Validation

Current study has developed two precise and direct RP-HPLC approaches for quantitative investigation of glimepiride (GLM) in both mass and pharmaceutical formulations. Glimepiride was analyzed using the RP-HPLC method with C-18 stationary phase and mobile phase of methanol and phosphate buffer (PBS) at pH 4.0 in equivalent volume ratio. The location was established at 239 nm wavelength, and the adaptable stage was extracted at a rate of 0.5 mL/mi. The retention time was observed at 2.470 minutes. Present approach was authenticated in terms of linearity, accurateness, precision, system applicability, limit of detection (LOD), limit of quantification (LOQ), robustness, and ruggedness. It has been demonstrated that the suggested approach is appropriate for monotonous examination of glimepiride in dose and bulk forms, yielding precise results. This method was employed to determine a compound's concentration in commercial pharmaceutical dosage forms. In comparison to alternative chromatographic techniques, this method is more direct, precise, and reproducible, rendering it a superior choice for routine quality control.

Artesunate (1) (ART), also called as dihydroartemisinin-12-α-succinate, it is a semisynthetic peroxide-bridged sesquiterpene lactone compound derived from Artemisinin, the bioactive component of the Chinese medicinal herb called Artemisia annua.  An accurate, simple and precise HPLC method was developed and validated for forced degradation studies of drug Artesunate .The column was used C18 column (150X4.6mmX5µm ) column  by isocratic elution. The mobile phase composition consisted of Acetonitrile Water and trifluoroacetic acid (TFA) in the ratio of 55:45:0.1 v/v. The analysis was performed at a 1ml/min flow rate. The analytical performance parameters such as linearity, precision, accuracy, specificity, limit of detection (LOD) and lower limit of quantification (LOQ) were determined according to International Conference for Harmonization ICH Q2 (R1) guidelines.

The present study describes the development and subsequent validation of Reverse phase HPLC (RP-HPLC) method for the analysis of Imeglimin hydrochloride. A novel economic, simple, rapid, accurate, reproducible, and precise Reverse Phase High Performance Liquid Chromatography (RP-HPLC) method for Imeglimin hydrochloride. The method was performed on a YOUNG LIN-HPLC system-ACME9000. The method developed for Imeglimin hydrochloride was quantitatively measured using an isocratic RP-HPLC methodology. The chromatographic separation of Imeglimin hydrochloride was achieved on RP-HPLC equipped with Hypersil BDS C18 (250mm x 4.6mm, 5 um) column using isocratic elution with a mobile phase consisting of MeOH: Buffer in a ratio of (70:30% v/v) at a flow rate of 1.0ml/min with an injection volume of 20µl, where detection was carried out by UV- 730D detector at 239nm The retention time for Imeglimin hydrochloride was found to be 3.47 min. The developed method was successfully with results falling within acceptable criteria validated for different validation parameters as per (ICH-Q2 (R1)) guidelines. The linear regression equation was found to be y = 27.83x - 8.512 with a correlation coefficient (R2) > 0.999 which shows excellent linear correlation. Accuracy, precision, specificity, system suitability, robustness, linearity, LOD and LOQ were determined for method validation. The results were found to be well within recommended limits as per ICH guidelines. Stability studies of Imeglimin hydrochloride were carried out under acidic, basic, peroxide, photolytic and thermal conditions. Degradation was observed in acidic, basic, and oxidative conditions, but not in photolytic and thermal conditions.  

A simple, precise, and accurate method was developed for the simultaneous estimation of Nivolumab (NVM) and Cabozantinib (CBZ) in tablet dosage form.  The chromatogram was analyzed through a Phenomenex C18 150 mm (4.6 x 150 mm, 5 m) for chromatogram processing. A mobile phase containing formic acid: methanol (50:50) was pumped through the column at a spurge flow of 1.0 mL/min.  The column temperature was upheld at 30°C.  The quantification was done at 260.0 nm.  The elution time of NVM and CBZ was found to be 2.243 min and 2.953 min, respectively. The validation for the developed method was performed and all the parameters were found within the specified limits. The standard curve results represent a correlation coefficient of more than 0.999.  The %RSD of NVM and CBZ were found to be 0.9 and 0.7, respectively.  % Recovery was achieved as 99.88% and 99.66% for NVB and CBZ, respectively. The NVM and CBZ regression equations yielded LOD and LOQ values of 0.63, 1.91, and 0.08, 0.24 respectively. The equation of Nivolumab y is 39306x + 12173 while the Cabozantinib y is 34894x + 1139.7. With all the parameters under the criteria, the developed method for the simultaneous estimation of Nivolumab and Cabozantinib can be successfully applied for regular quality control approaches.

In this review articles, the development, formulation, and manufacture of drugs, analytical method development & validation play a critical role. Methods are developed for ensuring purity, identity, potency, and performance of pharmaceutical products. Methods should be applied to the extent that they are sufficient for their intended purpose. Throughout the life cycle of a drug product and substance, a range of activities are associated with developing and validating methods. An objective of method validation is to prove that the procedure can be used as intended. Once the method is developed, validation is performed. Different national and international committees have defined the parameters for method validation. The International Conference on Harmonization attempted to harmonize pharmaceutical applications. In accordance with the ICH, other organizations define Linearity, Selectivity/Specificity, Range, Accuracy, Precision (repeatability, intermediate precision, and reproducibility), Limit of quantitation, Limit of detection, Ruggedness, and Robustness.

A novel Quality by Design methodology was used to develop and validate a rapid, accurate, precise, simple, efficient and reproducible isocratic Reversed Phase-High Performance Liquid Chromatographic (RP-HPLC) method for the estimation of Luliconazole in bulk and pharmaceutical dosage form. Luliconazole was separated using Kromasil C18 column (250mm×4.6 mm, 5µm particle size), Shimadzu LC2030 HPLC system having UV detector and the mobile phase contained a mixture of 0.01M Ammonium acetate buffer and Acetonitrile (35:65). The flow rate was set to 1.2 ml/min with the responses measured at 294nm. The retention time of Luliconazole was found to be 3.092 min. Central composite design employed for design of experiment and optimization. Desirability value was found to be 0.723 and overall model was found to significant. Linearity was established for Luliconazole in the range of 20-120 µg/ml with correlation coefficient (r2=0.9995). Limit of detection (LOD) and limit of quantitation (LOQ) were evaluated and found to be 1.1700 and 3.5455 respectively. The accuracy values were found to be in the range of 98 –102% and every parameter found with in limit. Validation parameters were evaluated for the method according to the International Conference on Harmonization (ICH) Q2 R1 guidelines. This method can be used for the estimation and analysis of Luliconazole drug in active pharmaceutical ingredients and pharmaceuticals.

To develop and validate simple, rapid, linear, accurate, precise and economical reverse phase-high-performance liquid chromatography (RP-HPLC) method for of stress degradation study of Teneligliptin. The separation and quantization were achieved on Inertsil C18(250 mm × 4.6ID,5 um ). The mobile phase selected was Methanol: Water (90:10) at a flow rate of 0.8 ml/min and detection of analytes was carried out at 248nm at pH 3. The method exhibited good linearity over the range of 10–50 µg/mL. The drug is freely soluble in organic solvents Methanol. The drug was identified in terms of solubility studies and on the basis of melting point done by capillary tube method. The drug which when subjected to thermal, photolytic, oxidative, and acidic stress degraded into many degradation products. In most of the cases, the degradation rate was seen to be directly proportional to the amount of stress applied. The thermal stress was increased by increasing the incubation temperature, the faster the degradation took place. The values of LOD were found to be  0.956  ug/ml for TNG  and the calculated LOQ values were found to be 0.171 ug/ml.

A sensitive and rapid HPLC method was developed and validated for the determination of potential genotoxic impurities i.e (Bromomethyl)biphenyl methyl ester and (Dibromomethyl)biphenyl methylester at trace level in Telmisartan drug substance by applying the concept of threshold of toxicological concern (TTC). The HPLC method was developed and optimized on Symmetry Shield RP18, 3.5 m (150mm × 4.6mm) column with oven temperature maintaining at 40°C and 0.02M Phosphate buffer pH 2.5 was chosen as mobile phase A and mixture of acetonitrile and Phosphate buffer (55:45) was selected as mobile phase B in gradient reverse phase mode in isocratic mode of composition. Chromatographic parameters i.e flow rate: 1.0 ml/min, wavelength detection: 205 nm, injection volume: 20µl and run time: 25 min were applied in this methodology.  Based on validation data, the method is found to be specific, sensitive, accurate and precise. The established limits of Limit of detection and Limit of quantification for subjected impurities are found to be 2.4 µg/g and 4.7 µg/g respectively for each impurity. The recovery at LOQ level obtained was 98.2% for (Bromomethyl) biphenyl methyl ester and 99.2% for (Dibromomethyl) biphenyl methyl ester. This method can be used as good quality control tool for quantization of these impurities at low level. The experimental results are discussed in detail in this research paper.

To method develop and validate simple, rapid, cost effective, linear, accurate, precise and economical for the simultaneous estimation of Pregabalin and Methylcobalamin in bulk and tablet dosage form by using UV-Spectrophotometry. The drug obeyed the Beer’s law and showed good correlation of concentration with absorption which reflect in linearity. The UV-Spectroscopic method was developed for estimation of Pregabalin and Methylcobalamin in bulk and tablet dosage form and also validated as per ICH guidelines. The method based on measurement of absorbance at two wavelengths 222 nm (λmax of Pregabalin) and 219 nm (λmax of Methylcobalamin) in Ethanol and distilled water. Linearity graph of Pregabalin and Methylcobalamin were found to be linear in the concentration ranges of 30-150 μg/ml and 0.6-3 μg/ml, respectively, with their correlation coefficient values (R2) 0.999. The low %RSD values indicate method to be accurate and precise. The % recovery of tablets found to be in range of 97-102% and other validation parameter were found to be within the limits as per ICH guidelines.

To develop and validate simple, rapid, linear, accurate, precise and economical UV Spectroscopic method for estimation of Fosfomycin in Fosfomycin for Injection. The drug is freely soluble in analytical grade water 1. The drug was identified in terms of solubility studies and on the basis of melting point done on melting point apparatus of Equiptronics 2. It showed absorption maxima were determined in analytical grade water. The drug obeyed the Beer’s law and showed good correlation of concentration with absorption which reflect in linearity 4, 5, 6. The UV spectroscopic method was developed for estimation of Fosfomycin in injection dosage form and also validated as per ICH guidelines. The drug is freely soluble in analytical grade water, slightly soluble in 95% Methanol and Ethanol. Almost insoluble in anhydrous acetone, ether and chlorinated solvent. So, the analytical grade water is used as a diluent in method. The melting point of Fosfomycin was found to be 94 - 95?C (uncorrected). It showed absorption maxima 254 nm in analytical grade water. On the basis of absorption spectrum the working concentration was set on 50µg/ml (PPM). The linearity was observed between 30-70 μg/ml (PPM). The results of analysis were validated by recovery studies. The recovery was found to be 98.75, 101.00 and 99.17% for three levels respectively. The % RSD for precision was found to be 0.41%. A simple, rapid, linear, accurate, precise and economical UV Spectroscopic method has been developed for estimation of Fosfomycin in Fosfomycin for Injection dosage form. The method could be considered for the determination of Fosfomycin in quality control laboratories.