Samples were not diluted

and measured at 25 °C. The measurement was done in triplicate and size d90 was reported. The osmolarity, zetapotential and pH of the prepared formulations of PM181104 were measured using an osmometer (Osmomat 030-3P, Gonotec, Germany), a Delsa Nano HC, Zeta Potential Particle Analyzer (Beckman Coulter Inc., USA) and pH meter (pH Tutor, Eutech Instruments Pte Ltd., Singapore), respectively. Each measurement was performed in triplicate at 25 °C. The particle morphology of formulation F5 and F6 were evaluated by Dolutegravir research buy transmission electron microscopy (TEM) (Model-CM200, Make-PHILIPS). The formulation drops were added to a 300 mesh copper grid, then dried using IR light and then examined by TEM. Male Balb/c mice (18–25 g) bred in-house at Piramal Enterprises Limited, Goregaon, Mumbai, India were used. Mice were

maintained in a temperature of (22±2 °C) and humidity (55±5%) controlled room with a 12 h light/dark cycle and free access to standard diet and water. All mice used in this study were not subjected to any form selleck compound of treatment/medication. Guidelines of Committee for the Purpose of Control and Supervision on Experts on Animals (CPCSEA), Government of India, were followed and the In-house Animal Ethics Committee approved all experimental procedures. Animals were provided food and water ad libitum. Mice were randomly and equally divided into eight groups containing 24 mice each. Animals in Groups 1–5 were administered with formulations F1–F5 intravenously at the dose of 2.5 mg kg−1. Animals in Groups 6–8 were administered Meloxicam with formulations F6–F8 intravenously

at the dose of 5.0 mg kg−1. The administration was done via tail vein using a 1.0 mL tuberculin syringe equipped with 27 G needle after dilation with ethanol solution (70%). Blood samples (0.5 mL) were collected on ice from three animals at each time point viz. 0.033, 0.083, 0.17, 0.25, 0.50, 0.75, 1 and 2 h post-dose using sodium citrate (10% v/v) as anticoagulant. Plasma was separated by centrifugation of blood samples at 10,000 rpm for 5 min at 4 °C and plasma samples were stored at approximately −70 °C until bioanalysis by LC-MS/MS method. One hundred micro-liters of mouse plasma sample was extracted with 2.5 mL ethyl acetate on a vortex mixer for 5 min followed by centrifugation for 5 min at 10,000 rpm at 20 °C. The organic layer was transferred into another test tube and was evaporated to dryness under a stream of nitrogen at 35 °C. The samples were reconstituted in 200 µL solution of formic acid (0.7%) in (acetonitrile: methanol: 1:1 v/v) and were vortexed, then transferred into polypropylene vials. From this vial, 10 µL of the sample was injected into the LC-MS/MS system for further analysis.