Method Validation

A simple, precise, and accurate UV spectrophotometric method was developed and validated for the simultaneous estimation of Lornoxicam and Thiocolchicoside in bulk and pharmaceutical formulations. The method employs methanol as solvent, and absorbance was measured at the respective wavelengths where both drugs showed maximum absorbance with minimal interference. Calibration curves were linear within the concentration ranges of 4–20 µg/mL for Lornoxicam and 5–25 µg/mL for Thiocolchicoside, with correlation coefficients (R²) of 0.9992 and 0.9995, respectively. Validation was carried out according to ICH Q2(R1) guidelines, including parameters such as linearity, accuracy, precision, LOD, LOQ, robustness, and ruggedness. Recovery studies at 50%, 100%, and 150% levels showed recoveries between 98.7–100.1%, indicating high accuracy. The proposed method is suitable for routine quality control of combined dosage forms of Lornoxicam and Thiocolchicoside.

To develop and validate a rapid, precise, and stability-indicating UPLC method for the simultaneous estimation of Sulfamethoxazole and Clindamycin in combined pharmaceutical dosage form, in accordance with ICH Q2 (R1) guidelines 5. A simple, precise, and robust UPLC method was developed and validated for the simultaneous estimation of Sulfamethoxazole and Clindamycin in a fixed-dose pharmaceutical formulation 1–3. The method was developed on a reverse-phase C18 column using a mobile phase of methanol and water. Detection was carried out at 254?nm. Validation followed ICH Q2 (R1) guidelines, including accuracy, precision, linearity, robustness, ruggedness, and sensitivity 4–6. Linearity was observed over the range of 10–200 μg/mL for both drugs (R² > 0.998). %Recovery was within 98–102% with % RSD < 0.5%. LOD and LOQ were 0.195 μg/mL and 0.592 μg/mL, respectively. The method was unaffected by small deliberate changes in analytical conditions. The validated method is stability-indicating, highly sensitive, and suitable for routine analysis and quality control of Sulfamethoxazole and Clindamycin in combined dosage form.

Albendazole (ALB), chemically known as methyl [5-(propylthio)-1H-benzimidazol-2-yl] carbamate is widely used as an anthelmintic having a wide spectrum of activity. Numerous numbers of analytical methods are there for the simultaneous estimation of bulk and in formulation, such as spectrophotometry and liquid chromatography. As the UV spectrophotometric method is rapid, simple, accurate and economical, the method has been developed for the assay of the albendazole in pharmaceutical formulation. The wavelengths selected for the method were at 291 nm. The results of analysis have been validated by recovery studies as per ICH guidelines. The developed method was rapid, simple, accurate and economical and it can be used for routine quality control analysis. It showed absorption maxima at 291 nm in analytical grade DMF. The drug obeyed the Beer’s law and showed good correlation of concentration with absorption which reflect in linearity.

A Novel stability-indicating method was developed and validated by Reverse phase Hplc method   for the estimation of Thiotepa in Lyophilized form. The simple, Accurate Assay method was developed by PDA Detector at 215nm and column(YMC,150mmx4.6mm, 3.5 μm,12nm).The mobile phase A consisted of (3.5g of potassium Dihydrogen phosphate/1liter: 4.2g of Disodium hydrogen Phosphate/2liters)Buffer : Acetonitrile (85:15v/v) and The mobile phase B consisted of (3.5g of potassium Dihydrogen phosphate/1liter: 4.2g of Disodium hydrogen Phosphate/2liters)Buffer : Acetonitrile (50:50v/v) gradient flow rate at 1.0 mL/min for 30minutes. Samples was stored at 5°c temperature in sample cooler and Column oven temperature was maintained  in the method at 30°c  Respectively. The retention time of Thiotepa was 11.313 minutes. The proposed method was developed and validated with respect to specificity, accuracy, precision, linearity, Analyte Stability. Specificity of the Assay method shows no interference from Diluent and degrading products formed by alkaline, acidic, oxidative, Water Hydrolysis conditions. At Each level, %Recovery of Thiotepa in formulations was obtained to be in a range of 98-102%. The linearity of the assay was established in the range of 159.915-479.744µg/mL with correlation coefficient (R2) > 0.9999. This method was shows simple, precise, more accurate, stability indicating and reliable determination of Thiotepa for drug stability assay in pharmaceutical studies.

A simple, rapid, precise LC-MS compatible method for the degradation impurities if any UHPLC method was developed for the determination of 1-deoxynojirimycin (DNJ) in Mulberry leaves. The DNJ in 200 mg dried leaves was extracted twice with 50 mL aqueous 0.05 mol/L HCl, then derivatized by 9 fluorenylmethyl chloroformate (9 FMOC-Cl) in Potassium Borate buffer, and analyzed using a high performance liquid chromatograph equipped with a fluorescence detector. Chromatographic separation of the same was performed by using a Shimpack XR ODS-III (150*3.0mm) 2.1µm as stationary phase with a mobile phase of 0.1% Acetic acid in water: Acetonitrile (60: 40, v/v) at the rate of 0.5 mL/min. Wavelengths used were 254 nm for excitation and 322 nm for emission, which were suitable for detecting the native fluorescence of all the pigments assayed. The derivatized DNJ was well dissolved from the hydrolysis products of 9 FMOC-Cl. The linearity ranged from 0.45 to 66 mg/L, and the detection limit was 0.2 mg/L (S/N = 3). The content of DNJ in Mulberry leaves was 0.09%, the recovery was 90.0%-110.0%.

The present study was undertaken to develop and validate a simple, accurate, precise, reproducible and cost effective UV-Visible spectrophotometric method for the estimation of cefpodoxime proxetil in bulk and pharmaceutical formulation. The solvent used throughout the experiment was methanol and water. Absorption maximum (λmax) of the drug was found to be 231 nm. The quantitative determination of the drug was carried out at 231 nm and Beer’s law was obeyed in the range of 5-25μg/mL. The method was shown linear in the mentioned concentrations having line equation y = 0.0331x + 0.0151 with correlation coefficient R2of 0.9998. The recovery values for cefpodoxime proxetil ranged from 99.80% - 100.02%.The percent relative standard deviation (RSD %) of interday precision range was 0.118 – 0.181% and intraday precision range was 0.139 – 0.446%. The limit of detection and limit of quantification was 0.081μg/mL and 0.121μg/mL. The percent relative standard deviation of robustness and ruggedness of the method was 0.126 – 0.313%.Cefpodoxime proxetil content in two pharmaceutical dosage forms (tablet and suspension) were determine which were in good agreement with the label claims with RSD value of 0.02% for tablet and 0.01% for suspension. Hence, proposed method was precise, accurate and cost effective. This method could be applicable for quantitative determination of the bulk drug as well as dosage formulation.

  First and second order derivative UV-Spectrophotometric methods have been developed and validated for the estimation of Iloperidone in bulk and its tablet formulations. The solutions of standard and sample were prepared in methanol. The Iloperidone solution was showed the maximum absorbance at 262nm and 248nm for the first and second order UV-Spectrophotometric methods respectively. Beer’s law was obeyed in the concentration range of 4- 12 μg / ml with r2 value 0.999 for both the methods. These methods were tested and validated for various parameters according to ICH guidelines. The precision expressed as relative standard deviation and was found within the range of 0.13 % to 1.7 % for the both methods. Limit of detection was 0.0133 μg/ml (first order), 0.0216 μg/ml (second order) and  limit of quantification was found to be 0.0403 μg/ml (first order), 0.0657 μg/ml (second order). Recovery of Iloperidone was found to be within the range of 99.51 – 100.16 % for the two methods. The proposed methods were successfully applied for the determination of Iloperidone in tablet formulations. In addition, the proposed methods are simple, easy to apply, low cost, and requires relatively inexpensive instruments.