|
Pickering Laboratories
specialize in manufacturing of cation-exchange
columns for amino acid analysis. No other
techniques, including reversed-phase
chromatography, have been shown to match
post-column ion-exchange methods in quantitation
and reproducibility. Advantages of this method,
such as absence of matrix interferences, are
especially important in analysis of native
samples.
Lithium columns and buffers
systems have high selectivity and are perfect for
physiological fluids and food analysis.
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 supercedes
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 Phosphoserine
2 Taurine
3 Phosphoethanolamine
4 Urea
5 Aspartic acid
6 Hydroxyproline
7 Threonine
8 Serine
9 Asparagine
10 Glutamic acid
11 Glutamine
12 Sarcosine
13
 -Aminoadipic
acid
14 Proline
15 Glycine |
16 Alanine
17 Citrulline
18
-Amino-n-butyric
acid
19 Valine
20 Cystine
21 Methionine
22 Cystathionine
23 Isoleucine
24 Leucine
25 Norleucine
26 Tyrosine
27 Phenylalanine
28 β-Alanine
29 β-Amino-i-butyric acid
30 Homocystine |
31
γ-Aminobutyric acid
32 Tryptophan
33 Ethanolamine
34 Hydroxylysines
35 Ammonia
36 Creatinine
37 Ornithine
38 Lysine
39 Histidine
40 3-Methyl-histidine
41 1-Methyl-histidine
42 Carnosine
43 Anserine
44
-Amino-β-guanidinopropionic
acid
45 Arginine
|
|
|
0 |
|
|
|
|
12 |
|
|
|
|
48 |
|
|
|
|
90 |
|
|
|
|
95 |
|
|
|
|
120 |
|
|
|
|
122 |
|
|
|
|
122.1 |
|
|
|
|
140 |
|
|
|
Flow rate: 0.35 mL/min, column
temperature 38°C, injection volume 10 µL of
0.25 µmole/mL Std. |
|
0 |
0 |
100 |
0 |
0 |
0 |
Inject
|
|
1 |
30 |
100 |
0 |
0 |
0 |
Isocratic |
|
2 |
30.1
|
0 |
100 |
0 |
0 |
Step
gradient |
|
3 |
57 |
0 |
100 |
0 |
0 |
Isocratic |
|
4 |
57.1
|
0 |
0 |
100 |
0 |
Step
gradient |
|
5 |
78 |
0 |
0 |
100 |
0 |
Isocratic |
|
6 |
78.1
|
0 |
0 |
80 |
20 |
Step
gradient |
|
7 |
96 |
0 |
0 |
80 |
20 |
Isocratic |
|
8 |
96.1
|
100 |
0 |
0 |
0 |
Step
gradient |
|
9 |
111 |
100 |
0 |
0 |
0 |
Re-equilibration |
|
|
0 |
32 |
|
13 |
32 |
|
30 |
61 |
|
67 |
61 |
|
80 |
70 |
|
90 |
70 |
|
92 |
32 |
|
1
Phosphoserine
2
Taurine
3
Phosphoethanolamine
4
Urea
5
Aspartic acid
6
Hydroxyproline
7
Threonine
8
Serine
9
Asparagine
10
Glutamic acid
11
Glutamine
12
Sarcosine
13
-Aminoadipic
acid
14
Proline
15
Glycine |
16
Alanine
17
Citrulline
18
-Amino-n-butyric
acid
19
Valine
20
Cystine
21
Methionine
22
Cystathionine
23
Isoleucine
24
Leucine
25
Norleucine
26
Tyrosine
27
Phenylalanine
28
β-Alanine
29
β-Amino-i-butyric acid
30
Homocystine |
31
γ-Aminobutyric
acid
32
Tryptophan
33
Ethanolamine
34
Hydroxylysines
35
Ammonia
36
Creatinine
37
Ornithine
38
Lysine
39
Histidine
40
3-Methyl-histidine
41
1-Methyl-histidine
42
Carnosine
43
Anserine
44
-Amino-β-guanidinopropionic
acid
45
Arginine |
|
0 |
0 |
0 |
86 |
14 |
0 |
Inject
|
|
1 |
25 |
11 |
73 |
27 |
0 |
Linear
gradient |
|
3 |
25.1
|
0.1 |
0 |
0 |
100 |
Step
gradient |
|
4 |
30 |
16.9
|
0 |
0 |
100 |
Isocratic |
|
7 |
30.1
|
0.1 |
86 |
14 |
0 |
Step
gradient |
|
8 |
42 |
14.9
|
86 |
14 |
0 |
Re-equilibration |
|
|
|
1 Phosphoserine
2 Taurine
3 Phosphoethanolamine
4 Urea
5 Aspartic acid
6 Hydroxyproline
7 Threonine
8 Serine
9 Asparagine
10 Glutamic acid
11 Glutamine
12 Sarcosine
13
-Aminoadipic
acid
14 Proline
15 Glycine |
| | | 
