Anticancer and Antioxidant activity of Tephrosia calophylla against cancer cell lines

To evaluate anticancer and antioxidant potentials of activity of methanol extract of Tephrosia calophylla. The methanol extract of Tephrosia calophylla was prepared and tested for in vitro anticancer activity by using human MCF 7, HCT-116, HEP-G2, A-549 cancer and vero normal cell lines. The antioxidant activity for the extract was evaluated by Superoxide scavenging, Lipid peroxidation and DPPH methods. IC50 values of extracts were parameters in both studies. The results of the study have shown that methanol extract of Tephrosia calophylla has shown significant IC50 values against MCF 7, HCT-116 and A-549 cell lines which shows its anticancer potentials but inhibitory effect of methanol extract against HEP-G2 and vero cell lines was not significant. The results also suggest that the methanol extract of Tephrosia calophylla has significant antioxidant properties against Superoxide scavenging, Lipid peroxidation and DPPH methods. The present study shows anticancer potentials of methanol extract of Tephrosia calophylla against MCF 7, HCT-116 and A-549 cell lines.

Study on influence of polymer and surfactant on in vitro performance of biodegradable aqueous-core nanocapsules of tenofovirdisoproxil fumarate by response surface methodology

The major objective of this study was to investigate the effect of biodegradable polymer type and surfactant concentration on various characteristics viz. particle size, entrapment efficiency and drug release rate constant of aqueous core nanocapsules (ACNs) containing tenofovirdisoproxil fumarate. In this study, the nanocapsules were prepared by modified multiple emulsion technique with biodegradable polymers viz. poly(lactide-co-glycolide) of two different grades (PLGA RG502H and PLGA RG503H) and poly lactic acid (PLA R203H); and the surfactant employed was span 80. The experiments were designed under response surface methodology by employing the Design Expert software. Entrapment efficiency, particle size and drug release rate constant were taken as response variables. The prepared nanocapsules were subjected to characterization studies and the obtained results were statistically analyzed by Analysis of Variance (ANOVA) for response surface 2-Factorial Interaction model. ANOVA studies showed that the influence of both factors on all the response variables were significant at p<0.05. The optimized formulation was found to have the entrapment efficiency of 71.58%, particle size of 252.41 nm and the drug release rate constant of 0.045 h-1; thus, indicating that the ACNs were obtained with finest characteristics. SEM studies showed that the particles were spherical

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

A sensitive bioanalytical method development and validation of cabozantinib in human plasma by LC-ESI-MS/MS

A simple, sensitive and specific liquid chromatography–tandem mass spectrometry (LC-MS/MS) method was developed for the quantification of cabozantinib (CZ) in human plasma using cabozantinib-d4 (CZD4) as an internal standard (IS). Chromatographic separation was performed on Xbridge C18, 50 x 4.6 mm, 5 mm column with an isocratic mobile phase composed of 10mM Ammonium formate and Methanol in the ratio of (20:80 v/v), at a flow-rate of 0.7 mL/min. CZ and CZD4 were detected with proton adducts at m/z 502.2 → 391.1 and 506.3 → 391.2 in multiple reaction monitoring (MRM) positive mode respectively. Liquid-Liquid extraction method was used to extract the drug and IS. The method was validated over a linear concentration range of 5.0-5000.0 pg/mL with correlation coefficient (r2 ) ≥ 0.9994. This method demonstrated intra and inter-day precision within 1.95 to 2.37 and 2.93 to 9.3 % and Accuracy within 101.4 to 102.4 and 99.5 to 104.8 %. Cabozantinib was found to be stable throughout freeze-thawing cycles, bench top and postoperative stability studies

Hepatoprotective studies of floral extracts of Gomphrena serrata L. and piperic acid on CCl4 induced hepatotoxicity

The present investigation aims to isolate, characterise and evaluate the phytoconstituents of Gomphrena serrata L. responsible for hepatoprotective activity in carbon tetrachloride-induced hepatotoxicity models both in vitro and in vivo. The plant species has not been explored for various therapeutic activities. HPLC analysis of subfraction of plant extract showed the presence of piperine, which was isolated and further hydrolysed to piperic acid. The results of the study indicate that the plant hydroalcoholic, acetone extracts at 500 mg/kg and compound piperic acid at 0.5 mg/kg exhibited better results in the regeneration of damaged hepatocytes and reduction of biochemical marker enzymes. The hepatoprotective activity might be due to inhibition of cytochrome P450 2E induced ER and oxidative stress. The present study reveals that the hepatoprotective activity of floral extracts might be due to in situ conversion of piperine into piperic acid. As piperic acid showed the equipotent potential to standard drug silymarin, it can be further developed as a hepatoprotective drug

Hepatoprotective studies of floral extracts of Gomphrena serrata L. and piperic acid on CCl4 induced hepatotoxicity

238-251The present investigation aims to isolate, characterise and evaluate the phytoconstituents of Gomphrena serrata L. responsible for hepatoprotective activity in carbon tetrachloride-induced hepatotoxicity models both in vitro and in vivo. The plant species has not been explored for various therapeutic activities. HPLC analysis of subfraction of plant extract showed the presence of piperine, which was isolated and further hydrolysed to piperic acid. The results of the study indicate that the plant hydroalcoholic, acetone extracts at 500 mg/kg and compound piperic acid at 0.5 mg/kg exhibited better results in the regeneration of damaged hepatocytes and reduction of biochemical marker enzymes. The hepatoprotective activity might be due to inhibition of cytochrome P450 2E induced ER and oxidative stress. The present study reveals that the hepatoprotective activity of floral extracts might be due to in situ conversion of piperine into piperic acid. As piperic acid showed the equipotent potential to standard drug silymarin, it can be further developed as a hepatoprotective drug

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

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