Preformulation Studies: Analysis of Honey

Pre look Studies Analysis of sweetenExperimental workPreformulation studiesAnalysis of honeyMaterialsMaterials selected for the analysis of honey were procured from College of Pharmacy, IPS Academy, and Indore. Samples of Honey (Dabur honey) of different batches were selected and analysed where (n=3).Parameters studiedSensory evaluationForeign matterAsh valuepHRefractive indexMoisture contentAcidity standardised procedure was followed for the analysis of different samples of honey.Determination of maxFor determination of max, stock radical of drug ( niggardliness 1000g/mL) in water was prepared by dissolving 10 mg curcumin in 10 mL of distilled water .The working root words in the concentration barf of 2-10 g/mL were prepared. Resulting roots were scanned in the range of 400 to 800 nm with encourage of UV-visible spectrophotometer, and the maximum wavelength was determined. The max of curcumin was found to be 420 nm.Preparation of Calibration convolute by UV-visible Spectroscop yA. Preparation of Calibration Curve in distilled water The stock solution of curcumin was prepared by dissolving 10 mg of curcumin in 10 mL methanol to produce concentration of 1000g/mL.Preparation of standard solutions Standard solutions were prepared in the concentration range of 2-10g/mL by suitable dilutions of the stock solution in methanol and absorbance were taken at 420nm in visible spectrum (Shimadzu 1800).B. Calibration curve in phosphate buffer solution 6.8Preparation of stock solution The stock solution of curcumin was prepared by dissolving 10 mg of curcumin in 10 mL Phosphate Buffer Saline to produce concentration of 1000g/mL.Preparation of standard solutions Standard solutions were prepared in the concentration range of 2-10g/mL by suitable dilutions of the stock solution in PBS 6.8 and absorbance were taken at 420 nm in visible spectrum (Shimadzu 1800).Formulation and optimization of gelMaterialsCarbopol 934p NF, triethanolamine, honey, glycerine, methyl and propyl Parabens and all other chemicals were procured from college of pharmacy IPS Academy, Indore are of analytical grade and used without further purification.Curcumin were procured as a gift sample from Ajmera Pharmaceuticals Pvt. Ltd., Indore, India.Preparation of gelThe topical gel was prepared by soaking the Carbopol 934 in water for 24 h. Drug was first dispersed in small quantity of glycerin with gentle heating and then preservatives were dissolved in glycerin and then added to Carbopol solution with stirring the remaining ingredients were added to it and triethanolamine was added to the neutralize the Carbopol gel base.Preparation of topical gel baseComposition for the medicated formulationEvaluation of gel formulationpHThe pH of prepared gel formulation was determined by employ digital ph meter. 1 g of gel was dissolved in 100 mL freshly prepared distilled water and stored it for 2 hours. The measurement of pH of for each one formulation was done in triplicate and average set were calculated.ViscosityBrookfield digital viscometer was used to measure the viscosity of prepared gel. The T shaped spindle was selected (T3) was rotated different ppm range. The reading, near to 100% crookedness was noted down. A sample was measured at 301C.SpreadabilitySpreadability was determined by wooden block and glass slide apparatus. Weight of about 2 g was selected and added to the pan and the epoch was noted for upper slide to separate completely from the fixed slide.Spreadability was calculated by the given formulaS= M.L/TWhereS= SpreadabilityM= weight tied to the movable upper slideL= length of a glass slideT= time taken to separate the slide completely from each other.HomogeneityAll the formulations were tested for this parameter by visual inspection after the gel maintain been set in the container. They are observed for any aggregation or their appearance.Drug contentA specific quantity of gel generally 1 g of gel was taken and dissolved completely in 100 ml of phosphate buffer 6.8. The volumetric flask containing gel was shaked for 2 h on a mechanical shaker in order to get uniform solution. The solution was filtered by 0.45m membrane filter and estimated spectrophotometrically at 420nm using phosphate buffer 6.8 as a blank solution.Invitro release profileIn- vitro release studies was performed by using a diffusion cell with a sense organ compartment capacity of about 20 ml. the egg membrane was mounted between the donor and receptor compartment of the assembly.The formulated preparation was weight up to 1g was placed oer the membrane and the receptor compartment of the diffusion cell was filled with phosphate buffer 6.8. the whole assembly was fixed on magnetic stirrer, and the solution in the receptor compartment was constantly and continuously stirred using magnetic beads at 50 rpm and the temperature was maintained at 370.50 C the samples of 1 ml was withdrawn at time interval of 15, 30, .60, 90, 120, 150, 180, 210, 240, 270 and 300 min., analysed for drug content spectrophotometrically at 420nm against blank. The receptor compartment was replaced with an equal volume of phosphate buffer at each time of the sample withdrawn. The cumulative graph was plotted against time.Determination of antimicrobial activityPreparation of inoculumsFor evaluation of antibacterial activity, 24 hours fresh culture of bacteria was suspended in sterile water to obtain a uniform suspension of microorganism.Determination of zone of inhibitionAntibacterial activity is checked by agar well diffusion method. in this method a previously liquefied medium was inoculated with 0.2ml of bacterial suspension having a uniform turbidity at temperature of 40C. 20 ml of culture medium was poured into the sterile petri dish having a internal diameter of 8.5 cm. care was taken for the uniform thickness of the tier of medium in different plates.After complete solidification of liquefied inoculated medium, the wells were made aseptically with cork bo rer having 6mm diameter. In each of the plates gel solution was placed carefully. Plates was kept for pre-diffusion for 30 min. after that plates were incubated at 37 C for 24 hr. after incubation period was over, the zone of inhibition was measured with the help of Hi-media.Stability studiesIt is the most important component of any formulation the acceptance and the rejection of the particular preparation depends on this study. The international conference on harmonization (ICH) guidelines titled stability testing of new drug substance and product (QIA) defines the stability test requirement for drug requirement for drug registration application in the European, USA and Japan.Long term stability testing 25 2 C /60 % RH 5 % for 12 months.Accelerated testing 40 2 C / 75 % RH 5%for 6 months.Stability studies were carried out at 40 2 C /75 5 % RH for the selected formulation for one month.