Although the analysis of glucosamine hydrochloride according to USP method should be used a column packed with L8 (aminopropylsilane chemically bonded to totally porous silica gel), silica-based amino columns are often highly durable. In this application, Asahipak NH2P-50 4D, a polymer based amino column, and a silica-based amino column from other manufacturer was compared for durability. In the case of the silica-based amino column, it began to deteriorate around 200 injections, and the column deteriorated sharply after 300 injections. On the other hand, NH2P-50 4D has no significant change in the theoretical plate numbers and tailing factor after 3,000 injections, proving its excellence in durability.
[Note] This analysis requires column equilibration.
For column equilibration: First introduce 60 mM Phosphoric acid aq. at 0.5 mL/min for 15 minutes, and then switch to the eluent at 0.9 mL/min over 3 hours.
Sample : 3.8 mg/mL Glucosamine hydrochloride (in H2O/CH3CN=1/1), 10 μL
Columns : Shodex Asahipak NH2P-50 4D (4.6 mm I.D. x 150 mm) Silica-based amino column from other manufacturer (4.6 mm I.D. x 150 mm) Eluent : *Buffer (pH7.5)/CH3CN=30/70 (NH2P-50 4D) *Buffer (pH7.5)/CH3CN=25/75 (Silica-based amino column) Flow rate : 0.9 mL/min (NH2P-50 4D) 1.5 mL/min (Silica-based amino column) Detector : UV (195 nm) Column temp. : 35 °C *Buffer ; In a 1-L volumetric flask, dissolve 3.5 g K2HPO4 in water. Add 0.25 mL Ammonium hydroxide (25 %), dilute with water to volume, and mix. Adjusted with H3PO4 to a pH7.5
Sample Name Index
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