Liquid chromatographic method for the analysis of tocopherols in malt sprouts with supercritical fluid extraction.

A simple, specific and sensitive high-performance liquid chromatographic method has been developed for the determination of tocopherols in malt sprouts. A supercritical fluid extraction (SFE) procedure was used to isolate tocopherols from the vegetal matrix before quantitative analysis. The analytes were separated on a Zorbax reversed-phase column using methanol-water as mobile phase and quantified by measuring its fluorescence at lambda(em)=328 nm after excitation of the analytes at lambda(exc)=303 nm. The limits of detection for alpha-, gamma- and delta-tocopherols were 0.04, 0.05, and 0.05 microg/ml, respectively. The calibration graphs of the method were linear from 0.1 to 1.5, 0.2 to 2.5, and 0.2 to 2.0 microg/ml, for alpha-, gamma- and delta-tocopherols, respectively. This SFE and HPLC procedure is simple, precise and accurate for the determination of tocopherols in malt sprouts.     

Simultaneous determination of carbaryl and azinphos-methyl in water by first-derivative synchronous spectrofluorimetry

A method is proposed for the simultaneous determination of the pesticides carbaryl (CBL) and azinphos-methyl (AZM) in water by first-derivative synchronous spectrofluorimetry. It is based on the alkaline hydrolysis of both pesticides to their metabolites 1-naphthol (from CBL) and anthranilic acid (from AZM). The constant wavelength difference chosen to optimize the determination is Δλ=λ_em?λ_ex=103 nm. CBL is measured at 302/405 nm and AZM at 333/436 nm. The calibration graphs are linear between 2.0 and 500.0 ng/ml for CBL and between 1.2 and 500.0 ng/ml for AZM with detection limits of 0.62 ng/ml and 0.35 ng/ml, respectively. The precision of the method (RSD) is 2.4% at the 80.0 ng/ml level for CBL and 2.5% at the 80.0 ng/ml level for AZM. The method is applied to the determination of both analytes in samples of natural waters.     

Study on the resonance Rayleigh scattering, second-order scattering and frequency doubling scattering spectra of the interactions of palladium(II)-ceftriaxone chelate with anionic surfactants and their analytical applications

In pH 1.8-2.9 Britton-Robinson (BR) buffer medium, ceftriaxone (CTRX) can react with palladium(II) (Pd(II)) to form 1:2 cationic chelate, which can further react with anionic surfactants (AS) such as sodium lauryl sulfonate (SLS), sodium dodecyl sulfate (SDS) and sodium dodecylbenzene sulfonate (SDBS) to form 1:3 ion-association complexes. As a result, the resonance Rayleigh scattering (RRS), second-order scattering (SOS) and frequency doubling scattering (FDS) were enhanced greatly. The maximum RRS, SOS and FDS wavelengths of three ion-association complexes were located at 335 nm, 560 nm and 390 nm, respectively. The increments of scattering intensity (DeltaI) were directly proportional to the concentrations of CTRX in certain ranges. The detection limits (3sigma) of CTRX for SLS, SDBS and SDS systems were 1.8 ng ml(-1), 2.3 ng ml(-1) and 2.3 ng ml(-1) (RRS method), 4.9 ng ml(-1), 7.4 ng ml(-1) and 4.7 ng ml(-1) (SOS method) and 6.8 ng ml(-1), 7.3 ng ml(-1) and 9.1 ng ml(-1) (FDS method), separately. The sensitivity of RRS method was higher than those of SOS and FDS methods. The optimum conditions of RRS method and the influence factors were investigated, and the composition of ion-association complexes and the reaction mechanism were discussed also. The effects of foreign substances were tested and it showed that the method has a good selectivity. Based on the ion-association reaction, the sensitive, simple and rapid methods for the determination of CTRX have been developed.     

Determination of nucleic acids using calcein-neodymium complex as a fluorescence probe.

A novel fluorometric method has been developed for rapid determination of DNA and RNA with calcein-neodymium complex as a fluorescence probe. The method is based on the fluorescence enhancement of calcein-Nd(III) complex in the presence of DNA or RNA, with maximum excitation and emission wavelength at 489 nm and 514 nm, respectively. Under optimal conditions, the calibration graphs are linear over the range 0.5 - 3.0 microg/ml for both DNA and yeast RNA, 0.4 - 2.0 microg/ml for fish sperm DNA (FS DNA) and 0 - 3.0 microg/ml for calf thymus DNA (CT DNA). The corresponding detection limits are 15.1 ng/ml for DNA, 21.2 ng/ml for yeast RNA, 10.5 ng/ml for FS DNA and 8.9 ng/ml for CT DNA. The interaction mechanism for the binding of calcein-Nd(III) complex to DNA is also studied. The results of absorption spectra, fluorescence polarization measurements and thermal denaturation experiments, suggested that the interaction between calcein-Nd(III) complex and DNA is an electrostatic interaction.     

Column-switching techniques for high-performance liquid chromatography of ibuprofen and mefenamic acid in human serum with short-wavelength ultraviolet detection.

Column-switching techniques for high-performance liquid chromatography of two acidic drugs, ibuprofen and mefenamic acid, in human serum with short-wavelength ultraviolet detection are described. The method involved extraction of the analyte from acidified serum followed by the chromatographic analysis using column switching. Three ODS columns were used each with different mobile phase, utilizing the difference of ion-pair formation or of ionization caused by pH change. The method offered high sensitivity and selectivity, with short-wavelength ultraviolet detection at 221 nm for ibuprofen and at 219 nm for mefenamic acid. The detection limits were 0.5 ng/ml (2.4 pmol/ml) for ibuprofen and 0.1 ng/ml (0.4 pmol/ml) for mefenamic acid using 1 ml of serum, both at a signal-to-noise ratio of 3. With some modifications, the principle of the method would be applicable to other acidic compounds in biological fluids.     

High-performance liquid chromatographic determination of penicillin G, penicillin V and cloxacillin in beef and pork tissues.

The objective was to develop confirmatory high-performance liquid chromatographic methods for penicillin residues in animal tissues with detection limits of less than or equal to 10 ng/g. A previously described procedure was modified by using a larger sample size and isocratic analysis. Tissues (15 g) were blended with 45 ml of water and 20 ml of homogenate were mixed with 40 ml acetonitrile and filtered. The filtrate (30 ml) was mixed with 10 ml of 0.2 M H3PO4 and extracted with methylene chloride. The combined methylene chloride layers were mixed with acetonitrile and hexane, washed with two 4-ml portions of water and then extracted with four 1-ml portions of 0.01 M phosphate buffer (pH 7). The combined buffer extracts were concentrated to 1 ml under reduced pressure. Analysis was isocratic during 0.01 M phosphate buffer (pH 7)-acetonitrile with proportions 85:15 (penicillin G), 82:18 (penicillin V) or 78:22 (cloxacillin). A polystyrene-divinylbenzene copolymer column, 150 x 4.6 mm I.D. (Polymer Labs. PLRP-S), was used with a flow-rate of 1 ml/min and detection at 210 nm. The presence of penicillins was confirmed by treating a duplicate sample with penicillinase. Recoveries were greater than 90% in most instances. Detection limits were 5 ng/g in muscle and higher in liver and kidney. The procedure is a simple and sensitive method for confirming the presence of penicillins in animal tissues.     

Simple high-performance liquid chromatography method for the simultaneous determination of ketoconazole and piperine in rat plasma and hepatocyte culture

Piperine, a major alkaloid of black and long peppers has been reported to act as bioavailability enhancer of several drugs by inhibiting drug metabolising enzymes and/or by increasing oral absorption. Ketoconazole is a well established potent inhibitor of CYP 3A4 and P-glycoprotein. A simple and rapid HPLC method has been developed for the simultaneous analysis of ketoconazole and piperine in rat plasma and hepatocyte culture. Analysis was performed using a Symmetry C18 column (150x4.6 mm, 5 microm) and isocratic elution with 25 mM KH2PO4 (pH 4.5)-acetonitrile (50:50) with a flow-rate of 1 ml/min. Photodiode array detection was used to simultaneously monitor piperine at 340 nm and ketoconazole at 231 nm in a single sample. Calibration plots in spiked plasma, hepatocytes and William's medium E were linear over the range studied (10-2000 ng for both drugs). The detection limits for piperine and ketoconazole are 2 and 4 ng, respectively, and the limits of quantitation are 10 and 12 ng, respectively. Intra- and inter-assay variations were less than 8%.     

Determination of dioxopiperazine metabolites of quinapril in biological fluids by gas chromatography-mass spectrometry.

The dioxopiperazine metabolites of quinapril in plasma and urine were extracted with hexane-dichloroethane (1:1) under acidic conditions. Following derivatization with pentafluorobenzyl bromide and purification of the desired reaction products using a column packed with silica gel, the metabolites were analysed separately by capillary column gas chromatography-electron-impact mass spectrometry with selected-ion monitoring. The limits of quantitation for the metabolites were 0.2 ng/ml in plasma and 1 ng/ml in urine. The limits of detection were 0.1 ng/ml in plasma and 0.5 ng/ml in urine, at a single-to-noise ratio of greater than 3 and greater than 5, respectively. The proposed method is applicable to pharmacokinetic studies.     

Determination of p-methylthiobenzamide and p-methylthiobenzamide-S-oxide from rat plasma using solid-phase extraction and high-performance liquid chromatography

p-Methylthiobenzamide (PMTB) is a thiocarbonyl compound exhibiting marked hepatotoxicity and nephrotoxicity. We describe a high-performance liquid chromatographic method for analyzing PMTB and a metabolite, p-methylthiobenzamide-S-oxide (PMTBSO), from rat plasma using a solid-phase extraction technique. In this way, PMTB and PMTBSO can be extracted from 0.5 ml of plasma and separation achieved by an ODS analytical column in as little as 9 min. The mobile phase used was methanol-water (55:45, v/v) and the wavelength for detection was 290 nm. The limits of detection in plasma were 15 ng/ml for PMTB and 33 ng/ml for PMTBSO; the absolute recovery from spiked plasma samples was greater than 84.4% for both compounds and the internal standard. The method was linear throughout the range used with correlation coefficients greater than 0.969. The intra-day accuracy ranged from 1.52 to 15.23% relative error for the PMTB concentration range 151-3025 ng/ml; accuracy of 4.97% or less was obtained for PMTBSO concentrations of 1672-20,068 ng/ml. The intra-day precision (coefficient of variation) of the procedure was found to be no greater than 5.28% for PMTB and 7.9% for PMTBSO. Inter-day accuracy and precision measurements were similar.     

Method for the determination of chromium (VI) as chromium (VI)-dibenzyidithiocarbamate

Dibenzyldithiocarbamic acid (DBDC) exhibits the ability to speciate between chromium(VI) and chromium(III), since only the chromium(VI) will form complexes with DBDC. The complex is then extracted into an organic solvent and assayed using an ultraviolet-visible (UV-VIS) spectrophotometer at 498.8 nm. Using 250 ml of aqueous sample detection limits less than 1 ng/ml are possible, while the linear range extends to 500 gmg/ml when working at 498.8 nm. Oxidation of the chromium(III) to chromium (VI) using cerium (IV) enables the determination of total chromium and subsequently the chromium (III) in solution. Evaluation of the method with a standard reference material produced only 4.81 part per thousand error in the determination of chromium(VI).     

Simultaneous determination of thioridazine and its S-oxidized and N-demethylated metabolites using high-performance liquid-chromatography on radially compressed silica

A method for simultaneously quantifying thioridazine, northioridazine, thioridazine-2-sulfoxide, thioridazine-2-sulfone and thioridazine-5-oxide in serum and plasma is described. Following solvent extraction these compounds were separated by high-performance liquid chromatography on radially compressed silica gel and detected by UV absorbance at 254 nm. Chromatography time is less than 7 min. The relative retention of these compounds as a function of the methanol and methylamine content of the mobile phase is discussed. Practical limits of detection, based upon on assayed plasma or serum volume of 1 ml, were 20 ng/ml for thioridazine-5-oxide and 10 ng/ml for the other compounds. The coefficient of variation for all compounds was less than 13%. The method is compared with more conventional high-performance liqiud chromatographic and gas chromatographic methodology.     

Determination of yohimbine and its two hydroxylated metabolites in humans by high-performance liquid chromatography and mass spectral analysis.

The existence of at least two metabolites of yohimbine (YO) in humans is demonstrated. Combined high-performance liquid chromatographic (HPLC), NMR and mass spectral analyses permitted them to be identified as hydroxylated metabolites at the C-10 and C-11 positions. A normal-phase HPLC method allowing the simultaneous determination of YO and its main metabolite, 11-hydroxyyohimbine (11-OHYO), in biological samples is described. This assay was performed using a LiChrosorb Si 60 column and a mobile phase consisting of 0.02 M sodium acetate (pH 5)-methanol (5:95, v/v) at a flow-rate of 1 ml/min. Detection was achieved by a fluorimetric method (excitation at 280 nm and emission at 320 nm). The extraction yields of YO, 10-OHYO and 11-OHYO from plasma were 91.8, 45.3 and 17.8%, respectively, and their respective within-day reproducibilities were 3.8, 1.4 and 5.9%. The between-day reproducibility for YO at the concentrations of 1 and 10 ng/ml were 8.9 and 6.4%, respectively. The accuracy of the method for YO at concentrations of 1 and 10 ng/ml were 5.1 and 2.3%, respectively. The limits of determination of YO, 10-OHYO and 11-OHYO were 0.1, 0.5 and 1 ng/ml, respectively. The method was used in bioavailability study of YO following oral and intravenous administration in humans.     

Determination of halides by microwave induced plasma and stabilized capacitive plasma atomic emission spectrometry after on-line continuous halogen generation

This paper describes a comparative study of the microwave induced plasma (MIP) and the stabilized capacitive plasma (SCP) for halide determinations. The MIP is generated in a Beenakker cavity TM(010) using a tangential flow torch and the SCP consists of a 27.12 MHz discharge sustained in a liquid-cooled, fused silica tube surrounded by two annular electrodes. Both discharges are operated in helium at atmospheric pressure and detection was carried out by Atomic Emission Spectrometry (AES). The halides (I(-), Br(-), Cl(-)) are converted to volatile halogens by continuous flow generation based on chemical oxidation and on-line separation from the aqueous phase, via a gas-liquid separator, to be finally introduced into the plasma. The different factors affecting the emission intensity of the volatile halogens generated are compared for both discharges and the analytical performance characteristics are also evaluated. Detection limits of 17 ng ml(-1), 24 ng ml(-1) and 55 ng ml(-1) are obtained for the determination of Cl(-), Br(-), and I(-), respectively, in the ultraviolet-visible (UV-VIS) region using the MIP-AES and 45 ng ml(-1), 135 ng ml(-1) and 400 ng ml(-1) for Cl(-), Br(-), and I(-) with the SCP-AES. Lines in the near infrared (NIR) region were also evaluated for the SCP-AES detection; improvements in detection limits higher than 30 times were observed in the NIR region as compared with the UV-VIS with detection limits in the NIR of 1.4 ng ml(-1) for Cl(-), 3 ng ml(-1) for Br(-) and 13 ng ml(-1) for I(-).     

Brilliant cresyl blue as a new red region fluorescent probe for determination of nucleic acids

A fluorescence quenching method was developed for determination of microamounts of nucleic acids by using brilliant cresyl blue (BCB) as a new red region fluorescent probe. In aqueous hexylmethylene tetramine solution, BCB showed maximum excitation and emission wavelengths at 626 and 670 nm, respectively, and the fluorescence of BCB could be greatly quenched by DNA (or RNA). Under optimal conditions, the calibration graphs are linear over the range of 0.02–0.80 μg/ml for SM DNA and 0.25–1.5 μg/ml for yeast RNA. The corresponding detection limits are 7 ng/ml for SM DNA and 25 ng/ml for yeast RNA, respectively. SM DNA can be determinated in the presence of 40% (w/w) RNA, and the relative standard deviation of six measurements is 2.5% for 500 ng/ml SM DNA. The result of the determination of golden staphylococcus DNA by this method was satisfactory.     

A biosensor for ferric ion

A new biosensor for monitoring iron has been developed. The active solid phase is pyoverdin, a natural fluorescent pigment biosynthesized by Pseudomonas fluorescens immobilized on controlled pore glass (CPG) and packed in a quartz flow-through cell. The biosensor is very selective for iron(III) and can be easily regenerated in about 2 min by passing 1M HCl through the cell. The optimum conditions and analytical characteristics (detection limit, precision and linear range) for the new sensor in solution (DL = 10 ng/ml) and in immobilized form (DL = 3 ng/ml) are reported. The biosensor has good stability and can be used continuously over a period for at least 3 months (over 1000 determinations). The sensor was successfully applied to determine iron in different water samples. There were no significant differences between the new method and the Inductively Coupled Plasma Atomic Emission Spectroscopy (ICP-AES) reference method at the 95% confidence level.     

The effects of magnetite particles encapsulated in dihexadecyl phosphate (DHP) vesicles on the luminescence characteristics of molecules

Luminescence of organic molecules is observed in DHP vesicles in the absence and presence of magnetite encapsulated in the vesicles. Room temperature phosphorescence calibration curves for carbazole were constructed at 297/440 nm and at 330/440 nm in the absence and presence of magnetite. The linear dynamic range was found to be over 2.5 orders of magnitude. At 297/440 nm, the limits LOD's were found to be 30 ng/ml and 9 ng/ml in the absence and presence of magnetite, respectively. At 330/440 nm, the LOD's were found to be 30 ng/ml both in the presence and absence of magnetite. The effect of the internal magnetic field on the luminescence properties of various molecules is discussed. The precision of the method ranged from ∼ 1 % to ∼ 40%.     

Study on the resonance light scattering spectrum of berberine-cetyltrimethylammonium bromide system and the determination of nucleic acids at nanogram levels

The interaction of berberine with nucleic acid in the presence of cetyltrimethylammonium bromide (CTMAB) in aqueous solution has been studied by spectrophotometry and resonance light scattering (RLS) spectroscopy. At pH 7.30, the RLS signals of berberine were greatly enhanced by nucleic acid in the region of 300-600 nm characterized by four peaks at 324.0, 386.5, 416.5 and 465.0 nm. The binding properties were examined by using a Scatchard plot based on the measurement of enhanced RLS data at 416.5 nm. Under optimum conditions, the increase of RLS intensity of this system at 416.5 nm is proportional to the concentration of nucleic acid. The linear range is 7.5 x 10(-9)-7.5 x 10(-5) g ml(-1) for calf thymus DNA, 7.5 x 10(-9)-2.5 x 10(-5) g ml(-1) for herring sperm DNA, and 5.0 x 10(-9)-2.5 x 10(-5) g ml(-1) for yeast RNA. The detection limits (S/N = 3) are 2.1 ng ml(-1) for calf thymus DNA, 6.5 ng ml(-1) for herring sperm DNA and 3.5 ng ml(-1) for yeast RNA, respectively. Three synthetic samples were analyzed satisfactorily.     

Spectrochemical Determination of Metallic Pollutants At Sub Ppm Levels In Fresh Water Samples

Abstract

A quantitative method to analyse fresh water samples for ultratrace pollutants such as Cd, Cr, Fe, Mn, Ni, Pb, V and Zn has been developed. the impurities in the water sample were preconcentrated by the method of slow evaporation and collected on specpure graphite which was subsequently analysed by d.c. are excitation using NaF as the carrier. the mean relative standard deviation of the method employed is + 15%. the accuracy of the method reported has been found to be quite satisfactory as borne out by the results of the Intercomparison Run conducted by the International Atomic Energy Agency, Vienna for the determination of trace elements in water. the detection limits for the various elements are as follows: Ni, Pb, V : 1 ng/ml; Cd, Cr, Mn: 2 ng/ml; Fe: 15 ng/ml and Zn : 20 ng/ml. Some of these detection limits have been compared with those obtained in some of the recent techniques such as AAS, Laser Induced Fluorescence (LIF) and Laser Enhanced Ionization (LEI) or Opto-Galvanic Spectrometry(òGS). It has been found that there is a tenfold increase in sensitivity for Pb in the present method as compared to the above techniques except LEI while the detection limits obtained for other elements are comparable with those in other techniques.     

Double resonance laser-induced fluorescence of Cadmium and Zinc in the inductively coupled plasma and electrothermal atomizer

Double resonance laser-induced fluorescence (LIF) spectrometry studies of Zn and Cd have been performed in the inductively coupled plasma (ICP) and electrothermal atomizer (ETA). Stepwise excitation of Zn is accomplished at 213.856 nm and 636.235 nm with fluorescence emissions observed near 334.5 nm. Excitation of Cd is accomplished at 228.802 nm and 643.847 nm with fluorescence emissions observed near 361.1 nm and 347.6 nm. Calibration studies demonstrate that the double resonance LIF approaches provide high sensitivity and linearity over several orders of magnitude in both atomizers. Limits of detection for Zn and Cd in the ICP are 1.7 ng/ml and 5 ng/ml, respectively. Excellent sensitivity is observed in the ETA resulting in limits of detection of 70 pg/ml (700 fg absolute mass) and 4 pg/ml (40 fg absolute mass) for Zn and Cd, respectively. The Zn content of a bovine serum standard reference material (NIST SRM #1598) has been determined by ETA-LIF and found to be 940±60 ng/g, which is in very good agreement with the certified value of 890±60 ng/g. Low-level ETA-LIF measurements of Zn in these studies are strongly limited by the high background level observed for this element.     

Spectrophotometric determination of manganese by using redox reaction of tris(2,2'-bipyridine)osmium(II) with Mn7+.

This paper reports on the analytical application of the oxidation reaction of [Os(bpy)3]2+ by Mn7+ (MnO4-). The present study developed a very simple, sensitive and selective spectrophotometric method for the determination of manganese in an acidic medium. Three analytical wavelengths were employed in the UV and visible regions at 290, 315 and 480 nm. Beer's law was obeyed up to a concentration of 330 ng ml(-1) of Mn7+ at different wavelengths with regression equations 0.001 - 0.0042C(Mn), 0.001 + 0.0021C(Mn), and 0.001 - 9.34 x 10(-4)C(Mn) at 290, 315 and 480 nm, respectively. Under the optimum conditions, the achieved detection limits were 0.72, 1.37 and 3.32 ng ml(-1) at 290, 315, and 480 nm, respectively. In addition to the high sensitivity of the method (0.24 ng cm(-2) at 290 nm, 0.45 ng cm(-2) at 315 nm and 1.0 ng cm(-2) at 480 nm), it can be used for the determination of manganese in the presence of a large number of anions and cations, since it tolerates most of the potential interferents. The relative standard deviation was 0.5% (n = 11) for 90 ng ml(-1) manganese. The method was successfully applied to the determination of manganese in water from different resources.