MA 380.1 / 30-MINUTE AMINO ACID ANALYSIS OF HYDROLYZED SAMPLES
Pickering Laboratories specializes in the manufacturing of cation-exchange columns and eluants for amino acid analysis. No other technique has been shown to match the reproducibility and selectivity of ion-exchange analysis with post-column Ninhydrin detection. Nor is there a chromatography technique that provides as much information; the 570/440 nm signal ratio for each amino acid is a constant and so offers information about peak purity.
Pinnacle PCX post-column derivatization system provides a unique opportunity to combine eluant gradient capabilities of modern HPLC instruments with column temperature gradients. As might be expected, this capability also reduces the analysis time.
We introduce an accelerated Amino Acid analysis method for hydrolyzed samples that utilizes temperature and eluant gradients.
METHOD
Analytical Conditions
Column: High-efficiency Sodium cation-exchange column, 4.6 x 110 mm, Catalog number 1154110T
Flow Rate: 0.6 mL/min
Mobile Phase: See method
Post-column Conditions
Post-column System: Pinnacle PCX
Reactor Volume: 0.5 mL
Temperature: 130 ˚C
Reagent: Trione Ninhydrin reagent
Flow Rate: 0.3 mL/min
Detection: UV/VIS 570 nm for primary amino acids and
440 nm for secondary amino acids
METHOD FOR OXIDIZED FEEDS HYDROLYSATE SAMPLES
|
TIME |
Na270 % |
Na425 % |
Na640 % |
RG011 % |
|---|---|---|---|---|
|
0 |
100 |
0 |
0 |
0 |
|
4.0 |
100 |
0 |
0 |
0 |
|
15.0 |
0 |
100 |
0 |
0 |
|
16.0 |
0 |
0 |
100 |
0 |
|
31.0 |
0 |
0 |
100 |
0 |
| 31.1 | 0 | 0 | 0 | 100 |
| 33.0 | 0 | 0 | 0 | 100 |
| 33.1 | 100 | 0 | 0 | 0 |
| 40 | 100 | 0 | 0 | 0 |
COLUMN OVEN PROGRAM
|
TIME |
TEMP ºC |
|---|---|
|
0 |
55 |
|
12 |
55 |
|
17 |
70 |
|
32 |
70 |
|
33 |
55 |
|
|
|
|
1 Aspartic Acid 2 Threonine 3 Serine 4 Glutamic Acid 5 Proline 6 Glycine |
7 Alanine 8 Cystine 9 Valine 10 Methionine 11 Isoleucine 12 Leucine |
13 Norleucine 14 Tyrosine 15 Phenylalanine 16 Histidine 17 Lysine |
18 Tryptophan 19 Ammonia 20 Arginine 21 Cysteic Acid 22 Methionine Sulfone |
METHOD FOR PROTEIN HYDROLYSATE SAMPLES
|
TIME |
Na315 % |
Na425 % |
Na640 % |
RG011 % |
|---|---|---|---|---|
|
0 |
100 |
0 |
0 |
0 |
|
4.0 |
100 |
0 |
0 |
0 |
|
15.0 |
0 |
100 |
0 |
0 |
|
16.0 |
0 |
0 |
100 |
0 |
|
31.0 |
0 |
0 |
100 |
0 |
| 31.1 | 0 | 0 | 0 | 100 |
| 33.0 | 0 | 0 | 0 | 100 |
| 33.1 | 100 | 0 | 0 | 0 |
| 40 | 100 | 0 | 0 | 0 |
COLUMN OVEN PROGRAM
|
TIME |
TEMP ºC |
|---|---|
|
0 |
46 |
|
4 |
46 |
|
9 |
70 |
|
32 |
70 |
|
33 |
46 |
|
|
|
|
1 Aspartic Acid 2 Threonine 3 Serine 4 Glutamic Acid 5 Proline 6 Glycine |
7 Alanine 8 Cystine 9 Valine 10 Methionine 11 Isoleucine 12 Leucine |
13 Norleucine 14 Tyrosine 15 Phenylalanine 16 Histidine 17 Lysine |
18 Tryptophan 19 Ammonia 20 Arginine 21 Cysteic Acid 22 Methionine Sulfone |
