| Abstract: |
V. SUMMARY AND CONCLUSIONDifferent chlorinated and brominated derivatives of sucrose were prepared at Biochemistry Department, Faculty of Agriculture, Zagazig University:These derivatives prepared were:1-6,6-dichlorosucrose.2- 6.6-dibromosucrose.3- 4,6-dichlorogalactosucrose.4- 4,6,6-trichlorogalactosucrose.5- 4,1,6-trichlorogalactosucrose.6- 4,6,1,6-tetrachlorogalactosucrose.7- 4,1,4,6-tetrachlorogalactosucrose.Various analysis were performed to study the structure of the modified compounds as well as their physical and chemical properties.1- chemical analysis of modified compounds.1.1. Analysis of elements composing the modified compounds (C, H, O, Cl, Br and OH group).1.1.1. 6,6-dichlorosucrose: 37.5% C, 5.17 % H, 38.5% O, 18% Cl and 26.1% OH group. C12H20O9Cl21.1.2. 6,6-dibromosucrose: 30.7% C, 4.6 % H, 30.5% O, 34.2% Br and 21.4% OH group. C12H120O9Br21.1.3. 4,6-dichlorosucrose: 37.9% C, 5.1 % H, 38.3% O, 18.7% Cl and 26.2% OH group. C12H20O9Cl21.1.4. 4,6,6-trichlorosucrose: 36.2% C, 4.8 % H, 33% O, 26% Cl and 21.18% OH group. C12H19O8Cl31.1.5. 4,1,6-trichlorosucrose: 36.2% C, 4.7 % H, 32% O, 27.1% Cl and 21.28% OH group. C12H19O8Cl31.1.6. 4,6,1,6-tetrachlorosucrose: 35% C, 4.3 % H, 26.5% O, 34.2% Cl and 16% OH group. C12H18O7Cl41.1.7. 4,1,4,6-tetrachlorosucrose: 35.4% C, 4.4 % H, 26% O, 34.2% Cl and 16.1% OH group. C12H18O7Cl41.2. HPLC analysis of modified compounds:HPLC had been used successfully to characterize modified sugars. Compounds have been separated clearly and each component appeared at defired retention time (Rt). Results showed that 6,6¢-dichlorosucrose appeared at 13.19 min, 6,6¢-dibromosucrose appeared at 17.8 min, 4,6¢-dichloro-galactosucrose appeared at 23.77 min, 4,6,6¢-trichlorogalacto-sucrose appeared at 23.83 min, 4,1¢,6¢-trichlorogalacto-sucrose appeared at 23.93 min, 4,6,1¢,6¢-tetrachlorogalactosucrose appeared at 31.81 min, 4,1¢,4¢,6¢-tetrachlorogalacto-sucrose appeared at 23.13 min and sucrose appeared at 10.18 min,1.3. Thin –Layer Chromatography analysis.TLC as a simple technique was also used to characterize modified compounds according to their retention factors (Rf ). Acetyl acetate: methanol: water (6:2:1, v/v/v) was used as solvent system on silica gel and Rf values were detected as follows: 0.43, 0.62, 0.71, 0.73, 0.75, 0.58, 0.52 and 0.82 for 6,6’-dichlorosucrose, 6,6’-dibromosucrose, 4,6’-dichlorogalacto-sucrose, 4,6,6’-tri-chlorogalactosucrose, 4,1’,6’-trichloro-galactosucrose, 4,6,1’,6’-tetrachloro-galactosucrose 4 and 1’,4’,6’- tetrachlorogalactosucrose, respectively.1.4. Infrared (IR) characterization of modified compounds.-IR absorbance value of C-Cl was 600-800 cm-1, while 500-750 cm-1 for C-Br- Hydroxyl group on position 4 in galactose moiety was absorbed at 3520cm-1.- Hydroxyl group in sucrose moiety was absorbed at 3335-3500cm-1.1.5. Mass spectroscopy of modified compounds.Applying mass spectroscopic analysis for characterization of components gave also useful information. Data from mass spectra showed that the molecular weight of 6,6’-dichlorosucrose, 6,6’-dibromosucrose, 4,6’-dichlorogalacto-sucrose, 4,6,6’-trichlorogalactosucrose, 4,1’,6’-trichlorogalacto-sucrose, 4,6,1’,6’-tetrachlorogalactosucrose4, 1’,4’,6’- tetrachloro-galactosucrose were 379, 468, 379, 397, 397, 416, 416 and 342, respectively.1.6. 1H-NMR spectrum:1- Signals from the protons of hydroxyl groups were observed at 4.2 – 4.7 ppm in the spectrum of sucrose also sucrose derivatives.2. Physical properties of modified compounds.2.1 Melting Point measurements:Different values for melting points were recorded for sucrose derivatives: 6,6’-dichlorosucrose 86°C, 6,6’-dibromosucrose 92°C, 4,6’-dichlorogalacto-sucrose 80°C, 4,6,6’-trichlorogalactosucrose 117°C, 4,1’,6’-trichlorogalacto-sucrose 106°C, 4,6,1’,6’-tetrachlorogalactosucrose 62°C, 4,1’,4’,6’- tetrachlorogalactosucrose 60°C, and sucrose 170°C.2.2. Optical Rotation:Solution of 2% concentration at 20C from sucrose derivatives were subjected to measure their optical rotation and the values were 6,6’-dichlorosucrose +60°, 6,6’-dibromosucrose +57°, 4,6’-dichlorogalactosucrose +71°, 4,6,6’-trichlorogalactosucrose +87°, 4,1’,6’-trichlorogalactosucrose +68.2°, 4,6,1’,6’-tetrachlorogalactosucrose +79°, 4,1’,4’,6’- tetrachlorogalactosucrose 72°, and sucrose +81.5°.2.3. Viscosity:At different temperatures (20, 30, 40, 50°C) and different concentrations (10, 20, 30, and 40%), viscosity of sucrose derivatives was lower then sucrose (15mPa/s. 10% for20°C) while derivatives was range (1-3 mPa/s. 105 for 20°C) for 40% 20°C give sucrose 35.28 mPa/s while sucrose derivatives range (2.4-4.2 mPa/s).2.4. Refractive Index.At different concentrations (4, 8, 12, 16 and 20%) refractive index values of sucrose and its derivatives were compared. The data showed that of sucrose were (1.343-1.354-1.366-1.371-1.380 respectively) while values of sucrose derivatives were range (1.331-1.328).2.5. Surface Tension.Comparison between compounds at different concentrations (0.1- 0.5- 1.0- 2.0%) have been done according to their surface tension values. The results showed that sucrose derivatives had range (70.2-63.3) while sucrose had 70.5 -71.3-71.9-73.4 Dyane /cm, respectively).2.6. Solubility.Solubility of different modified compound at different concentration (10, 20, 30 %) at different temperatures (20, 30, 40, 50°C) was measured. Sucrose was completely soluble (100%), while other derivatives recorded range (80-100 %) for 10% and temp. 20°C as compounds galactosucroses, while compounds 6,6-dichlorosucrose and 6,6-dibromosucrose range (66-70 %) the same temp. and concentration.2.7. Stability of modified compounds.At different pH values of 5% solution of modified compounds and sucrose, stability under microwave and ultrasonic conditions was recorded.2.7.1. Stability under microwave condition.Data showed that the stability of sucrose derivatives were higher than normal sucrose and the stability values for sucrose derivatives were (65-95%), (82-97%) and (89-98%) at pH3, pH5, pH7 respectively. and for sucrose were 58-71-82.5 at pH3, pH5, pH7 respectively.2.7.1. Stability under ultrasonic condition.Data showed that the stability of sucrose derivatives were higher than normal sucrose and the stability values for sucrose derivatives were (81.7-96.8%), (88.8-98.6%) and (92.3-99.3%) at pH3, pH5, pH7 respectively. and for sucrose were 78.8-88.5-94.5 at pH3, pH5, pH7 respectively.2.8. pH value.Solution 10% of modified compounds and sucrose was measured for pH values. The data showed that sucrose derivatives had acidic effect more than sucrose and the values were pH 5.7 for 6,6-dichlorosucrose, 5.9 for 6,6’-dibromosucrose, 6.4 for 4,6’-dichlorogalactosucrose, 6.3 for 4,6,6’-trichloro-galactosucrose, 6.1 for 4,1’,6’-trichlorogalactosucrose and 6.4 for sucrose.2.9. Energy measurement.Values of energy for different components were recorded and there were as follow: zero kilocalorie for derivatives (4,6’-dichlorogalactosucrose, 4,1’,6’-trichlorogalactosucrose, 4,6,1’,6’-tetrachlorogalactosucrose and 4,1’,4’,6’- tetra-chlorogalactosucrose) while give 4 kilocalorie other derivatives and sucrose .2.10. Sweetness.Values of sweetness for modified compounds were recorded at different concentration. Generally, sucrose derivatives recorded higher values of sweetness as compound to sucrose compounds chlorogalactosucrose derivatives recorded higher than from derivatives chlorosucrose.3. Biological evaluation of some sucrose derivativesChlorinated sucrose products (4,1,6-trichlorogalacto-sucrose; 4,6,1,6-tetrachlorogalactosucrose and 4,1,4,6-tetra-chlorogalactosucrose) were administered to Albino rats at different levels (0.05, 0.1, 0.2; 0.01, 0.05, 0.1 and 0.005, 0.01, 0.02 % respectively), for 2 months. Serum samples were withdrawn at different intervals during feeding to follow the changes in the biochemical indicators. At the end of the experiment, the rats were killed, and body weight gain and organs weights were recorded. The results indicated that body weight was reduced in all rats receiving the chlorinated sugars, while the organ weights remained significantly unchanged by the different treatments.3.1. Serum analysis of rats fed on diets sucrose derivatives:Also serum total protein and urea concentrations were reduced, while creatinine did not show any significant changes. Significant decreases in the activities of AST (GOT) and ALT (GPT) were recorded in all rats receiving chlorinated sugars. Liver glycogen content tended to increase in the rats administered 4,1,6-trichlorogalactosucrose; 4,6,1,6-tetra-chlorogalactosucrose and 4,1,4,6-tetrachlorogalactosucrose at all used levels for 2 months while liver protein level did not show significant changes from the controls. Feeding on both 4,1,6-trichlorogalactosucrose; 4,6,1,6-tetrachlorogalacto-sucrose and 4,1,4,6-tetrachlorogalactosucrose has generally reduced serum glucose level. This reduction in serum glucose level was not sweetener concentration dependent but rather it was an indirect effect of eliminating sucrose in the meal. Serum total lipids, triglyceride and cholesterol were diminished in all experimental rat groups compared to control.3.2. Histological examination of some organs (Liver, Kidney, brain and heart).(A)- 4,1,6-trichlorogalactosucrose:-The brain: normal while few cases showed perineural edema.-The heart: was appeared normally in shape and size.-The kidney: the severity was dose dependent, the doses causes: moderate number of renal tubules are cystic.-The live: the liver was congested and fatty.(B)- 4,6,1,6-tetrachlorogalactosucrose:-The brain: normal while few cases showed perineural edema.-The heart: was appeared normally in shape and size.-The kidney: the doses revealed cystic dilatation of many renal tubules.-The liver: showedvaculation and fatty changes of moderate number of hepatocytes.(C)- 4,1,4,6-tetrachlorogalactosucrose:-The brain: normal-The heart: normal-The kidney: normal except some renal tubules were dialated-The live: normal in same cases
|
|
|