pyridine, chloroform and metahnol as a solvent for chemopreventive agents - Is there anyone who has used these solvent for cell culture experiment (Aug/09/2007 )
When i am feelinng any doubt about my experiment or confused , i simply put my query here on this forum and, my problem is gone....... thanx to all who are really dedicating to make science live on this forum....
i am going to see the proliferative effect of these chemical on the cells. here are the solvent which i am to going to use acc to supplier information...
Hesperidin---------------------1 % pyridine
trans-Chalone-----------------soluble in chloroform
Kaempferol -------------------slightly soluble in water, but more soluble in ether or ethanol
delphinium chloride-----------soluble in methanol.
Now, i want to know minimum toxicity of limit of these solvent on the cells. or i want to want to whether can i use pyridine, chloroform and methanol as a solvent for the cell culture experiment.
iknow the minimum toxicity limit of the DMSO for cell culture experiment is .1 % and and aslo we take .1% for control experiment.
but dont have any idea about other solvent, if anyone who has even very little information kindly share with me as i am new in this field.
Really looking with very much expectation.
thanx in advance.
Most of these solvents are fairly toxic to cells. Pyridine is basic. Chloroform evaporates very fast and does not dissolve in water. The net result will be leaving your drug in an insoluble form. MeOH is OK if used in low volume in vitro. In vivo application would be problematic. You may use DMSO for these drugs, although it is not a inert solvent itself.
yeah, DMSO makes membranes rather permeable to non-polar subtances. thus handle your DMSO solutions with care, you can poison your self with aren't normally toxic.
You could use ethanol to dissolve and try to make something like a 1000x stock solution for use in cell culture.
Before you do try and dissolve the substances in these reagents, try to calculate what concentration stock you are planning, important to make sure the amount which will be added to the cell culture media will be as little as possible. Then try adding the same amount of these solvents to some cells and compare them to cells which didnot receive any solvent. Once you are certain that these solvents can be used for cellculture, go ahead by prepapring the solutions according to twhat you calculated.
thanx for all the replies,
i prepared the stock by dissolving in DMSO, and when i added medium and filter, then i found media was clear. it sees that drug is retained by the filter.
what should i do ?
should i dont have to filter the drug ?
but in such case contamination can come.
suggest me something.
thanx for cooperating me.
this means you didn't let the drug dissolve enough time and/or you saturated your contentration.
so try longer time dissolution at first, and then dilute the drug to see if dissolution is better
Or, the filter nature binds per se the drug ?
Because of DMSO, you should use a nylon filter else it might not be optimal for filtering. The conc. of the substance might decrease as some amount of the chemical might bind to the filter.