A Modified Enzymatic Method for the Estimation of Dihydroxyacetone

Abstract

A two stage assay procedure has been developed for the estimation of dihydroxyacetone (DHN). DHA is at first phosphorylated to dihydroxyacetone phosphate (DHAP) by glycerokinase (GK) and ATP at pH 8.5. The DHAP formed is then measured enzymatically using glycerol-3P dehydrogenase (GDH) and NADH at pH 7.6. The sensitivity of this method is found to be adequate for both routine assays and kinetic studies.     

High-Performance Liquid Chromatography Analysis of Hexamethylmelamine and its Demethylated and Hydroxylated Metabolites in Mice Plasma

Abstract

A selective assay of hexamethylmelamine and its metabolites in mice plasma has been developed. The method uses protein precipitation with acetonitrile in glass centrifuge tubes and analysis of the resulting supernatant by reverse-phase high performance liquid chromatography on a Lichrocart RP 18 column with a mobile phase acetonitrile-phosphate buffer containing n-propylamine (pH 7,5). The influence of various parameters on the separation (solvent composition, pH, n-propylamine content, temperature) has been examined.     

Optimization of a Direct Competitive Enzyme-Linked Immunoassay for Carbofuran and Application to Water Samples

Abstract

A direct competitive enzyme-linked immunosorbent assay (ELISA) for carbofuran was developed, which was based on the anti-BFNB IgG monoclonal antibody (McAb) and a homologous enzyme tracer (BFNB-HRP). The influence of several physicochemical factors (salt, pH, detergent, and solvent) on the immunoassay was studied. For the standard curve, an I₅₀ of 2.98 µg/l and a limit of detection (I₁₀) of 0.27 µg/l was obtained in a high salt concentration buffer (0.08 M PBS, pH 7.0) with 0.03% BSA. A common challenge for immunoassay, time-dependent drift, was effectively circumvented in our study. The optimized ELISA has been used to quantify carbofuran in water samples spiked at different amounts. The excellent recoveries (71% to 130%) achieved confirmed the potential of the immunoassay for monitoring of carbofuran in waters without purification steps.     

Rapid Spectrophotometric Assay of Cephradine and its Dosage Forms

Abstract

A new spectrophotometric method to determine cephradine based on its reaction with acetylacetone-formaldehyde reagent (pH 4.3) is proposed. The yellow chromophore shows an absorption maxima at 400 nm, having a molar absorptivity coefficient of 2.59. 10³ L mol^?1 cm^?1. Optimization of the reaction conditions has been investigated. Obediance to Beer's law (0–110 mog per ml) permitted the assay of cephradine in its dosage forms. The method has been compared with the official method and found to be simple, artwrate (t-test) and reproducible (F-test).     

Atomic Absorption Determination of Some Amino Acids Containing a Sulphur Group

Abstract

A simple and rapid atomic absorption method for the determination of some amino acids containing a sulphur group is described. The studied compounds are cysteine, cystine and methionine. The method is based on the addition of an excess of mercury (II) chloride in phosphate buffer, pH 9, forming a white precipitate; the unreacted mercury is separated by centrifugation and the mercury ions in both precipitate and filtrate are determined by atomic absorption spectrophotometry at 253.6 nm. The concentration of the studied amino acids is then indirectly determined from a calibration curve for standard mercury (II) chloride solution.

The procedure has been successfully applied to the assay of the pharmaceutical preparations of the studied drugs after thin-layer chromatographic separation. The results of the studied compounds compare favourably with the official methods.     

Application of Central Composite Design to the Optimization of HPLC Analysis of Nitroimidazoles

Abstract

A 2 factor central composite design is used to study the effects of acetonitrile concentration and eluent pH on the retention and resolution of metronidazole and tinidazole on a C-18 column. The equations obtained have been used to predict the composition of an eluent for an optimum separation. The assay developed has been applied to the quantitation of metronidazole in human plasma following simple protein precipitation. The detection limit at 320nm for metronidazole is 0.01μg/mL. Using this assay a relative bioavailability study of two commercial metronidazole tablets in ten healthy volunteers has been performed.     

Spectrophotometric Determination of Ceftazidime in Pharmaceutical Preparations Using Neocuproin as a Complexing Agent

Abstract

A simple and reproducible spectrophotometric method for the assay of ceftazidime with neocuproin-copper(II) reagent has been developed. The procedure is based on the drug in an acidic medium, subsequent formation of yellow ternary complex in citrate buffer solution (pH 4.2), and measurement at 454 nm. Beer's law is obeyed in the range 15.0–40.0 µg mL^?1 with correlation coefficient r ² = 0.9995. The procedure holds good accuracy and precision when applied to the analysis of ceftazidime in powder for injection with good recovery percent ranging from 100.17±1.0 without interference from additives.     

High Performance Liquid Chromatographic Analysis of Desipramine in Human Plasma Using a Microbore Column Technique

Abstract

A reversed-phase, isocratic HPLC method has been developed for the quantitation of desipramine in human plasma. the method involved the use of cloimpramine as an internal standard. the chromatographic separation was accomplished with a mobile phase comprising acetonitrile-aqueous solution (60:40. v/v) containing 10 mM disodium hydrogenphosphate and 80 mM sodium dodecyl sulfate adjusted to pH 2. the mobile phase was pumped at a flow rate of 0.5 ml/min. the column used was a microbore column (2 mm I.D. × 100 mm) packed with a C18 reversed-phase material (5μm ODS Hypersil). Plasma samples were extracted at basic pH with diethyl ether followed by back-extraction into 0.1 N sulfuric acid. Using UV detection at 250 nm, the lower limit of sensitivity was 10 ng/ml. the inter- and intra-assay coefficients of variation were found to be less than 10%. the assay procedure was applied to a long term oral dosing study in patients to monitor the plasma concentration of desipramine.     

Quantitation of Chlorpropamide and Tolbutamide in Tablets by Stability-Indicating Reverse Phase High-Performance Liquid Chromatography

Abstract

A stability-indicating assay method for the quantitation of chloropropamide and tolbutamide based on reverse phase high-performance liquid chromatography has been developed. The method is accurate, reproducible and precise with a percent relative standard deviations based on 5 readings of 0.52 and 0.46 for chlorpropamide and tolbutamide, respectively. There is no interference from the products of hydrolysis of chlorpropamide or tolbutamide (they elute before the intact drug) and th excipients present in the tablets. The hydrolysis process proceeds very fast in acidic pH as compared with basic pH. The products of hydrolysis, p-chlorobenzenesulfonamide (from chlorpropamide) and p-toluenesulfonamide (from tolbutamide) can also be quantified if required. The hydrolysis of both chlorpropamide and tolbutamide in 0.09N H₂SO₄ followed first order law with K values of 0.0036 and 0.0047 per day (at 50°), respectively.     

Determination of Ketotifen Fumarate in Raw Material and Pharmaceutical Products Using Ion‐pair Formation

Abstract

A simple and sensitive extractive spectrophotometric method has been described for the determination of Ketotifen. The developed method is based on the formation of a colored ion‐pair complex (1∶1 drug/dye) of Ketotifen and bromocresol green in buffer pH 3 and extraction in chloroform. The extracted complex shows absorbance maxima at 423 nm. Beer's law is obeyed in the concentration range of 5.15–61.91 µg/ml. The proposed method has been applied successfully for the determination of drug in commercial tablets and syrup dosage form. No significant interference was observed from the excipients commonly used as pharmaceutical aids with the assay procedure.     

Synthesis of a New pH-Dependent Ligand: Conformational and Complexation Studies

A new macrocyclic ligand, 3, which exhibits pH-induced conformational changes, has been prepared. This ligand consists of a crown ether derived from a trans-anti-trans 1,2,4,5-tetrasubstituted cyclohexane. Due to the stereochemistry of the substituents on the carbocyclic ring, two different low-energy conformations of the crown ether are possible. Ligand 3 has been studied in solution by ¹H NMR spectroscopy at different values of pH and temperature, showing that the conformation of the crown ether, and thus its complexing ability, is strongly pH-dependent. The solid-state structure of the ligand has been determined by X-ray diffraction.     

Spectrofluorimetric Determination of Gallium with 5-Bromosalicylidene-o-Aminophenol

Abstract

A method for the determination of trace amounts of gallium has been developed, based on the formation of a fluorescent complex between Ga (III) and 5-Bromosalicylidene-o-aminophenol (5-BrSOAPh). With excitation at 425 nm the chelate has an emission maximum at 520 nm. The reaction is carried out at apparent pH (pH?) 5.40 in an aqueous-ethanol medium (60% V/V ethanol). The influence of the reaction variables is discussed.     

Reverse Phase HPLC Determination and Murine Pharmacokinetics of Arabinosyl-5-azacytosine

Abstract

A sensitive and specific reverse phase HPLC assay has been developed to measure the new antitumor agent arabinosyl-5-aza-cytosine (ara-AC) in biological fluids at concentrations as low as 50 ng/ml (0.2 μM). This assay also detects arabinosyl-N-formyl-guanylurea (AGU-CHO), the initial hydrolytic metabolite of ara-AC. 2′-Deoxy-5-azacytidine, an analogue with similar chemical stability, is used as an internal standard. Chromatographically interfering plasma ribosides are removed by solid phase extraction on a phenyl boronic acid cartridge. Separation of ara-AC, AGU-CHO and internal standard is then accomplished isocratically (1% CH₃CN in 10 mM pH 6.8 phosphate buffer) on fully carbon loaded and end-capped C₈ and C₁₈ columns connected in tandem. The compounds of interest are detected by UV absorption at 240 nm and total analysis time is 20 min. This assay has been used to determine bolus dose plasma kinetics in male BDF₁ mice given 200 mg/kg ara-AC as a tail vein injection. Plasma elimination of the ara-AC is triphasic with a terminal phase half-life of 52 min and the elimination of the AGU-CHO metabolite parallels that of the parent drug. Analysis of ara-AC in human plasma indicates that this method is suitable for determining drug disposition and pharmacokinetics in human subjects.     

Stability Indicating Hplc Method for the Determination of Sparfloxacin (Spar)

ABSTRACT

A rapid, sensitive and stability indicating method for the determination of sparfloxacin (SPAR) by RP - HPLC has been developed on a Merck RP - Select B (5 μm; 12.5 cm x 4.0 mm) column using a mobile phase of water: acetonitrile: triethylamine (80 : 20 : 0.2 v/v) pH of which was adjusted to 2.6 with orthrophosphoric acid. The flow rate was 1 ml / min. and the detection was carried out at 304 nm using Waters 486 variable wavelength detector. The retention time for SPAR was 7.2 min. Linearity range was from 8 - 1000 ppm. The method showed good precision and accuracy when applied to two brands of tablets containing SPAR. In alkaline media SPAR is stable where as it undergoes degradation in acidic and oxidising conditions generating different degradation products the nature of which is required to be established. The proposed method nicely separates the degraded products from SPAR and hence can be used as stability indicating method for the assay of SPAR.     

Flow Injection Manifold for Simultaneous Spectrophotometric Determination of Bi (III), Pb (II)

Abstract

A FIA assembly producing a carrier with pH which can be adjusted to the desired pH value is propposed. It is based on the merging of two different solutions, one of them at constant flow-rate and the other at variable flow-rate. This manifold has been used for the simultaneous determination of Bi(III) and Pb(II) with Arsenazo III. Calibration curves are linear in the 1.0-11.0 ppm Bi (III) range at pH 0.25 and 1.0-12.1 ppm Pb (II) at pH 2.15. The effect of foreign ions is also reported.     

A new procedure for determining copper in igneous rock and alloys

The influence of cationic surfactants, cetylpyridinium chloride (CPC), cetylpyridinium bromide (CPB), and cetyltrimethylammonium bromide (CTMAB), on the complexation of copper(II) with thenoyl-(2-oxopropyl)-N-(2-sulfo-4-nitro-5-oxyphenyl) azomethine (R) has been investigated. It has been found that monoligand compounds are formed at pH 4, while at pH 3 the formation of mixed-ligand compounds is observed. The detection limit for copper decreases and the stability constants of the complexes increase in the row Cu-R-CPC > Cu-R-CPB > Cu-R-CTMAB, with AN increase in the stability of the associates (Rh-CPC > R-CPB > R-CTMAB) in the complexation reaction. The ratio of components in monoligand (1: 1) and mixed-ligand (1: 1: 1) compounds has been determined. The limits of obedience to the Beer law have been found. A procedure has been developed for the photometric determination of copper in igneous rock and alloys.     

Untersuchung Der Kcmplexbildung Von Thallium(III) Mit 1-Phenyl-3-Methyl-4-Benzoylpyrazolon-5

Abstract

A study has been made of the method used for extracting the complex formed between thallium(III) and PhMBP. It has been demonstrated that thallium(III) can be readily extracted as a complex with PhMBP by organic solvents at low pH. The extraction of thallium(III) has been studied as a function of pH, concentration of reagent, nature of solvent, duration of contact of the phases and concentration of thallium(III). It has been established that thallium(III) forms a complex with PhMBP of the type MA_n, where n=3. The stability constants and distribution constant of the neutral complex has been calculated by the method of Leden-Rydberg.     

Preliminary Evalution of Fluorscamine As a Clorimetric Reagent for Primary Amines

Abstract

Fluorescamine reacts efficiently with primary amines to form pyrrolinones with long wavelength absorption at 390–410nm. A procedure has been worked out which allows the use of this reaction for the colorimetric assay of primary amines.     

Acid Phosphatase Assay with a Wireless Magnetoelastic Biosensor

Abstract

A wireless remote‐query disposable magnetoelastic (ME) biosensor was developed for the assay of acid phosphatase (ACP). The sensor was fabricated by applying a magnetoelastic ribbon with a layer of pH‐sensitive polymer and upon it a sensing film containing bovine serum albumin (BSA) and adenosine‐5′‐monophosphate (5′‐AMP). The ACP‐catalyzed hydrolysis of 5′‐AMP decreases the solution pH, resulting in the polymer shrinking and consequently the resonance frequency of the magnetoelastic sensor increasing. The kinetic parameters were measured to be 1.64×10^?3 M (Michaelis constant) and 130 Hz/min (maximum initial rate). The proposed sensor can determine 0.2 to 1.2 U/ml of ACP.     

Stabilization of Sensing Assay within Polyelectrolyte-Coated Alginate Microspheres for Optical Urea Sensing

Abstract

A new absorbance-based enzymatic biosensor for determination of urea (in the range 0.01 to 6.7 mM) is described. Quantification using cresol red dye, immobilized in the nanofilm coatings assembled on alginate microspheres to immobilize the urease enzyme, has been accomplished using ratiometric absorbance measurements. The effect of salt concentration in polyelectrolyte nanofilms (on the stability of dye molecules) and buffer pH (on the enzyme stability) are reported. The results demonstrate excellent stability of sensing assay within alginate microspheres. Urea-sensing experiments demonstrate the potential to develop an optical urea sensor that is stable over a month.