TABLE OF CONTENTS - BGB Analytik AG

LC and LC/MS
BioPharmaceutical Applications
Proteins: Effect of Bonded Phase, RP
Column A: ZORBAX 300SB-C8
883995-906
4.6 x 150 mm, 5 µm
Column B: ZORBAX 300SB-CN
883995-905
4.6 x 150 mm, 5 µm
Mobile Phase: A = 0.1% TFA in Water,
B = 0.1% TFA in 50/50 ACN/Water
Flow Rate: 1.0 mL/min
Gradient: 1. 46-96% B in 60 min. 23-48% ACN
2. 50-86%B in 60 min. 25-43% ACN
Proteins: Effect of Flow Rate in SEC
Column: ZORBAX GF-250
884973-901
9.4 x 250 mm, 4 µm
Mobile Phase: 200 mM Sodium Phosphate, pH 7.0
Flow Rate: As indicated
Temperature: Ambient
Detector: UV 225 nm
Sample: 1 mg/mL each protein, 0.1 mg/mL azide, 10 µL
ω-gliadins
1. BSA - dimer
2 3
2. BSA
3. Ovalbumin
4
4. Lysozyme
5. Sodium Azide 1
5
738 www.agilent.com/chem/supplies
Absorbance, 210 nm, mv x 10 3
0.21
0.15
0.09
0.03
A
1.
Temperature: 50°C
Detector: UV 210 nm
Sample: Wheat Proteins, including w-gliadins
10
0 20 40 60
Time (min)
9
5
7*
2*
0.50
0.35
0.20
0.05
B
2.
0 20 40 60
Time (min)
LCBP040
10 9
5
7*
ω-gliadins
2*
Rs = 2.3
N = 15,680
Run Time = 60 min
Flow = 0.25 mL/min
Rs = 2.1
N = 11,939
Run Time = 30 min
Flow = 0.5 mL/min
Rs = 1.9
N = 8,913
Run Time = 15 min
Flow = 1 mL/min
Rs = 1.7
N = 6,623
Run Time = 7.5 min
Flow = 2 mL/min
Rs = 1.4
N = 3,588
Run Time = 3.5 min
Flow = 5 mL/min
LCBP038