Determination of dissociation constants of sparingly soluble phenothiazine derivatives by second-derivative spectrophotometry

The dissociation constants (pK_a) for sparingly soluble phenothiazines (promazine, chlorpromazine, trifluoropromazine) in water were measured by second-derivative spectrophotometry. The intense background signals in the absorption spectra due to the turbidity caused by the precipitation of insoluble free base of the phenothiazine derivatives were eliminated in the second-derivative spectra, and the solubilities of the phenothiazine derivatives could be easily determined from the peak-to-trough lengths (D values) of the second-derivative signals. The pK_a values were calculated from the pH dependence of the D values. The pK_a values obtained agreed well with reported values and the standard deviations for 6–10 determinations were ? 0.02. The solubilities of the free bases of the phenothiazines were also determined.     

Indirect Potentiometric Determination of Chlorpromazine with an Oxidative Column In a Flow Injection System

Abstract

A flow-injection method is proposed for the determination of chlorpromazine and other N-substituted phenothiazines. The procedure is based on the oxidation of analyte with lead dioxide entrapped into polymeric material in a packed-bed reactor. The oxidation of the drug yields soluble Pb^2-, which is monitored by means of the lead ion selective electrode in the wall-jet configuration. The calibration graph is linear over the range of 0.01 – 2 μg ml^?1 of chlorpromazine with relative standard deviation of 1.4% and sample throughput 20 h^?1. The developed method was applied to the determination of chlorpromazine in pharmaceutical preparations.     

Determination of Silicon in Lubricant Oil by High-Resolution Continuum Source Flame Atomic Absorption Spectrometry Using Least-Square Background Correction and Internal Standardization

The least-squares background correction (LSBC) and internal standardization procedures were combined to eliminate spectral interferences caused by the CS molecular band (251.602 nm) and transport effects for determining Si in sulfuric acid digests of lubricant oil by high-resolution continuum source flame atomic absorption spectrometry. Aluminum, Ba, Ti, V, and W were tested as internal standard (IS) candidates, and W provided the best results. For absorbance measurements of solutions containing 0.5–5.0 mg L^?1 Si in the presence of 25 mg L^?1 W (at the wavelength integrated absorbance equivalent to 3 pixels), the correlation coefficient for the ratio of absorbance of Si to absorbance of W vs. analyte concentration was 0.9978. Fluctuations in analytical signals due to variations in sulfuric acid concentrations or acetylene/nitrous oxide flow-rate ratios were corrected by using this calibration plot. Relative standard deviations varied from 1.9 to 7.2% and 2.1 to 5.4% (n = 12) with and without LSBC/IS, respectively. Recoveries for samples spiked with 2.0 mg L^?1 Si in 5.0% (v/v) sulfuric acid were within the 72.5–82.5% and 94.0–99.0% ranges without correction and by LSBC associated with internal standardization procedure, respectively. Accuracy of the proposed method was checked for the determination of Si in commercial lubricant oils and results obtained with internal standardization were better than those without correction.     

UV-Derivative Spectrophotometric Determination of Ritonavir Capsules and Comparison with LC Method

Abstract

An ultraviolet-derivative spectrophotometric method (UV-D) has been proposed as an alternative to a previously described liquid chromatographic (LC) method for the quantitative determination of ritonavir in soft gelatin capsules. The spectrophotometric method is based on recording the second-derivative spectra for ritonavir at 222.3 nm of its solutions in methanol. The linear dynamic range was 10.0–30.0 µg · mL^?1 with a correlation coefficient of 0.9995. Mean recoveries were between 99.2% and 100.2% for the tested capsules samples. Mean intra- and interassay relative standard deviations (RSDs) were less than 2.0%. The statistic analysis showed that LC and UV-D methods were equivalent to assay ritonavir capsules.     

Selection of internal standards for quantitative analysis by matrix-assisted laser desorption-ionization (MALDI) time-of-flight mass spectrometry

 The selection of an appropriate internal standard (IS) for quantification by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry is critical for the successful application of quantitative MALDI. Selection of the IS depends on the chemical similarity of the analyte and IS and the mass separation of the analyte and IS as a function of instrumental peak resolution. For the quantification of bovine insulin, a series of internal standards including horse heart cytochrome C, bovine insulin chain B, des-pentapeptide human insulin, and des-octapeptide porcine insulin was investigated. Des-pentapeptide human insulin was found to be the most appropriate internal standard (relative standard deviation of the standard curve slope=2.99%, correlation coefficient=0.988 in the range of 0.5–0.4 μmol/L). Two methods for measuring of the MALDI signal intensity were evaluated, direct peak integration following subtraction of a linear background and non-linear least squares curve fitting. The results obtained with these methods were equivalent. Received: 10 November 1995 / Revised: 4 March 1996 / Accepted: 6 March 1996     

Use of the internal standardization for difficult sampling by graphite furnace atomic absorption spectrometry

This work shows the potentiality of As as internal standard to compensate errors from sampling of sparkling drinking water samples in the determination of selenium by graphite furnace atomic absorption spectrometry. The mixture Pd(NO₃)₂/Mg(NO₃)₂ was used as chemical modifier. All samples and reference solutions were automatically spiked with 500 μg l⁻¹ As and 0.2% (v/v) HNO₃ by the autosampler, eliminating the need for manual dilutions. For 10 μl dispensed sample into the graphite tube, a good correlation (r=0.9996) was obtained between the ratio of analyte absorbance by the internal standard absorbance and the analyte concentrations. The relative standard deviations (R.S.D.) of measurements varied from 0.05 to 2% and from 1.9 to 5% (n=12) with and without internal standardization, respectively. The limit of detection (LD) based on integrated absorbance was 3.0 μg l⁻¹ Se. Recoveries in the 94-109% range for Se spiked samples were obtained. Internal standardization (IS) improved the repeatability of measurements and increased the lifetime of the graphite tube in ca. 15%.     

Internal standardization and least-squares background correction in high-resolution continuum source flame atomic absorption spectrometry to eliminate interferences on determination of Pb in phosphoric acid

In this work the least-squares background correction (LSBC) and internal standardization (IS) techniques were combined to eliminate spectral and transport interferences in the determination of Pb in phosphoric acid by high-resolution continuum source atomic absorption spectrometry (HR-CS AAS). Blanks, samples and reference solutions [0.10–1.00 mg L^− 1 Pb in 1% (v/v) HNO₃] were spiked with 4.00 mg L^− 1 Co used as internal standard. For absorbance measurements at the wavelength integrated absorbance equivalent to 9 pixels, correlations between the ratio of absorbance of Pb to absorbance of Co and the analyte concentration were close to 0.9992. Relative standard deviations of measurements varied from 0.6 to 4% and 1 to 7% (n = 12) without and with IS/LSBC techniques, respectively. Recoveries for Pb spikes were in the 96–104% and 76–180% range with and without IS/LSBC, respectively. The limit of detection improved with IS/LSBC techniques. Accuracy of the proposed method was checked for the determinations of Pb in commercial phosphoric acid samples and results obtained with IS were better than those without IS.     

Investigating the post-chemiluminescence behavior of phenothiazine medications in the luminol–potassium ferricyanide system: molecular imprinting–post-chemiluminescence method for the determination of chlorpromazine hydrochloride

A new post-chemiluminescence (PCL) phenomenon was observed when phenothiazine medications were injected into the reaction mixture after the chemiluminescence (CL) reaction of luminol and potassium ferricyanide had finished. A possible reaction mechanism was proposed based on studies of the kinetic characteristics of the CL, CL spectra, fluorescence spectra, and on other experiments. The feasibility of determining various phenothiazine medications by utilizing these PCL reactions was examined. A molecular imprinting–post-chemiluminescence (MI-PCL) method was established for the determination of chlorpromazine hydrochloride using a chlorpromazine hydrochloride-imprinted polymer (MIP) as the recognition material. The method displayed high selectivity and high sensitivity. The linear range of the method was 1.0×10⁻⁸∼1.0×10⁻⁶, with a linear correlation coefficient of 0.9985. The detection limit was 3×10⁻⁹ g/ml chlorpromazine hydrochloride, and the relative standard deviation for a 1.0×10⁻⁷ g/ml chlorpromazine hydrochloride solution was 4.0% (n=11). The method has been applied to the determination of chlorpromazine hydrochloride in urine and animal drinking water with satisfactory results.      

Electrothermal Vaporization of Trace Gallium via In SITU Alkylation for Inductively Coupled Plasma Atomic Emission Spectrometry

Abstract

A sample introduction method based on alkylation of the analyte was proposed for introducing trace gallium into inductively coupled plasma for atomic emission spectrometry. By using 10 μl of a sample solution with 15 μl of a 0.9 mol 1^-1 solution of ethylmagnesium bromide in tetrahydrofuran as alkylating reagent, a detection limit of 1.9 ng of gallium was obtained and the calibration graph was linear with the amount of gallium up to 100 ng. The relative standard deviation (n = 10) was 2.0% when 10 ng of gallium was present in the sample.     

Selection of internal standards for quantitative analysis by matrix-assisted laser desorption-ionization (MALDI) time-of-flight mass spectrometry

The selection of an appropriate internal standard (IS) for quantification by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry is critical for the successful application of quantitative MALDI. Selection of the IS depends on the chemical similarity of the analyte and IS and the mass separation of the analyte and IS as a function of instrumental peak resolution. For the quantification of bovine insulin, a series of internal standards including horse heart cytochrome C, bovine insulin chain B, des-pentapeptide human insulin, and des-octapeptide porcine insulin was investigated. Des-pentapeptide human insulin was found to be the most appropriate internal standard (relative standard deviation of the standard curve slope = 2.99%, correlation coefficient = 0.988 in the range of 0.5-0.4 μmol/L). Two methods for measuring of the MALDI signal intensity were evaluated, direct peak integration following subtraction of a linear background and non-linear least squares curve fitting. The results obtained with these methods were equivalent.     

Flow-Injection Spectrophotometric Determination of Oxalate,Citrate and Tartrate Based on Photochemical Reactions

Abstract

A flow-injection configuration for the spectrophotometric determination of oxalate, citrate and tartrate is proposed. The procedure is based on the photochemical decomposition of the complexes formed between iron(III) and these anions. The iron(II) produced in the photochemical reactions was detected by measuring the absorbance after complexation with ferrozine (λ_max=562 nm). Linear calibration graphs were obtained over the concentration ranges 5.0 × 10^?6 - 1.0 × 10^?4 M, 8 × 10^?6 - 1.8 × 10^?4 M and 1.0 × 10^?6 - 2 × 10^?5 M for oxalate, citrate and tartrate, respectively. The relative standard deviations at the 1x10^?5 M concentration level were within the range 1.29 - 1.47 %. The sampling frequency was about 40 samples h^?1. The usefulness of the method was tested in the determination of oxalate in urine and spinach, of citrate in pharmaceuticals and soft drinks and of tartrate in pharmaceuticals. For the determination of oxalate in urine samples a prior separation of the analyte by precipitation with calcium chloride is recommended.     

Spectrophotometric Multicommutated Flow System for the Determination of Hypochlorite in Bleaching Products

Abstract

A multicommutation flow system for the spectrophotometric determination of hypochlorite in bleaching products is proposed. In this system, N,N-diethyl-p-phenylenediamine (DPD) reacts with hypochlorite, and the product was monitored at 515 nm. The analytical curve for hypochlorite was linear in the concentration range from 2.68 × 10^?5 to 1.88 × 10^?4 mol L^?1 (2–14 mg L^?1) with a detection limit of 6.84 × 10^?6 mol L^?1 (0.51 mg L^?1). The sampling rate was 45 h^?1, and a relative standard deviation of 1.4% (n = 10) was obtained. The recovery of this analyte ranged from 97.2% to 102.5%, and the results found using the proposed spectrophotometric multicommutated flow system agreed with the data obtained using a reference method (iodometric titration) at the 95% confidence level.     

Simultaneous Determination of Carbamazepine,Phenobarbital and Their Major Metabolites in Serum,Brain Tissue and Urine by High-Performane Liquid Chromatographya

Abstract

A high-performance liquid chromatographic method is described for the simultaneous measurement of carbamazepine, phenobarbital and their major metabolites in small samples of serum, brain tissue and urine. This involves solvent extraction, reversed-phase chromatography and ultraviolet detection at 195 nm. Glucuronides in urine are hydrolyzed by enzymatic cleavage with β-glucuranidase. Quantitation is based on the peak-height ratio of the analyte to its internal standard (10, 11-dihydrocarbamaze-pine or p-methylphenobarbital). The results obtained show that the method is precise and reproducible.     

Liquid chromatographic–electrospray mass spectrometric determination of cyclosporin A in human plasma

A rapid, sensitive and selective liquid chromatography–mass spectrometry (LC–MS) assay has been developed for determination of cyclosporin A (CyA) in human plasma; cyclosporin B (CyB) was used as internal standard (IS). The method utilized a combination of a column-switching valve and a reversed-phase symmetry column. The mobile phase was a 25:75 (v/v) mixture of 10% aqueous glacial acetic acid and acetonitrile. Running time per single run was less than 10 min. Sample preparation included C₈ SPE of human plasma spiked with the analyte and internal standard, evaporation of the eluate to dryness at 50°C under N₂ gas, and finally reconstitution in the mobile phase. Detection of cyclosporin A and the IS was performed in selected ion-monitoring mode at m/z 601.3 and 594.4 Da for CyA and IS, respectively. Quantitation was achieved by use of the regression equation of relative peak area of cyclosporin to IS against concentration of cyclosporin. The method was validated according to FDA guideline requirements. The linearity of the assay in the range 5.0–400.0 ng mL^–1 was verified as characterized by the least-squares regression line Y=(0.00268±1.9×10^–4)X+(0.00078±1.8×10^–3), correlation coefficient, r=0.9986±1.1×10^–3 (n=48). Intra and inter-day quality-control measurements in the range 5.0–350.0 ng mL^–1 revealed almost 100% accuracy and 9% CV for precision. The mean absolute recovery of CyA was found to be 84.01±9.9% and the respective relative recovery was 100.3±9.19. The limit of quantitation (LOQ) achieved was 5 ng mL^–1. Eventually, stability testing of the analyte and IS in plasma or stock solution revealed that both chemicals were very stable when stored for long or short periods of time at room temperature or –20°C.     

Development of a Spectrofluorimetric Method for Determination of Albendazole in Tablets

Abstract

In this study a spectrofluorimetric method was developed to determine drug active compound in the tablets for albendazole (ABZ). For this aim, fluorescence spectra of albendazolee drug active compound in various solvents were taken, it was determined that the most suitable solvent was chloroform and excitation (λ_ex) and emission (λ_em) wavelength in a row was 360 nm and 440 nm in this solvent. Calibration graphics were drawn and shown linear at 0–2.5 mg/l concentration range (R²=0.9993). Albendazole quantity in the andazol tablets was determined from directly calibration graphs and with standard addition method. Results obtained with developed spectrofluorimetric method were compared with standard USP method and it was found no difference between two methods within 95% confidence limits. It was determined that proposed method was easy and highly accuracy method to be able to use in routine albendazole analyze for the quality control.     

Spectrofluorimetric Flow Injection Determination of Glycerol & Ethylene Glycol with Alizarin Navy Blue

ABSTRACT

A simple and accurate procedure for indirect spectrofluorimetric determination of glycerol and ethylene glycol in aqueous media was developed. Alizarin Navy Blue can be oxidized by potassium periodate to produce a fluorescent compound, that can be detected fluorimetrically (λ_ex = 370 nm, λ_em = 516 nm) by a flow through method. Glycerol and ethylene glycol react with periodate decreasing its concentration and thus the forming the basis for an indirect determination of these compounds. The influence of acid concentration, reagent concentration and manifold variables were studied. For glycerol and ethylene glycol linear calibration curves were obtained in the ranges of 4.8 × 10^-7 – 8.7 × 10^-5 M and 6.4 × 10^-7 – 8.7 × 10^-5 M, respectively. The limit of detection (defined as the concentration that gives a signal three times the standard deviation of the background signal) for glycerol and ethylene glycol was 3.2 × 10^-7 M and 4.3 × 10^-7 M, respectively. The proposed method was applied for determination of glycerol and ethylene glycol in synthetic and real samples. The results obtained were satisfactory.     

Rapid Liquid Chromatographic – Tandem Mass Spectrometric Method for the Quantification of Pentoxifylline in Human Plasma

A simple, rapid, sensitive and selective liquid chromatography / tandem mass spectrometry method was developed and validated for the quantification of pentoxifylline, a haemorheological agent. The analyte and internal standard, tamsulosin were extracted by liquid-liquid extraction with ethyl acetate and were separated using an isocratic mobile phase on a reverse phase C₁₈ column. The analytes were analyzed by mass spectrometry in the multiple reaction monitoring mode using the respective [M+H]⁺ ions, m/z 279/138 for pentoxifylline and m/z 409/228 for the IS. The assay exhibited a linear dynamic range of 2–1000 ng mL⁻¹ for pentoxifylline in human plasma. The lower limit of quantification was 2 ng mL⁻¹ with a relative standard deviation of less than 10%. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. A run time of 1.5 min for each sample made it possible to analyze more than 400 human plasma samples per day. The validated method has been successfully used to analyze human plasma samples for application in pharmacokinetic, bioavailability or bioequivalence studies. Revised: 4 and 20 October 2005     

Optimization,Validation and Application of a Capillary Zone Electrophoresis Method for the Assay of Sparfloxacin in Pharmaceutical Formulation

Abstract

A capillary zone electrophoresis (CZE) method has been developed for the determination of the antibiotic sparfloxacin in tablets. The CZE separation was performed using 75 µm×35 cm fused-silica capillary under the following conditions: 25°C; applied voltage, 12 kV; 25 mM H₃PO₄-NaOH running buffer (pH 8.5). The detection wavelength was 254 nm. Flumequine was used as internal standard (IS). The method was suitably validated with respect to linearity, limit of detection and quantification, accuracy, precision, specificity, and robustness. The calibration was linear from 10 to 60 µg mL^?1 and the limit of detection and quantification were 5.38 and 9.46 µg mL^?1, respectively. Recoveries ranging from 95.68%–102.4% were obtained in the determination of sparfloxacin that were spiked to placebos. Excipients in the commercial tablets and degraded products from different stress conditions did not interfere in the assay. The method was successfully applied to the determination of sparfloxacin in pharmaceutical tablets.     

Flow injection spectrophotometric determination of fosfestrol, following on-line thermal induced digestion and using an orthophosphate calibration graph

A novel flow injection (FI) system for the spectrophotometric determination of diethyl stilbestrol diphosphate (fosfestrol) in pharmaceutical formulations is described. On-line thermal induced digestion of the analyte by peroxodisulfate ion was performed and the orthophosphate ion generated was determined spectrophotometrically (λ_max=690 nm) using a molybdenum blue based FI approach. As the achieved conversion of the analyte was quantitative, an orthophosphate calibration graph can be used for its determination as well. The chemical and FI variables affecting the digestion were investigated. A linear calibration graph was obtained in the range 5.0×10⁻⁷-1.0×10⁻⁴ mol l⁻¹ fosfestrol. The relative standard deviation was very good (s_r=0.8% at 5.0×10⁻⁵ mol l⁻¹ fosfestrol, n=12) and the 3σ detection limit was 2.5×10⁻⁷ mol l⁻¹. The sampling rate was 60 injections h⁻¹. The average accuracies for the determination of the analyte in a pharmaceutical formulation evaluated by comparison of the results with those obtained by the supplier (Asta Medica) and the method recommended by the US Pharmacopoeia were also very good (e_r of +0.8 and −0.3%, respectively). Average recoveries of known amounts of the analyte ranging between 97.9 and 100.8% were also obtained.     

Determination of limonin in dog plasma by liquid chromatography–tandem mass spectrometry and its application to a pharmacokinetic study

A highly sensitive liquid chromatography–tandem mass spectrometry method was developed and validated for the determination of limonin in beagle dog plasma using nimodipine as internal standard. The analyte and internal standard (IS) were extracted with ether followed by a rapid isocratic elution with 10 mm ammonium acetate buffer–methanol (26:74, v/v) on a C₁₈ column (150 × 2.1 mm i.d.) and subsequent analysis by mass spectrometry in the multiple reaction monitoring mode. The precursor to product ion transitions of m/z 469.4 → 229.3 and m/z 417.2 → 122.0 were used to measure the analyte and the IS. The assay was linear over the concentration range of 0.625–100 ng/mL for limonin in dog plasma. The lower limit of quantification was 0.312 ng/mL and the extraction recovery was >90.4% for limonin. The inter‐ and intra‐day precision of the method at three concentrations was less than 9.9%. The method was successfully applied to pharmacokinetic study of limonin in dogs. Copyright © 2012 John Wiley & Sons, Ltd.