Stability Indicating HPLC Method For Determination Research Articles

Stability-Indicating HPLC Method for Determination of Ibuprofen and Famotidine Degradation Products.

A new stability-indicating high-performance liquid chromatographic method for the quantitative determination of ibuprofen and famotidine degradation products in combined pharmaceutical products was developed and validated. The current aim of this study is to develop a rapid, accurate and robust analytical stability indicating impurity method that can separate ibuprofen, famotidine and their related impurities by using a reversed-phase high-performance liquid chromatography. A Zorbax SB-Phenyl column (4.6 × 150mm2, 5-μm particle size) with mobile phase containing phosphate buffer solution with a pH value of 3.0 and acetonitrile was used. The flow rate was 0.8mL/min and the analytes were detected by UV detector at 265nm. The retention times of ibuprofen and famotidine were 18.43 and 5.14min, respectively. This method was validated to confirm specificity, linearity, sensitivity (limit of detection and limit of quantitation), precision, accuracy, robustness and sample stability according to the International Conference on Harmonization guidelines. Studies have been completed and reported with two active substances in the combined dosage form and seven impurities in total. There is no method in the literature that simultaneously distinguishes and quantitatively analyzes both active substances and degradation products.

Development and Validation of a Stability-indicating HPLC Method for Determination of Insulin Detemir and Preservatives in their Injection Products

Insulin detemir (ID) is a long-acting form of insulin that is characterized by the covalent attachment of a lipophilic tail of myristate, and commercially available as Levemir®. No satisfactory simple isocratic stability-indicating HPLC method has been reported for its quality control. A novel simple and isocratic reversed-phase HPLC method was developed and validated for the simultaneous determination of ID along with its dosage form additives in the available commercial preparations. The method employed C4 column (5μm, 250 × 4.6 mm), a mobile phase consisting of 50 mM phosphate buffer pH 2.7, acetonitrile, triethylamine (62:37:1) and 0.02 g/ml sodium sulfate, that was delivered isocraticaly at a flow rate of 1.5 ml/min, and detection performed at 214nm. The method was properly validated and was shown linear over the range 80–120% of the assay concentration for ID, phenol, and m-cresol. The method was also selective, specific, precise, and accurate. Furthermore, the validated method was applied to separate the major degradation products in those preparations. Forced degradation studies in different pH values, which are the first to be reported for ID, showed that the degradation products were baseline separated from ID itself and/or other formulation additives. Thus, the method has been demonstrated to be a stability-indicating assay.

Stability-indicating HPLC method for determination of amiodarone hydrochloride and its impurities in tablets: a detailed forced degradation study

Resumo Amiodarone hydrochloride is one of the most important drugs used to treat arrhythmias. The USP monograph for amiodarone hydrochloride describes an HPLC method for the quantification of seven impurities, however, this method shows problems that result in unresolved peaks. Moreover, there is no monograph for tablets in this compendium. Thus, a stability indicating HPLC method was developed for the determination of amiodarone, its known impurities and degradation products in tablets. A detailed forced degradation study was performed submitting amiodarone API, tablets and placebo to different stress conditions: acid and alkaline hydrolysis, oxidation, metal ions, heat, humidity, and light. Amiodarone hydrochloride API was susceptible to degradation in all stress conditions. The tablets also showed degradation in all environments, except in acidic condition. The analytes separation and quantification were achieved on an Agilent Zorbax Eclipse XDB-C18 column (100 x 3.0 mm, 3.5 µm). The mobile phase was composed of 50 mM acetate buffer pH 5.5 (A) and a mixture of methanol-acetonitrile (3:4, v/v) (B) in gradient elution. The method was validated in the range of 350-650 µg/mL for assay and 10-24 µg/mL for impurities determination. Therefore, this method can be used both for stability studies and routine quality control analyses.

Stability-Indicating HPLC Method for Determination of Spironolactone in Oral Liquids and Its Application to Stability Study

Stability-Indicating HPLC Method for Determination of Spironolactone in Oral Liquids and Its Application to Stability Study

Stability-indicating HPLC Method for Determination of Naproxen in an Extemporaneous Suspension

Naproxen is a nonsteroidal anti-inflammatory drug (NSAID) used for treatment of fever and inflammation. It is mostly available as tablet dosage form. Hospital pharmacists always prepare the drug in an orally liquid preparation such as suspension for pediatric use. To determine the amount of naproxen in an extemporaneous preparation and during stability studies, a HPLC method was developed and validated by following the Asean guideline for validation of the analytical procedure. The method used a C18 column as the stationary phase whereas the mobile phase was a mixture of 50 mM sodium phosphate buffer pH 7.8 and acetronitrile (70:30). The flow rate was 0.7 ml/min and injection volume was 20 µl. The drug was measured at 230 nm. The result showed that the retention time of naproxen was about 8.18 min, and the method was specific to the drug in the presence of other common excipients in the preparation. It was accurate (97.45 ± 0.63%) and precise (1.33-2.33% RSD). It was linear in the concentration range of 2.56 ng/ml- 50 µg/ml (R2= 0.9999). The limit of detection (LOD) and limit of quantification (LOQ) were 0.19 and 0.59 µg/ml, respectively. Minor changes of flow rate, mobile phase composition, wavelength and injection volume slightly affected the drug analysis. Forced degradation studies (hydrolysis, oxidation and temperature) showed that the method was served as a stability-indicating procedure. It demonstrated that the validated method was simple, reliable, accurate and robust to determine naproxen in the extemporaneous suspension. In addition, it could use for analysis of the drug in the stability studies.

A validated stability-indicating HPLC method for determination of brimonidine tartrate in BRI/PHEMA drug delivery systems

BackgroundA simple, rapid and accurate stability-indicating reverse phase high performance liquid chromatography (RP-HPLC) was developed and validated for the determination of brimonidine tartrate in brimonidine tartrate/poly(2-hydroxyethyl methacrylate) (BRI/PHEMA) drug delivery contact lenses and pharmaceutical formulations.ResultsOptimum chromatographic conditions for separating brimonidine tartrate from other impurities in the leaching liquor of BRI/PHEMA drug delivery contact lenses or pharmaceutical formulations have been achieved by using a Diamonsil C18 column (150 mm × 4.6 mm, 5 μm) as a stationary phase and a mixture solution of phosphate buffer (10 mM, pH3.5) containing 0.5% triethlamine and methanol (85:15, v/v) as a mobile phase at a flow rate of 1 mL/min. The theoretical plates for the brimonidine tartrate measurement were calculated to be 8360 when detection was performed at 246 nm using a diode array detector. The proposed method was validated in accordance with ICH guidelines with respect to linearity, accuracy, precision, robustness, specificity, limit of detection and quantitation. Regression analysis showed a good correlation (R2 > 0.999) for brimonidine tartrate in the concentration range of 0.01–50 μg/mL. The peak purity factor is ≥980 for the analyte after all types of stress tests, indicating an excellent separation of brimonidine tartrate peak from other impurities. The measurement course could be completed within 10 min, which was very quick, effective and convenient.ConclusionsOverall, the proposed stability-indicating method was suitable for routine quality control and drug analysis of brimonidine tartrate in BRI/PHEMA drug delivery contact lenses and other pharmaceutical formulations.

A comprehensive stability-indicating HPLC method for determination of chloroquine in active pharmaceutical ingredient and tablets: Identification of oxidation impurities

Malaria is the most common parasitic disease in humans. It is estimated that 3 billion people live under the risk of contracting this disease in the world. Chloroquine (CQ) is the drug of choice to treat cases of non-complicated malaria. Forced degradation studies are important to know the drug’s potentials degradation products and to develop a stability indicating method. Thus, chloroquine active pharmaceutical ingredient (API), chloroquine tablets and placebo were submitted to a detailed forced degradation study, using several stressing agents. The results were used on the development of a stability indicating method, using high performance liquid chromatography. The method was validated showing selectivity, precision, accuracy, robustness and linearity in the range of 30–360μg/mL of chloroquine. Chloroquine API and tablets were susceptible to alkaline hydrolysis with NaOH 1mol/L, and to oxidation with H2O2 3.0%. Two degradation products were formed in oxidative test. Kinetics of chloroquine degradation in alkaline hydrolysis was performed for both API and tablets. The calculated decay constant (k1) was 0.223days−1 for API and 0.182days−1 for tablets, while the half-life (t1/2) was 3.1days for API and 3.8days for tablets. Chemical structures have been proposed for the two degradation products formed in the presence of H2O2, using an UHPLC-UV-MS/MS approach.

Development and Validation of a Rapid Stability-Indicating HPLC Method for Determination of Carbamazepine in Pure and Dosage Forms

Development and Validation of a Rapid Stability-Indicating HPLC Method for Determination of Carbamazepine in Pure and Dosage Forms

A fast stability-indicating HPLC method for determination of sotalol hydrochloride in bulk powder and in dosage form

A fast stability-indicating HPLC method for determination of sotalol hydrochloride in bulk powder and in dosage form

Development, Optimization, and Validation of a Green and Stability-Indicating HPLC Method for Determination of Daptomycin in Lyophilized Powder.

Daptomycin is an antimicrobial that plays an important role in clinical practice today because it is considered a promising drug to combat resistant strains, such as methicilin and vancomycin-resistant Gram-positive bacteria. Considering the analysis of daptomycin in a pharmaceutical dosage form, the only method found in literature uses potentially toxic organic solvents. Therefore, the objective of this work was to develop a green and stability-indicating HPLC method for determination of daptomycin in lyophilized powder. The mobile phase was ethanol-water (55+45, v/v) at pH 4.5 pumped at a flow rate of 0.6 mL/min. A C18 column was used, and UV detection was performed at 221 nm. Stress degradation studies were conducted in order to demonstrate the specificity and stability-indicating capability of the method. The method was validated according to International Conference on Harmonization guidelines, proving to be linear (r=0.9996), precise, accurate, robust (demonstrated by the Plackett-Burman model), and specific within the range 20-70 μg/mL. The retention time of daptomycin was 5.8 min. It can be concluded that the validated method can be a fast, safe, and environmentally friendly alternative for the analysis of daptomycin.

Development and Validation of a Stability-Indicating HPLC Method for Determination of Clorazepate Dipotassium and Its Main Impurities in Bulk Drug and Capsules

Development and Validation of a Stability-Indicating HPLC Method for Determination of Clorazepate Dipotassium and Its Main Impurities in Bulk Drug and Capsules

A Stability-Indicating HPLC Method for the Determination of Memantine Hydrochloride in Dosage Forms through Derivatization with 1-Fluoro-2,4-dinitrobenzene.

Memantine is chemically a tricyclic amine and is used for Parkinson’s disease and movement disorders. Although several HPLC methods with different derivatization reagents have been developed for the determination of memantine in biological fluids, there are some complications which limit the use of these methods in routine analysis of memantine in in vitro tests. We established a simple, sensitive, precise, and accurate HPLC method for the quantification of memantine in dosage forms. Pre-column derivatization of memantine was performed with 1-fluoro-2,4-dinitrobenzene and the reaction product was separated on a Nova-Pak C18 column. A mixture of acetonitrile and sodium dihydrogenphosphate (pH 2.5; 0.05 M) (70: 30, v/v) was used as the mobile phase. UV detection was performed at 360 nm. Forced degradation studies were performed on a powdered tablet sample of memantine hydro-chloride using acidic (0.1 M hydrochloric acid), basic (0.1 M sodium hydroxide), oxidative (10% hydrogen peroxide), thermal (105°C), photolytic, and humidity conditions. Good linearity (r2=0.999) was obtained over the range of 1–12 μg mL−1 of memantine hydrochloride with acceptable within-day and between-day precision values in the range of 0.05–0.95%. The proposed method was used for the assay determination and dissolution rate study of memantine dosage forms with excellent specificity.

Development and Validation of a Stability-Indicating HPLC Method for Determination of Bromocriptine Mesylate in Bulk Drug and Tablets

Development and Validation of a Stability-Indicating HPLC Method for Determination of Bromocriptine Mesylate in Bulk Drug and Tablets

Development and Validation of a Stability-Indicating HPLC Method for Determination of Bromocriptine Mesylate in Bulk Drug and Tablets

Development and Validation of a Stability-Indicating HPLC Method for Determination of Bromocriptine Mesylate in Bulk Drug and Tablets

Validation and Uncertainty Estimation of an Ecofriendly and Stability-Indicating HPLC Method for Determination of Diltiazem in Pharmaceutical Preparations

A green, simple, and stability-indicating RP-HPLC method was developed for the determination of diltiazem in topical preparations. The separation was based on a C18 analytical column using a mobile phase consisted of ethanol: phosphoric acid solution (pH = 2.5) (35 : 65, v/v). Column temperature was set at 50°C and quantitation was achieved with UV detection at 240 nm. In forced degradation studies, the drug was subjected to oxidation, hydrolysis, photolysis, and heat. The method was validated for specificity, selectivity, linearity, precision, accuracy, and robustness. The applied procedure was found to be linear in diltiazem concentration range of 0.5–50 μg/mL (r 2 = 0.9996). Precision was evaluated by replicate analysis in which % relative standard deviation (RSD) values for areas were found below 2.0. The recoveries obtained (99.25%–101.66%) ensured the accuracy of the developed method. The degradation products as well as the pharmaceutical excipients were well resolved from the pure drug. The expanded uncertainty (5.63%) of the method was also estimated from method validation data. Accordingly, the proposed validated and sustainable procedure was proved to be suitable for routine analyzing and stability studies of diltiazem in pharmaceutical preparations.

Stability-Indicating HPLC Method for Determination of Naftazone in Tablets. Application to Degradation Kinetics and Content Uniformity Testing

A simple, sensitive, stability-indicating HPLC method was developed and validated for the quantitative determination of the vasoprotective drug, naftazone in presence of its degradation products. The analysis was carried out on a Nucleosil 100-5 phenyl column (250 mm × 4.6 mm, 5 μm) using a mobile phase consisting of methanol-0.02 M sodium dihydrogen phosphate mixture (60:40, v/v) of pH 6.0. The analyses were performed at ambient temperature with a flow rate of 1.0 mL/min and UV detection at 270 nm. The method showed good linearity over the concentration range of 0.1-10.0 μg/mL with a lower detection limit of 0.032 and quantification limit of 0.096 μg/mL. The suggested method was successfully applied for the analysis of naftazone in its commercial tablets. Moreover, it was utilized to investigate the kinetics of alkaline, acidic and oxidative degradation of the drug. The apparent first-order rate constants, half-life times, and activation energies of the degradation process were calculated. The pH-rate profile curve was derived. Furthermore, the proposed method was successfully applied to the content uniformity testing of naftazone tablets.

A validated stability-indicating HPLC method for determination of varenicline in its bulk and tablets

A simple, sensitive and accurate stability-indicating HPLC method has been developed and validated for determination of varenicline (VRC) in its bulk form and pharmaceutical tablets. Chromatographic separation was achieved on a Zorbax Eclipse XDB-C8 column (150 mm × 4.6 mm i.d., particle size 5 μm, maintained at ambient temperature) by a mobile phase consisted of acetonitrile and 50 mM potassium dihydrogen phosphate buffer (10:90, v/v) with apparent pH of 3.5 ± 0.1 and a flow rate of 1.0 ml/min. The detection wavelength was set at 235 nm. VRC was subjected to different accelerated stress conditions. The degradation products, when any, were well resolved from the pure drug with significantly different retention time values. The method was linear (r = 0.9998) at a concentration range of 2 - 14 μg/ml. The limit of detection and limit of quantitation were 0.38 and 1.11 μg/ml, respectively. The intra- and inter-assay precisions were satisfactory; the relative standard deviations did not exceed 2%. The accuracy of the method was proved; the mean recovery of VRC was 100.10 ± 1.08%. The proposed method has high throughput as the analysis involved short run-time (~ 6 min). The method met the ICH/FDA regulatory requirements. The proposed method was successfully applied for the determination of VRC in bulk and tablets with acceptable accuracy and precisions; the label claim percentages were 99.65 ± 0.32%. The results demonstrated that the method would have a great value when applied in quality control and stability studies for VRC.

A validated stability-indicating HPLC method for the determination of PEGylated puerarin in aqueous solutions

The aim of this study was to develop a validated specific stability-indicating HPLC method for the quantitative determination of PEGylated puerarin (PEG-PUE) in aqueous solutions. The method was validated by subjecting PEG-PUE to forced degradation under stress conditions of acid, alkali, water hydrolysis, and oxidation. Both PEG-PUE and puerarin (PUE) were simultaneously determined and separated on CAPCELL PAK C18 column by gradient elution with 0.2% aqueous phosphoric acid and acetonitrile as the mobile phase. The flow rate was 1.0 mL min −1 and detection wavelength was set at 250 nm. Both calibration curves showed good linear regression ( r ≥ 0.9998) within test ranges. The LOD and LOQ of PEG-PUE were determined to be 3 and 9 μg mL −1 respectively. Degradation of PEG-PUE followed pseudo-first-order kinetics with t 1/2 of 59 min at pH 9.0 and 17.79 h at pH 7.4. However, at pH 5.0 and 2.0, there was no significant degradation of PEG-PUE over time. In conclusion, the method was observed to have the necessary specificity, precision, and accuracy, and to be suitable for quantity monitoring the degradation process of PEG-PUE during stability studies. The degradation studies may give insight into useful information for formulation development of PEG-PUE.

Principles into Practice Setting the Bar for Green Chemistry

Recent years have seen a disheartening string of revelations in which everyday items once considered safe—food packaging, toys, clothes, furniture, electronic components, and many more products—are found to contain carcinogens, endocrine disruptors, and other harmful chemicals.1 Growing demand for healthier alternatives, already seen in food production and housing construction,2 is also happening at the building-block level of manufacturing, where so-called green chemistry represents a revolutionary change in preventing pollution and health problems starting at the chemical design stage. Many industry and government entities are beginning to espouse the principles of green chemistry on their websites and in public statements. Now comes the task of crafting policy to put those principles into action. The U.S. Environmental Protection Agency (EPA) defines green chemistry as “the design of chemical products and processes that reduce or eliminate the use or generation of hazardous substances. Green chemistry applies across the life cycle of a chemical product, including its design, manufacture, and use.”3 Green chemistry also aims to mitigate the type of uncertainty Alan Gold-berg, a professor of toxicology at the Johns Hopkins Bloomberg School of Public Health, recently described to The New York Times: “I can get [toxicity] information on only 20 percent of chemicals we interact with on a daily basis.”4 Of that 20%, he now says, he may be able to find information on overt toxicity for about half, but for details on specific effects such as developmental neurotoxicity, the figure shrinks toward zero. So what does green chemistry look like? Consider the example of pregabalin, the active ingredient in the neuropathic pain drug Lyrica®. Pfizer developed an alternative green-chemistry process that converted several steps of pregabalin synthesis from use of organic solvents to water. That reduced both health hazards and production heating requirements. With the new synthesis, waste from the process dropped from 86 kg of waste per kg product to 17 kg, and energy use dropped by 82%.5 Proponents say that’s how the field can offer a win–win–win solution: good performance, lower cost, and less environmental impact—what Richard Engler, program manager of the EPA Green Chemistry Program, calls the “triple bottom line.” For many, a standard is a logical next step. “At some point you have to go beyond a definition and principles,” says Engler. “I think that’s something the standard will enable.”

Development and Validation of a Stability-Indicating HPLC Method for Determination of Voriconazole and Its Related Substances

An isocratic reversed-phase high performance liquid chromatographic (RP-HPLC) method has been developed and validated for the determination of voriconazole and its related substances. The drug substance was subjected to stress conditions of UV light, water hydrolysis, acid, base, oxidation, and deoxidization to observe the degradation products. The successful separation of voriconazole from its synthetic impurities and degradation products formed under stress conditions was achieved using an Agilent Zorbax SB-C18 (250mm x 4.6 mm i.d., 5 microm) column maintained at 25 degrees C with a mobile phase of a mixture of ammonium phosphate dibasic buffer (pH adjusted to 6.0 using diluted orthophosphoric acid; 50 mM)-acetonitrile (52:48, v/v). The mobile phase flow rate was 1.0 mL/min, and the detection wavelength was 250 nm. The stress sample solutions were assayed against the qualified reference standard of voriconazole and the mass balance in each case was close to 99.7%, confirming its stability-indication capacity. The developed HPLC method was validated with respect to linearity, accuracy, precision, specificity, and robustness. The developed HPLC method to determine the related substances and assay determination of voriconazole can be used to evaluate the quality of regular production samples. It can be also used to test the stability samples of voriconazole.