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Method Abstract 382

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

chart-m-382
 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

10

100

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