Tests for alkaloids a. Morquies test: For detecting the alkaloids 2-3 gms of the sample was ground with sufficient chloroform to make slurry. Ammonical chloroform was added and the mixture was stirred for one minute. Extraction of alkaloids from choloroform was accomplished by shaking the solution with 0.5 ml of 2 N-H2SO4 and separation of the acid layer by means of a dropper. A few drops of drug solution were tested with the following alkaloidal reagents A small quantity of the drug solution was placed in a glass plate and allowed to evaporate to dryness. A drop of water and Morquies reagent (HgCl2 + KCN) was added and the colour was observed.
Briefly, 1.5 ml of DPPH solution (0.1 mM, in 95 % ethanol) was incubated with different concentrations of unirradiated and irradiated CMCS solutions. The reaction mixture was shaken well and incubated for 15 min at room temperature and the absorbance of the resulting solution was read at 517 nm against a blank (control). Ascorbic acid was used for comparison as antioxidant materials. The radical scavenging effect was measured as a decrease in the absorbance of DPPH and can be calculated using the following
Figure 1 shows the diluted and actual concentrations of salicylic acid, the concentration and log value of aspirin at various times. Time / min C1 / mol L-1 C2 (actual) / mol L-1 [Aspirin] / mol L-1 ln([Aspirin]) 0 1.996*10-5 9.98*10-4 0.0225 -3.79 10th 6.925*10-5 3.46*10-3 0.0200 -3.91 20th 1.135*10-4 5.68*10-3 0.0178 -4.03 30th 1.372*10-4 6.86*10-3 0.0166 -4.10 40th 1.653*10-4 8.27*10-3 0.0152 -4.18 50th 1.828*10-4 9.14*10-3 0.0144 -4.24 60th 1.953*10-4 9.77*10-3 0.0137 -4.29 Figure 1. A graph of ln([aspirin]t) against time (min) was plotted. The gradient gave the value of K, the rate constant for the reaction. Figure 2 shows the plotted graph Figure 2.
The anticancer activity of samples on HepG2 was determined by the MTT assay (Mosmann 1983). Cells (1 × 105/well) were plated in 1ml of medium/well in 24-well plates (Costar Corning, Rochester, NY). After 24, 48 and 72 hours incubation the cell reaches the confluence. Then, cells were incubated in the presence of various concentrations of the samples in 0.1% DMSO for 48h at 37°C. After removal of the sample solution and washing with phosphate-buffered saline (pH 7.4), 200µl/well (5mg/ml) of 0.5% 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl--tetrazolium bromide cells (MTT) solution was added.
L.H2O yields [ZnL3](ClO4)2.0.75THF.5H2O but L gives [ZnL3](ClO4)2.4H2O the same complex obtained by aminolysis of [ZnL3](ClO4)2.2H2O by 4-methoxyaniline in water. Such selectivity is not common in inorganic synthesis [9.10]. We have found that two diastereomers are obtained when L.H2O is reacted with Ru(phen)2Cl2.2H2O in equimolar proportion in 1:1 (v/v) methanol-water mixture under refluxing condition to synthesise [Ru(phen)2L](ClO4)2.2H2O. This conclusion is based on the observation of two methyl signals in the
• Injection volume: 20μl • Injection concentration: 1mg /ml • Detection wavelength: 280nm 2.3.6-Isolation and purification of Flavonoids (Genistien, Rutin, Quercetin) by: 18.104.22.168- Preparative TLC plate: Isolation and purification of genistein,rutin, quercetin were carried out by preparative TLC. On a glass plates (20 cm x20 cm) a slurry of 60 gm of silica gel GF 254 suspended in 120 ml of distilled water was applied in 0.75 mm thickness manually by using Jobling laboratory division plate coater. The freshly coated plates were left until the transparency of the layer disappears. After 10 minutes, the plates heated for 1 hour at 110ºC. The completely dried and activated plates are kept in a dry place for use.
Later, the hydrolyzed samples were allowed to cool. The sample was filtered using Whatman filter paper; No.42 into a round bottom flask and evaporated till the acid content is completely removed. Finally, the volume was made upto 5 ml with 0.05N HCL and derivatized to phenyl thiocarbomyl amino acid before injected to HPLC. Fatty acid
The extract filtered and filtrate evaporated or concentrated by heating at 55º C on water bath to get a paste. Then it was taken up in distilled water till it dissolved. Each of the extracts was successively extracted with equal amount of petroleum ether (40°-60°C) (fraction-I), ethyl ether (fraction-II) and ethyl acetate (fraction-III). Each of the steps was repeated three times to ensure complete extraction in each case. Fraction I was discarded due to the presence of high fatty substances, whereas fraction II was analysed for the free flavonoids in each of the samples.
The acidified solution was steam distilled until no more oily drops were collected. The distillate was extracted with ether (3x30mL). Most of ether was removed by distillation. The residue, which contains thymol and thymolaldehyde was transfered to a small glass stoppered flask and about twice the volume of saturated sodium metabisulphite solution was added. The solution was stirred vigoursly for ½ h and allowed to stand for 1 h. The paste of bisulphite compound was filtered and washed with little ethanol and finally with little ether.
Synthesis of oleyl 9,(12)-oleoyloxy-10,(13)-oleioxyoctadecanoate (OLOLOODT) (5) OLHYOODT 4 (2.5g; 0.003 mol), pyridine (1.66 g; 0.002 mol) and CCl4 (10 mL) were mixed and heated at 60 °C. OLC (16.2 g; 0.013 mol) was adding during 1 h, and the reaction mixture was refluxed for (5.5 h). The mixture was washed with the water and was dried by using anhydrous sodium sulphate