UPLC

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.

The present study is carried out using the UPLC as the analytical technique in developing and validating an accurate, precise, linear and robust analytical method for the simultaneous estimation of Pitofenone hydrochloride, Diclofenac potassium and Fenpiverinium bromide in tablets. The method is optimized with a mixture of 0.01M phosphate buffer (PH4.8) and Acetonitrile in the ratio of 40:60 (V/V) as mobile phase and Agilent SB C18 (250 X 4.6) mm, 5µm as stationary phase. The Chromatographic peaks were detected and measured at 215nm. The retention times of Pitofenone hydrochloride, Diclofenac potassium and Fenpiverinium bromide were found to be 1.0, 1.66 and 2.0 respectively. The developed method was demonstrated to access its suitability for meeting its intended purpose by the Validation with a set of validation parameters as per ICH and USP guidelines.  The method is found to be precise with %RSD - 0.48, 0.69, 0.66 for Pitofenone hydrochloride, Diclofenac potassium and Fenpiverinium bromide respectively: accurate with the recoveries of 99.6 to 101.6, 100.51 to 101 and about 100% for Pitofenone hydrochloride, Diclofenac potassium and Fenpiverinium bromide respectively. The method is proved to be linear from the conc.12.5 to 75ppm for Pitofenone, 250 to750 ppm for Diclofenac and 0.5 to 1.5ppm for Fenpiverinium bromide with the correlation coefficients of 0.999, 0.999 and 0.999 respectively. Hence the developed method could be used for the routine analysis purpose in the evaluation of Pitofenone, Diclofenac potassium and Fenpiverinium bromide Tablets.

A novel, simple, rapid and stability-indicating reversed-phase ultra performance liquid chromatographic and mass spectrometric method was developed and subsequently validated for quantitation of Efavirenz (EFV) from drug substance matrix. The separation was achieved in 2.5 minutes on Waters ACQUITY UPLC BEH C18 (50 x 2.1) mm, 1.7µm column in isocratic mode with flow rate 0.4 mL/min. Mobile phase used was 0.01 M ammonium acetate buffer pH 7.5 and acetonitrile in ratio 50:50 v/v. Detection was carried out at the maximum wavelength of 247 nm using a photodiode array detector. The retention time of Efavirenz was found 1.8 minutes. Specificity of the method was established on drug substance by hydrolytic and oxidative stress conditions. Validation of analytical method was carried out as per the current ICH guidelines for linearity, recovery, precision, limit of detection, limit of quantification and robustness parameters.

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%.

A simple, selective, linear, precise and accurate UPLC Method was developed and validated for rapid assay of Nordette in tablet Formulation. Isocratic elution at a flow rate of 0.4ml/min was employed on C8 1.7 µm (2.1 mm x 100 mm) Column at ambient temperature40 °C. Injection Volume was found to be 5.0 µl. The mobile phase consisted of Acetonitrile : Water  60:40 v/v which is filter through a 0.2 µm filter The UV detection wavelength was 220nm and 2µl sample was injected. The retention time for Ethinylestradiol, Levonorgestrel is found to be ± 1.4 minutes and ± 2.1 minutes respectively. A linear regression curve was constructed, and the correlation coefficients (R2) and assessment values calculated. The percentage RSD for both Ethinylestradiol, Levonorgestrel was found to be 1.5%.The Accuracy of method ranges between 97.0 – 102.8%.  The method was validated as per the ICH guidelines. The method was successfully applied for routine quality control analysis of pharmaceutical formulation.