Methods for Purifying Polysaccharides and Pharmaceutical Compositions and Medical Devices Containing the Same

摘要

Methods for removing endotoxin from naturally occurring materials, such as polysaccharides (e.g., agarose and/or carrageenan) are described herein. Polysaccharides that are substantially free of endotoxins and uses thereof are also described. The polysaccharide materials can be isolated from microorganisms, multicellular organisms, such as, algae, plants, seaweed, etc. The method involves the use of acidic and basic solutions to hydrolyze the lipid-polysaccharide bond in endotoxins. Cleaving the fatty acid from the polysaccharide reduces the water-solubility of the fatty acid and enables its removal with an organic solvent such as ethanol. The polysaccharide component can also undergo acidic or basic hydrolysis due to the weak glycosidic bond between the sugar rings.

主权项

1. A method for isolating and purifying a naturally occurring agarose or derivative thereof produced from a biological source, the method comprising:
(i) dispersing the agarose in one or more aliphatic alcohols to disrupt the bacterial wall to solubilize the lipid portion of endotoxins; (ii) removing the aliphatic alcohol to remove the lipid portion of the endotoxins and obtain the agarose or derivative thereof in solid form; (iii) dispersing the solid agarose or derivative thereof in a basic solution to hydrolyze lipid-inner core bonds of the endotoxins and solubilize the polysaccharide component of the endotoxins; (iv) washing the solution from step (iii) with an aliphatic alcohol to remove free lipids; (v) removing the basic solution in step (iii) to obtain the agarose or derivative thereof in solid form; (vi) dispersing the solid agarose or derivative thereof in an acidic solution to hydrolyze lipid-inner core bonds of the endotoxins not cleaved in step (iii); (vii) removing the acidic solution in step (vi) to obtain the agarose or derivative thereof in solid form; (viii) dispersing the solid agarose or derivative thereof from step (vii) in a second basic solution to further cleave lipid-inner core bonds and to neutralize the acid from step (vi); (ix) removing the basic solution in step (viii) to obtain the agarose or derivative thereof in solid form; and (x) dispersing or suspending or dissolving the solid agarose or derivative thereof from step (ix) in sterile water to solubilize the polysaccharide component of the endotoxin and to neutralize any residual acid and base; and (xi) removing the sterile water in step (x) to obtain the agarose or derivative thereof in solid form, wherein the agarose or derivative thereof is substantially free of endotoxin.