MA 382 / 70-MIN AMINO ACID ANALYSIS OF PHYSIOLOGICAL SAMPLES
Pickering Laboratories specializes in the manufacturing of cation-exchange columns and eluants for Amino Acid Analysis. Post-column derivatization with Ninhydrin offers unmatched selectivity and reproducibility of the analysis for most challenging matrixes.
Pinnacle PCX post-column derivatization system allows analysts to combine eluant gradient capabilities of modern HPLC instruments with column temperature gradients. We introduce new accelerated Amino Acids Analysis method for physiological samples that utilizes temperature and eluant gradients.
METHOD
Analytical Conditions
Column: High-efficiency Lithium cation-exchange column,
4.6 x 75 mm, 5 μm Catalog number 0354675T
Flow Rate: 0.55 mL/min
Mobile Phase: 1700-1125, Li365, Li375, RG003
Post-column Conditions
Post-column System: Pinnacle PCX
Reactor Volume: 0.5 mL
Temperature: 130 ˚C
Reagent: Trione
Flow Rate: 0.5 mL/min
Detection: UV/VIS 570 nm for primary amino acids
440 nm for secondary amino acids
Injection Volume: 10 μL of 0.25 μ mole/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 1 8 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 Ornitine 36 Lysine 37 1-Methylhistidine 38 Histidine 39 3-Methylhistidine 40 Anserine 41 Carnosine 42 Arginine 43 Glucosaminic Acid* |
COLUMN OVEN PROGRAM
|
TIME |
TEMP ºC |
|---|---|
|
0 |
34 |
|
6 |
34 |
|
17 |
65 |
|
25 |
70 |
|
70 |
70 |
|
71 |
34 |
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 |
