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1.
Fluorescence and terbium-sensitised luminescence properties of new quinolone garenoxacin have been studied. The fluorimetric method allows the determination of 0.060-0.600 μg ml−1 of garenoxacin in aqueous solution containing HCl/KCl buffer (pH 1.5) with λexc=282 nm and λem=421 nm. Micellar-enhanced fluorescence was also studied, leading to a higher than 400% increase in analytical signal in presence of 12 mM sodium dodecyl sulphate (SDS), allowing the determination of 0.020-0.750 μg ml−1 of garenoxacin. The terbium-sensitised luminescence method allows the determination of 0.100-1.500 μg ml−1 of garenoxacin in 12 mM SDS solution containing 0.08 M acetic acid/sodium acetate buffer (pH 4.1) and 7.5 mM Na2SO3 (chemical deoxygenation agent), with λexc=281 nm and λem=546 nm. Relative standard deviation (R.S.D.) values for the three methods were in the range 1.0-2.0%. The proposed procedures have been applied to the determination of garenoxacin in spiked human urine and serum.  相似文献   

2.
In this study the development, validation and application of a new chromatographic method for the determination of glutathione (GSH) in wine samples is presented. The separation of the GSH was carried out using a sulfobetaine-based hydrophilic interaction chromatography (HILIC) analytical column whereas its detection was carried out spectrofluorimetrically (λext/λem = 340/455 nm) after post-column derivatization with o-phthalaldehyde. GSH was separated efficiently from matrix endogenous compounds of wines by using a mobile phase of 15 mmol L−1 CH3COONH4 (pH = 2.5)/CH3CN, 35/65% (v/v). The parameters of the post-column reaction (pH, amount concentration of the reagent and buffer solution, flow rate, length of the reaction coil) were investigated. The linear determination range for GSH was 0.25–5.0 μmol L−1 and the LOD was 19 nmol L−1. No matrix effect was observed, while the accuracy was evaluated with recovery experiments and was ranged between 89% and 108%.  相似文献   

3.
A simple, precise, and accurate hydrophilic interaction liquid chromatographic (HILIC) method has been developed for the determination of five aromatic amines in environmental water samples. Chromatography was carried out on a bare silica column, using a mixture of acetonitrile and a buffer of NaH2PO4–H3PO4 (pH 1.5, containing 10 mM NaH2PO4) (85:15, v/v) as a mobile phase at a flow rate of 1 mL min−1. Aromatic amines were detected by UV absorbance at 254 nm. The linear range of amines was good (r2 > 0.998) and limit of detection (LOD) within 0.02–0.2 mg L−1 (S/N = 3). The retention mechanism for the analytes under the optimum conditions was determined to be a combination of adsorption, partition and ionic interactions. The proposed method was applied to the environmental water samples. Aromatic amines were isolated from aqueous samples using solid-phase extraction (SPE) with Oasis HLB cartridges. Recoveries of greater than 75% with precision (RSD) less than 12% were obtained at amine concentrations of 5–50 μg L−1 from 100 mL river water and influents from a wastewater treatment plant (WWTP). The present HILIC technique proved to be a viable method for the analysis of aromatic amines in the environmental water samples.  相似文献   

4.
A sensitive and effective micellar electrokinetic capillary chromatography with laser‐induced fluorescence detection approach was described for the determination of low molecular‐mass thiols using 1,3,5,7‐tetramethyl‐8‐phenyl‐(4‐iodoacetamido) difluoroboradiaza‐s‐indacene as the labeling reagent. After precolumn derivatization, baseline separation of six thiol compounds including cysteine, glutathione, N‐acetylcysteine, homocysteine, 6‐mercaptopurine, and penicillamine were achieved within 18 min. The optimal running buffer was composed of mixtures involving 25 mM sodium dodecyl sulfate, 25% (v/v) acetonitrile and 15 mM sodium phosphate buffer, pH 7.5. The detection limits (S/N = 3) were found as low as 40 pM under argon ion laser‐induced fluorescence detector (λex/λem = 488/520 nm), which were much better than the reported approaches. The accuracy and specificity of this assay for real samples were assured by a standard addition method. The proposed method has been applied to the analysis of thiols both in human plasma and plum flower samples with recoveries of 92.0–109.4%.  相似文献   

5.
We describe here a method for detecting and quantifying the highly carcinogenic polycyclic aromatic hydrocarbon (PAH) benzo[a]pyrene (BaP) in water, based on a flow-trough optical sensor. The technique is fast (response time of 40 s) and simple and at the same time meets the standards of sensitivity and selectivity required by the European Guidelines on Water for Human Consumption. The optosensor is based on the on-line immobilization of BaP on a non-ionic resin (Amberlite XAD-4) solid support in a continuous-flow system. BaP was analyzed in a 15 mM H2PO4/HPO42− buffer solution with 25% (v/v) 1,4-dioxane at pH 7. Fluorescence intensity was measured at λex/em=392/406 nm. The experimental conditions (reagent phase, pH, type and concentration of buffer solution and organic solvent) and flow-injection values (flow rate and injection volume) were carefully controlled. Under these conditions the optosensor was sensitive to a linear concentration range of between 3.0 and 250.0 ng l−1 with a detection limit of 3.0 ng l−1 and a standard deviation of 1.5% at 150 ng l−1. The optosensor was applied to the quantification of BaP in drinking and waste water samples (95-105% recovery) in presence of the other 15 EPA PAHs at 1000 ng l−1 concentration level. The influence of other fluorescent polycyclic aromatic hydrocarbons and potential interference from ions usually present in water was also evaluated.  相似文献   

6.
An analytical strategy micelle to trapping solution stacking (MSS) was developed in acidic buffer in micellar electrokinetic chromatography (MEKC). The stacking mechanism is based on the transport, release, capturing of molecules bound to micelle carriers that are made to collapse into trapping solution (TS) to serve as the medium to contain and stacking the analytes. Tetrandrine and fangchinoline were selected as model mixture using sodium dodecyl sulfate (SDS) micelles as carrier to demonstrate this stacking method. The experiments by MSS-MEKC were carried out and further compared with those by normal MEKC. The results reveal that 113–123-fold improvements in the detection sensitivity was obtained for the analytes, and separation and determination of tetrandrine and fangchinoline in Stephaniae tetrandrae S. Moore and Fengtongan capsules were finished under optimum conditions using the sample matrix containing 8.0 mM SDS and TS containing 50 mM H3PO4–55% (v/v) ethanol.  相似文献   

7.
Microemulsion electrokinetic chromatography (MEEKC) was developed for quantitative analysis of avermectins, such as abamectin, doramectin and ivermectin, in commercial formulations, using the microemulsion buffer containing a 50 mM phosphate buffer at pH 2.5, 1.1% (v/v) n-octane as oil droplets, 180 mM sodium dodecylsulphate as surfactant, 890 mM 1-butanol as co-surfactant and 30% (v/v) ethanol as organic co-solvent. High accuracy and precision of the method were obtained. The contents of avermectins in commercial formulations determined by MEEKC were found to be insignificantly different with those determined by high performance liquid chromatography (HPLC). Therefore, MEEKC can be used an alternative method to HPLC for quantitative determination of avermectins.  相似文献   

8.
A simple and reliable co-electroosmotic capillary electrophoresis system for the fast determination of aromatic acids has been developed by employing poly (1-vinyl-3-butylimidazolium bromide) as the background electrolyte modifier. The polymeric ionic liquid was synthesized by the conventional radical polymerization. The reversed electroosmotic flow was obtained by adding a small amount of the polymeric ionic liquid (0.0006%, w/v) to the electrolyte. To further improve the resolution of aromatic acids, conditions including the concentration of polymeric ionic liquid and pH of background electrolytes were optimized. All eight aromatic acids were baseline resolved in one measurement in a short time (less than 3.5 min) under optimized conditions, 100 mM NaH2PO4 buffer containing 0.006% (w/v) polymeric ionic liquid, pH 6.0. Separation efficiencies were in the range from 355,000 to 943,000 (plates/m). Satisfactory reproducibility on the basis of the migration time of analytes was achieved. RSDs (n = 3) were less than 0.33% except the p-aminobenzoic acid (0.9%). The applicability of the present method has been demonstrated for the determination of water-soluble aromatic acids in a common drug for external use.  相似文献   

9.
In the present work, an analytical method for determination of risedronate, a member of bisphosphonates, is described for the routine analysis in rat plasma. Sample pre-treatment involves protein precipitation, co-precipitation with calcium at alkaline pH, hydrolysis of possible derivatives of pyrophosphate and reprecipitation. A good separation was obtained by using a reversed-phase column (Hypersil ODS-2 C18, 4.6 mm × 250 mm, 5 μm). The mobile phase was an aqueous solution of buffer (contained 1.5 mM EDTA-2Na, 1 mM sodium etidronate, 11 mM sodium phosphate and 5 mM tetrabutylammonium bromide as ion-pair reagent) - methanol (88:12, v/v) adjusted to pH 6.75 using 1 M NaOH. The flow rate was 1 ml min−1. UV detection (λ = 262 nm) was used to quantitate risedronate in the concentration range of 10-500 ng ml−1. The limit of detection and quantitation for risedronate were 7 and 10 ng ml−1, respectively. The method was applied successfully to plasma samples from Wistar rats undergoing oral administration of risedronate mini-pills. Precision, extraction recoveries, as well as accuracy results, were satisfactory and no interference was found at the retention time of risedronate. Hence, the method is suitable for monitoring risedronate in rat plasma.  相似文献   

10.
The use of menadione (MD) as a pre-column reagent for high performance liquid chromatography (HPLC) analysis of aliphatic thiols is proposed. The reaction was carried out for 5 min at room temperature and pH 8.5. The developed method was applied to the N-acetylcysteine (NAC) analysis of alimentary supplements and pharmaceutical formulations. The effect of the complex matrix was evaluated by the study of the thiol derivatization reaction both in standard and in placebo solutions. The yield of NAC-MD adduct was found to be quantitative at a reagent to thiol molar ratio of about 4 in comparison with an authentic specimen of synthesized NAC adduct, which was characterized by 1H NMR, IR and UV. The routine chromatographic separations were performed on a Synergi MAX-RP column using a mobile phase consisting of methanol/triethylammonium (TEA) phosphate buffer (pH 3; 0.05 mol L−1) 70:30 (v/v) at a flow-rate of 0.4 mL min−1. UV-diode array detection was used setting the wavelength at λ = 260 nm. The validation parameters such as linearity, sensitivity, accuracy, precision, selectivity and ruggedness were found to be highly satisfactory. Similar linear responses were observed by standard and placebo solutions (determination coefficient: 0.9996). Limit of detection was about 0.019 μg g−1. Intra-day precision (relative standard deviation, R.S.D.) was ≤0.81% for NAC to internal standard (IS) peak area ratio, ≤0.28% and ≤0.32%, respectively, for NAC and IS retention times (tR), without significant differences between intra- and inter-day data. NAC recovery studies gave good results (100.12%) with R.S.D. = 1.05%.  相似文献   

11.
A method for simultaneous determination of seven benzodiazepines (BZPs) (flunitrazepam, clonazepam, oxazepam, lorazepam, chlordiazepoxide, nordiazepam and diazepam using N-desalkylflurazepam as internal standard) in human plasma using liquid-liquid and solid-phase extractions followed by high-performance liquid chromatography (HPLC) is described. The analytes were separated employing a LC-18 DB column (250 mm × 4.6 mm, 5 μm) at 35 °C under isocratic conditions using 5 mM KH2PO4 buffer solution pH 6.0:methanol:diethyl ether (55:40:5, v/v/v) as mobile phase at a flow rate of 0.8 mL min−1. UV detection was carried out at 245 nm. Employing LLE, the best conditions were achieved with double extraction of 0.5 mL plasma using ethyl acetate and Na2HPO4 pH 9.5 for pH adjusting. Employing SPE, the best conditions were achieved with 0.5 mL plasma plus 3 mL 0.1 M borate buffer pH 9.5, which were then passed through a C18 cartridge previously conditioned, washed for 3 times with these solvents: 3 mL 0.1 M borate buffer pH 9.5, 4 mL Milli-Q water and 1 mL acetonitrile 5%, finally the BZPs elution was carried with diethyl ether:n-hexane:methanol (50:30:20). In both methods the solvent was evaporated at 40 °C under nitrogen flow. The validation parameters obtained in LLE were linearity range of 50-1200 ng mL−1 plasma (r ≥ 0.9927), limits of quantification of 50 ng mL−1 plasma, within-day and between-day CV% and E% for precision and accuracy lower than 15%, and recovery above 65% for all BZPs. In SPE, the parameter obtained were linearity range of 30-1200 ng mL−1 plasma (r ≥ 0.9900), limits of quantification of 30 ng mL−1 plasma, within-day and between-day CV% and E% for precision and accuracy lower than 15% and recovery above 55% for all BZPs. These extracting procedures followed by HPLC analysis showed their suitable applicability in order to examine one or more BZPs in human plasma. Moreover, it could be suggested that these procedures might be employed in various analytical applications, in special for toxicological/forensic analysis.  相似文献   

12.
The coupling of Ru(bpy)32+ based electrochemiluminescence (ECL) detection with capillary electrophoresis (CE) was developed for the simultaneous determination of the two major active ingredients (atropine and scopolamine) in Flos daturae. Parameters related to the separation and detection were discussed and optimized. It was proved that 20 mM phosphate buffer at pH 8.48 could achieve the most favorable resolution, and the high sensitivity of detection was obtained by maintaining the detection potential at 1.2 V. Under the optimized conditions: ECL detection at 1.2 V, 20 mM phosphate buffer at pH 8.48, 5 mM Ru(bpy)32+ and 50 mM phosphate buffer at pH 7.48 in the detection reservoir, detection limits of 5 × 10−8 mol/l for atropine and 1 × 10−6 mol/l for scopolamine were obtained. Relative standard derivations of the ECL intensity and the migration time were 5.16 and 0.71% for atropine and 5.07 and 1.22% for scopolamine, respectively. Developed method was successfully applied to determine the amounts of both alkaloids in Flos daturae. A baseline separation for atropine and scopolamine was achieved within 11 min.  相似文献   

13.
A procedure for the determination of Imidacloprid and its main metabolites was set up by means of liquid chromatography with an electrochemical detector and post-column photochemical reactor (LC--ED). Sample clean-up was developed for bees, filter paper and maize leaves. Chromatographic conditions were based on a reversed-phase C-18 column operated by phosphate buffer 50 mM/CH3CN (80/20, v/v) at pH 2.9. Detection of Imidacloprid and its metabolites was performed at a potential of 800 mV after photoactivation at 254 nm. Compared to conventional techniques such as gas chromatography/mass spectrometry (GC/MS) or LC coupled to other detectors, the present method allows simultaneous trace-level determination of both Imidacloprid (0.6 ng ml−1) and its main metabolites (2.4 ng ml−1).  相似文献   

14.
A fluorimetric liquid chromatographic method for the determination of 5-hydroxyindoles based on the benzylamine derivatization process mediated through an online photocatalytic oxidation has been developed. In this study, we used a photocatalytic column comprising tefzel tubing packed with TiO2-coated glass beads, as a pre-column derivatization reactor. The fluorescence derivatization of 5-hydroxyindoles using benzylamine proceeded during their passage through the reaction column under near-UV irradiation. The 5-hydroxyindole derivatives were separated continuously on a reversed-phase liquid chromatography within 50 min, using 100 mM acetate buffer (pH 4.6)-acetonitrile (72:28, v/v; isocratic elution) containing 3 mM sodium octanesulfonate; the samples were detected fluorimetrically at 465 nm upon excitation at 350 nm. The detection limits (signal-to-noise ratio = 3) of the 5-hydroxyindoles were in the range from 160 to 360 fmol per 5 μL injection. We have applied this method, which requires minimal sample pre-treatment, to the determination of 5-hydroxyindole-3-acetic acid in human urine.  相似文献   

15.
A bithiophene-substituted poly(p-phenyleneethynylene) derivative (CP1) having water-solubilizing side chains was prepared and characterized. Copper(II)-induced quenching of CP1 emission was quantified in H2O, MeCN/H2O (90:10), and pH 7.4, 50 mM HEPES, 100 mM KCl buffer. In buffer, treatment of CP1-Cu(II) with nitroxyl (HNO) produces an immediate 2.1-fold increase in emission, whereas exposure to NO(g) effects no fluorescence restoration. The ability to distinguish HNO from NO chemically at physiological pH represents a productive step towards the development of selective, fluorescence-based biosensors for HNO.  相似文献   

16.
Melatonin was determined in pharmaceutical preparations by means of two simple and reliable analytical methods based on micellar electrokinetic chromatography (MEKC) and spectrofluorimetry. The fluorescence emission values were measured at λ=350 nm when exciting at λ=275 nm. The MEKC analysis was achieved using a system consisting of 40 mM SDS in phosphate buffer (20 mM, pH 7.5). The extraction of melatonin from the tablets was achieved by means of a simple one-step dissolution with methanol/water. Both methods were applied for the determination of melatonin in commercial formulations and galenic preparations. The MEKC procedure allows the quantitative determination of melatonin in all pharmaceutical preparations tested. On the contrary, the spectrofluorimetric method is not suitable for tablets which also contain tryptophan; this interference can be eliminated by a suitable liquid-liquid extraction procedure. The results obtained with the two methods are in good agreement and satisfactory in terms of precision and accuracy.  相似文献   

17.
A family of 6-mono(3-alkylimidazolium)-β-cyclodextrins with one primary hydroxyl group replaced by an alkylimidazolium cation has been developed. The effect of alkyl substitutents on the enantioresolution ability of these single-isomer cyclodextrins towards dansyl amino acids has been studied by capillary electrophoresis. Systematical investigations on the effect of buffer pH and selector concentration on the enatioseparation show that chiral selectors with a shorter alkyl chain (R = CnH2n+1, n ≤ 4) presented more powerful chiral recognition ability. These newly introduced single-isomer β-cyclodextrin derivatives proved to be effective chiral selectors for most selected dansyl amino acids at low buffer pH (e.g. pH 5.0) with selector concentration no less than 3 mM. The apparent complex stability constants between alkylimidazolium β-CDs and dansyl amino acids were also theoretically determined by using the mobility difference model proposed by Wren and Rowe. The side alkyl chains from both dansyl amino acids and alkylimidazolium β-CDs displayed significant effect on the apparent complex stability constants. Both the optimum selector concentrations calculated according to the model, however, were much lower than the experimental values giving the maximum chiral resolution of enantiomers.  相似文献   

18.
Sema Demirci Çekiç 《Talanta》2009,79(2):344-1658
Proteins are not considered as true antioxidants but are known to protect antioxidants from oxidation in various antioxidant activity assays. This study aims to investigate the contribution of proteins, especially thiol-containing proteins, to the observed overall antioxidant capacity measured by known methods. To determine the antioxidant properties of thiol-containing proteins, the CUPRAC method of antioxidant assay using the oxidizing reagent Cu(II)-neocuproine previously used for simultaneous analysis of cystine and cysteine was adopted. While the CUPRAC method is capable of determining all antioxidant compounds including thiols in complex sample matrices, the Ellman method of thiol quantitation basically does not respond to other antioxidants. The antioxidant quantities in the selected samples were assayed with the ABTS and FRAP methods as well as with the CUPRAC method. In all applied methods, the dilutions were made with a standard pH 8 buffer used in the Ellman method by substituting the Na2EDTA component of the buffer with sodium citrate. On the other hand, the standard CUPRAC protocol was modified by substituting the pH 7 ammonium acetate buffer (at 1 M concentration) with 8 M urea buffer adjusted to pH 7 by neutralizing with 6 M HCl. Urea helps to partly solubilize and denaturate proteins so that their buried thiols be oxidized more easily. All methods used in the estimation of antioxidant properties of proteins (i.e., CUPRAC, Ellman, ABTS, and FRAP) were first standardized with a simple thiol compound, cysteine, by constructing the calibration curves. The molar absorptivities of these methods for cysteine were: ?CUPRAC = 7.71 × 103, ?Ellman = 1.37 × 104, ?ABTS = 2.06 × 104, and ?FRAP = 2.98 × 103 L mol−1cm−1. Then these methods were applied to various samples containing thiols, such as glutathione (reduced form:GSH), egg white, whey proteins, and gelatin. Additionally, known quantities of selected antioxidants were added to these samples to show the additivity of responses.  相似文献   

19.
A procedure based on BIA Separations CIM DEAE anion-exchange chromatography was developed to separate double-stranded (ds) RNA of hypovirus infecting phytopathogenic fungus Cryphonectria parasitica. Using a linear gradient of 25 mM 4-morpholinepropanesulfonic acid (MOPS), pH 7.0 as a binding buffer, and 25 mM MOPS, 1.5 M NaCl, 0.1 mM EDTA, 15% isopropanol (v/v), pH 7.0 as an elution buffer, hypoviral dsRNA was additionally purified from nucleic acid species present in preparations partially purified by standard CF-11 cellulose chromatography. Moreover, crude phenol/chloroform extracts of the fungal tissue were also applied to monolithic supports and CIM DEAE chromatograms revealed clear evidence for hypoviral presence without CF-11 chromatography, nucleic acid precipitation, and electrophoresis.  相似文献   

20.
A capillary electrophoresis (CE) and a high performance liquid chromatography (HPLC) method are described for the simultaneous determination of ethylenediaminetetraacetic acid (EDTA), S,S′-ethylenediaminedisuccinic acid (EDDS) and R,S-iminodisuccinic acid (IDS) complexing agents as their Fe(III) complexes in cosmetics like shower cream and foam bath. The non-biodegradable EDTA is used in combination with biodegradable analogues like EDDS and IDS in many commercial products. The HPLC method involves separation by reversed-phase ion pair chromatography on a C18 column using methanol-formate buffer (20 mM tetrabutylammonium hydrogen sulfate, 15 mM sodium formate adjusted to pH 4.0 with formic acid) (10:90, v/v) as mobile solvent at a flow rate of 0.8 mL min−1 at 24 °C using UV detection at 240 nm. The CE separation was performed in a fused silica capillary of 50 μm i.d. with the total length of 50 cm with a 10 mM MES and MOPSO (pH 5.5) at an applied voltage of −25 kV. The samples were introduced by applying a 50 mbar pressure for 2 s. Absorbances at 215 and 225 nm were monitored for the detection of the complexes. The methodology performance of the two methods was evaluated in terms of linearity, limit of detection (LOD), limit of quantitation (LOQ) and reproducibility. The LOD values obtained from HPLC are low when compared with CE. The applicability of both the methods was demonstrated for the analysis of cosmetic products such as shower cream and foam bath. The results obtained by both CE and HPLC were found to be comparable and in good agreement.  相似文献   

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