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Sulfated polysaccharides constitute a large and complex group ofmacromolecules which possess a wide range of important biological properties.Many of them hold promise as new therapeutics, but determination of their bloodlevels during pharmacokinetic studies can be challenging. Heparin Red, acommercial mix-and-read fluorescence assay, has recently emerged as a tool inclinical drug development and pharmacokinetic analysis for the quantificationof sulfated polysaccharides in human plasma. The present study describes theapplication of Heparin Red to the detection of heparin, a highly sulfatedpolysaccharide, and fucoidan, a less sulfated polysaccharide, in spiked mouseand rat plasmas. While the standard assay protocol for human plasma matrix gaveless satisfactory results, a modified protocol was developed that provideswithin a detection range 0 to 10 micrograms per mL better limits ofquantification, 1.1 to 2.3 micrograms per mL for heparin, and 1.7 to 3.4micrograms per mL for fucoidan. The required plasma sample volume of only 20microliters is advantegous in particular when blood samples need to becollected from mice. Our results suggest that Heparin Red is a promising toolfor the preclinical evaluation of sulfated polysaccharides with varyingsulfation degrees in mouse and rat models.

Quantification of sulfated polysaccharides in mouse and rat plasma by the Heparin Red mix-and-read fluorescence assay