84
RPC
APPLICATIONS OF TSKgel ODS-100V /ODS-100Z
Organic Acids
Organic acids play an important role in many metabolic processes,
fermentation and food products.
Figure 7
shows a baseline separation
of 15 organic acids in less than 25 minutes using a simple 0.1%
phosphoric acid mobile phase.
Polymer Additives
A baseline separation of 26 well known polymer additives is shown
in
Figure 8
. Note that while a simple linear acetonitrile gradient was
used, the column temperature was increased to 50°C to achieve the
required baseline separation on a TSKgel ODS-100V column.
Nucleotides
The analysis of mono-, di-, and tri-phosphorylated nucleotides on a
TSKgel ODS-100V column is shown below
(Figure 9)
. The separation is
accomplished by adding a short chain ion pairing agent, t-butylamine,
and adjusting the mobile phase pH to 6.8.
Column:
TSKgel ODS-100V (4.6mm ID × 15cm)
Mobile phases: A) H
2
O
B) ACN
Gradient:
0 min (B: 60%) -- 20 min (B: 100%)
Flow rate:
1.0mL/min
Temp:
50°C
Detection:
UV (225nm)
Inj. Volume: 10µL
Concentration: 10mg/L each
Samples:
1. Cyasorb UV-24, 2. BHA, 3. Ionox 100,
4. Seesorb 101, 5. Tinuvin P, 6. Yoshinox SR,
7. Seesorb 202, 8. BHT, 9. Noclizer M-17,
10. Yoshinox 2246R, 11. Topanol CA,
12. Yoshinox 425, 13. Cyanox 1790,
14. Cyasorb UV-531, 15. Ionox 220,
16. Nonflex CBP, 17. Tinuvin 326,
18. Tinuvin 120, 19. Irganox 3114,
20. Uvtex OB, 21. Tinuvin 327, 22. Tinuvin 328,
23. Irganox 1010, 24. Irganox 1330,
25. Irganox 1076, 26. Irgafos 168
0
50
100
150
0
10
20
30
40
50
Retention time (min.)
mV
1
2 3
4
5
6
7 8 9
10
11
12
13
14
15
16,17
18
19,20
21
22
23
24
25
26
figure 8
Analysis of polymer additives with TSKgel ODS-100V
Column:
TSKgel ODS-100V (4.6mm ID × 25cm)
Mobile phase: 0.1% H
3
PO
4
, pH 2.3
Flow rate:
1.0mL/min
Temp:
40°C
Inj. Volume: 10µL
Samples:
1. Oxalic acid (0.1mg/mL)
2.
L
-Tartaric acid (0.5mg/mL)
3. Formic acid (1.0mg/mL)
4.
L
-Malic acid (1.0mg/mL)
5.
L
-Ascorbic acid (0.1mg/mL)
6. Lactic acid (1.0mg/mL)
1
2
3
4
5
6
7
8
9 10
11
12
13
14
15
0
10
20
30
40
0
5
10
15
20
Retention time (min.)
mV
25
figure 7
Analysis of organic acids with TSKgel ODS-100V
Column: TSKgel ODS-100V (4.6 mm ID × 25 cm L)
Mobile phase: 0.1 % H
3
PO
4
, (pH 2.3); Flow rate: 1.0 mL/min;
Temp: 40 °C; Inj. Volume: 10 μL; Samples: 1. Oxalic acid (0.1 mg/mL) 2. L-Tar-
taric acid (0.5 mg/mL) 3. Formic acid (1.0 mg/mL) 4. L-Malic acid (1.0 mg/mL)
5.L-Ascorbicacid(0.1mg/mL)6.Lacticacid(1.0mg/mL)7.Aceticacid(1.0mg/mL)
8.Maleicacid(0.01mg/mL)9.Citricacid(1.0mg/mL)10.Succinicacid(1.0mg/mL)
11. Fumaric acid (0.025 mg/mL) 12. A rylic acid (0.1 mg/mL) 13. Propionic
acid (2.0 mg/mL) 14. Glutaric acid (1.0 mg/mL) 15. Itaconic acid (0.025 mg/mL)
Column: TSKgel ODS-100V (4.6mm ID × 15 cm L);
Mobile phases: (A) H
2
O (B) ACN; Gradient: 0 min (B: 60%) - 20 min (B: 100%);
Flow rate: 1.0 mL/min; Temp: 50 °C; Detection: UV@225nm;
Inj. Volume: 10 μL; Concentration: 10 mg/L each; Samples: 1. Cyasorb UV-24,
2. BHA, 3. Ionox 100, 4. Seesorb 101, 5. Tinuvin P, 6. Yoshinox SR,
7. Seesorb 202, 8. BHT, 9. Noclizer M-17, 10. Yoshinox 2246R, 11. Topanol CA,
12. Yoshinox 425, 13. Cyanox 1790, 14. Cyasorb UV-531, 15. Ionox 220,
16. Nonflex CBP, 17. Tinuvin 326, 18. Tinuvin 120, 19. Irganox 3114, 20. Uvtex
OB, 21. Tinuvin 327, 22. Tinuvin 328, 23. Irganox 1010 24. Irganox 1330,
25. Irganox 1076, 26. Irgafos 168
Column:
TSKgel ODS-100V (4.6mm ID × 25cm)
Mobile phases: A) 20 mmol/L t-butylamine + H
3
PO
4
(pH 6.8)
B) A/MeOH (90/10)
Gradient:
0 min (B: 0%) -- 35 min (B: 100%)
Flow rate:
1.0mL/min
Temp:
25°C
Detection:
UV (260nm)
Inj. Volume: 2µL
Concentration: 0.3g/L each
Samples:
1. CMP, 2. UMP, 3. CDP, 4. dUMP, 5. GMP,
0
5
10 15 20 25 30 35 40 45
Retention time (min)
1
2
3
4
5
6
7
8 9
10
11
12
13
14
15
16
17
18
19
20
21
figure 9
Analysis of nucleotices with TSKgel ODS-100V
Column: TSKgel ODS-100V (4.6 mm ID × 25 cm L)
Mobile phases: (A) 20 mmol/L t-butylamine + H
3
PO
4
(pH 6.8) (B) A/MeOH
(90/10); Gradient: 0 min (B: 0%) - 35 min (B: 100%); Flow rate: 1.0 mL/min;
Temp: 25 °C; Detection: UV@260nm; Inj. Volume: 2μL; Concentration: 0.3 g/L
each; Samples: 1. CMP, 2. UMP, 3. CDP, 4. dUMP, 5. GMP, 6. IMP, 7. UDP,
8. CTP, 9. TMP, 10. GDP, 11. IDP, 12. AMP, 13. UTP, 14. dGMP, 15. TDP,
16. GTP, 17. ITP, 18. ADP, 19. TTP, 20. dAMP, 21. ATP
