In a 217 total volume of 1.5 mL, the reaction mixture contained 1 mL of the eluate, 400 mL of 218 12.5 mM 3-(dimethylamino) benzoic acid in 0.375 M phosphate buffer (pH 6.5), 80 mL 219 of 3-methyl-2-benzothiazoline hydrazone and 20 mL of peroxidase (0.25 unit). The 220 reaction was started by the addition of peroxidase and the increase in absorbance was
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.
5-aminotetrazole monohydrate: In a 250 ml round-bottom flask equipped with a condenser for refluxing (90 °C) and a magnetic stirring bar, 5.00 g (5.95 mmol) dicyandiamide (three times crystallized), 7.47 g (11.9 mmol) sodium azide and 11.00 g (17.8 mmol) boric acid and 100 ml of water is added and allowed to reflux for 24 hours, after the completion of the reaction, until the solution pH to about 2 to 3 as hydrochloric acid 37% is added (about 12 ml) Then the reaction mixture was cooled in a refrigerator for 18 hours and the white crystals formed. The mixture was filtered and washed three times with 10 ml of water and and dried in 60 °C for 5 hours and finally 45.8 g of product by it will be obtained. 5-Aminotetrazol monohydrate: Yield:,
MDA level was determined by thiobarbituric acid reactive substances (TBARS) in serum, based on the reaction between MDA and TBARS. Standard Malondialdehyde solution in 5 mL of volume was processed along with test samples. 1.5 mL of 0.8% of TBA was added to 1 mL of the serum sample. Then 0.4 mL of 8.1% sodium dodecyl sulphate and 1.5 mL of acetic acid was added. 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.
In the round-bottom flask (100 mL), we placed p-aminobenzoic acid (1.2 g) and ethanol (12 mL). We swirled the mixture until the solid dissolved completely. We used Pasteur pipet to add concentrated sulfuric acid (1.0 mL) to the flask. We added boiling stone and assembled the reflux. Then, we did reflux for 75 minutes.
In this experiment, racemic 2-methylcyclohexanone was reduced using sodium borohydride as a nucleophile to give a diastereomeric mixture of cis and trans secondary alcohols. The products were analyzed for purity using IR spectroscopy and gas chromatography. 1.2 g of 2-methylcyclohexanone and 10 mL of methanol were combined in a flask and cooled in an ice bath. Two 100 mg portions of sodium borohydride were added to the flask and stirred. 5 mL of 3M sodium hydroxide, 5 mL of de-ionized water, and 15 mL of hexane were added to the reaction flask and stirred.
Isolation of Ibuprofen: Fifteen 200 mg ibuprofen tablets were used, which was 3.37g (16.34 mmol, 112.3%) ibuprofen. This amount of ibuprofen was submerged in 25 mL of acetone and stirred vigorously for 5 minutes to dissolve. Red coatings of the tablet were separated and vacuum filtration was utilized to dissolve all insoluble components. The melting point range of the crude ibuprofen was 72.4-73.9°C. The major bands from the IR are, FTIR: sp3 O-H stretch, about 3200-2500 cm-1 (b, m); sp3 C-H stretch, 2991-2868 cm-1 (sh, m); sp2 =C-H stretch, 3100-3000 cm-1 (sh, w); sp2 C=O stretch, 1701 cm-1 (sh, s); and sp2-sp2, aromatic, C=C stretch (in ring), 1507 cm-1 (s, m).
After the evaluation of stomach for ulcers, the gastric mucosa of glandular portion was scrapped with the help of two glass slides, weighed (100 mg) and homogenized in 1 mL of a 0.15 M, ice cold potassium chloride (KCl) solution and centrifuged at 3,000 RPM for 10 minutes (REMI centrifuge). 1 mL of suspension was taken from the above tissue homogenate in test tube and 0.5 mL of 30% w/v TCA (trichloroacetic acid) was added to it, followed by 0.5 mL of 0.8% w/v TBA (thiobarbituric acid) reagent. The tubes were then covered with aluminium foil and kept in water bath for 30 minutes at 80 °C. After 30 minutes, tubes were taken out and kept in ice-cold water for 30 minutes and centrifuged at 3000 rpm for 15 minutes (R-BC DX REMI centrifuge). The absorbance of the supernatant was read in spectrophotometer (UV-1601, SHIMADZU) at 540 nm against blank.
A sample of fish oil (50 mg) was transferred into screw-cap vial. 2 ml benzene and 10 ml sulfuric acid (1%) in absolute methanol were added. The vial was covered under a stream of nitrogen before heating in an oven 90 °C for 90 minutes. Ten ml of distilled water were added to the cooled vial and the methyl esters in each vial were extracted with 5 ml of petroleum ether for three times. The three petroleum ether extracts were combined and concentrate to its minimum volume by using a stream of nitrogen.
Rose Bengal-(bis(aminoethyl)ethylene glycol) (2) from Rose Bengal disodium salt (1) The synthesis was done following procedure from . Rose Bengal Na+ salt (915 mg, 0.90 mmol) was dissolved in DMF (2ml) and DIPEA (0.312 ml, 1.80 mmol), HATU (308 mg, 0.81 mmol) were added. After activation for 15 min, the mixture was added to O-Bis-(aminoethyl)ethylene glycol trityl resin (309 mg, 0.31 mmol) preswollen in DMF for 2 hours. The coupling reaction wrapped in aluminum foil was allowed to proceed overnight on a nitrogen bubbler apparatus. The resulting red-burgundy coloured resin was filtered and washed well with DMF.