The total amount of phenolic compounds was calculated using propyl gallate (1 mg/ml) as standard. 2.4. Total triterpenoid content Total triterpenoid content (TTC) was estimated by the method of Chang and Lin (2011), using ursolic acid (1 mg/ml in methanol) as standard. 2.5. Total flavonoid content To determine the total flavonoid content (TFC), the aluminium chloride method was followed (Zhang et al., 2011).
Determination of total phenolic content The total phenolic content in extracts was measured by UV spectrophotometry based on a Colorimetric oxidation/reduction reaction. The oxidizing agent used was Folin-Ciocalteu reagent (AOCS,1990). To 0.2 ml of extract,7.5 ml water was added in a test tube and after that 0.5 ml of Folin-Ciocalteu reagent (diluted 2 times with water) was added and, then, 1 ml of Na2CO3 (40 %) were added. The sample was incubated for 30 min at room temperature . For a control sample, 0.2 ml of distilled water was used.
SDS – PAGE (10%) PREPARATION OF GELS: RESOLVING GEL PREPARATION: 1. Pipette out 2.66 ml of 30% Acrylamide and pour in beaker. 2. Add 2 ml of Lower Tris (L-Tris, 1.5 M, 8.8 pH) to this. 3.
A substitute we used the thermometer. Methods Trial Test In a 100ml beaker 30mls of water was placed the temperature of the water was recorded. 1 teaspoon of Ammonium Nitrate was added to the water and stirred until dissolved. The temperature was then recorded again. This was to see the difference between the initial temperature and the final temperature.
1. Estimation of Aceclofenac and re-leated substances. (IP 2010) • Liquid chromatographic sys-tem: • Column: a stainless steel col-umn 25 cm x 4.6 mm packed with spherical end-capped octadecylsilane bonded to porous silica (5µm), with a pore size of 10 µm and carbon loading of 19 per cent • Mobile phase: A. a 0.112 per cent w/v solu-tion of orthophosphoric acid adjusted to pH 7.0 using a 4.2 per cent solution of sodium hydroxide, B. 1 volume of water and 9 volumes of acetonitrile • Flow rate: 1 ml per minute • UV detection: 275 nm • Injection volume:10 µL 23 2. Assay of aceclofenac (IP 2010) • Weigh accurately about 0.3 g and dissolve in 40 ml of me-thanol.
By calculating the Rf values for each compound and comparing them to the known lipids, we are able to distinguish the lipids within the grounded nutmeg. II. Procedures Add 5 g crushed nutmeg and 50 mL hexane-isopropanol into a flask and warm for 15 minutes. Remove the extra solvent on a steam bath under a hood while flushing the flask with N2 gas, leaving the crude extract. Weigh extract.
After the calibration process, few drops of each solution were transferred from each test tube without removing the test tubes from their temperatures. These drops were added into the first row of wells on the spot plate, then add two drops of iodine to the wells on the spot plate and wait 1 minute. Use a color- coding scheme to convert the data to qualitative data into quantitative data this will serve as the control. Add 0.5 ml of amylase to the appropriate test tube containing starch and wait two minutes. Then add two drops of the starch-amylase mixture from each tube to the third row of
Now heat to 80-85°C over 10 minutes and hold there for at least two minutes but not more than five. Cool in water to room temperature. Transfer the solution to a 100 mL beaker, keep some (~2-3 mL) in the Conical flask. Stir at medium speed on a magnetic stirrer. Now add 30% hydrochloric acid drop by drop till pH of the solution become 7 and filter it.
Appearance of violet ring at the junction indicated the presence of carbohydrates. Fehling’s Test: To 1 ml of aqueous extract, 1 ml of Fehling’s A and 1 ml of Fehling’s B solutions were added in a test tube and heated in the water bath for 10 minutes. Appearance of red precipitate indicated the presence of reducing sugar. Benedict’s Test: Equal volume of Benedict’s reagent and extract were mixed in a test tube and heated in the water bath for 5-10 minutes. Appearance of green, yellow or red solution depending on the amount of reducing sugar present in the test solution which indicated the presence of reducing sugar.
(1999). Freeze-dried avocado mesocarp (0.05 to 0.10 g) was mixed with 10 mL 80% (v/v) ethanol and homogenized for 1 minute. Thereafter, the mixture was incubated in an 80oC water bath for 60 min to extract the soluble sugars. Subsequently the mixture was kept at 4 ºC overnight. After centrifugation at 12000 g for 15 min at 4 ºC, the supernatant was filtered through glass wool and taken to dryness in a vacuum concentrator.