Overview
Lithium Amino Acid Analysis Columns
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: λ=570 nm for primary amino acids
λ=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: λex 330 nm, λem 465 nm
The recommended gradient conditions are subject to change without notice. This may happen because of lotspecific changes in the columns, or improvements in the overall method.
The recommended gradient for the column will always be included in the column package, and it supersedes the information in this catalog. 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 a temperature gradient allows the user to improve separation, shorten analysis time and fine-tune the method for detecting compounds of interest.
70-Minute
70-minute High-efficiency Lithium Cation-exchange Column
Catalog number 0354675T
Use With Cation-Exchange Gard™ Column Protection System 1700 – 3102
Use For Physiological Samples
Temperature Gradient
HPLC PROGRAM
| TIME | 1700-1125 % | Li365 % | Li375 % | RG003 % |
|---|---|---|---|---|
|
0 |
100 |
0 |
0 |
0 |
|
10 |
100 |
0 |
0 |
0 |
|
19 |
40 |
60 |
0 |
0 |
|
32 |
0 |
100 |
0 |
0 |
|
43 |
0 |
100 |
0 |
0 |
|
43.1 |
0 |
0 |
100 |
0 |
|
57 |
0 |
0 |
100 |
0 |
|
57.1 |
0 |
0 |
70 |
30 |
|
72 |
0 |
0 |
70 |
30 |
|
72.1 |
100 |
0 |
0 |
0 |
HPLC FLOW RATE: 0.55 mL/min, INITIAL TEMP.: 34 ºC
COLUMN OVEN PROGRAM
| TIME | TEMP ºC |
|---|---|
| 0 | 34 |
| 6 | 34 |
| 17 | 65 |
| 25 | 70 |
| 70 | 70 |
| 71 | 34 |
REAGENT FLOW RATE: 0.5 mL/min ;
For more information see Method Abstract MA 382 (page 90) and Pickering Amino Acids Brochure.
| 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 α-Aminoadipic acid 14 Proline 15 Glycine 16 Alanine 17 Citrulline 18 α-Amino-n-butyric acid 19 Valine 20 Cystine 21 Methionine 22 Allo-Isoleucine 23 Cystathionine |
24 Isoleucine 25 Leucine 26 Tyrosine 27 Phenylalanine 28 β-Alanine 29 β-Amino-i-butyric acid 30 Homocystine 31 γ-Aminobutyric acid 32 Tryptophan 33 Ethanolamine 34 Ammonia |
35 Hydroxylysines 36 Ornitine 37 Lysine 38 1-Methylhistidine 39 Histidine 40 3-Methylhistidine 41 Anserine 42 Carnosine 43 Arginine 44 Glucosaminic Acid* 45 2-Aminoethyl-cysteine (AEC)* |
*Internal Standard
NOTE: This method utilizes column temperature gradient. Use Pinnacle PCX column oven
or HPLC column oven with temperature gradient capabilities.
High-efficiency
High-efficiency Lithium Cation-exchange Column
Catalog number 0354100T
Use with Cation-Exchange Gard™ Column Protection System 1700 – 3102
Use for Physiological Samples and Complex Matrices
Temperature Isocratic
| 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 α-Aminoadipic acid 14 Proline 15 Glycine 16 Alanine 17 Citrulline 18 α-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 α-Amino-β- guanidinopropionic acid 45 Arginine |
CONDITIONS
| Step | Time | Interval | Li275 % | Li750 % | RG003 % | Comment |
|---|---|---|---|---|---|---|
|
0 |
0 |
0 |
100 |
0 |
0 |
Inject |
|
1 |
8 |
8 |
100 |
0 |
0 |
Isocratic |
|
2 |
46 |
38 |
65 |
35 |
0 |
Linear Gradient |
|
3 |
86 |
40 |
0 |
100 |
0 |
Linear Gradient |
|
4 |
90 |
4 |
0 |
100 |
0 |
Isocratic |
|
5 |
115 |
25 |
0 |
94 |
6 |
Linear Gradient |
|
6 |
122 |
2 |
0 |
94 |
6 |
isocratic |
|
7 |
122.1 |
0.1 |
100 |
0 |
0 |
Step Change |
|
8 |
140 |
17.9 |
100 |
0 |
0 |
Re-equilibration |
Flow rate: 0.35 mL/min, column temperature 37°C, injection volume 10 µL of 0.25 µmole/mL Std.
For more information see Pickering Amino Acids Brochure.
Temperature Gradient
CONDITIONS
| Step | Time | 1700-1125 % | Li365 % | Li375 % | RG003 % | Comment |
|---|---|---|---|---|---|---|
|
0
|
0
|
100
|
0
|
0
|
0
|
Inject |
|
1
|
15
|
100
|
0
|
0
|
0
|
Isocratic
|
|
2
|
27
|
40
|
60
|
0
|
0
|
Linear Gradient |
|
3 |
45 |
0 |
100 |
0 |
0 |
Linear Gradient |
|
4 |
60 |
0 |
100 |
0 |
0 |
Isocratic |
|
5 |
60.1 |
0 |
0 |
100 |
0 |
Step Change |
|
6 |
78 |
0 |
0 |
100 |
0 |
Isocratic |
|
7 |
78.1 |
0 |
0 |
70 |
30 |
Step Change |
|
8 |
95 |
0 |
0 |
70 |
30 |
Isocratic |
|
9 |
95.1 |
100 |
0 |
0 |
0 |
Step Change |
|
10 |
115 |
100 |
0 |
0 |
0 |
Equilibration |
HPLC FLOW RATE: 0.4 mL/min, INITIAL TEMP.: 34 ºC INJECTION VOLUME: 10 μL of 0.25 μmole/mL Std.
COLUMN OVEN PROGRAM
| Time (Min) | Temperature ºC |
|---|---|
| 0 | 34 |
| 13 | 34 |
| 30 | 65 |
| 67 | 65 |
| 80 | 70 |
| 97 | 70 |
| 98 | 34 |
| HPLC FLOW RATE: 0.4 mL/min, INITIAL TEMP.: 34 ºC INJECTION VOLUME: 10 μL of 0.25 μmole/mL Std. |
| 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 α-Aminoadipic acid 14 Proline 15 Glycine 16 Alanine 17 Citrulline 18 α-Amino-n-butyric acid 19 Valine 20 Cystine 21 Methionine 22 Cystathionine 23 Isoleucine 24 Leucine |
25 Tyrosine 26 Phenylalanine 27 β-Alanine 28 β-Amino-i-butyric acid 29 Homocystine 30 γ-Aminobutyric acid 31 Tryptophan 32 Ethanolamine 33 Ammonia 34 Hydroxylysines 35 Ornithine |
36 Lysine 37 1-Methyl-histidine 38 Histidine 39 3-Methyl-histidine 40 Anserine 41 Carnosine 42 Anserine 43 Glucosaminic Acid* |
*Internal Standard
NOTE: This method utilizes column temperature gradient.
Use Pinnacle PCX column oven or HPLC column oven with temperature gradient capabilities.
For more information see Pickering Amino Acids Brochure.
Temperature Isocratic
CONDITIONS
| Step | Time (Min) |
Interval | Li275 % | Li750 % | RG003 % | Comment |
|---|---|---|---|---|---|---|
|
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 |
HPLC FLOW RATE: 0.35 mL/min, COLUMN TEMP.: 38 ºC
INJECTION VOLUME: 10 μL of 0.25 μmole/mL Std.
Standard
Standard Lithium Cation-Exchange Column
Catalog Number 0393250
Use With Cation-Exchange Gard™ Column Protection System 1700 – 3102
Use for Physiological Samples and Complex Matrices
Temperature Isocratic
| 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 α-Aminoadipic acid 14 Proline 15 Glycine 16 Alanine 17 Citrulline 18 α-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 α-Amino-β- guanidinopropionic acid 45 Arginine |
CONDITIONS
| Time | Li275 % | Li750 % | RG003 % |
|---|---|---|---|
|
0 |
100 |
0 |
0 |
|
17 |
100 |
0 |
0 |
|
65 |
65 |
35 |
0 |
|
128 |
0 |
100 |
0 |
|
145 |
0 |
100 |
0 |
|
185 |
0 |
94 |
6 |
|
185.1 |
100 |
0 |
0 |
|
210 |
100 |
0 |
0 |
FLOW RATE: 0.3 mL/min, COLUMN TEMP.: 40 ºC,
INJECTION VOLUME: 10 µL OF 0.25 µmole/mL STD.
GARD™
GARD™ Column Protection System
The new GARD™ uses a proprietary material to prevent matrix compounds from passing through (and thereby protecting the analytical column), but allows the analytes of interest to pass unimpeded through the GARD™ and onto the analytical column.
The new GARD™ significantly prolongs column life without band spreading or added pressure. We demonstrated, by means of a performance comparison for Amino Acid Analysis, that the use of a GARD™ will effectively protect the analytical column, will be more cost-effective for the laboratory, is easy to change, is universal to cation-exchange applications, and most importantly has zero band spreading.
Wth the new GARD™ Column protection system from Pickering Laboratories the same cation-exchange GARD™ can be used for nearly all cation-exchange applications.
Pricing & Ordering
Pricing & Ordering
| |||
Catalog # | Description | ||
|---|---|---|---|
0354675T | Lithium Ion-exchange Column, 4.6 x 75 mm (with test mixture 1700-0070) | ||
0354100T | Lithium Ion-exchange Column, 4.0 x 100 mm (with test mixture 1700-0070) | ||
0393250 | Standard Lithium Cation-exchange Column, 3.0 x 250 mm, includes Amino Acid Test Mixture 1700-0070 | ||
| |||
Catalog # | Description | ||
|---|---|---|---|
1700-3102 | Cation-exchange GARD™ assembly: Holder w/ 2 replaceable GARD™ | ||
1700-3101 | Replacement Cation-exchange GARD™ (2/PK) | ||
1700-3100 | GARD™ Holder | ||
