Flow-injection spectrophotometric determination of methyldopa in pharmaceutical formulations

A flow-injection spectrophotometric procedure is proposed for methyldopa determination in pharmaceutical preparations. The determination is based on formation of a yellow product (measured at 410 nm) after complexation of methyldopa with molybdate. Under optimal conditions, Beer's law is obeyed in a concentration range of 50-200 mg l⁻¹ methyldopa. Typical correlation between absorbance and analyte concentration was 0.9999. Usual excipients used as additives in pharmaceuticals do not interfere with the proposed method. The analytical frequency was 210 h⁻¹ and the relative standard deviation (R.S.D.) was ≤2% for sample solution containing 150 mg l⁻¹ methyldopa (n = 11). The analytical results obtained in commercial formulations by applying the proposed FIA method were in good agreement with labeled values and those obtained by the Brazilian Pharmacopoeia procedure at 95% confidence level.     

Enzymatic determination of vitamin A in pharmaceutical formulations with spectrophotometric detection

An enzymatic method for the determination vitamin A (retinol) is reported using soluble and immobilized alcohol dehydrogenase, isolated from rabbit liver. The reaction is based on the oxidation of retinol and simultaneous reduction of NAD⁺ to NADH followed by spectrophotometric detection at 340 nm. The calibration graph was linear over the range of 2.0–10 μM with correlation coefficients of 0.9967 and 0.9992 (n = 5) for soluble and immobilized alcohol dehydrogenase respectively, with relative standard deviations (n = 3) in the range of 0.5–1.2%. The limit of detection was lower than 1.0 μM. The proposed method was applied to determine vitamin A in pharmaceuticals, and the results obtained were in reasonable agreement with the amount labeled. The results were compared using spectrophotometric reference method, and no significant difference was found between the results of the both methods.     

Simultaneous determination of Folpet and Metalaxyl in pesticide formulations by flow injection Fourier transform infrared spectrometry

Fourier transform infrared (FTIR) spectrometric methodology has been developed for the simultaneous determination of Folpet and Metalaxyl in pesticide formulations. The method involves the extraction of both active principles by sonication of the sample with CHCl₃ and direct measurement of the absorbance using peak height values at 1798 cm⁻¹ corrected with a baseline defined at 1810 cm⁻¹ for Folpet, and peak area data established from 1677 to 1667 cm⁻¹ corrected using a baseline from 1692 to 1628 cm⁻¹ for Metalaxyl. Limits of detection (3 s) values of 17 and 16 μg g⁻¹ were found for Folpet and Metalaxyl, respectively, with results comparable with those found by liquid chromatography with UV detection. The new method involves a considerable decrease in solvent consumption. The automation of the procedure by incorporating on-line dissolution and filtration of the samples allows complete mechanisation of the method and improves the safety of operation.     

Electrochemical studies and square-wave voltammetric determination of fenofibrate in pharmaceutical formulations

The electrochemical reduction of fenofibrate at a hanging mercury drop electrode (HMDE) was investigated by cyclic voltammetry, square-wave voltammetry, and chronoamperometry. Different buffer solutions were used over a wide pH range (3.0–10.0). The best definition of the analytical signals was found in borate buffer (pH 9.0)–tetrabutylammonium iodide mixture containing 12.5% (v/v) methanol at –1.2 V (versus Ag/AgCl). According to cyclic voltammetric studies, the reduction was irreversible and diffusion controlled. The diffusion coefficient was 2.38×10^–6 cm² s^–1 as determined by chronoamperometry. Under optimized conditions of square-wave voltammetry, a linear relationship was obtained between 0.146–4.96 g mL^–1 of fenofibrate with a limit of detection of 0.025 g mL^–1. Validation parameters such as sensitivity, accuracy, precision, and recovery were evaluated. The proposed method was applied to the determination of fenofibrate in pharmaceutical formulations. The results were compared with those obtained by a published high-performance liquid chromatography method. No difference was found statistically.     

Fluorimetric Determination of Gatifloxacin in Aqueous,Pure and Pharmaceutical Formulations

Abstract

A spectrofluorimetric method was developed for the determination of gatifloxacin. The emission peak for gatifloxacin was recorded at 495 nm upon excitation at 291 nm. The fluorescence process was pH dependent. The dynamic range for the method was 16–80 ng ml^?1with detection limit of 3.97 ng ml^?1. A linear relationship between the fluorescence intensity and the concentration of gatifloxacin solution was obtained with r ² of 0.9968. The method has successfully applied to the determination of gatifloxacin in pure, authentic and aqueous samples.     

Chromatographic determination of caffeine in pharmaceutical formulations using micellar mobile phases

Summary An HPLC procedure is described for the determination of caffeine in pharmaceutical preparations. A Spherisorb octadecylsilane ODS-2 C₁₈ analytical column and spectrophotometric detection at 273 nm were used. The chromatographic behaviour of caffeine with different micellar eluents containing sodium dodecyl sulphate (SDS) is described. The determination of caffeine in pharmaceutical preparations was performed by use of a mobile phase containing 0.05 M sodium dodecylsulphate (SDS) and 1.5% propanol at pH7. At a 6.0 g mL^–1 concentration level the peak area and peak height repeatability were 2.6 and 2.4%, respectively. The application of the proposed method to the analysis of five pharmaceutical formulations, using peak heights as the dependent variable, gave recoveries between 85 and 104% of the values declared by the manufacturers. The proposed procedure for the determination of caffeine is rapid (15 min per sample), reliable and free from interferences.     

Electrochemical determination of prolintane in pharmaceutical formulations and in human urine

The oxidative behavior of 1-[1-(phenylmethyl)butyl]pyrrolidine, prolintane, was studied at a glassy carbon electrode using linear-sweep and differential-pulse voltammetry. The oxidation process was shown to be irreversible using 0.04 M Britton–Robinson buffer and was diffusion-adsorption controlled. Two voltammetric methods were developed for the determination of prolintane using different techniques: linear-sweep and differential-pulse voltammetry. The peak current varied linearly with prolintane concentrations in the range of 1.0 × 10⁻⁵ −2.5 × 10⁻⁴ M, with a detection limit of 8.5 × 10⁻⁶ and 4.0 × 10⁻⁶ M, and with relative standard deviations of 2.1 % and 3.1 %, respectively. The methods were applied to commercial preparations, giving relative errors less than 3.1 % and relative standard deviations lower than 4.8 % (n = 10). Determination of prolintane (down to the 8.5 × 10⁻⁸ M level) can be performed by using a preconcentration step prior to the determination by differential-pulse voltammetry in 0.04 M Britton–Robinson buffer (pH 8.0) with preconcentration potential of 0.0 V. The detection limit was found to be 6.2 × 10⁻⁸ M (4 min preconcentration) and the relative standard deviation for 2.5 × 10⁻⁷ M prolintane (n = 5) was 4.6 %. Applicability to human urine analysis is illustrated (recovery 98 ± 2 %). Standard additions method can be used to determine prolintane in real samples of urine.     

Determination of fenoterol and salbutamol in pharmaceutical formulations by electrogenerated chemiluminescence

Fenoterol and salbutamol were determined by electrogenerated chemiluminescence (ECL) coupled with flow injection analysis (FIA), using Ru(bpy)₃²⁺ as the luminescent substance. Fenoterol and salbutamol oxidize together with the ruthenium 2,2-bipyridyl at a platinum electrode, which leads to an increase in the luminescent intensity, and this increase is proportional to the analyte concentration. For fenoterol a linear calibration curve within the range from 1.0 × 10⁻⁵ to 1.0 × 10⁻⁴ mol l⁻¹ was obtained with a correlation coefficient of 0.998 (n = 5) and for salbutamol the linear analytical curve was also obtained in this range with a correlation coefficient of 0.995 (n = 5). The relative standard deviation was estimated as ≤2.5% for 3 × 10⁻⁵ mol l⁻¹ for fenoterol solution and as ≤1.3% for 5.0 × 10⁻⁵ mol l⁻¹ salbutamol solution for 15 successive injections. The limit of detection for fenoterol was 2.4 × 10⁻⁷ mol l⁻¹ and for salbutamol was 4.0 × 10⁻⁷ mol l⁻¹. Fenoterol and salbutamol were successfully determined in drug tablets and the soluble components of the matrix did not interfere in the luminescent emission. The results obtained using the luminescent methodology were not statistically different from those obtained by UV-spectrophotometry at 95% confidence level.     

Simultaneous determination of olmesartan medoxomil and irbesartan and hydrochlorothiazide in pharmaceutical formulations and human serum using high performance liquid chromatography

 A simple, selective, sensitive, precise, simultaneous high performance liquid chromatographic analysis of serum samples and commercial tablet formulation containing hydrochlorothiazide, olmesartan medoxomil and irbesartan are reported. Good chromatographic separation was achieved using a μ-Bondapak, C18 column (15 cm×4.6 mm, 5 μm), and a mobile phase consisting of acetonitrile-0.2% acetic acid aqueous solution (50∶50, v/v) at a flow rate of 1.0 mL/min. The ultraviolet detector was set at a wavelength of 260 nm. Hydrochlorothiazide, olmesartan medoxomil, and irbesartan were eluted at 1.2, 3.8, and 4.4 min, respectively. No extraneous materials were found to interfere. The method uses protein precipitation with acetonitrile for the preparation of serum sample. The linear ranges for hydrochlorothiazide, olmesartan medoxomil, and irbesartan were 6.25-18.75, 20-60, and 75-225 ng/mL, respectively. The recoveries of hydrochlorothiazide, olmesartan medoxomil, and irbesartan in spiked samples were all greater than 98%, and their relative standard deviations were less than 2.0%. The limits of detection were 1, 2, and 2 ng/mL for hydrochlorothiazide, olmesartan medoxomil, and irbesartan, respectively, and the limits of quantification were 3 ng/mL, which allow their determination at the expected serum concentration levels.     

FIA-Fluorimetric assembly for the determination of noradrenaline hydrochloride by a solid-phase reactor with immobilized hexacyanoferrate(III)

An FIA assembly provided with immobilized hexacyanoferrate(III) is proposed for the fluorimetric determination of noradrenaline hydrochloride. The oxidative reagent is immobilized by means of a strong anion-exchange resin. The FIA manifold is very simple and the calibration graph is linear over the range 0.5–75mgl^–1 noradrenaline hydrochloride with an r.s.d of 0.88% (17 replicates) and a sample throughput of 84h^–1. Foreign compounds such as NaCl, sucrose, lactose and sodium sulfate caused no significant errors. The procedure is applied to the determination of noradrenaline in a medicinal formulation.     

Terbium sensitized luminescence for the determination of fexofenadine in pharmaceutical formulations

A sensitive and specific luminescence method for the determination of Fexofenadine (FEX), in pharmaceutical formulations is reported. The method is based on the sensitization of terbium (Tb³⁺) by complex formation with FEX. The luminescence signal for Tb–FEX complex is greatly enhanced by the addition of triethylamine (ET₃N) and zinc nitrate in methanol solution. Monitoring of the signal is accomplished when the instrument is in the phosphorescence mode with the excitation and emission wavelengths set at λ_ex = 220 nm and λ_em = 550 nm respectively. Optimum conditions for the formation of the complex in methanol were 2.25 × 10^?6 M of Tb³⁺, 5.00 × 10^?6 M of Et₃N and Zn²⁺ which allows for the determination of 10–800 ppb of FEX in the batch mode with a detection limit of 0.3 ppb. The proposed method was successfully applied for the determination of FEX in pharmaceutical formulations.     

Determination of fluoxetine enantiomers in pharmaceutical formulations by electrokinetic chromatography–counter current technique

In this work, an electrokinetic chromatography–counter current procedure for the separation of fluoxetine enantiomers using highly sulfated β‐cyclodextrin was optimized and applied to the determination of the enantiomers in three pharmaceutical formulations according to the matrix features. Quality criteria were applied to facilitate its transferability to testing laboratories. Fluoxetine was used therapeutically as the racemate, although a stereospecificity associated with its interactions with the neuronal serotonin‐uptake carrier was demonstrated. In this context, the development of enantioselective methods for the chiral analysis of pharmaceuticals allowing stereoisomer ratio estimations has increasing interest in pharmaceutical industry. The proposed method allows the quantification of both enantiomers in less than 2 min with high resolution (R_s = 2.4). Copyright © 2012 John Wiley & Sons, Ltd.     

Simultaneous densitometric analysis of amlodipine,hydrochlorothiazide, lisinopril,and valsartan by HPTLC in pharmaceutical formulations and human plasma

A simple, accurate, and precise high-performance thin layer chromatographic (HPTLC) method has been developed and validated for the simultaneous quantification of antihypertensive drugs, amlodipine (AML), hydrochlorothiazide (HCTZ), lisinopril (LIS), and valsartan (VAL) in their pharmaceutical formulations and human plasma. Separation of the drugs was performed on aluminum-backed layer of silica gel 60?F₂₅₄ using a mixture of methanol–dichloromethane–glacial acetic acid (9.0:1.0:0.1, v/v/v) as the mobile phase. Densitometric determination of the separated spots was done at 215?nm. The retention factors (R_f) obtained under the optimized conditions were 0.56, 0.75, 0.29, and 0.67 for AML, HCTZ, LIS, and VAL, respectively. Linearity of the method was established in the range of 200–1,500?ng/band for AML, 300–1,500?ng/band for HCTZ, 400–2,000?ng/band for LIS, and 1,000–7,000?ng/band for VAL. The limit of detection/limit of quantitation of the method found were 54.21/164.28, 77.27/234.15, 83.45/252.87, and 156.48/474.19?ng/band for AML, HCTZ, LIS, and VAL, respectively. To determine the drugs in spiked plasma samples, solid phase extraction was performed, which provided highly consistent and quantitative recovery for all four drugs. The method was satisfactorily applied for the analysis of different tablet formulations and proved to be specific and accurate for the quality control of these drugs.     

p-Aminobenzoate ion determination in pharmaceutical formulations by using a potentiometric sensor immobilized in a graphite matrix

The characteristics, performance, and application of an electrode, namely, Pt|Hg|Hg₂(PABzt)₂| graphite, where PABzt stands for p-aminobenzoate ion, are described. This electrode responds to PABzt with sensivity of (58.1±1.0) mV per decade over the range 1.0×10⁻⁴ to 1.0×10⁻¹ mol l⁻¹ at pH 6.5-8.0 and a detection limit of 3.2×10⁻⁵ mol l⁻¹. The electrode shows easy construction, fast response time (within 10-30 s), low-cost, and excellent response stability (lifetime greater than 6 months, in continuous use). The proposed sensor displayed good selectivity for p-aminobenzoate in the presence of several substances, especially, concerning carboxylate and inorganic anions. It was used to determine p-aminobenzoate in pharmaceutical formulations by means of the standard additions method. The results obtained by using this electrode compared very favorably with those given by an HPLC procedure.     

Flow-injection turbidimetric determination of homatropine methylbromide in pharmaceutical formulations using silicotungstic acid as precipitant reagent

A flow-injection turbidimetric procedure exploiting merging zones is proposed for determining homatropine methylbromide (HMB) in pharmaceutical preparations. The determination is based on the precipitation reaction of homatropine methylbromide with silicotungstic acid in acidic medium to form a precipitate, which was measured at 410 nm. The analytical curve was linear in the HMB concentration range from 8.1 × 10⁻⁵ to 2.2 × 10⁻⁴ mol l⁻¹, with a detection limit of 5.0 × 10⁻⁶ mol l⁻¹. The recoveries ranged from 96 to 103%, the sampling frequency was 70 determinations per hour and relative standard deviations were less than 1.5% (n = 10). The results obtained for commercial formulations using the FIA procedure were in good agreement with those obtained by using a comparative method.     

Validated spectrophotometric method for quantitative determination of simvastatin in pharmaceutical formulations and human serum

A simple ultraviolet spectrophotometric method for the determination of simvastatin in methanol has been devised and compared with the existing pharmacopoeial RP-HPLC method for the estimation of the drug. Analytical parameters such as stability, selectivity, accuracy, and precision have been established for the method, using SIMS® tablets and human serum samples, and evaluated statistically to assess the application of the method. The method was validated under ICH and USP guidelines and was found to comprise the advantages for simplicity, stability, sensitivity, reproducibility, and accuracy for use as an alternative to existing nonspectrophotometric methods for the routine determination of the drug in pharmaceutical formulations and for pharmaceutical investigations involving simvastatin.     

HPLC‐DAD method for the simultaneous determination of zofenopril and hydrochlorothiazide in oral pharmaceutical formulations

An HPLC method with DAD detection was developed and validated for the simultaneous determination of zofenopril and hydrochlorothiazide in tablets. The separation was carried out through a gradient elution using an Agilent LiChrospher C18 column (250×4.0 mm id, 5 μm) and a mobile phase consisting of (A) water–TFA (99.9:0.1 v/v) and (B) acetonitrile–TFA (99.1:0.1 v/v) delivered at a flow‐rate of 1.0 mL/min. 8‐Chlorotheophylline was used as internal standard. Calibration curves were found to be linear for the two drugs over the concentration ranges of 5.0–40 and 1.0–20 μg/mL for zofenopril and hydrochlorothiazide, respectively. Linearity, precision, accuracy, specificity and robustness were determined in order to validate the proposed method, which was further applied to the analysis of commercial tablets. The proposed method is simple and rapid, and gives accurate and precise results.     

Development and validation of high-performance liquid chromatography method for determination of clarithromycin in pharmaceutical tablets

Clarithromycin is a very important macrolide antibiotic used to treat bacterial infections in human and veterinary medicine. This study reports the development and validation of cost-effective, simple, precise, accurate, and robust high-performance liquid chromatography (HPLC) for the determination of clarithromycin (CLA) in tablets. Reversed-phase chromatography was conducted using a standard column at 55°C with ultraviolet detection at 215 nm. A mobile phase consisting of acetonitrile –2-methyl-2-propanol –potassium phosphate buffer was used at a flow rate of 1.0 mL/min. The proposed method displayed good linearity, precision, accuracy, robustness, and specificity. The present HPLC was compared with capillary electrophoresis and bioassay methods and the results indicated that there was no significant difference between these methods. Moreover, the obtained results demonstrated the validity of the isocratic HPLC, which allows reliable quantitation of CLA in pharmaceutical samples. Thus, it can be used as a substitute alternative methodology for the routine quality control of this medicine, in situations where other methods are less accessible in the laboratory.     

Enantioseparation and quality control of biperiden in pharmaceutical formulations by capillary electrophoresis

An original capillary electrophoretic method has been developed and applied for the enantioselective analysis of the antiparkinson drug biperiden in pharmaceutical formulations, using a modified cyclodextrin as the chiral selector. Baseline enantioseparation of the racemic compound was achieved in less than 7 min using an uncoated fused silica capillary (50 μm i.d. and 48.5, 40.0 cm, total and effective length, respectively), filled with a background electrolyte consisting of a 50 mM phosphate buffer at pH 3.5 supplemented with 3% (w/v) β-cyclodextrin sulphate and applying a voltage of 20 kV, reversed polarity. Samples were injected by pressure (50 mbar, 90 s) at the cathodic end of the capillary and detection wavelength was 195 nm (bandwidth: 10 nm). A simple and fast pre-treatment procedure allowed the complete extraction of the drug from commercial formulations (sustained release tablets and ampoules for injections) without any interference from the matrix. Good linearity was found in the 1–50 μg/mL concentration range; the limit of quantitation was 1 μg/mL and the limit of detection was 0.4 μg/mL. Precision and accuracy were good, with R.S.D. values always lower than 2.8% and a mean recovery value of 101.1%. The method was suitable for the quality control of biperiden in commercial formulations.     

Stability-indicating high performance thin layer chromatography determination of Paroxetine hydrochloride in bulk drug and pharmaceutical formulations

A simple selective precise and stability-indicating high performance thin layer chromatographic method of analysis of Paroxetine hydrochloride both as a bulk drug and in formulations was developed and validated. The method employed TLC (Thin Layer Chromatography) aluminum precoated with silica gel 60F-254 as the stationary phase. The solvent system consisted of butanol:acetic acid:water (8:2:0.5, v/v/v). This system was found to give compact spots for Paroxetine HCl (Rf, retardation factor, value-0.48 ± 0.02). Paroxteine HCl was subjected to acid and alkali hydrolysis, oxidation and photodegradation, where the degraded product was well separated from the pure drug. Densitometric analysis of Paroxetine hydrochloride was carried out in the absorbance mode at 295 nm. The linear regression analysis data for the calibration spots showed good relationship with (regression) r² = 0.9903 in the amount range of 300-1500 ng (nanogram) per spot. The mean value of co-relation co-efficient, slope and intercept were 0.9903 ± 0.001, 5.38 ± 0.058 and 182.5 ± 2.16 respectively. The method was validated for precision, recovery and robustness. The limits of detection and quantitation were 50 and 150 ng, respectively. The drug doesnot undergo degradation with oxidation, but gets affected in acidic and alkaline conditions. The acid and alkali degradation showed extra peaks at 0.4 and 0.08 Rf, respectively. This indicates that the drug is susceptible to acidic and alkaline medium. As the method could effectively separate the drug from its degradation products, it can be employed as a stability-indicating one.