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IsoTopics™ – July 2016

Stable-Isotope Labeling of Algal Biomass Carbohydrates | Cambridge Isotope Laboratories, Inc.

Given the growing interest in algal biomass in biological and biofuel applications, Antoniewicz M et al. developed a new analytical method to more conveniently quantify carbohydrates in C. vulgaris.  The method involved two-step HCl hydrolysis, chemical derivatization of monomer sugars by aldonitrile propionate, and GC-MS analysis. The use of 13C-labeled carbohydrates (e.g., glucose) enabled their composition and metabolic flux to be determined in C. vulgaris media grown autotrophically and heterotrophically.

Abstract

We have developed a method to measure carbohydrate composition and stable-isotope labeling in algal biomass using gas chromatography/mass spectrometry (GC/MS). The method consists of two-stage hydrochloric acid hydrolysis, followed by chemical derivatization of the released monomer sugars and quantification by GC/MS. Fully (13)C-labeled sugars are used as internal standards for composition analysis. This convenient, reliable, and accurate single-platform workflow offers advantages over existing methods and opens new opportunities to study carbohydrate metabolism of algae under autotrophic, mixotrophic, and heterotrophic conditions using metabolic flux analysis and isotopic tracers such as (2)H2O and (13)C-glucose. 

McConnell BO, Antoniewicz MR.

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stable isotope, stable isotope labeled compounds, environmental contaminant standards
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