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Analysis of Glucosamine According to USP Method (5NH 4D)


According to USP 39 method, glucosamine hydrochloride should be analyzed using a column packed with L8. It is necessary for the system suitability to satisfy tailing factor of ≤ 2.0, theoretical plate number of ≥ 1,500 and relative standard deviation (RSD) of ≤ 2.0%. It was confirmed that the all conditions were satisfied when Silica 5NH 4D was used for analysis.

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

Column       : Shodex Silica 5NH 4D (4.6mmI.D. x 150mm)
Eluent       : *Buffer(pH7.5)/CH3CN=30/70 
               *Buffer ; in a 1-L volumetric flask, dissolve 3.5g K2HPO4 in water. 
               Add 0.25mL Ammonium hydroxide (25%), dilute with water to volume, and mix. 
               Adjusted with H3PO4 to a pH7.5
Flow rate    : 1.1mL/min
Detector     : UV(195nm)
Column temp. : 35°C

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