance-in-series model was employed for the modeling of the permeate flux. The cake resistance (R(c), induced by the concentration polarization of microspheres) and the fouling resistance (R(f), induced by the adsorption of BSA inside the membrane pore) must be considered simultaneously for the modeling. These modeling results were in good agreement with the experimental data. These can be applied to the special system considering both R(c) and R(f) as well as the general filtration systems using a stirred cell. Copyright 2000 Academic Press.