Development and Validation of the Rp-HPLC Method for Estimation of Cilnidipine in Rat Plasma

Objective: Pemigatinib is an active component in treatment of cholangiocarcinoma, but the low solubility and bioavailability of Pemigatinib limit its wide application. The aim of the present study was to prepare and evaluate supersaturable self-nanoemulsifying drug delivery systems (sSNEDDS) followed by investigating and comparing the pharmacokinetic profiles of Pemigatinib and Pemigatinib sSNEDDS in rat plasma by HPLC. Methods: Pemigatinib loaded SNEDDS were obtained by dissolving drug in the isotropic mixture of oil, surfactant, and co-surfactant. The conventional SNEDDS were converted to sSNEDDS by precipitation method by using an experimented polymer. An appropriate high sensitivity and selectivity was applied to the comparison of plasma pharmacokinetics in Pemigatinib and Pemigatinib sSNEDDS using Entrectinib as an internal standard (IS). Results: The droplet of sSNEDDS ranges from 166.78±3.14 to 178.86±1.24 nm with PDI 0.212–0.256, transmission electron microscopy images revealed the spherical shape of the nanodroplets, emulsification time was 15 secs when added to physiological fluids, percent transmittance of the diluted formulation was 99.12±0.46, and viscosity was 574±26 centipoises indicating the good flow ability. FTIR and DSC studies indicated the amorphization of the drug. The dissolution profile of sSNEDDS indicated the faster release of drug compared to both pure drug suspension and SNEDDS formulation. Cmax of the sSNEDDS 3.52±0.13ng/ml was significant (P<0.05) as compared to the pure drug suspension formulation 2.82±0.42 ng/ml. The AUC0-t, AUC0–∞ of sSNEDDS was increased, while the Tmax and t1/2 was decreased. Moreover, the AUC value in the sSNEDDS group was significantly increased and the relative bioavailability was calculated to be 69% when compared with that of the Pemigatinib group. Conclusion: These results concluded that Pemigatinib sSNEDDS when compared with pure drug after a single oral administration and the formulation modification of Pemigatinib into sSNEDDS can effectively enhance gastrointestinal absorption and relative bioavailability by improving solubility and dissolution rate.

Section: Discussionmentioning

confidence: 99%

“…Consisted of mobile phase 0.1% OPA pH 2.5 buffer (60%):Acetonitrile(40%) pumped at a flow rate of 1.06 ml/min gave the highest desirability. The retention time of the drug and IS was found to be 3.258 and 4.43 min respectively [22][23][24][25].…”

Section: Methodsmentioning

confidence: 96%

Formulation and in Vivo Evaluation of Pemigatinib Super Saturable Self-Nano Emulsifying Drug Delivery System

REDDY,

GUBBIYAPPA

2023

“…Further, the mixed solution was neutralized with 2N HCl and again diluted to obtain a solution of 100, 100 and 4 µg/ml for Clarithromycin, Amoxicillin, and Vonoprazan correspondingly. After injecting a volume of 10 µl of the prepared solution above, chromatograms were obtained and then assessed to determine the amounts of Clarithromycin, Amoxicillin, and Vonoprazan that had been degraded by comparing them to the control solution [24].…”

Section: Alkali Degradationmentioning

confidence: 99%

A Reliable Rp-Uplc-Tuv Method for Simultaneous Estimation of Clarithromycin, Amoxicillin, and Vonoprazan in Co-Packed Pharmaceutical Dosage Forms: Method Development and Validation With Stability Indicating Properties

ANUSHA,

SOWJANYA

2024

Objective: The study aims to develop a reliable RP-UPLC-TUV method for simultaneous estimation of Clarithromycin, Amoxicillin, and Vonoprazan in bulk and combined dosage. Methods: A simple, specific, and reliable method for determining Clarithromycin, Amoxicillin, and Vonoprazan has been developed using the RP-UPLC method. In order to successfully separate Clarithromycin, Amoxicillin, and Vonoprazan, 1.0 µl of a 100 % level solution was injected into a Hibar C18 (100 x 2.1 mm and 2 µm) column. The mobile phase consisted of Ammonium Acetate and Acetonitrile in equal volumes, and the flow rate was kept at 0.3 ml/min while the detection wavelength was set to 210 nm. Both the column and the injection port were kept at a temperature of 30 °C at all times. Results: The retention time (RT) of Clarithromycin, Amoxicillin, and Vonoprazan was observed at 1.24 min, 0.97 min and 1.66 min, correspondingly with accepted system suitability. The linear responses were observed for Clarithromycin, Amoxicillin, and Vonoprazan in the range of 25 to 150 µg/ml, 25 to 150 µg/ml and 1 to 6 µg/ml, respectively. The LOD and LOQ values were calculated to 0.07 µg/ml and 0.22 µg/ml for Clarithromycin, 0.81 µg/ml and 2.45 µg/ml for Amoxicillin and 0.03 µg/ml and 0.09 µg/ml for Vonoprazan. The % RSD values of both precision were assessed in the range of 0.8-1.4. The mean recovery of Clarithromycin, Amoxicillin, and Vonoprazan was in the range of 99.66 %-100.88 %. The statistical analysis of the validation parameters confirmed that the approach was reliable in terms of its accuracy, sensitivity, and precision while also exhibiting a high degree of sensitivity. The study of analytes in a variety of stressful situations guarantees the stability of the substances, ensuring that they represent the method's stability indication. Conclusion: The newly established technique is quite effective in separating Clarithromycin, Amoxicillin, and Vonoprazan from one another. Also separated with excellent resolution were the degradation products that were formed as a result of the stress conditions. The study concluded that the developed method has considerable adoption in the pharmaceutical sector.

“…Bioanalytical methods are employed for quantitative estimation of drugs and metabolites in biological fluids [10]. Method validation will provide insight into the identification, characterization, and estimate of the therapeutic drug molecule [11]. Reversed-phase HPLC assembled with a PDA detector is widely used in industries and research-based organizations for the quantification of drugs present in the plasma sample [12].…”

Section: Introductionmentioning

confidence: 99%

Development of Quantitative Method of Tulobuterol Hydrochloride in Rat Plasma: Validation and Application to Preclinical Pharmacokinetics

NAZ,

SUBRAHMANYAM,

RACHAMALLA

2023

Objective: A robust, simple, accurate, rapid, and selective bioanalytical high-performance liquid chromatography (HPLC) method was established and validated to determine the tulobuterol hydrochloride in rat plasma. Methods: The protein precipitation method deproteinated analyte from rat plasma using acetone. The analysis of tulobuterol hydrochloride from rat plasma was accomplished using a mobile phase comprising of methanol: potassium dihydrogen orthophosphate buffer (0.05M; pH 4.0) in 90:10 (v/v) ratio run at 1.0 ml/min flow rate. Separation was carried on BDS hypersil C18 column (4.6 mm × 250 mm; 5 µ) at ambient temperature employing a 996 photodiode array (PDA) detector at 228 nm. Results: The linearity model was exhibited from 100-500 ng/ml with a good correlation of 0.999. Tulobuterol hydrochloride was efficiently separated at a retention time of 7.281 min. The percent recovery rate was between 100.21-100.46 %. The accuracy, precision, robustness, and ruggedness study showed relative standard deviation (%RSD) was within 2% (acceptable limit), and that revealed the method was efficient, precise, reliable, and reproducible. Conclusion: A simple, accurate, suitable method to quantitate tulobuterol hydrochloride in rat plasma was established using HPLC employed with a PDA detector that overcomes the increased cost for analysis. The developed method was successfully validated in rat plasma.