Page 59 - Application Handbook - Liquid Chromatography

Application No.L432
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mV
22
2
2
200
200
150
150

100
100
1
3
1 2 13 15
12
5
1
10 11
1 1 1 0 1 1 1 18 2 0
20
8
17
4 4 7 7 9 9 1 7 1 9
19
50
50 3 3 5 5
2 2 8 8
1
2
6 6 1 4 16 21
14
0 0
0.0 2.5 5.0 7.5 min
Peaks
1. Aspartic Acid 2. Glutamic Acid 3. Asparagine 4. Serine 5. Glutamine 6. Histidine 7. Glycine 8. Threonine 9. Citruline
10. Arginine 11. Alanine 12. GABA 13. Tyrosine 14. Cys-Cys 15. Valine 16. Methionine 17. Tryptophan 18. Phenylalanine
19. Isoleucine 20. Leucine 21. Lysine 22. Proline
Fig. 2 Chromatogram of 22 Amino Acids (10 μmol/L Each, 1 μL Injection)
n Linearity and Repeatability n Analysis of Actual Samples
Using calibration curves generated with a concentration Fig. 3 shows a chromatogram of analysis of a
range from 1-100 μmol/L for each amino acid, excellent commercially available soft drink using this method. The
2
linearity was obtained, with a ratio of contribution (R sample was analyzed after filtering it through a 0.2 μm
value) greater than 0.999 in all cases. Table 3 shows the membrane filter.
peak area repeatability for all 22 amino acids obtained
in repeat analysis (n = 6).
Table 3 Repeatability
mV
Area%RSD Area%RSD 450 Peaks
Asp 0.50 GABA 0.41 1. Aspartic Acid 2. Glutamic Acid 3. Serine
Glu 0.48 Tyr 0.55 400 4. Histidine 5. Glycine 6. Threonine 7. Arginine 8. Alanine
Asn 0.51 Cys-Cys 0.46 350 9. Tyrosine 10. Valine 11. Methionine 12. Tryptophan
13. Phenylalanine 14. Isoleucine 15. Leucine 16. Lysine
Ser 0.41 Val 0.71 17. Proline
300
Gln 0.56 Met 0.71
His 0.57 Trp 0.70 250
Gly 0.29 Phe 0.73 200 5 17
Thr 0.55 Ile 0.63
Cltruline 0.46 Leu 0.55 150 11 14
Arg 0.45 Lys 0.56 100 8 9 10 15
Ala 0.46 Pro 2.35 6 7 13
50 2 3 12 16
1 4
0
0.0 2.5 5.0 7.5 min
Fig. 3 Chromatogram of Soft Drink

First Edition: March, 2012

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