Development and validation of a rapid HPLC method for the determination of ketotifen in pharmaceuticals

In the present study, a simple, sensitive, rapid, and stability-indicating high performance liquid chromatographic (HPLC) method with ultraviolet detection for the analysis of ketotifen was developed and validated. The method was applied to the determination of ketotifen in pharmaceutical formulations (tablets and syrups). The HPLC method utilized isocratic elution technique with a reversed phase C8 column, detection at 297 nm and a mixture of methanol, triethylamine phosphate buffer (pH 2.8; 0.04 M), and tetrahydrofuran (43: 55: 2, v/v/v) as mobile phase at a flow rate of 1.2 mL/min. Total analysis time was about 7 min with typical retention time of ketotifen of about 5 min. The method was validated for selectivity, linearity, accuracy, and precision following International Conference of Harmonization, 1996 (ICH) recommendations. Due to its simplicity and accuracy, the method can be used for routine quality control analysis.     

A validated HPLC method for separation and determination of promethazine enantiomers in pharmaceutical formulations

A simple, rapid, and validated method for separation and determination of promethazine enantiomers was developed. Promethazine was separated and quantitated on a Vancomycin Chirobiotic V column (250 x 4.6 mm), using a mixture of methanol, acetic acid, and triethylamine (100:0.1:0.1%, by volume) as a mobile phase at 20 degrees C and at a flow rate of 1 mL/min. The UV-detector was set to 254 nm. Acetyl salicylic acid (Aspirin) was used as an internal standard. The applied HPLC method allowed separation and quantification of promethazine enantiomers with good linearity (r > .999) in the studied range. The relative standard deviations (RSD) were 0.29 and 0.36 for the promethazine enantiomers with accuracy of 100.06 and 100.08. The limit of detection and limit of quantification of promethazine enantiomers were found to be 0.04 and 0.07 microg/mL, respectively. The method was validated through the parameters of linearity, accuracy, precision, and robustness. The HPLC method was applied for the quantitative determination of promethazine in pharmaceutical formulations.     

Pharmaceutical Analysis of β-Methyldigoxin in Dosage Forms Using RP-HPLC

Abstract

A reversed-phase high-performance liquid chromatographic method has been developed for the assay of β-methyldigoxin in Dimekor® tablets (0.1 mg) and ampules (0.2 mg/2 mL). Quantitation of cardiac glycoside in mentioned dosage forms was carried out by the incorporation of phenacetin as an internal standard. A Varian HPLC configured with a Paltisil P10 ODS1 column was used for the separation and quantitation of β-methyldigoxin in pharmaceutical preparations. The mobile phase was acetonitrile-water 38: 62 v/v with flow rate 1.6 ml/min and UV detection was set at 220 nm. The range of linearity extended from 0.01 to 0.11 mg/mL. For the quantitative analysis of β-methyldigoxin in tablets the recovery was 100.16% and for ampules 99.50%. The excipients did not interfere with the determination of the analysed substance. The proposed method is precise and sensitive for the examination of examine the content uniformity of tablets and is in a good agreement with PH.JUG.IV (1). A spectrofluorimetric method was used for the dissolution test by the method described in USP XXII (2).     

A Validated Stability Indicating LC Method for Nateglinide

ABSTRACT

A simple, isocratic, rapid and accurate reverse phase high-performance liquid chromatography (RP-HPLC) method was developed for the quantitative determination of Nateglinide. The developed method is also applicable for determination of related substance in bulk drugs. The chromatographic separation was achieved on a Hypersil C18 (250 × 4.6 mm 5 μm) column using aqueous mixture of 0.025 M potassium hydrogen phosphate and 0.1% triethyl amine, v/v (pH 3.0 with dilute phosphoric acid)—methanol (25:75, v/v) as a mobile phase. Solution concentrations were measured on a weight basis to avoid the use of an internal standard. The chromatographic resolutions between Nateglinide and its potential impurities A and B were found to be greater than four. Forced degradation studies were performed for Nateglinide using acid (0.5 N hydrochloric acid), base (0.5 N sodium hydroxide), oxidation (3% hydrogen peroxide) heat (60°C) and UV light (254 nm). The limit of detection and limit of quantification of Nateglinide, impurities A and B were found to be 0.05 and 0.15 μg /mL, respectively for 20 μL injection volume. The percentage recovery of Nateglinide was ranged from 98.4 to 100.9. The percentage recovery of impurities in Nateglinide sample was ranged from 96.8 to 103.5. The developed RP-HPLC method was validated with respect to linearity, accuracy, precision, robustness, and forced degradation studies prove the stability indicating power of the method.     

Determination of hydroxytyrosol in plasma by HPLC

Hydroxytyrosol (2-(3,4-dihydroxyphenyl)ethanol), a phenolic compound present in extravirgin olive oil, has been reported to contribute to the prevention of cardiovascular disease. The present study describes an accurate and reproducible reversed-phase HPLC method to measure hydroxytyrosol in plasma. This compound was extracted from acidified plasma by solid-phase extraction using an Oasis HLB copolymer. The plasma sample was rinsed with water and methanol in water (5:95; v/v). Hydroxytyrosol was eluted with methanol, which was subsequently evaporated under a nitrogen stream. Analysis by HPLC with diode array-UV detection was carried out using a C18 column and a gradient elution with acidified water and methanol/acetonitrile (50:50; v/v). The method was validated by the analyses of plasma samples spiked with pure hydroxytyrosol, obtaining a linear correlation (0.9986) and precision with a coefficient of variation ranging from 0.79 to 6.66%. The recovery was approximately 100%, and the limit of detection was 37 ng/mL. The oral administration of hydroxytyrosol to rats and its subsequent detection in plasma showed that the method is suitable for pharmacokinetic studies.     

Simultaneous high-performance liquid chromatographic analysis for famotidine, ranitidine HCl, cimetidine, and nizatidine in commercial products

A high-performance liquid chromatographic (HPLC) procedure for the simultaneous determination of famotidine (FMT), ranitidine HCl (RNT), cimetidine (CMT), and nizatidine (NZT) was developed using a two-level, full-factorial design with three variables (volume of methanol, percentage of triethylamine, and concentration of phosphate buffer) to select an acceptable mobile phase. A column (15cm × 4.6 mm ID) of Inertsil ODS-2 (5 μm) was used, and 0.04M aqueous sodium dihydrogen phosphate/acetonitrile/methanol/TEA at a proportion of 345/20/35/0.7 (v/v/v/v) was the selected mobile phase (1 ml/min). The detection wavelength was set at 230 nm, and procaine HCl was used as the internal standard. Precision and linearity of the method were assessed. None of the commercial samples was found to be outside the compendial limits of 90.0% to 110.0% of the claim amount.     

A Stability-Indicating HPLC Method to Determine Celecoxib in Capsule Formulations

ABSTRACT

A simple and accurate high-performance liquid chromatographic (HPLC) method was developed to determine Celecoxib in capsule formulations. The drug was chromatographed on a reversed-phase C-18 column. Eluents were monitored at a wavelength of 251 nm using a mixture (85:15) of methanol and water. Solution concentrations were measured on a weight basis to avoid the use of an internal standard. The method was statistically validated for linearity, accuracy, precision, and selectivity. Due to its simplicity and accuracy, we believe that the method will be useful for routine quality control analysis.     

A Stability Indicating Method for Dipyridamole

An HPLC method for the assay and chromatographic purity assessment of dipyridamole raw material and capsule product was developed. A mobile phase composing of methanol: aq 200 mM pentane sulphonic acid, sodium salt, [70:30 v/v], had triethylamine added [2ml/L] and was then adjusted to pH 3.0 with phosphoric acid. The mobile phase was pumped through an octadecylsilated-silica column, being held at 60° C, at a flow rate of 1.5ml/min. Detection was made at 288nm.

The resolution of degradation products, along with precision and accuracy were investigated for the system.     

Simultaneous determination of methocarbamol and aspirin in presence of their pharmacopeial-related substances in combined tablets using novel HPLC-DAD method

Objective and Significance: Methocarbamol (MET) and aspirin (ASP) are widely used as a muscle relaxant combination. The USP reports guaifenesin (GUA) and salicylic acid (SAL) as related substances and hydrolytic products of MET and ASP, respectively. This work aimed at developing and validating a simple and sensitive RP-HPLC method for the determination of both drugs as well as their related substances (at their pharmacopeial limits) in their bulk powders, laboratory prepared mixtures, and MET-ASP combined tablets. Methods and Results: Chromatographic separation was achieved in less than 9?min with the required resolution, peak symmetry, and accuracy on C₁₈ column using isocratic elution system of diluted acetic acid (pH 3.2): acetonitrile at the ratio of 79: 21, v/v, at a flow rate of 1?mL/min. Detection was achieved with photodiode array at 233?nm for MET, GUA, and SAL and at 273?nm for ASP. The developed method has been validated as per ICH guidelines and the calibration plots were linear over the concentration ranges of 2–150, 0.4–30, 25–450, and 0.2–27?μg/mL for MET, GUA, ASP, and SAL, respectively. Conclusion: The optimized method proved to be specific, robust and precise for the quality control of the studied drugs in pharmaceutical preparations to ascertain that their related substances are not exceeding the permitted pharmacopeial limits.     

Pharmaceutical Analysis of β-Methyldigoxin in Dosage Forms Using RP-HPLC

A reversed-phase high-performance liquid chromatographic method has been developed for the assay of β-methyldigoxin in Dimekor® tablets (0.1 mg) and ampules (0.2 mg/2 mL). Quantitation of cardiac glycoside in mentioned dosage forms was carried out by the incorporation of phenacetin as an internal standard. A Varian HPLC configured with a Paltisil P10 ODS1 column was used for the separation and quantitation of β-methyldigoxin in pharmaceutical preparations. The mobile phase was acetonitrile-water 38: 62 v/v with flow rate 1.6 ml/min and UV detection was set at 220 nm. The range of linearity extended from 0.01 to 0.11 mg/mL. For the quantitative analysis of β-methyldigoxin in tablets the recovery was 100.16% and for ampules 99.50%. The excipients did not interfere with the determination of the analysed substance. The proposed method is precise and sensitive for the examination of examine the content uniformity of tablets and is in a good agreement with PH.JUG.IV (1). A spectrofluorimetric method was used for the dissolution test by the method described in USP XXII (2).     

Stability Indicating HPTLC Determination of Linezolid as Bulk Drug and in Pharmaceutical Dosage Form

Abstract

A simple, selective, precise, and stability-indicating high-performance thin layer chromatographic method of analysis of Linezolid both as a bulk drug and in formulations was developed and validated in pharmaceutical dosage form. The method employed TLC aluminium plates precoated with silica gel 60F-254 as the stationary phase. The solvent system consisted of toluene–acetone (5:5, v/v). This system was found to give compact spots for Linezolid (R_f value of 0.29 ± 0.01). Linezolid was subjected to acidic, alkali hydrolysis, oxidation, and photodegradation. The degraded products also were well separated from the pure drug. Densitometric analysis of Linezolid was conducted in the absorbance mode at 254 nm. The linear regression data for the calibration plots showed good linear relationship with r² = 0.997 ± 0.001 in the concentration range of 300–800 ng/spot. The mean value of correlation coefficient, slope, and intercept were 0.998 ± 0.003, 0.15 ± 0.009, and 19.52 ± 1.66 respectively. The method was validated for precision, accuracy, ruggedness, and recovery. The limits of detection and quantification were 20 ng/spot and 50 ng/spot, respectively. The drug undergoes degradation under acidic and basic conditions, oxidation and photo degradation. All the peaks of degraded product were resolved from the standard drug with significantly different R_f values. This indicates that the drug is susceptible to acid–base hydrolysis, oxidation, and photo degradation. Statistical analysis proves that the method is reproducible and selective for the estimation of the said drug. Because the method could effectively separate the drug from its degradation products, it can be used as a stability indicating one.     

Analysis of Nifedipine and Related Compounds in Soft Geletin Capsules by Liquid Chromatography

Abstract

A simple, sensitive, highly specific, stability-indicating, reversed-phase HPLC method for the quantitation of nifedipine and its related compounds such as nitrophenyl pyridine, nitrosophenyl pyridine analogs with average recoveries greater than 100% were obtained using a mobile phase methanol-water (55:45, v/v) at 265 nm.     

The Social Impact of Pharmaceutical Innovation

Abstract

A new simple, precise, rapid, and selective reversed-phase ion pair high-performance liquid chromatography (RP-HPLC) method has been developed for the simultaneous determination of pseudoephidrine (PSE) and terfanidine (TER) from tablets using 60:15:25 acetontrile:methanol:water (v/v) containing 2.9 g sodium lauryl sulfate/liter, pH adjusted to 3.1 using phosphoric acid as a mobile phase and C₁₈ Spherisorb ODS 2 (3 µm, 5 cm × 4.6 mm i.d.) as stationary phase. Detection was carried out using a UV detector at 254 nm. A constant flow of 1.0 ml/min was maintained throughout the analysis. Retention times for PSE and TER were 1.90 and 7.35 min, respectively. Linearity range and percentage recoveries for PSE and TER were 24–1200 and 12–600 µg/ml, and 100.01 and 100.4%, respectively.     

Validation of a Simple and Rapid HPLC Method for Determination of Metronidazole in Dermatological Formulations

A rapid and simple method using an isocratic high-pressure liquid chromatography (HPLC) and UV detection for the determination of metronidazole in dermatological formulations is presented. Metronidazole samples were extracted with a solution composed of 60% methanol and 40% mobile phase by a procedure that can be completed in less than 10 min. Subsequent separation and quantification was accomplished in less than 20 min using reversed-phase HPLC with isocratic elution with 0.01% trifluoroacetic acid/acetonitrile (85:15%, vol/vol). Validation experiments confirmed the precision and accuracy of the method. When applied to a commercial metronidazole cream and gel formulation, recoveries of 100.4% for cream formulations and 102.3% for gel formulations were obtained. The method should facilitate studies of the formulation compatibility of metronidazole topical formulations with agents that may improve its clinical tolerability for treatment of rosacea.     

Formulation,Quality Control and Shelf Life of the Experimental Cytostatic Drug Cyclopentenyl Cytosine

ABSTRACT

This paper describes the formulation and quality control of an aqueous sterilized formulation of the experimental cytostatic drug cyclopentenyl cytosine (CPEC) to be used in Phase I/II clinical trials. The raw drug substance was extensively tested. A High Pressure Liquid Chromotography (HPLC) method was validated for the quality control of the formulated product. The aqueous formulation was found to be stable for at least 2 years at 2–8°C. Sterilization (15 min at 121°C) showed no influence on drug stability. The results show that CPEC can be formulated in an aqueous solution. The described HPLC method is a useful tool in the pharmaceutical quality control.     

Enantiomeric separation and quantitative determination of atenolol in tablets by chiral high-performance liquid chromatography

The separation and quantitative determination of atenolol isomers by chiral high-performance liquid chromatography (HPLC) are described. Atenolol isomers were separated using a Chiralcel OD® column (250 × 4.6 mm,10 μm); the mobile phase was hexane-ethanol-diethylamine (75:25:0.1 v/v/v); ultraviolet detection was at 276 nm; and flow rate was 0.7 ml/min. The coefficient of variation and average recovery of (R)-isomer were 0.60% and 100.37%, respectively, for sample A and 0.69% and 100.63%, respectively, for sample B. The coefficient of variation and average recovery of (S)-isomer were 0.59% and 100.33%, respectively, for sample A and 0.63% and 99.78%, respectively, for sample B.     

Enantiomeric Separation and Quantitative Determination of Atenolol in Tablets by Chiral High-Performance Liquid Chromatography

The separation and quantitative determination of atenolol isomers by chiral high-performance liquid chromatography (HPLC) are described. Atenolol isomers were separated using a Chiralcel OD® column (250 × 4.6 mm,10 μm); the mobile phase was hexane-ethanol-diethylamine (75:25:0.1 v/v/v); ultraviolet detection was at 276 nm; and flow rate was 0.7 ml/min. The coefficient of variation and average recovery of (R)-isomer were 0.60% and 100.37%, respectively, for sample A and 0.69% and 100.63%, respectively, for sample B. The coefficient of variation and average recovery of (S)-isomer were 0.59% and 100.33%, respectively, for sample A and 0.63% and 99.78%, respectively, for sample B.     

Validation of a simple and rapid HPLC method for determination of metronidazole in dermatological formulations

A rapid and simple method using an isocratic high-pressure liquid chromatography (HPLC) and UV detection for the determination of metronidazole in dermatological formulations is presented. Metronidazole samples were extracted with a solution composed of 60% methanol and 40% mobile phase by a procedure that can be completed in less than 10 min. Subsequent separation and quantification was accomplished in less than 20 min using reversed-phase HPLC with isocratic elution with 0.01% trifluoroacetic acid/acetonitrile (85:15%, vol/vol). Validation experiments confirmed the precision and accuracy of the method. When applied to a commercial metronidazole cream and gel formulation, recoveries of 100.4% for cream formulations and 102.3% for gel formulations were obtained. The method should facilitate studies of the formulation compatibility of metronidazole topical formulations with agents that may improve its clinical tolerability for treatment of rosacea.     

High Performance Liquid Chromatographic Determination of Sodium Cromoglycate

Abstract

A simple and precise liquid chromatographic method was developed for the estimation of sodium cromoglycate (SC) in pharmaceuticals. The drug was chroma-tographed on a reverse phase C18 column. The eluants were monitored at a wavelength of 325 nm utilizing a mixture (80:20) of solution A: 0.025 M 1-octane sulfonic acid sodium salt in acetic acid (1:100), and solution B: methanol, aceto-nitrile (3:2). Solution concentrations were measured on a weight basis to avoid the use of an internal standard. The method was statistically validated for its linearity, accuracy, precision, selectivity, limit of detection and limit of quantitation. Due to its simplicity and accuracy, the authors believe that the method can be used for routine quality control analysis. It does not require any specific sample preparation except for the use of a column guard before the analytical column and a suitable profiler attached to the syringe prior to injection.     

Quantitative Determination of Troventol from Aerosols by Reversed Phase Liquid Chromatography

Abstract

Troventol, an anticholinergic agent and a derivative of atropine was estimated quantitatively from the samples of aerosols. An isocratic, reversed-phase liquid chromatographic method was developed using a C6, 5-um column with mobile phase acetonitrile-water-diethylamine-phosphoric acid (40:60:0.1:0.1, v/v) filtered through 0.45-micron and degassed before use. Salbutamol in combination with troventol from an aerosol preparation was also determined simultaneously by the present method. Recoveries obtained for troventol as well as for salbutamol were in the range 98.0 to 102.0% from aerosols, monitored at wavelength 215 ran. A flow rate of 1.0 mL/min was maintained throughout the analysis.