Hi,
I am looking for Fluorescence Labeling of saponins, which contains a free carboxylic group, a terpene ring, and various sugars attached by glycosidic linkage. I supposed to bind a dye preferably by covelent bond, so that I can quantify the intact drug in cell lysate. Any suggestion please.
Regards,
I need to know a little more about what you are doing. Merely labeling the saponin with a fluorescent tag will not let you determine the amount of material present. You still need to run some type of separation of the derivatized saponin and your matrix. For example, if you derivatize the carboxylic acid groups of Glycyrrhizin you will get a molecule with three fluorescent tags--one on each glucuronic acid and one on the terpenoid carboxylate. Of course, in your sample you will also have some enoxolone (18-β-glycyrrhetinic acid) which will get derivatized on its terpenoid carboxylate and some 3-β-D-(Monoglucuronyl)-18-β-glycyrrhetinic acid which will be derivatized on the glucuronic acid and the terpenoid carboxylates. To complicate matters, there will likely be other saponin carboxylates that can be derivatized.
The separation of these components by HPLC is not necessarily difficult if you have an HPLC. What equipment do you have? Are you looking only to quantitate or do you want to run subsequent analyses on the saponin?
Dear Olechno! Thank You very much for your informative response. In details, I want to load the saponins onto polymeric nanoparticles by emulsification method, and afterwards washing the formulation by dialysis, which necessarily eliminate the non-saponin derivatives, since they mostly are water soluble of course, and have no surface active potential enough to be loaded onto particles.
Actually, I want to check the delivery of saponins into cells , while carrying through the nanoparticles,
I do think.that I need to remove non-saponins to be removed! I need you input also. thankyou again Joe Olechno
Regards,
Waqas Ahamd,
I agree that many of the water-soluble non-saponins will be removed via washing. Depending upon how the sample was collected, you may have binding of some more hydrophobic compounds to your nanoparticles including alkaloids and terpenes and terpenoids.
Labeling of the saponins with a covalently-linked fluorescent dye might be difficult. Not because the chemistry is hard but because of 1.) the interactions of the activated dyes with other components, 2.) incomplete derivatization of the saponins and 3.) changes in the properties of the saponins after they are derivatized.
What saponins do you expect to be loading? How heterogeneous is the original mixture of saponins? Do these particular saponins contain nitrogen?
I look forward to continuing the conversation.
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