Experimental Methods: 1. SYNTHESIS OF 4-BENZOYL BUTYRIC ACID METHYL ESTER Materials required * 5 oxopentanoic acid : 2 gm (Aldrich) * Methanol : 50 ml * Acetic Acid (Rankem) Procedure: * 2 grams of 5 oxopentanoic acid was weighed and placed in a round bottom flask and then to it 50 ml of methanol was added. It was placed on a hot plate and the temperature was increased to 50 degrees under ambient air conditions. * To the RB, 2 ml of acetic acid was added and then by attaching a condenser the entire reaction was put on refluxing at 70 degrees Celsius in an oil bath. * For work up: * The reaction media was concentrated till about 10 ml and then dry silica gel was added.
Higuchi model94,95 [Q = KH t½] 4. Korsmeyers-Peppas model:96,97 F = (Mt/M) = Km tn 6.4.7. Drug content: The drug content from pellet formulations was investigated; in which the quantity of pellets equivalent to 6.25 mg of dose of zolpidem tartarate was weighed and dissolved in 0.01 N HCl, sonicated for 15 minutes to dissolve it completely and then the solution of 14 ppm was prepared which was considered as a working level for the complete analysis. The absorbance was determined at the 294.5nm by UV spectrophotometer. Then the drug content was determined by comparing the absorbance of this solution with standard solution having same concentration.
The mixture was finally made upto 5 mL with distilled water and placed in hot water bath at 95ºC for 1 h. After cooling, 1 mL of distilled water and 5 mL of the mixture of n-butanol and pyridine (15:1, v/v) was added. The mixture was vortexed and after centrifugation at 4000 rpm for 10 minutes, the absorbance of the organic layer (upper layer) was measured in UV-Vis spectrophotometer (Shimatzu) at 532 nm against blank using distilled water. TBA when allowed to react with MDA aerobically formed a colored complex [MDA-(TBA) 2 complex] which was measured with spectrophotometer. MDA concentration (measured as TBARS) was calculated as
As many as 1 mL hydrogel preparations added in 1 mL of growth medium with stratified so that dilution dilution series made were 50%, 25%, 12.5%, 6.25%, 3.12%, 1.56%, 0.78%, 0.39%, 0.02%, and 0.01% by volume end of the tube was 1 mL. Then as much as 1 mL of bacterial culture equal Mc. Farland 0.5 added into the test tubes so that the final volume of the tubes were 2 mL. All test tubes were incubated at 37 °C for 18 h. Turbidity test observed in the media and determined MIC value preparations. Tube with negative results or does not indicate the presence of growth, then conducted subculture with solid growth media each bacteria as test assertion MIC value chloramphenicol preparations hydrogel.
2. Add this solution to the cooled suspension of diazotized sulhanilic acid in the beaker. Stir the mixture vigorously. In a few minutes, a red precipitate of helianthin should form. Keep the mixture cooled in an ice bath for about 15 minutes to ensure completion of the coupling
All the test tubes contained in total 3 mL of solution. The following solutions’ concentrations in a tube were .1 mL of dye and 2.9 mL of water, .25 mL of dye and 2.75 mL of water, .5 mL of dye and 2.5 mL of water, 1.0 mL of dye and 2.0 mL of water, 2.0 mL of dye and 1.0 mL of water, and 3.0 mL of dye and 0 mL of water. These samples were tested by the spectrophotometer, and the absorbencies recorded. This whole process was completed twice and the absorbencies were averaged. Lastly, final concentrations and dilution factors were calculated by using the appropriate formulas.
 Appropriate concentrations of SBP hydrolysate were mixed 1: 4 (v/v) with 200 M DPPH solution in anhydrous methanol for 30 min in the dark. The absorbance of the samples was recorded spectrophotometrically on a microplate reader (Multiskan Go, Thermo Fisher Scientific) at 517 nm. A control group containing DPPH solution without sample was also prepared. Ascorbic acid was used as a positive control. ABTS radical scavenging
The volume was made with 6.8 pH Phosphate buffer to get a concentration of 1000µg/ml (SS-I). UV Absorption Maxima (λ max) of Itraconazole sample in pH 6.8 Phosphate buffer • Stock II: 10ml of above solution was then further diluted to 100ml with 6.8 pH Phosphate buffer to get a stock solution of 100µg/ml. UV scanning was done for 100 µg/ml drug solution from 200-400 nm using pH 6.8 Phosphate buffer as a blank in Shimadzu, UV 2450 spectrophotometer. The wavelength maximum was found to be at 262
During the experiment, the solution in the receptor side was maintained at 370C and stirred at 800 rpm with Teflon coated magnetic stirrer bar. At fixed time intervals, 3ml of the sample was withdrawn from the receiver compartment through side tube and analyzed spectrophotometrically at 353 nm by the UV-VIS spectrophotometer. A graph of % cumulative drug release against time was plotted as depicted in Table 5.6 and shown in Fig 5.5
Zeta potential: The zeta potentials (Z-potential) were measured at 25°C using a streaming potential analyzer Values of the Z-potential were calculated from the Helmholtz-Smoluchowski equation Determination of in vitro drug release: Five milliliters of uniformly dispersed suspension was taken and placed straight into a dissolution container. Then, the solution was agitated and withdrawn at prearranged interval to evaluate for drug release by HPLC