Comparison of Analysis of Glucosamine Hydrochloride between Polymer-based Amino Columns and Silica-based Amino Columns

Although the analysis of glucosamine hydrochloride according to USP method should use a column packed with L8 (aminopropylsilane chemically bonded to totally porous silica gel), silica-based amino columns often have problems in their durability. In this application, Asahipak NH2P-50 4D, a polymer based amino column, and a silica-based amino column from other manufacturer was compared for their durability. Silica-based amino column began to show deterioration around 200 injections, and the column deteriorated significantly after 300 injections. On the other hand, NH2P-50 4D did not show much theoretical plate number nor tailing factor changes even after 3,000 injections, proved its excellent durability.

（Note）
・Asahiak NH2P-50 4D is a L82 column.
・This analysis requires column equilibration.
    For column equilibration: First introduce 60 mM aqueous phosphoric acid at 0.5 mL/min for 15 minutes, and then switch and introduce the eluent at 0.9 mL/min over 3 hours.

*The version at the time of the application acquisition.

Sample: 10 µL
3.8 mg/mL Glucosamine hydrochloride (in H2O/CH3CN=1/1)

  1. 1.Glucosamine
Column
：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)
*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
Flow rate
：0.9 mL/min (NH2P-50 4D)
1.5 mL/min (Silica-based amino column)
Detector
：UV (195 nm)
Column temp.
：35 ℃

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