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Fractionation of Forage Grass Carbohydrates Using Liquid (Water) Chromatography¹

Because of simplicity and minimum cost, one of the most promising approaches to carbohydrate analysis uses high performance liquid chromatography (HPLC) to separate neutral sugars on cation exchange resins, with only water as the mobile phase. The purpose of this research was to evaluate this technique for the analysis of carbohydrates in forage grasses.

Stem bases from field-grown tall fescue (Festuca arundinacea Schreb.) were sequentially extracted with 95% ethanol and de-ionized water to remove free sugars and fructosans, respectively. The fructosan extract was hydrolized with 1.0 N H₂SO₄. Tall fescue leaf tissue was treated with neutral detergent and the resulting neutral detergent fiber (NDF) was treated with 1.0 N H₂SO₄ to remove and hydrolize hemicellulose. The neutralized extracts were chromatographed on a column containing Aminex Ql5-S, Ca⁺⁺, with water as mobile phase.

Neutral sugars identified in 95% ethanol extracts were glucose, sucrose, and fructose. Fructosan hydrolizates contained glucose and fructose, and hemicellulose hydrolizates of leaf tissue contained xylose, arabinose, and a small amount of glucose. Baseline separation of all sugars was achieved within 39 min and injections could be made every 20 min. Although retention times were longer than those obtained using other methods, ease of operation, minimum cost, and column stability over time make this technique suitable for use with automatic sampling devices and data acquisition systems.

Key Words: HPLC • Cation exchange resins • Forage quality • Sugar analysis • Fructosan • Hemicellulose • Festuca arundinacea Schreb

² Assistant professor, Univ. of Georgia Experiment Stations, G5orgia Station, Experiment, GA 30212.

³ Professors of agronomy and assistant professor of agronomy, respectively, Agronomy Dep., Purdue Univ., West Lafayette, IN 47907.

⁴ Assistant professor and research associate, Laboratory for Renewable Resources Engineering, Purdue Univ., West Lafayette, IN 47907.

Received for publication October 29, 1981.