Determination of methyl ethyl ketone and its metabolites in urine using capillary gas chromatography

A method is described for the determination of the concentration of methyl ethyl ketone and its metabolites: 2-butanol, 3-hydroxy-2-butanone and the meso- and d,l-isomers of 2,3-butanediol in urine. The analytes were isolated from urine by solid-phase extraction and analysed by capillary gas chromatography. The recovery rates were 50-70% for the 2,3-butanediol isomers and 88-96% for the other analytes. The precision of the method ranged from 5 to 12% (S.D.%). The detection limit was 1.0 and 1.4 mg/l for meso- and d,l-2,3-butanediol, respectively, and ranged from 0.1 to 0.15 mg/l for the other analytes.     

Determination of lead in the urine of exposed and unexposed adults by extraction and flow-injection/atomic absorption spectrometry

Lead is extracted from urine with ammonium pyrrolidien dithiocrbamate into methyl isobutyl ketone, and 20 μl of the extract is injected into a water carrier stream in a flow-injection/atomic absorption system. The procedure is simple, quick, accurate and reproducible. Recoveries are 96–105%. The mean urine lead contents of 20 unexposed and 20 printing-press lead exposed workers was found to be 39.0 ± 8.3 and 71.7 ± 26.3 μg Pb l^?1, respectively.     

Rapid high-performance liquid chromatographic determination of urinary N-(1-methylethyl)-N'-phenyl-1,4-benzenediamine in workers exposed to aromatic amines.

A procedure has been developed for determining N-(1-methylethyl)-N'-phenyl-1,4-benzenediamine in urine by using high-performance liquid chromatography. The method uses chloroform extraction for partial clean-up of the urine sample. The separation is carried out on a reversed-phase column using 65 mmol/l aqueous ammonium acetate in acetonitrile (30:70, v/v) as the mobile phase. The column effluent is monitored at 290 nm with an ultraviolet detector. The analyte is separated from other normal urine constituents in less than 4 min. Peak height and concentration are linearly related. Coefficients of variation assessed for within-day reproducibility were 5.9 and 3.7% at concentrations of 22.3 and 92.1 micrograms/l, respectively. The mean analytical recovery from urine samples spiked with known amounts of amine was 89.7 +/- 6.8%. The request of only a small volume of urine and the simple pre-treatment procedure makes it suitable for the routine monitoring of the exposure of rubber vulcanization workers to aromatic amines.     

Determination of thiazolidine-4-carboxylates in urine by chloroformate derivatization and gas chromatography-electron impact mass spectrometry

The derivatization method of thiazolidine-4-carboxylic acid (TZCA) and methyl-thiazolidine-4-carboxylic acid (Me-TZCA) in urine with alcohol/chloroformate was achieved. TZCA and Me-TZCA were derivatized in one step in urine with ethyl chloroformate in 1 min at room temperature. The derivatives of TZCA and Me-TZCA had very good chromatographic properties and offered very sensitive response for gas chromatography-electron impact ionization-mass spectrometry (GC-EI-MS). On the basis of derivatization, the method for simultaneous determination of TZCA and Me-TZCA in human urine was developed. Deuterated Me-TZCA (Me-TZCA-d(4)) was synthesized as the internal standard (IS) for the analysis of urine samples. TZCA and Me-TZCA were derivatized and extracted from urine at pH 9.5 with toluene, and then the dried extract was dissolved with 100 microl ethyl acetate and injected in GC/MS system. The recoveries of TZCA and Me-TZCA were about 102 and 103%, respectively, at the concentration of 0.05 mg/l. The method detection limits (MDL) were 1.0 and 0.5 microg/l, respectively, for TZCA and Me-TZCA in 1 ml human urine. The coefficients of variation of TZCA and Me-TZCA were less than 6% at the concentrations of 0.05 and 0.2 mg/l, respectively. To assess the formation of TZCA during inhalation with formaldehyde (FA) (about 3.1 and 38.1 ppm FA in air), urine samples from rats were taken during 3 days after initiation of treatment. The mean amount of TZCA determined was 0.07 mg/l in control group and 0.18 mg/l during treatment with 3.1 ppm. The TZCA levels increased up to about 1.01 mg/l during treatment with 38.1 ppm. It is planned to study whether urinary TZCA can be used as an indicator in the biological monitoring of exposure to FA.     

High performance liquid chromatographic determination of methyl ethyl ketone in urine as its 3-methyl-2-benzothiazolinone hydrazone derivative

Summary A HPLC method for the determination of methyl ethyl ketone (MEK) in urine after derivatization with 3-methyl-2-benzothiazolinone hydrazone is proposed. The calibration curve for the ketone was linear, ranging between 0.23–10 mg/L, with a detection limit of 0.025 mg/L. The results were compared to those obtained by GC-MS, coupled to the headspace technique. MEK derivatization and the derivative purification processes were verified with respect to the main variables such as reaction temperature, reagent concentration, probable interferences and enrichment phase. The method is simple and reliable and shows a good sensitivity.     

Stereospecific high-performance liquid chromatographic assay of acebutolol in human plasma and urine

A sensitive high-performance liquid chromatographic technique is described for the separation of R- and S-acebutolol in human plasma and urine. The procedure involves derivatization with the chiral reagent S-(+)-1-(1-naphthyl)ethyl isocyanate. The resulting diastereoisomers are quantified using normal-phase high-performance liquid chromatography with fluorescence detection (220/389 nm). Virtual baseline separation, free from interference, with achieved (resolution factor = 1.45). Excellent linearity (r greater than 0.998) was observed throughout the range 10-500 ng/l and 2-100 mg/l in plasma and urine, respectively. Inter-assay variability was less than 5% for each enantiomer at concentrations of 10 ng/ml. This method is applicable for the determination of the pharmacokinetics, in man, of acebutolol enantiomers in plasma and urine.     

Concentration profiles of cocaine, pyrolytic methyl ecgonidine and thirteen metabolites in human blood and urine: determination by gas chromatography-mass spectrometry

When cocaine is smoked, a pyrolytic product, methyl ecgonidine (anhydroecgonine methyl ester), is also consumed with the cocaine. The amount of methyl ecgonidine formed depends on the pyrolytic conditions and composition of the illicit cocaine. This procedure describes detection of cocaine and 10 metabolites--cocaethylene, nor-cocaine, nor-cocaethylene, methyl ecgonine, ethyl ecgonine, benzoylecgonine, nor-benzoylecgonine, m-hydroxybenzoylecgonine, p-hydroxybenzoylecgonine and ecgonine--in blood and urine. In addition, the detection of pyrolytic methyl ecgonidine and three metabolites--ecgonidine (anhydroecgonine), ethyl ecgonidine (anhydroecgonine ethyl ester) and nor-ecgonidine (nor-anhydroecgonine)--are included. The newly described metabolites, ethyl ecgonidine and nor-ecgonidine, were synthesized and characterized by gas chromatography-mass spectrometry (GC-MS). All 15 compounds were extracted from 3 mL of blood or urine by solid-phase extraction and identified by a GC-MS method. The overall recoveries were 49% for methyl ecgonine, 35% for ethyl ecgonine, 29% for ecgonine and more than 83% for all other drugs. The limits of detection were between 0.5 and 4.0 ng/mL except for ecgonine, which was 16 ng/mL. Linearity for each analyte was established and in all cases correlation coefficients were 0.9985-1.0000. The procedure was applied to examine the concentration profiles of analytes of interest in post-mortem (PM) blood and urine, and in urine collected from living individuals (LV). These specimens previously were shown to be positive for the cocaine metabolite, benzoylecgonine. Ecgonidine, the major metabolite of methyl ecgonidine, was present in 77% of PM and 88% of the LV specimens, indicating smoking as the major route of cocaine administration. The new pyrolytic metabolites, ethyl ecgonidine and nor-ecgonidine, were present in smaller amounts. The urine concentrations of nor-ecgonidine were 0-163 ng/mL in LV and 0-75 ng/mL in PM specimens. Ethyl ecgonidine was found only in PM urine at concentrations 0-39 ng/mL. Ethanol-related cocaine metabolites, ethyl ecgonine or cocaethylene, were present in 69% of PM and 53% of cocaine-positive LV specimens, implying alcohol consumption with cocaine use. The four major metabolites of cocaine--benzoylecgonine, ecgonine, nor-benzoylecgonine and methyl ecgonine--constituted approximately 88 and 97% of all metabolites in PM and LV specimens, respectively. The concentrations of nor-cocaine and nor-cocaethylene were consistently the lowest of all cocaine metabolites. At benzoylecgonine concentrations below 100 ng/mL, ecgonine was present at the highest concentrations. In 20 urine specimens, benzoylecgonine and ecgonine median concentrations (range) were 54 (0-47) and 418 ng/mL (95-684), respectively. Therefore, detection of ecgonine is advantageous when benzoylecgonine concentrations are below 100 ng/mL.     

The effect of water on the rate and direction of the oxidation of organic substances

The effect of the polarity of the medium on the rate and direction of oxidation processes has been studied. It has been shown that the rate of oxidation of methyl ethyl ketone in aqueous solutions diminishes with increase in the water concentration. A considerably greater reduction in the rate of decomposition of the RO ₂ ^. radical is observed than in the rate of the interaction of this radical with methyl ethyl ketone, which results in an increase in the selectivity of the oxidation process. When methyl ethyl ketone is diluted with water in the molar ratio C₄H₈O H₂O=120, the quantity of acetic acid in the reaction products amounts to 98–99% of the methyl ethyl ketone consumed. The suggestion is made that the high selectivity of the oxidation of methyl ethyl ketone in aqueous solutions is due to the effect of the dielectric constant of the medium on the ratio between the two parallel reactions of the peroxide radical RO ₂ ^. .     

Significance of solvation in the effectiveness of inhibitor action

It is shown that the effectiveness of inhibitors in the oxidation of methyl ethyl ketone is significantly less than in a hydrocarbon medium; this is due to the formation of hydrogen bonds between ketone and inhibitor. The elementary rate constants of the interaction of the RO ₂ ^. radical of methyl ethyl ketone with -naphthol and phenol derivatives were measured. In aqueous solutions of methyl ethyl ketone, the k _i ^exp value for all inhibitors drops with increasing H₂O concentration. The interaction constants of solvated RO ₂ ^. ...H₂O peroxide radicals with the inhibitor and the equilibrium constants for the formation of H-bonds between inhibitor and water were studied. The k _i ^exp with hydroquinone in the oxidation of C₄H₈O in acetic acid were measured.     

The determination of antimony in natural waters with particular reference to sea water

A procedure is described for the determination of antimony in natural waters at concentrations down to 0.1 μg/l or less. The element is concentrated by coprecipitation with hydrous manganese dioxide (produced by the reaction of permanganate with ethanol). It is separated from manganese, iron and interfering elements by extraction from 5 M sulphuric acid, 0.01 M with respect to iodide, using methyl isobutyl ketone. After back-extraction with 0.4 M hydrochloric acid, it is determined photometrically using rhodamine B. The overall chemical yield of the process is measured radiochemically and amounts to ca. 80%. Sea water samples from the Irish Sea were found to contain 0.13–0.40 μg Sb/l.     

High-performance liquid chromatographic determination of iothalamic acid in human plasma and urine.

A simple, rapid and sensitive method for the determination of iothalamic acid (IA) in both plasma and urine is reported. After extraction with ethyl acetate, IA was determined by strong anion-exchange high-performance liquid chromatography with ultraviolet detection at 254 nm. The lower limit of detection was 0.5 micrograms/ml. The average recovery was 73 and 57% from plasma and urine, respectively. Linearity was found over the investigated concentration range (up to 500 micrograms/ml for plasma and up to 10.0 mg/ml for urine). The reproducibility of the technique was good (coefficient of variation less than 6%) as was the precision and accuracy (coefficient of variation less than 2.5%). No interference from endogenous substances or any of the common drugs tested was found.     

Determination of Phenols and Cresols in Urine by Gas Chromatography

Abstract

A rapid, simple, and sensitive method was developed for the determination of three cresol isomers and phenol in urine. The sample was spiked with m-chlorophenol, as internal standard, and hydrolyzed with sulfuric acid at ambient temperature. The hydrolyzed solution was saturated with ammonium sulfate and extracted with ethyl acetate. The extract was injected without derivatization into a gas chromatograph fitted with a flame ionization detector and a glass column packed with 0.1% SP 1000 on Carbopack C. All four analytes, which eluted within 13 minutes, were quantified from standard curves.

The method was evaluated by analysis of spiked urine pools from laboratory workers and samples of urine from workers in a coal gasification plant.     

Coulometric generation of hydrogen ions by oxidation of mercury in methyl ethyl ketone, methyl isobutyl ketone and cyclohexanone

Mercury(II)-chloride reacts with anhydrous methyl ethyl ketone, methyl isobutyl ketone and cyclohexanone in a precise stoichiometry ratio (1:2), and weakly ionized compounds of mercury with ketones are formed and equivalent quantity of HCl is released. The application of a mercury anode for the quantitative generation of H⁺ ions in 0.25 M sodium perchlorate in anhydrous methyl ethyl ketone, methyl isobutyl ketone and cyclohexanone has been investigated. Current/potentials curves for the solvents, titrated bases, indicator and mercury showed that in these solvents mercury is oxidized at potentials much more negative than those for the titrated bases and other components present in the solution. The protons generated in this way have been used for the titration of some organic bases, with either visual or potentiometric end-point detection. The oxidation of mercury in methyl ethyl ketone, methyl isobutyl ketone and cyclohexanone and the reaction of mercury ions with these solvents have been found to proceed with 100% current efficiency.     

Evaluation of a radioimmunoassay of alpha 1-microglobulin (alpha 1-m) with simplified procedures

A newly established double antibody radioimmunoassay (RIA) was fundamentally and clinically evaluated. Original procedures were partially modified as follows: Sample volume for serum and urine was changed to 25 microliters, and thus 200 mg/l of alpha 1-m standard was prepared using 50 microliters of original standard solution (100 mg/l). The results were satisfactory in sensitivity (0.3 mg/l obtained from -2SD method), intra-assay precision with its coefficient variation (CV) ranging from 3.0 to 7.4%, interassay precision with its CV ranging from 3.0 to 10.7%, and recovery with the mean value of 102.4% in serum and 108.2% in urine respectively. There were no changes about alpha 1-m value between diluted (2 times) and undiluted with high concentration samples. Normal levels of alpha 1-m were less than 25 mg/l in serum and less than 10 mg/l in urine. The present results indicate that the determination of alpha 1-m could be very simple and useful for the most sensitive screening test for the evaluation of renal function.     

Gas-liquid chromatographic method for the measurement of methyl urea in blood and urine.

A technique for the measurement of methyl urea in biological fluids is described based upon gas-liquid chromatography of its trifluoroacetyl derivative. The method requires 10 ml of either blood or urine and is capable of measuring methyl urea at concentrations of less than 5 mg/l.     

顶空–气相色谱/质谱法测定餐垫纸中16种挥发性有机物残留

采用顶空–气相色谱/质谱法同时测定餐垫纸中16种挥发性有机化合物(无水乙醇、异丙醇、正丁醇、丙酮、丁酮、己醛乙酸乙酯、乙酸异丙酯、乙酸丁酯、丙二醇甲醚、乙酸正丙酯、苯、甲苯、间二甲苯、对二甲苯、邻二甲苯)的含量。采用恒流模式,流速设定为2.5 mL/min,程序升温,16种挥发性有机化合物可以很好地分离。各成分工作曲线的相关系数r2均大于0.999 5,加标回收率在86.6%~104.8%之间,测定结果的相对标准偏差均不大于6.9%(n=6);各成分的检出限均低于0.01 mg/m2。该法具有进样简单,分离效果好,精密度高等特点,能够满足餐垫纸中挥发性有机物检测要求。     

钛硅分子筛/H2O2 体系催化合成丁酮肟的反应行为

 钛硅分子筛/H2O2 体系催化酮氨肟化反应合成肟的过程中, 产物肟的反应行为直接影响氨肟化过程的反应效率和操作稳定性. 在 Ti-MWW/H2O2 体系催化丁酮氨肟化反应中研究了产物丁酮肟的反应行为, 发现 Ti-MWW 和 H2O2 的共同作用促使丁酮肟发生水解, 生成丁酮和氮氧化物, 使得反应体系从碱性变成了酸性. TS-1/H2O2 体系及产物环己酮肟均存在类似的现象.     

Amberlyst-15催化丁酮氧化制备过氧化甲乙酮

作为一种新型绿色催化剂,离子交换树脂能够代替某些有机反应中的传统催化剂。本文以离子交换树脂Amberlyst-15为催化剂,由30%的双氧水、丁酮和稀释剂邻苯二甲酸二丁酯合成过氧化甲乙酮。在双氧水与丁酮的摩尔比为1.0,离子交换树脂与丁酮的质量比为0.06,反应温度为27℃,反应时间55 min的条件下,过氧化甲乙酮的产率接近86%,活性氧含量为12.9%。Amberlyst-15在循环使用8次后,依然保持良好的稳定性。研究表明:Amberlyst-15用于过氧化甲乙酮的制备具有催化活性高、可重复使用、清洁无腐蚀等优点,为工业环保生产有机过氧化物提供了新途径。     

Determination of 8-hydroxy-2'-deoxyguanosine in untreated urine by capillary electrophoresis with UV detection

A capillary electrophoresis method with UV detection was developed for the determination of 8-hydroxy-2'-deoxyguanosine (8-OHdG) in untreated urine samples. The calibration graph for 8-OHdG in urine is linear in the concentration range 10-500 mg/l. and the detection limit is 5 mg/l (17 microM). 8-OHdG was determined in urine from oncological patients treated by radiation therapy. Its concentrations relative to creatinine were found to be in the range 10-47 microg 8-OHdG/l mg creatinine (4-19 micromol 8-OHdG/mmol creatinine). The overall time of the analysis of a urine sample was less than 15 min.     

Ruthenium-Lewis acid catalyzed asymmetric Diels-Alder reactions between dienes and alpha,beta-unsaturated ketones

The complex [Ru(Cp)(R,R-BIPHOP-F)(acetone)][SbF(6)], (R,R)-1 a, was used as catalyst for asymmetric Diels-Alder reactions between dienes (cyclopentadiene, methylcyclopentadiene, isoprene, 2,3-dimethylbutadiene) and alpha,beta-unsaturated ketones (methyl vinyl ketone (MVK), ethyl vinyl ketone, divinyl ketone, alpha-bromovinyl methyl ketone and alpha-chlorovinyl methyl ketone). The cycloaddition products were obtained in yields of 50-90 % and with enantioselectivities up to 96 % ee. Ethyl vinyl ketone, divinyl ketone and the halogenated vinyl ketones worked best and their reactions with acyclic dienes consistently provided products with >90 % ee. alpha-Chlorovinyl methyl ketone performed better than alpha-bromovinyl methyl ketone. The reaction also provided a [4.3.1]bicyclic ring system in 95 % ee through an intramolecular cycloaddition reaction. Crystal structure determinations of [Ru(Cp)((S,S)-BIPHOP-F)(mvk)][SbF(6)], (S,S)-1 b, and [Ru(Cp)((R,R)-Me(4)BIPHOP-F)(acrolein)][SbF(6)], (R,R)-2 b, provided the basis for a rationalization of the asymmetric induction.