L-Glutamic Acid α-Benzyl Ester CAS 13030-09-6 (H-Glu-OBzl) Assay >98.5% (HPLC)

1. Purity (HPLC)
1.1 Instruments
High performance liquid chromatography, UV3000 detector.
Electronic analytical balance
1.2 Reagents
Acetonitrile (chromatographic grade), TFA (chromatographic grade)
1.3 Chromatographic conditions
1.3.1 Column: C18 column, 5 μl * 250 * 4.6 mm
1.3.2 Detection wavelength: 220nm
Flow rate: 1.0 mL/min
Injection volume:10 μL(for reference)
Diluent: acetonitrile
Run time: 20 minutes
1.4 Mobile phase preparation
Mobile phase A (0.1% Trifluoroacetic acid Water): accurately absorb 1.0 ml TFA diluted to 1000 ml with water, mix and degas.
Mobile phase B (0.1% Trifluoroacetic acid Acetonitrile): accurately absorb 1.0 ml TFA, diluted to 1000 ml with acetonitrile, mix and degas.
1.5 Gradient Program
Time (min) A% B%
0:00 90 10
10.00 10 90
15.00 10 90
15.01 90 10
20.00 90 10
1.6 Preparation of sample solution
Weigh 0.1 g sample and dissolve it with acetonitrile and dilute it to 100ml. Shake well for future use, or the same concentration. Prepare two samples in parallel.
1.7 Sample Determination
Analyze the sample according to the following sampling procedure:
1) More than 1 injection of blank solution
2) 1 injection of sample solution 1#
3) 1 injection of sample solution 2#
1.8.0 Calculations
1.8.1 The peak area normalization method was used to calculate the purity of HPLC.
1.8.2 The relative mean deviation of the purity of two needles shall not be greater than 1%
1.8.3 If the results of both injections meet the acceptance criteria, take the average purity as the final result.