validation

Teneligliptin hydrobromide hydrate is a new FDA approved drug for treatment of Diabetes Mellitus. Very few methods have been reported for its identified degradation products and their effects on human. A simple, rapid, precise and accurate stability indicating RP-HPLC method was developed and validated for identification of Teneligliptin hydrobromide hydrate and its degradants on Kromacil C18 column using pH 5.5 phosphate buffer and methanol (75:25v/v) as a mobile phase in isocratic mode of elution at a flow rate 1.2 ml/min. The column effluents were monitored by a variable wavelength UV detector at 270 nm. The method was validated as per ICH guidelines. Forced degradation studies of Teneligliptin hydrobromide hydrate were carried out under acidic, basic, neutral, peroxide, photo and thermal conditions. Degradation was observed in basic and neutral stress samples, but not in acid, peroxide, photo and thermal stress samples.

The present work deals with the development of reliable method for the estimation of pioglitazone hydrochloride by using UV spectroscopy. The pioglitazone hydrochloride showed absorption maxima at wavelength 268nm respectively. The linearity range for pioglitazone hydrochloride was in the range of 10-50μg/ml with correlation coefficient of 0.999. The precision was carried out for pioglitazone hydrochloride and value was found to be less than 2. The proposed method’s results were found satisfactory and are suitable for determination of pioglitazone hydrochloride for routine quality control of drug in bulk and formulation. This method is validated according to ICH guidelines Q2R1. Keywords: Pioglitazone hydrochloride, ICH guidelines, validation, method development.  

To develop a simple, precise, accurate, and stability indicating a UV-method for estimation of Clotrimazole. In bulk and formulated dosage form. The method was under subjected to stress degradation at different conditions recommended by the International Conference on Harmonization (ICH). The drug samples are generated and used for the degradation studies. The λmax of the Clotrimazole was found to be 220 nm. The linearity of calibration curve (Absorbance Vs Concentration) in pure solution was checked over the concentration ranges of about 5-30μg/ml for Clotrimazole respectively, with the correlation coefficient higher than 0.999. The regression equation of the curve was Y = 0.0168x + 0.0041. The % RSD was found to be within the limit as per ICH guidelines. The obtained percentage recovery of Clotrimazole was found to be within the limit 100% ± SD. The proposed method can be successfully applied for the method development, validation and stress degradation studies of Clotrimazole. The percentage degradation limit should be 5-20%. The drug Clotrimazole was found to be within the limit.

A stable, simple, accurate, precise, robust and highly selective Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method was developed and validated using ritonavir. Chromatographic separation was achieved using cyber labs, LC 100 separation module, Agilent C18 column at temperature 30°C. Flow rate selected was 1ml/min. Both drugs are identified with UV detector at 256nm. Mobile phase employed was Methanol: Water (50:50), which resulted best   sensitivity. Developed method was validated in terms of linearity, range (25-150µg/ml), precession (correlation coefficient is less than 0.999), robustness, accuracy(recovery was 101.96%) and  under stress conditions drug degradation was less than 10%.The validation of proposed stability indicating method was verified by recovery studies and can be applicable in routine pharmaceutical analysis.

Both Qualitative and Quantitative analysis plays a significant role in promising the safety and therapeutic efficacy of drugs in variety dosage forms. A bioanalytical method is a set of procedures involved in the collection, processing, storage, and analysis of a biological matrix for a chemical compound. Bioanalytical studies are employed to obtain a quantitative measure of the drug or its metabolites for the study of pharmacokinetics, toxicokinetic, bioequivalence and exposure-response like pharmacokinetic/ pharmacodynamic studies. Leukotriene receptor antagonists are widely used for the treatment and management of bronchial asthma and allergic rhinitis in different dosage forms. Drugs of this class are Zafirlukast, Montelukast and Pranlukast which are being potent drugs and are more than 99% bound to plasma proteins presenting special challenges in the development and validation of analytical methods from a variety of matrices. The main objective of this review is discussion on various analytical methods used, different solvents used as mobile phase and their retention times to understand final optimized chromatographic method which could be useful for the assessment of Pharmacokinetic parameters. Among different analytical methods, HPLC, LC-MS, UV-Visible spectroscopy and spectroflourimetric techniques are the most widely preferred techniques applied by the researchers worldwide. This review article gives information about various types of extraction procedures of the drug in plasma matrices to create an optimized method for method development and to offer practical approaches for determining validation parameters like specificity, selectivity, recovery, lower limit of quantitation (LOQ), limit of detection (LOD), linearity, range, accuracy, precision, stability, ruggedness and robustness of High performance liquid chromatographic methods (HPLC)  to support pharmacokinetic studies. Accurate and sensitive analytical methods for quantitation of drugs and their metabolites are very important for the successful conduct of preclinical and clinical pharmacology studies..

A simple, specific, quick, isocratic Reversed Phase High Performance Liquid Chromatographic method was developed and validated for the analysis of Noscapine. RP-HPLC method was developed on a Symmetry C-8 (4.6 × 150 mm), 3.5 µm particle, reversed-phase column. The mobile phase was 0.1% octane sulphonic acid (pH- 3): acetonitrile, 40:60 (v/v) at a flow rate of 0.8 ml/min. and the eluate was monitored at 260 nm. The retention time of the drug was found to be 2.314 min. The method was linear over the range of 4-8 μg/ml with a regression coefficient of 0.999 and validated with respect to accuracy, precision, linearity, and specificity, limit of detection and limit of quantization as per the guidelines of International Conference for Harmonization (ICH). This method can be used in the industries for determination of Noscapine to analyze the quality of formulation without interference of the excipients.

The objective of the present research work is to develop and validated analytical method for the determination of Darunavir in pure and tablet dosage form. A simple, rapid, precise, selective and accurate novel RP-HPLC method was developed and validated for separation and determination for the Darunavir in pure and tablet dosage form. Darunavir was analyzed by Zodiac C18, (250×4.6mm, 5µ), Shimadzu LC-20AT Prominence Liquid Chromatograph and mobile phase constituted of Triethylamine buffer: Acetonitrile (60:40 v/v). The pH of the buffer was adjusted to 4.5 with diluted ortho-phosphoric acid. The flow rate of mobile phase was 1.0 mL/min and the analysis was performed using UV-Visible detector at 260nm. The Darunavir was eluted with in 6 min and retention time was showed 2.753 min. The developed assay method was validated by the guidelines of ICH Q2R1.The method was found to be linear in the drug concentration range of 20 µg/mL -100 µg/mL. The value of correlation coefficient was found to be 0.999. Method was found good percentage recovery it indicate the method is highly accurate. The method has been successfully applied for determination of Darunavir in pharmaceutical dosage form in regular quality control analysis.

A rapid, sensitive, precise, and accurate HPLC method coupled with a photodiode array detector was developed, optimized, and validated for the estimation of everolimus in bulk and tablet dosage forms. The chromatographic separation was achieved using a kromosil C8 column (200 mm × 4.6 mm i.d., 5 μm particle size) at 30°C temperature. The isocratic mobile phase consisted of 0.1M dipotassium hydrogen phosphate and methanol (60:40, v/v). The mobile phase was delivered at 1.0 ml/min and the analyte was monitored at 277 nm. The method was successfully validated in accordance to International Conference on Harmonization. The retention time of everolimus was found to be 3.496 min, and the calibration curve was linear in the concentration range of 50-150 μg/ml (R2 = 0.9992). The limit of detection and the limit of quantitation were found to be 0.109 μg/ml and 0.364 μg/ml, respectively.  The proposed method is accurate (percent recoveries were in the range of 99.56-100.09%) and precise (percent relative standard deviation - 0.047 %). No chromatographic interferences from the tablet excipients and components of mobile phase were found. The proposed method was found to be suitable for the quantitative determination of everolimus in tablet dosage forms.

Arbutin and quercetin are anti-cancer agents, extracted from leaves and flowers of Origanum majorana L which can be simultaneously estimated using a rapid, specific and sensitive high – performance thin layer chromatographic technique using silica gel G60F254 as the stationary phase and butyl acetate: methanol: formic acid: toluene (8: 1.5: 5: 0.5) as mobile phase for the separation. The method was validated for linearity, precision and recovery. Linearity was established in the range 300-900 ng/spot for arbutin and 150-450 ng/spot for quercetin, respectively. The % RSD values of repeatability of application, repeatability of measurement, intra-day and inter day precision studies of arbutin and quercetin were found to be below 1 for 500 and 250 ng/spot of arbutin and quercetin, respectively. The recovery of the drugs were found to be 99.8 and 100.2%, proves that the developed method was highly accurate. Hence the proposed validated method could be applied for the simultaneous analysis of arbutin and quercetin in methanolic extract of Origanum majorana L as well as for its marketed formulation.

An accurate, simple, rapid and sensitive HPLC method has been developed for the determination of mebhydroline napadisylate in the tablet. The Chromatography was performed on a reversed phase C-18 column(150 mm × 4.6 mm id, 5μm) by isocratic elution, using a mobile phase of acetonitrile : ammonia 25% (80 : 20 v/v) at ambient temperature 25±5 °C and UV detection operates at 320 nm in an overall analysis time of about 10 min.The total retention time was 1.612 min with a flow rate of 1.0 ml/min. % 0f RSD values for precision is found to be 0.293 (< 2).The limits of detection (LOD) and quantification (LOQ) were 0.03µg/ml and 0.096133µg/ml, respectively. The correlation coefficient for Mebhydroline Napadisylate 0.9972 indicates linearity of the methods within the limits. The linear range of determination for Mebhydroline Napadisylate was 100-500μg/ml. However, the change in flow rate and column temperature also did not show any significant variance. The % recovery was found to be 99.70%-99.41%.  As per ICH guidelines the proposed method is fully validated and found to be linear over a workable drug concentration, accurate, precise and robust.  This HPLC method is selective, precise, and accurate and can be used for routine analysis of pharmaceutical preparation in industrial quality-control laboratories.