I need sterilize a sucrose solution (5% y 10%). I prepared them the first time with sterile water and then I used the filters of 0.2 um. Would it be necessary to autoclave the solutions? Would the sucrose be affected by the autoclave?
Thanks!
Please check the following links.
https://www.researchgate.net/post/Is_it_really_suitable_to_autoclave_glucose_solution
https://www.researchgate.net/post/Has_anyone_filtered_the_high_percentage_of_sucrose_solution_for_a_sucrose_gradient_experiment
http://molecularbiology.forums.biotechniques.com/viewtopic.php?f=5&t=30857
Hi, please read the attach discussion
https://www.researchgate.net/post/Could_somebody_recommend_how_to_sterilize_polysaccharides
https://www.sartorius.com/fileadmin/fm-dam/DDM/Bioprocess-Solutions/Filtration_Technologies/General/Application_Notes/Appl_Filtration_of_Concentrated_Sugar_Solutions_SP-4001-e.pdf
Please be careful with autoclaving sucrose.
It is a disaccharide composed of glucose and fructose which can hydrolyze when heated.
As for the hydrolysis products glucose and fructose: They also degrade during autoclaving, releasing carbon, although they are likely more stable than the disaccharide sucrose.
According to my own experience,
different sugars react much different to higher temperatures:
You can autoclave fructose and glucose for 20 min (solutions turn yellow indicating slight degradation), but you cannot autoclave, e.g., ribose (a pentose which disintegrates quickly at high temperatures).
Fructose is less stable in the autoclave than glucose (turns yellow quicker).
Sigma-Aldrich has this to say about autoclaving Sucrose:
Storage/Stability
Solutions can be autoclaved for 15-20 minutes at a
maximum of 121 °C. There will be some hydrolysis to
glucose and fructose, depending in part on how rapidly
the autoclave comes to the required temperature and
pressure. Care must be taken to prevent solutions
from caramelizing
Hope that’s helpful,
Best regards.
Filtering it through filter paper of 0.2um size would be good to proceed with, because autoclaving may affect sucrose
Each chemically prepared composition has a best quality method to be treated. Generally, low concentration solutions are highly subjected to heat effect than high concentration ones. If sterilization is a must for your solution, radiation or filtration are the best methods to be used. For your case, and to avoid any possible dissociation to glucose and fructose, filtration with 0.2um size filters is enough. Duplicates of post-filtration culturing of about 250-500uL amounts of your preparation into an appropriate bacterial culture medium (peptone water, nutrient broth, Malachite green media, etc) followed by incubating for a week, one at room temp and the other 37oC while guarding, is the best routine laboratory practice especially in virology to ensure sterility.
I think filtration is the optimum method for sterilization of such solution. If you doubt it, you can test it by agar culture, but I'm sure it'll work
you can do a tyndilization..it is recommended for sugars....put in a boiling water bath for 20-30 minutes for three successive days(in between put in a refrigerator)
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