|
Ion-exchange
chromatography followed by post-column
derivatization has been a method of choice for
amino acid analysis for many years. Pickering
ion-exchange columns allow you to obtain
consistent results with sensitivity, stability,
selectivity and speed. Sodium columns and buffers
systems are designed for amino acids analysis of
hydrolyzed samples.
Post-column
conditions for amino acids analysis:
Reagent: TRIONE®
Reactor: 130 °C, 0.5 mL
Reagent flow rate: 0.3 mL/min
Detection:
UV-Vis detector:  =570
nm for primary amino acids,
=440 nm for secondary
amino acids
or
Reagent: 300 mg of OPA, 2 g Thiofluor™, 3
mL of 30% Brij® 35 solution in 950 mL of OD104
Reactor: 45 °C, 0.15 mL
Reagent flow rate: 0.3 mL/min
Detection:
Fluorometer: ex
330 nm, em 465
nm
The recommended
gradient conditions are subject to change without
notice. This may happen because of lot changes in
the columns, or improvements in the overall
method. The recommended gradient for the column
will always be in the column package, and it
supersedes the information in this catalog.
Pickering
Laboratories recommends six different gradient
conditions depending on the column and type of
sample. Use the program recommended on the
column data sheet for the initial testing.
The column oven temperature programming gives
additional flexibility when optimizing methods.
Using temperature gradient allows to improve
separation, shorten analysis time and fine-tune
the method for detecting compounds of interest.
|
 |
 |
|
1 |
Methionine-D,L,- Sulfoxide |
|
2 |
trans-4-Hydroxy-LProline |
|
3 |
Aspartic Acid |
|
4 |
Threonine |
|
5 |
Serine |
|
6 |
Glutamic Acid |
|
7 |
Proline |
|
8 |
Glycine |
|
9 |
Alanine |
|
10 |
Cystine |
|
11 |
Valine |
|
12 |
Methionine |
|
13 |
Isoleucine |
|
14 |
Leucine |
|
15 |
Tyrosine |
|
16 |
Phenylalanine |
|
17 |
D,L &
allo-Hydroxylysine |
|
18 |
Lysine |
|
19 |
Ammonia |
|
20 |
Histidine |
|
21 |
Tryptophan |
|
22 |
Arginine |
|
|
0 |
|
|
|
|
12 |
|
|
|
|
34 |
|
|
|
|
53 |
|
|
|
|
53.1 |
|
|
|
|
55 |
|
|
|
|
55.1 |
|
|
|
|
67 |
|
|
|
Flow rate: 0.4 mL/min, column
temperature 48 °C,
injection volume: 10 µL of 0.25 µmole/mL Std. |
|
0 |
100
|
0 |
0 |
|
14 |
100
|
0 |
0 |
|
42 |
0 |
100
|
0 |
|
56 |
0 |
100
|
0 |
|
56.1
|
0 |
0 |
100
|
|
58 |
0 |
0 |
100
|
|
58.1
|
100
|
0 |
0 |
|
70 |
100
|
0 |
0 |
Flow rate:
0.4
mL/min, column temperature 50 °C,
injection volume: 10 μL of 0.25 μmole/mL Std. |
|
|
|
 |
3 Aspartic Acid
4 Threonine
5 Serine
6 Glutamic Acid
7 Proline
8 Glycine
9 Alanine
11 Valine
13 Isoleucine
14 Leucine
16 Phenylalanine
18 Lysine
19 Ammonia
20 Histidine
22 Arginine
23 Cysteic Acid
24 Methionine Sulfone |
|
1154150 |
High efficiency Sodium Cation-exchange Column
4.0 × 150 mm |
|
1193250 |
Standard Sodium Cation-exchange Column 3.0 ×
250 mm |
|
1193020 |
Sodium Guard Column, 3.0 × 20 mm (for use with
1154150 Column) |
|
1192020 |
Sodium Guard Column, 2.0 × 20 mm (for use with
1193250 Column) |
|
0 |
0 |
0 |
100 |
0 |
0 |
Inject |
|
1 |
11 |
11 |
100 |
0 |
0 |
Isocratic
|
|
2 |
25 |
14 |
30 |
70 |
0 |
Linear
gradient |
|
3 |
25.1 |
0.1 |
0 |
100 |
0 |
Step
gradient |
|
4 |
42 |
16.9 |
0 |
100 |
0 |
Isocratic
|
|
5 |
42.1 |
0.1 |
0 |
0 |
100 |
Step
gradient |
|
6 |
45 |
2.9 |
0 |
0 |
100 |
Isocratic
|
|
7 |
45.1 |
0.1 |
100 |
0 |
0 |
Step
gradient |
|
8 |
60 |
14.9 |
100 |
0 |
0 |
Re-equilibration |
|
0 |
46 |
|
5 |
46 |
|
20 |
55 |
|
40 |
70 |
|
42 |
46 |
|
1. Methionine sulfoxide
2. Hydroxyproline
3. Aspartic acid
4. Threonine
5. Serine
6. Glutamic acid
7. Proline |
8. Glycine
9. Alanine
10. Cystine
11. Valine
12. Methionine
13. Isoleucine
14. Leucine |
15. Tyrosine
16. Phenylalanine
17. Hydroxylysines
18. Lysine
19. Ammonia
20. Histidine
21. Tryptophan |
22. Arginine
23. Cysteic acid
24. Methionine Sulfone
25. Norleucine |
|
0 |
0 |
0 |
100 |
0 |
0 |
Inject |
|
1 |
12 |
12 |
100 |
0 |
0 |
Isocratic
|
|
2 |
28 |
16 |
50 |
50 |
0 |
Linear
gradient |
|
3 |
28.1 |
0.1 |
0 |
100 |
0 |
Step
gradient |
|
4 |
46 |
17.9 |
0 |
100 |
0 |
Isocratic
|
|
5 |
46.1 |
0.1 |
0 |
0 |
100 |
Step
gradient |
|
6 |
49 |
2.9 |
0 |
0 |
100 |
Isocratic
|
|
7 |
49.1 |
0.1 |
100 |
0 |
0 |
Step
gradient |
|
8 |
64 |
14.9 |
100 |
0 |
0 |
Re-equilibration |
|
0 |
50 |
|
14 |
50 |
|
30 |
70 |
|
44 |
70 |
|
46 |
50 |
|
1. Methionine sulfoxide
2. Hydroxyproline
3. Aspartic acid
4. Threonine
5. Serine
6. Glutamic acid
7. Proline |
8. Glycine
9. Alanine
10. Cystine
11. Valine
12. Methionine
13. Isoleucine
14. Leucine |
15. Tyrosine
16. Phenylalanine
17. Hydroxylysines
18. Lysine
19. Ammonia
20. Histidine
21. Tryptophan |
22. Arginine
23. Cysteic acid
24. Methionine Sulfone
25. Norleucine |
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