The carbohydrate fraction of microalgal biomass is complex and consists of a combination of neutral, acidic, amino sugars, and sugar alcohols. These carbohydrate amalgamations can be difficult to accurately quantify with the most analytical carbohydrate determination techniques. The method described here provides a straightforward two-step sulfuric acid hydrolysis followed by soluble carbohydrate quantification by either a spectrophotometric method (based on aldehyde functional group derivatization), for an overall combined monomeric sugar concentration determination, or liquid chromatography, specifically high-performance anion exchange method, for a more specific monomeric sugar profile.