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:
Na315, Na740, RG011
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 440
nm for secondary amino acids
Runtime:
30 min
Equilibration Time:
9 min
COLUMN OVEN
PROGRAM
TIME
TEMP
ºC
0
46
3
46
10
55
22
70
27
70
28
46
HPLC PROGRAM
TIME
Na315 %
Na740 %
RG011 %
0
100
0
0
7.3
100
0
0
15
53
47
0
15.1
0
100
0
28
0
100
0
28.1
0
0
100
30
0
0
100
Chromatogram of
Protein Hydrolysate Standard
1
Aspartic
Acid
8
Cystine
15
Lysine
2
Threonine
9
Valine
16
Histidine
3
Serine
10
Methionine
17
Ammonia
4
Glutamic
Acid
11
Isoleucine
18
Tryptophan
5
Proline
12
Leucine
19
Arginine
6
Glycine
13
Tyrosine
7
Alanine
14
Phenylalanine
30-Minute Amino Acid Analysis
CATALOG INFORMATION
CATALOG
NO.
DESCRIPTION
1154110T
30-minute Sodium Cation-exchange Column 4.6 x
110 mm & 1700-0070 amino acid test mixture
