Rapid analytical method for the determination of 220 pesticide with their isomers by GCMS-TIC

This paper presents a cost-effective and validated multi residue modified and miniaturized method for the determination of 220 chemically different groups of pesticides and their isomers. This determination method is performed with single Quaid Gas Chromatography Mass Spectrometry -Total Ion Chromatogram GCMS-TIC. Two methods was experimented and modified with different GCMS parameters to analyses most common used pesticide and their residues in the standers solution and can be applied for real environmental samples. The results showed by single Quaid GCMS-TIC it can analyze 220 pesticides including their isomers within 49.6 min and low detection limit by using modified method 2 as described in this research. Limit of detection (LOD) was ranged from 0.78 to 14.74 ng/ml (ppb) with good separation and resolution. Limit of quantification (LOQ) was ranged between 2.34 and 44.22 ng/ml (ppb). Method 2 was more accurate, shorter, and clear separation rather than method 1. This method can be successfully applied in real environmental samples proven to be a good option for routine analysis of pesticide within the maximum residue limits (MRL) referenced to European commission especially with the most common GCMS-TIC which exists in most of labs and low income countries.     

Determination of triamterene and its main metabolite hydroxytriamterene sulfate in human urine by capillary electrophoresis using ultraviolet absorbance and laser-induced fluorescence detection

Two capillary electrophoresis methods have been developed for the direct determination of triamterene and its main metabolite hydroxytriamterene sulfate in human urine. Analytes were detected using conventional UV detection as well as laser-induced fluorescence (LIF) detection with an HeCd-laser operating at a wavelength of 325 nm. The results of both detection techniques were compared. Indeed, the limit of quantification was eightfold lower using LIF detection (50 ng/ml) in comparison to UV detection (400 ng/ml). As no interference due to endogenous urine compounds was observed, direct urine analysis was feasible. Analysis was very simple and fast-one run could be performed within less than 10 min (CE-UV method) and 2.5 min (CE-LIF method), respectively. Both assays were fully validated and applied to urine samples from a human volunteer. The results of the application of the CE-LIF method to human urine samples are presented in this publication.     

Investigation of volatile biomarkers in lung cancer blood using solid-phase microextraction and capillary gas chromatography-mass spectrometry

In the present work, solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) was developed for investigation of lung cancer volatile biomarkers. Headspace SPME conditions (fiber coating, extraction temperature and extraction time) and desorption conditions were optimized and applied to determination of volatiles in human blood. To find the biomarkers of lung cancer, investigation of volatile compounds in lung cancer blood and control was performed by using the present method. Concentrations of hexanal and heptanal in lung cancer blood were found to be much higher than those in control blood. The two molecules of hexanal and heptanal were regarded as biomarkers of lung cancer. By comparison of volatiles in breath and in blood, it is demonstrated that hexanal and heptanal in breath were originated from blood and screening of lung cancer by breath analysis be feasible. These results show that SPME/GC-MS is a simple, rapid and sensitive method very suitable for investigation of volatile disease markers in human blood.     

Determination of piretanide and furosemide in pharmaceuticals and human urine by high-performance liquid chromatography with amperometric detection

A high-performance liquid chromatographic method with electrochemical detection (ED) has been developed for the determination of two diuretics: 4-phenoxy-3-(1-pyrrolidinyl)-5-sulfamoylbenzoic acid (piretanide) and 4-chloro-2-furfurylamino-5-sulfamoylbenzoic acid (furosemide). The chromatographic separation was performed on a μBondapak C₁₈ column with a mobile phase of acetonitrile-water (40:60) containing 5 mM KH₂PO₄/K₂HPO₄ and with a flow-rate of 1 ml/min (69 bar). The temperature was optimized at 30 ± 0.2°C. The amperometric detector equipped with a glassy carbon electrode was operated at + 1200 mV versus Ag/AgCl in the direct current mode. The method was applied to the determination of these compounds in two concentration ranges (ppm and ppb), obtaining a reproducibility in terms of relative standard deviations lower than 1% for within-day and 4% for day-to-day and determination limits of 15 ppb for both compounds. Recoveries greater than 90% were obtained for spiked urine samples, using a liquid-liquid extraction method in the sample clean-up procedure. The LC-ED method was applied to commercially available pharmaceuticals (Seguril, furosemide 40 mg, and Perbilén, piretanide 6 mg) and urine samples obtained from healthy volunteers and hypertensive patients.     

Simultaneous determination of inosine, hypoxanthine, xanthine, and uric acid and the effect of metal chelators

We describe a sensitive, reproducible method for the simultaneous determination of the ATP catabolites inosine, hypoxanthine, xanthine, and uric acid in biological samples and organ perfusate using reverse-phase chromatography and multiwavelength detection at 254, 270, and 292 nm. Sample preparation includes precipitating proteins with perchloric acid, neutralizing the sample, passing the supernatant over a polyethyleneimine column, and analyzing the collected fractions by high-performance liquid chromatography. Addition of metal chelators to the perchloric acid resulted in increased values for xanthine, hypoxanthine, and uric acid. The method was sensitive (limit of detection, 0.08 nmol on column; S/N = 4) and linear over the range 0.5-30 microM. Precision and accuracy of the method were evaluated for lung tissue and lung perfusate. Coefficients of variation ranged from 2.8 to 6.1% for perfusate and from 1.7 to 12.6% for tissue. Recoveries for all compounds exceeded 90%. We applied this method to rat lung tissue, lung perfusate, and rat and human blood. Advantages of this method are simultaneous quantitation with excellent sensitivity of all compounds, simplified peak identification by using multiwavelength detection, and improved accuracy by preventing loss of compounds with metal chelators.     

Development of headspace solid-phase microextraction with on-fiber derivatization for determination of hexanal and heptanal in human blood

Hexanal and heptanal in human blood have been regarded as potential biomarkers of lung cancer. Owing to their high volatilities and activities, it is difficult to accurately measure the two biomarkers. In the current work, headspace solid-phase microextraction (HS-SPME) with on-fiber derivatization technique was developed for quantitative analysis of hexanal and heptanal in human blood. In the proposed method, the two aldehydes in blood were headspace extracted by using a poly (dimethylsiloxane)/divinylbenzene (PDMS/DVB) fiber with O-2,3,4,5,6-(pentafluorobenzyl) hydroxylamine (PFBHA) at 60 degrees C for 8 min. The aldehyde oximes formed on the fiber were desorbed and analyzed by gas chromatography-mass spectrometry (GC-MS). The method validations including detection limit, recovery and precision were studied. It was found that the method provided low detection limits of 0.006 nM for hexanal and 0.005 nM for heptanal, recoveries from 89% to 95% and R.S.D. values less than 8.5%. The present method was applied to quantitative analysis of hexanal and heptanal in normal blood and lung cancer blood. Hexanal concentrations from 7.33 to 15.23 microM and heptanal concentrations from 2.47 to 9.23 microM were found in the lung cancer blood, while both hexanal and heptanal in the control blood were lower than 0.6 microM. This further demonstrated that hexanal and heptanal might be the biomarkers of lung cancer. The experimental results showed that GC-MS and HS-SPME with on-fiber derivatization is a simple, rapid, sensitive and solvent-free method for determination of in hexanal and heptanal human blood.     

High-performance liquid chromatography with amperometric detection applied to the screening of β-blockers in human urine

A high-performance liquid chromatographic method with electrochemical detection has been developed for the determination of six β-blockers; atenolol, nadolol, timolol, metoprolol, oxprenolol, and alprenolol.The chromatographic separation was performed using a μBondapack C₁₈ column, a mobile phase of acetonitrile-water (40:60), containing 5 mM KH₂PO₄/K₂HPO₄ proved to be optimal at a 1.3 ml/min flow-rate, and a pH of 6.5. The temperature was optimized at 30±0.2°C. The amperometric detector, equipped with a glassy carbon electrode, was operated at 1300 mV versus Ag/AgCl in the direct current mode. The method was applied to the determination of these compounds at two concentration levels: ppm and ppb (ng/ml), obtaining relative standard deviations lower than 5% at ppm levels and lower than 10% at ppb levels, and quantitation limits ranging from 15 ppb to 500 ppb.The method was applied to the screening of β-blockers in spiked urine samples, with a total elution time lower than 12 min, obtaining the best recoveries for timolol and metoprolol (never greater than 93%). These recoveries together with the low limits of quantitation achieved, allows its application to doping analysis in human urine.     

Identification of incurred sulfonamide residues in eggs: methods for confirmation by liquid chromatography-tandem mass spectrometry and quantitation by liquid chromatography with ultraviolet detection

Two complementary methods for identifying and measuring sulfonamide residues in eggs were developed for use in surveying eggs for potential drug residues. The first method uses liquid chromatography-tandem mass spectrometry (LC-MS-MS) to confirm the presence of sulfonamide residues in eggs. During its validation the limit of confirmation was estimated to be 5-10 ng/g (ppb) depending on the drug. Also, a method for measuring residue level by liquid chromatography with ultraviolet detection (LC-UV) was validated using the same extraction procedure as the confirmatory method. The determinative method was validated over the 50-200 ppb range. Samples were prepared by homogenizing whole egg, extracting with acetonitrile, and cleaning up with a C(18) solid-phase extraction cartridge. For confirmation, analytes were separated by gradient LC on a C(18) column, ionized by electrospray ionization (ESI), and detected by MS-MS with an ion trap mass spectrometer. For determination, analytes were separated by a different gradient LC procedure and detected by UV at 287 nm. Fifteen drugs were dosed individually in laying hens, and residues of parent drug and/or metabolites were found in eggs for all the drugs. Validation was based on repetitive analyses of control samples, control samples fortified at 100 ppb sulfonamides, and samples of blended incurred eggs.     

Determination of zearalenone and its metabolites in urine and tissue samples of cow and pig by LC-MS/MS

For the sensitive and selective determination of zeranol, taleranol, α-zearalenol, β-zearalenol and zearalenone in animal urine and tissue a LC-MS/MS method has been developed. Sample preparation included extraction of meat samples and enzymatic digest of urine samples followed by solid-phase extraction with RP-18 columns for sample clean-up. Mass spectrometric determination was carried out with an atmospheric pressure chemical ionisation interface (APCI) in the multi-reaction monitoring mode (MRM). Using the negative ion mode detection limits between 0.1 and 0.5 ppb and determination limits between 0.5 and 1 ppb could be achieved. With zearalanone as internal standard, a linear range between 0.5 (1.0) and 100 ppb in urine samples (cow; pig) and between 1 and 100 ppb in meat samples (cow, calf, pig) could be established. Depending on the biological matrix and analyte standard deviations were below 8.2 %, with recovery rates between 86 and 102 % in spiked samples. The applicability of the method was demonstrated via several contaminated cow and pig urine samples.     

Development and validation of a method for the quantitative determination of aflatoxin contaminants in Maytenus ilicifolia by HPLC with fluorescence detection

A method for the quantification of aflatoxins B1, G1, B2 and G2 in the medicinal herb Maytenus ilicifolia was developed and validated. The method used immunoaffinity columns for sample clean-up and HPLC with fluorescence detection without any derivatisation step. The method showed good inter-day accuracy (bias values in the range 4.5-10.7%) and precision (5-16% RSD) when applied to the determination of levels of aflatoxins ranging from 7 to 20 ppb in the plant material. The detection limits for samples of the plant material spiked with aflatoxins were 3.5 ng/g for B1 and G1 and 0.1 ng/g for B2 and G2. The method was successfully applied to commercial samples of Maytenus ilicifolia for the screening of aflatoxin contaminants.     

Solid phase microextraction for analysis of alkanes and aromatic hydrocarbons in human breath

In this work, solid phase microextraction-gas chromatograph (SPME-GC) was applied to analyze alkanes and aromatic hydrocarbons in human breath, providing a potential non-invasive method to screen lung cancer. This method has been optimized and evaluated. It provided quantification limits ranging from 0.04 to 4.2 ng/mL, linear correlations ranging from 0.9845 to 0.9966 and R.S.D. values less than 9.8%. Total 30 breath samples, from 15 lung cancer patients and 15 healthy persons, were analyzed, and the alkanes and aromatic hydrocarbons were detected in 73.3% lung cancer patients and in 13.3% healthy persons by this method. Above all, It was demonstrated that this SPME-GC method provided a sensitive and non-invasive measure means to analyze alkanes and aromatic hydrocarbons in human breath, and brought forward a potential application for screening lung cancer.     

Headspace solid-phase microextraction in combination with gas chromatography and tandem mass spectrometry for the determination of organochlorine and organophosphorus pesticides in whole numan blood

A method for the determination of several organochlorine and organophosphorus pesticides in human whole blood samples was developed. The combination of solid-phase microextraction in headspace mode with gas chromatography with tandem mass spectrometry allowed the determination of 11 selected pesticides at ppb levels, minimizing the sample treatment. Quantitation was carried out by means of calibration curves prepared in blood using labelled surrogate/internal standards. The method showed good linearity between 1 and 50 ng ml(-1) (0.5-25 ng ml(-1) for HCB) using second-order calibration curves. Precision was found to be better than 20% at the three concentration levels assayed in the range of ng ml(-1). The detection limits obtained were in the range 0.02-0.7 ng ml(-1), except for p,p'-DDT (3 ng ml(-1)). The developed procedure was applied to blood and serum samples obtained from agricultural workers. HCB. beta-HCH and p,p'-DDE were most frequently detected in the samples analyzed.     

肺癌诊断方法的比较与研究进展

近年来,随着周围环境的恶化,肺癌的发病率最高并且呈现逐年上升的趋势,且其早期表现隐匿,很容易被忽视,很多患者确诊时已属晚期,丧失去了治疗的最佳时机。.本文主要总结和比较了几种传统的肺癌诊断方法如X线片、CT、MRI、PET-CT,并介绍了几种肺癌的最新的诊断技术,比如肺泡灌洗液或血清肿瘤标记物的联合检测、呼出气中的有机化合物的构成分析以及纤维支气管镜镜检技术等。上述方法诊断肺癌的敏感性和特异性各有优势,临床医生在临床工作中应合理应用上述检测方法,必要时联用多种检测手段,尤其要重视支气管肺泡灌洗液癌胚抗原如Cyfra21-1和CEA的联合检测,利用其较高的敏感性以提早发现和诊断肺癌。     

Bioelectric recognition assay (BERA)

A novel biosensory method has been developed for the determination of various chemical and biological molecules by assessing their electrophysiological interactions with a group of cells and cell components immobilized in a gel matrix that preserves their 'physiological' functions. The method was applied for the detection of: (i) hepatitis C virus in human blood samples; (ii) plant viruses; and (iii) a herbicide (glyphosate) in aqueous solutions. It was able to rapidly (assay time 3-5 min) and specifically detect the molecules in question at a concentration lower than 100 pg/ml, among other compounds f similar structure. The potential use of BERA biosensors for a rapid and cost-efficient molecule determination without prior knowledge of a specific receptor-molecule interaction is discussed.     

Selective photo-reduction of N-nitroamines combined with micellar electrokinetic chromatography and laser fluorimetric detection

N-nitroso compounds (NOC) are potent carcinogens. Reliable methods for the analysis of volatile carcinogenic NOC are well established; however selective and sensitive methods for routine analysis of thermally unstable, ionic or non-volatile NOC are still needed. For this purpose, a method based on micellar electrokinetic chromatography (MEKC) with laser induced fluorescence (LIF) detection is described for the simultaneous determination of a broad range of N-nitroso compounds. In this procedure, the nitroso group is photolytically cleaved from the NOC to yield the corresponding amine. The amines are then derivatized with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl), identified and quantified using MEKC-LIF. For the standard mixture of NOC, this method has good sensitivity and a large dynamic range. The detection limit provided by the method is 9 ppb for N-nitrosopyrrolidine.     

Positive and negative electrospray LC-MS-MS methods for quantitation of the antiparasitic endectocide drugs, abamectin, doramectin, emamectin, eprinomectin, ivermectin, moxidectin and selamectin in milk

Avermectin endectocides are used for the treatment of cattle against a variety of nematode and arthropod parasites, and consequently may appear in milk after normal or off-label use. The compounds abamectin, doramectin, and ivermectin, contain only C, H and O and may be expected to be detected by LC-MS in negative ion mode. The others contain nitrogen in addition and would be expected to be preferentially ionized in positive mode. The use of positive ion and negative ion methods with electrospray LC-MS-MS were compared. Using negative ion the compounds abamectin, doramectin, ivermectin, emamectin, eprinomectin, and moxidectin gave a curvilinear response and were quantified in raw milk by LC-MS-MS with a triethylamine-acetonitrile buffer over the concentration range 1-60 ppb (microg/kg) using selamectin as the internal standard. The limits of detection (LOD) were between 0.19 ppb (doramectin) and 0.38 ppb (emamectin). The compounds gave maximum sensitivity with positive ionisation from a formic acid-ammonium formate-acetonitrile buffer and were detected in milk (LC-MS-MS) also with a curvilinear response over the range 0.5-60 ppb. Although the positive ion signals were larger, with somewhat lower limits of detection (LOD between 0.06 ppb (doramectin) and 0.32 ppb (moxidectin) the negative ion procedure gave a more linear response and more consistent results. Comparison of spiked samples in the range 2-50 ppb showed a high degree of correlation between the two methods.     

Spectrofluorimetric determination of aluminium using 2‐hydroxy‐1‐naphthylidene‐(8‐aminoquinoline)

A sensitive and selective spectrofluorimetric method has been developed for the rapid determination of aluminium. This method is based on the complex formation between aluminium and 2‐hydroxy‐1‐naphthylidene‐(8‐aminoquinoline) (HNAQ). The optimum conditions for the complex formation were a metal‐to‐ligand (M : L) stoichiometric ratio of 1:1, a pH of 5.5 and a 0.20 m acetate buffer. The fluorescence of the complex was monitored at an emission wavelength of 502 nm with excitation at 438 nm. Under these conditions, linear calibration curves were obtained in the ranges 0.05–1 and 1–5 ppm. The detection limit was 3.4 ppb for the former and 13.5 ppb for the latter. The maximum relative standard deviation of the method for an aluminium standard of 200 ppb was 1.5% (n = 5). This method was successfully applied for the determination of aluminium in drinking water, pharmaceutical antacid tablets and suspension samples. Copyright © 2010 John Wiley & Sons, Ltd.     

An optimized method for determination of benzene in exhaled air by gas chromatography-mass spectrometry using solid phase microextraction as a sampling technique

The determination of benzene in exhaled air has contributed for the increase in the use of breath analysis in biological monitoring. This paper describes SPME as a sampling technique for determining benzene in exhaled air by GC-MS. A system was developed to generate a gaseous benzene standard by a permeation method to accomplish the breath analyses. The method presented good resolution, repeatability (the mean of %RSD values for intra-day measurements was 6.3), sensitivity (2.4 and 3.1 ppb for LOD and LOQ, respectively), and linearity of response (R(2)=0.994). After optimizing the conditions, analyses of real samples were performed on two groups (exposed and not exposed to benzene). The results presented an average of 8.2 ppb for the control group and 25.3 ppb for the exposed group.     

Determination of grepafloxacin and clinafloxacin by capillary zone electrophoresis

A simple and rapid capillary zone electrophoresis determination method with UV detection of grepafloxacin and clinafloxacin has been developed. The separation was performed in 35 mM borate-35 mM phosphate buffer solution (pH 8.6), containing 6% (v/v) of acetonitrile. Analyses were realised using fused-silica capillaries (57 cm length x 75 microm I.D.) and the operating conditions were: 15 kV applied voltage, 30 degrees C and detection at 279 nm. Piromidic acid was used as an internal standard. The linear concentration range of application was 1.0-120.0 microg ml(-1) for both compounds, with a detection limit of 0.2 microg ml(-1) for grepafloxacin and 0.3 microg ml(-1) for clinafloxacin. The analysis yielded good reproducibility (RSD between 3.37 and 1.74%). It was applied to the determination of grepafloxacin and clinafloxacin in human and rat urine samples. The method was validated using HPLC as a reference method. Recovery levels were between 94.5 and 103%.     

Detection of synthetic corticosteroids in bovine urine by chemiluminescence high-performance liquid chromatography.

The development of a black market of chemical cocktails for illegal growth promotion in food-producing animals includes substances that are potentially dangerous for human health, such as synthetic corticosteroids. The potential presence of these residues in food makes it necessary to develop rapid and sensitive analytical methodologies to detect such substances, preferably in live animals before they arrive at the market. A chemiluminescence (CL) detection method for the determination of four synthetic corticosteroids (prednisolone, betamethasone, dexamethasone and flumethasone) in bovine urine has been developed. The proposed system, which does not need any derivatization procedure, offers an easy method well suited for routine research. Urine samples were homogenized with methanol:water (50:50, v/v) and centrifuged. The upper layer was collected and Strata X cartridges were used for cleaning up. The purified residues were evaporated to dryness and then redissolved in the mobile phase. Analysis of the extracts was performed using high-performance liquid chromatography with chemiluminescence detection, employing luminol as the CL reagent. The recovery curves, obtained at four spiking levels (different for each corticosteroid), showed that recoveries of at least 70% could be obtained for urine. The chemiluminescence detection procedure afforded satisfactory results with respect to sensitivity and the LOD and LOQ, taken as the first point of the regression curve, ranged from 4 ppb to 65 ppb. The maximum mean RSD was below 13% and below 15% for intra- and inter-day assay, respectively, in all cases.