Acetonitrile

The method was developed on a Novapak C18 (250×4.6mm, 5µ particle size) column using phosphate buffer (pH7.4) and acetonitrile in the ratio of 60:40, v/v as a mobile phase which was pumped at flow rate of 1.0m/min and detection was done at 302nm.the retention time of drug was found to be 7.71min.A prominence isocratic HPLC system (HPLC YL9000 series) with autochro 3000 software an uv detector. A 20 µl hamitton injection syringe was used for sample injection. Preparation of standard and stock solutions 40 mg of Omeprazole was weighed and transferred to 100ml volumetric flask containing 30 ml of 0.1NNaoH. The drug was allowed to dissolved and volume was made up to 100ml with 0.1 Noah. The above solution was diluted with mobile pause to obtain a working standard solution of 40 µg/ml.

In the donor compartment, proliposomal formulation and control (drug suspended in sodium CMC) equivalent to 10mg of KTZ were placed separately. 30% v/v acetonitrile containing phosphate buffer saline (pH 7.4) was taken as receptor medium and kept under constant stirring upto 24 h. In order to avoid evaporation of the contents, donor compartment and sampling port are covered with aluminium foil. 500 µL of aliquots are drawn at predetermined time intervals and equal volumes have been replaced so as to maintain receptor phase volume. All the samples withdrawn were estimated for drug content using HPLC and the data was fitted into mathematical equations (zero order, first order and Higuchi models) (Szuts et al., 2010) to derive the kinetics and mechanism of drug release from proliposomal gel

6). RP-HPLC program: 0-5 min 5-30 min 30-35 min 20% MeCN 20-85% MeCN 85-99% MeCN 5-{3-(3-N-polyethyleniminemaleimide)propylamidophenyl}-10,15,20-tris(4-sulfonato-phenyl)porphyrin trisodium salt (TPPS-PEI) (8b) TPPS-Mal (20 mM) in 500 μL of dry DMF was titrated to 500 μL of 40 mM polyethylenimine in dry DMF. After stirring at room temperature for 8 h additional 0.5 ml of 40 mM PEI were added. The solution mixture was kept with stirring overnight. The solvent was removed under reducing

In addition, phenolphthalein was added as an indicator. The aliquots were titrated against sodium hydroxide (NaOH) solution until end point was reached, after which volume of NaOH consumed was recorded. The value of the rate constant, k, obtained was 0.0002 s-1. The experiment was then repeated with 40/60 V/V isopropanol/water mixture and a larger value of k = 0.0007 s-1 was obtained. We concluded that the rate of hydrolysis of (CH3)3CCl is directly proportional to water content in the solvent mixture.

The solution homogeneity expelled, by centrifugation for 10 min. The sample was centrifuged and separated into two layers, and took the top of the sample is injected for HPLC (11,12). Measured concentration in total lipid: The total fats balanced concentration of the pesticide getting by dividing the measured pesticide residue concentration in the overalll tissue sample by the decimal fraction of the sample that consisted of ether-extractable lipid. The total lipid content of each specimen was estimated from its total cholesterol & triglycerides levels by using a summation method. Analytical results for organochlorine pesticides were reported on a lipid-adjusted basis (nanograms per gram or parts per billion) (14).

The reaction mixture was kept at room temperature for 30 min. Absorbance was read at 517 nm in spectrophotometer. The percentage of the radical scavenging activity was calculated as follows. % of inhibition = Control – Sample ×

Standard Preparation: 100 mg of standard ascorbic acid was weighed precisely and transferred to a 100 ml volumetric flask, added 70 ml of 0.5% sodium metabisulphite and dissolved by shaking. The volume was made up to the mark with 0.5% sodium metabisulphite for getting a concentration of 1 mg/ml. 2 ml of this solution was taken into another 100 ml volumetric flask and made the volume up to the mark with 0.5% sodium metabisulphite which resulted in concentration of 0.02 mg/ml. The solution was filtered through 0.45 µ nylon syringe filter. Sample Preparation: 2.5 g of sample was weighed accurately and transferred to a 100 ml volumetric flask.

The solvent system used for analysis was 10 mM NaOAc/ 150 mM NaOH and the kestoses were eluted at a flow rate of 1 mL /min. 3.2.2. Investigation of reaction parameters There are certain reaction conditions that may favour the production of one kestose isomer over the other. Therefore, several parameters (pH, temperature and time) were chosen and investigated to develop the optimum reaction conditions for each of the kestoses. The experiments for each parameter was carried out in triplicate.

Approximately 1µg Mbgl was used with 5 mM 4-nitropheny-β-D-glucopyranoside (PNPG) in the reaction mixture of either 2 ml or 1 ml. The reaction was stopped by adding an equal volume of 0.2M Na2CO3and the released product 4-nitrophenol was quantified based on the millimolar extinction coefficient of 18.1 mM-1cm-1at 400 nm (Workman and Day 1982). The optimum pH was determined using the same assay in the 100 mM phosphate-citrate buffer in the pH range of 3.0 to 7.0 and for pH 8.0; 100 mMtris buffer was used. Similarly, the temperature optimum was determined in 100mM citrate buffer of pH-6.0. Thermal stability was determined by incubating the protein solution at 50°C and 55°Cfor different times followed by standard PNPG assay, as described earlier.

The analysis was carried on C18 shim- pack GIST (150mmx 4.6mm 5µ) column used as stationary phase. A freshly prepared mobile phase consisting of methanol: potassium dihydrogen phosphate buffer in ratio of (30:70 v/v), PH-3 adjusted using ortho phosphoric acid (OPA) these were filtered by 0.45µM Whatmann filter paper and sonicated before use. The flow rate of mobile phase was 1ml/min. The detection was carried out at 220 nm and run time was around 10 minutes. Selection of wavelength A UV spectrum of drotaverine hydrochloride, ethamsylate, tranexamic acid in water was noted by scanning the solution in the range of 200-400nm.