SIMULTANEOUS REVERSE-PHASE ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY METHOD DEVELOPMENT AND VALIDATION FOR ESTIMATION OF GRAZOPREVIR AND ELBASVIR

 Objective: The objective of this study was to develop and validate rapid, specific, sensitive, and precise reverse-phase ultra performance liquid chromatography (RP-UPLC) method for the quantitative determination of grazoprevir and elbasvir, as there are no official monograph and no analytical method by UPLC.Methods: Chromatographic separation was achieved on a Waters Acquity UPLC HSS C18 (2.1 mm × 100 mm, 1.8 micron) column with a 45:55 (v/v) mixture of 0.1% orthophosphoric acid (pH 2.8) and acetonitrile as a mobile phase, thermostated at 30°C with a short run time of 3.0 min.Results: The retention times were 0.73 and 1.29 min for grazoprevir and elbasvir, respectively. Quantification is achieved with TUV detection at 254 nm over the concentration range of 25–150 μg/ml for grazoprevir and for elbasvir 12.5–75 μg/ml, with a correlation coefficient of 0.999 and 0.999, respectively. The developed method was validated according to the International Conference on Harmonization (ICH) guidelines with respect to linearity, accuracy, precision, specificity, and robustness. Forced degradation study was extended out under acidic, alkaline, oxidative, photolytic, and thermal conditions to demonstrate the stability-indicating capability of the developed UPLC method. The degradation products were well resolved from the main peak, thus proved the stability-indicating power of the method. The results of the analysis were validated statistically.Conclusion: The method is precise, accurate, linear, robust, and fast. The short retention time allows the analysis of a large number of samples in a short period of time and, therefore, should be cost-effective for routine analysis in the pharmaceutical industry

IN VIVO AND IN VITRO EVALUATION OF TEPHROSIA CALOPHYLLA FOR ANTI-DIABETIC PROPERTIES

Objective: The objective the present work was to investigate in vivo and in vitro anti-diabetic potentials of methanol extract of Tephrosia calophylla against alloxan-induced diabetes in albino rats.Methods: For in vivo evaluation, diabetes was induced in albino rats by administering a single dose of alloxan. The study was designed to test the acute effect of methanol extract of Tephrosia calophylla (TCME) to reduce blood glucose in OGTT. The chronic study of 21 d was performed against diabetic rats and blood glucose was determined at 1st, 7th, 14th and 21st day. In chronic in vivo study, serum concentrations of insulin, urea, creatinine, total cholesterol, triglycerides, ALT and AST were also estimated at 21st day. The in vitro α-glucisidase inhibitory activity and α-amylase inhibitory activity were performed and IC50 values of the extract was determined. The glucose uptake by rat hemidiaphragm model was also used test potentials of the extract to increase utilization of the glucose by tissues.Results: In OGTT, standard glibenclamide and TCME at 400 mg/kg treated animals have shown significant reduction in blood glucose at 90 min but at 120 min, blood glucose level (BGL) was significantly reduced in glibenclamide and TCME at 200 mg/kg and 400 mg/kg treated animals compared to diabetic control group. In chronic modelthe methanol extract effectively reduced blood glucose levels (P<0.001) at 14 th and 21st day of study in therapeutic groups and effect was comparable to that of standard. The extract could also significantly (P<0.001) reduce concentrations of SGOT, triglycerides (TGs), Total cholesterol (TC) and urea in serum and significantly (P<0.001) increased the insulin level in blood which proves beneficial effects of the extract in diabetes. The change in concentrations of SGPT and urea were less significant (P>0.01). In vitro studies, against both glucosidae and  amyalase inhibitory activities, extract has shown significant IC50 values and it also enhanced glucose utilization by rat hemidaphragm.Conclusion: The results obtained from the present study suggest that, the methanol extract of Tephrosia calophylla possess significant in vivo anti-diabetic properties against alloxan induced diabetes in rats. The results also suggests that, TCME also possess the significant in vitro anti-dabetic potentials

DEVELOPMENT AND VALIDATION OF LIQUID CHROMATOGRAPHY COUPLED WITH TANDEM MASS SPECTROMETRY METHOD FOR ESTIMATION OF LENVATINIB IN HUMAN PLASMA

Objective: This study was to develop and validate a liquid chromatography–tandem mass spectrometry (LC–MS/MS) for the quantification of lenvatinib (LT) in human plasma.Methods: A simple, sensitive and specific LC–MS/MS method was developed for quantification of LT in human plasma using LTD4 as internal standard (IS). The analytical method consists of liquid–liquid extraction of plasma sample followed by the determination of LT by a LC–MS/MS. The analyte was separated on a Zorbax Eclipse XDB-C18 (150×4.6 mm, 5 μ) column with an isocratic mobile phase of acetontrile:0.1% formic acid (80:20 v/v) at a flow rate of 0.6 mL/minutes. The protonated ions were formed by a turbolon spray in a positive mode were used to detect analyte and IS. The MS/MS detection was made by monitoring the fragmentation of m/z 427.10→370.10 for LT and m/z 430.30→370.10 for IS on a MS.Result: The method was validated with the correlation coefficients of (r2) ≥0.995 over a linear concentration range of 10.20-501.60 pg/mL. This method demonstrated intra- and inter-day precision within 1.06-2.42% and 0.03-0.55% and accuracy within 95.64-100.08% and 97.16-100.07%.Conclusion: This method is suitable and convenient to pharmacokinetics and bioavailability studies for estimation of LT in biological samples by LC–MS/MS

Gastro-protective effects of methanol extract of Tephrosia calophylla

Objective: The present research work was designed to investigate gastro protective potentials of methanol extract of Tephrosia calophylla. Methods: The aerial parts of Tephrosia calophylla were dried under shade, powdered and deffated with petroleum ether and then marc left over was subjected to methanol extraction using soxhlet apparatus. Antiulcer activity of methanol extract was determined against stress induced and aspirin induced ulcers in experimental animal models. The total number of ulcers formed, ulcer index, percentage inhibition, ulcerated area, protected area, pH and Total acidity were parameters in the study. Results: Methanol extract of Tephrosia calophylla have significantly reduced the total number of ulcers formed, ulcer index, ulcerated area and total acidity in therapeutic groups compare to vehicle control and there by significantly increased percentage inhibition of ulcers and protected area which was evident by significant rise in pH of gastric content. The effect of extracts was dose dependent and results were comparable to that of standard drug omeprazole. Conclusion: The results obtained from the present work suggest that the methanol extract of Tephrosia calophylla possess significant anti-ulcer potentials against experimentally induced ulcers in albino rats. Keywords: Tephrosia calophylla, Anti ulcer activity, Ethanol, Aspirin Ulcer index, pH, total acidity, Percentage inhibition and percentage of protected area

BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF ENTRECTINIB IN RAT PLASMA BY LIQUID CHROMATOGRAPHY–TANDEM MASS SPECTROMETRY

Objective: The objective of the study was to develop and validate the bioanalytical liquid chromatography–mass spectrometry (LCMS/MS) method for the estimation of entrectinib in bulk and pharmaceutical drugs in rat plasma. Methods: Chromatographic separation of entrectinib with D4-entrectinib as internal standard (IS) was achieved using Waters Alliance high-performance liquid chromatography system, quaternary gradient pump of e2695, using Luna, 250×4.6 mm, 5 μm column and the mobile phase containing 0.1% formic acid and acetonitrile (ACN) within the ratio of 70:30% v/v. The flow was 1.0 ml/min; detection was carried out by absorption at 294 nm using a photodiode array detector at ambient temperature. Results: The peak of entrectinib was eluted at retention times of 5.225 min. The multiple reaction monitoring was 560.6/475.1 (m/z) for entrectinib and 580.6/496.3 (m/z) for IS entrectinib (D4). The linearity range was 1–20 ng/ml with a regression coefficient of 0.999. % relative standard deviation of peak areas of all measurements always <2.0. Conclusion: The method was successfully validated and it had been found to be within limits for accuracy, precision, and linearity and it is stable under analytical conditions used

BIOAVAILABILITY ENHANCEMENT BY FLOATING MICROBALLOONS OF DIPYRIDAMOLE AND CLOPIDOGREL: IN VIVO PHARMACOKINETIC STUDY

Objective: In vivo pharmacokinetic studies of clopidogrel and dipyridamole floating microballoons to check their bioavailability enhancement. Methods: The bioanalytical method development was carried by using HPLC with column Poroshell 120 EC-C 18; 4.6x100 mm. The in vivo pharmacokinetic studies were performed in Wistar male rats and the obtained data from the pharmacokinetic parameters were analyzed using PK Solver software. Results: The developed bioanalytical method was found to be linear in the concentration range of 1-100 ng/ml for clopidogrel bisulfate and 0.02-4µg/ml for dipyridamole with correlation coefficient of 0.9993 and 0.9987 respectively. The study results showed that the method was simple, linear, accurate and precise. The in vivo studies indicated that the AUC was found to be increased by 33.3% and 154.5% for clopidogrel and dipyridamole micro balloons, respectively, when compared to their pure drugs. Conclusion: The bioanalytical methods development and their validation parameters indicated that the methods are accurate, precise and linear in the studied range of concentrations. In vivo test results infer to the effective, sustained release of both the drugs when formulated as micro balloons and increase in the absorption, thereby enhancing the bioavailability of the drugs. The pharmacokinetic studies also confirmed the increase in the mean residence time of the drugs when formulated as floating microballoons

scope of sorafenib tosylate in renal cell carcinoma

Renal cell carcinoma  is a kidney cancer that originates in the lining of the proximal convoluted tubule. It  is the most common type of kidney cancer in adults, responsible for approximately 80% of cases. It is also known to be the most lethal of all the genitourinary tumors. Sorafenib received FDA regular approval on December 20, 2005 for the treatment of advanced RCC. Sorafenib belongs to the class of diaryl ethers. Sorafenib is a kinase inhibitor that decreases tumor cell proliferation in vitro. The long-term treatment with sorafenib is associated with continued efficacy and a well-tolerated safety profile. It can be used in combination with other antineoplastic drugs effectively. Based on its mechanism of action and findings in animals, sorafenib may cause fetal harm when administered to a pregnant woman. Greater  sensitivity of some older individuals was observed. However, treatment  with sorafenib prolongs progression-free survival in patients with advanced  renal cell carcinoma in whom previous therapy has failed. Â

Assessing Software Reliability Using Exponential Imperfect Debugging Model

Software reliability is one of the most important characteristics of software quality. As the usage of software reliability is growing rapidly, accessing the software reliability is a critical task in development of a software system. So, many Software Reliability Growth Models (SRGM) are used in order to decide upon the reliable or unreliable of the developed software very quickly. The well known software reliability growth model called as Exponential Imperfect Debugging model is a two parameter Non Homogeneous Poisson Process model which is widely used in software reliability growth modeling. In this paper, we propose to apply Statistical Process Control (SPC) to monitor software reliability process. A control mechanism is proposed based on the cumulative observations of failures which are grouped using mean value function of the Exponential Imperfect Debugging model. The Maximum Likelihood Estimation (MLE) approach is used to estimate the unknown parameters of the model. The process is illustrated by applying to real software failure data

BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE DETERMINATION OF SOFOSBUVIR FROM HUMAN PLASMA

Objective: This study points to build up and validate a simple methodology to quantify the most used drug sofosbuvir for the treatment of hepatitis C virus (HCV) infection, in human plasma by using atazanavir as an Internal Standard (IS) for preclinical studies and validate as per USFDA guidelines.Methods: Sofosbuvir was isolated from plasma samples by liquid-liquid extraction method using acetonitrile; good chromatographic separation was achieved on Kromasil Column (250 mm ×4.6 mm, 5 µm). The mobile phase consisted of 0.1 % orthophosphoric acid (OPA) buffer pH 2 and acetonitrile in the ratio of (68:32, v/v), respectively. The analysis time was 7 min at a flow rate 1 ml/min. The photodiode array detector (PDA) detection was carried out at 228 nm. The suggested method was validated by performing linearity, system suitability, specificity and sensitivity, accuracy and precision, recovery, ruggedness, stability studies. The method was validated as per USFDA guidelines.Results: The developed method resulted in retention times of sofosbuvir and IS were found out to be 4.7 and 4.2 min respectively. The calibration curves are linear (r2 = 0.999) over the concentration range of 0.050-2.0 µg/ml of plasma analytes concentration. LOQ value was found to be 0.050 µg/ml with precision and accuracy. Within-batch % mean accuracy of the method ranged between 96.00% and 109.09%, and within-batch and total precision, expressed as the coefficient of variation, was 1.40–10.33%. Overall percentage mean recovery of sofosbuvir from spiked plasma was 84.14%. All the validated parameters were found to be within the limit.Conclusion: A simple, accurate, precise, linear, rugged and rapid RP-HPLC method was developed for quantitative estimation of sofosbuvir in human plasma and should be suitable for conducting pharmacokinetics studies and therapeutic drug monitoring

Influence of Kollidon SR on Ondansetron HCl pH Independent Drug Release from Hydroxypropyl Methyl Cellulose Matrix Tablets

The Polymers are tools used in novel drug delivery system to modify the drug release of pharmaceutical dosage form. Ondansetron HCl is a weakly basic drug belongs to BCS class-II; it is showing distinct pH dependent solubility. The major intention of the current study was to develop a pH independent controlled released system for pH dependent poorly soluble Ondansetron HCl. The effect of combination of polymers on parameters like release pattern, release mechanism of the drug were studied. A 32 full factorial design was used to study the effect of Kollidon SR on Ondansetron HCl Drug Release from Hydroxypropyl Methyl Cellulose Matrix Tablets. The release rate from formulated matrix tablets was studied at both SGF (pH 1.2) and SIF (pH 6.8).  Drug release from  Kollidon SR and Methocel (1:1) based  matrix system based tablets was found to  pH independent controlled  drug release up to 24h with  >90% drug release. Kollidon SR has a distinctive character of maintaining tablets geometric shape until the end of dissolution test, this is mainly due to the water insoluble content, polyvinyl acetate, forming 80% (w/w) of Kollidon SR, while the remaining content 20% (w/w) is the water soluble, polyvinylpyrrolidone, responsible for pore formation causing a diffusion controlled release The similarities in Release profiles were evaluated by applying the model independent (f2) similarity factor. The Optimized formulation characterized by DSE, X-RD and FT-IR studies was found not having any interaction with polymer and drug. The Optimized Formulation followed zero order with non-Fickian diffusion method. In conclusion, Kollidon SR and Methocel k100 were found to be novel potential candidates for the development of pH independent controlled delivery system of Ondansetron HCl