Characterization of the Antibacterial Agent in Honey
Samples of honey will be subjected to various temperatures from RT to boiling; then cooled and "disked" and placed on lawns of Ec and Se.
Samples of honey will be subjected to various pH's, then neutralized and "disked" and placed on lawns of Ec and Se. This is a bit tricky because you cannot just mix in some acid or base to some honey until the desired pH's are attained. Picture this: you add a drop of acid to the honey. All the agent near the interface between the honey and the acid experiences extreme acidity until you mix the viscous batch. You need to think of how to let the desired pH diffuse into the honey so that no "over-shoot" occurs. How might dialysis/osmosis be of help?
mix in some meat tenderizer and allow the mix to sit overnight, then disk it and place disks on lawns of Ec and Se.
Perhaps you have seen how on CSI various DNA samples are added to a gel and then electricity moves the fragments of DNA through a gel. You are going to make a petri plate filled with gel of starch. A small well will be made in the center of the gel into which you will place your honey. One one side of the plate you will have a positive electrode, and on the other side a negative one. If molecules of the agent have a net positive charge, they will move towards the negative electrode, and vice versa.
This might an easy experiment, but it will take time - at least overnight. You will put some honey into a dialysis tube (a sausage made of semipermeable cellophane). This will be placed in a large volume of water. If the agent is a small molecule (less than 10,000), it will diffuse out of the tubing. What else will happen? Osmosis: huge amounts of water will go into the tube to dilute the hypertonic honey! How do you intend to get the volume back down to what you started with?
Perhaps dialyze small samples of honey against various concentrations of alcohol.
Add enough AS to make the honey saturated with it. If the "agent" is "salted-out," it should precipitate and this can be enhanced by centrifugation. When disks made from the precipitate are placed on bacterial lawns, do zones of exclusion develop?
Remembering that honey has lots of saccharides in it (particularly lots of fructose), your periodate will surely start reacting with the great bulk of fructose. But if half of the fructose is degraded, then also half of any other saccharide will also be degraded, and your zone of inhibition will be only half as large as the untreated sample. Add more periodate, and the zone should be smaller still. And so on. BUT periodate is known to degrade saccharides into possibly toxic things. Happily, E. coli shouldn't be affected by the formic acid (HCOOH) produced. Perhaps you should consider running this periodate oxidate experiment via dialysis - but then you will want to watch the molecular weight group's results closely.