Steric Exclusion Chromatography Results

Extract

[...] It varies from one equipment to another. From fraction N°3 to fraction N°9, proteins are detected, with a maximum for fraction N°6 (for an absorbance of 0.714 AU). From fraction N°10 onwards, there is no significant amount of protein in the solutions (low absorbance). It can therefore be confirmed that the proteins initially present in the sample were collected only in fractions N°3 to N°9. Determination of NaCl Reminder of the experimental part: - 1 ml of each collected fraction is mixed with 5 ml of distilled water and in the presence of 2 drops of colored indicator. [...]

[...] If there are no proteins, the tube is blue. The complementary color of rose-violet being green, it is consistent to measure the absorbance of the solutions at 555 nm (wavelength of green). The higher the value of the absorbance, the higher the amount of proteins in the solution will be. The results obtained are as follows: N° fraction Absorbance measured at 555 nm 1 0,002 2 0,002 3 0,020 4 0,177 5 0,445 6 0,714 7 0,501 8 0,218 9 0,048 10 0,010 11 0,006 12 0,002 13 0,001 14 0,001 15 0,001 The results obtained show that no protein is detected in fractions 1 and 2 (sample elution time). [...]

[...] Synthesis of the results obtained and conclusions The graph below synthesizes the results obtained: It is observed that the proteins were recovered progressively up to a maximum in fraction N°6 and then less and less with each subsequent fraction. As for NaCl, it was collected progressively from fraction N°10. These curves demonstrate that the desalination process was very effective. Steric exclusion chromatography allowed for a complete separation of proteins and NaCl. In fact, the proteins were isolated in fractions N°3 to while the salt was recovered in fractions N°10 to 15. No chloride ions were detected in the protein fractions. Salts are mostly present at the end of elution. [...]

[...] The results obtained are as follows: N° fraction Volume of titrant AgNO3 1 2 3 4 5 6 7 8 9 10 0,05 11 0,40 12 1,25 13 1,80 14 1,95 15 2,00 The results obtained show that no Cl ion- no Cl ion is detected in fractions N°1 to N°9 since the solution was immediately colored brown-red. This means that AgNO3 was in direct excess, so there was no chloride ion to react with. From fraction N°10 to fraction N°15, the quantities of chloride ions increase until reaching a maximum in fraction N°15, with a titrant volume of 2.00 ml. Therefore, we can confirm that the initially present sodium chloride NaCl in the sample was collected only in fractions N°10 to 15, in a progressive manner. [...]