83
rpc
RPC
SRM 870
Standard Reference Material SRM 870 was developed by NIST
(National Institute of Standards and Technology) as a means to classify
the many commercially available reversed phase columns into closely-
related groups. Amitriptyline, a tertiary amine, and quinizarin, a strong
chelating compound, are included in the SRM 870 mixture, together
with more traditional compounds. As shown in
Figure 5
,
symmetrical
peaks are obtained on TSKgel ODS-100V and TSKgel ODS-100Z for the
compounds in this test mixture, clearly demonstrating the superior
performance of these columns for the analysis of basic and chelating
compounds.
Vitamins
Simple and fast analysis of water- and lipid-soluble vitamins is possible
on the TSKgel ODS-100V and TSKgel ODS-100Z columns, as shown in
Figure 6
. Clearly the TSKgel ODS-100Z column provides better overall
resolution for the polar compounds in the mixture, while much shorter
analysis time was obtained on TSKgel ODS-100V for the late eluting
non-polar compounds.
APPLICATIONS OF TSKgel ODS-100V / ODS-100Z
200
400
600
0
0
10
20
30
50
40
3.
4.
5.
6.
7.
2.
1.
B)
8.
9.
10.
11.
Retention time (min)
Signal int. (mV)
Columns:
A) TSKgel ODS-100V (4.6mm ID x 15cm)
B) TSKgel ODS-100Z (4.6mm ID x 15cm)
Eluent:
A) 0.1% TFA in H
2
O
B) 0.1% TFA in ACN
Gradient:
0 min (B: 0%) -- 20 min (B: 40%) --
22min (B: 100%) -- 50min (B: 100%)
Flow rate: 1.0mL/min.
Temp.:
40˚C
Detection: UV @ 280nm
Inj. volume: 5µL
Samples:
1.
L
-Ascorbic acid, 2. Nicotinic acid,
3. Thiamine, 4. Pyridoxal, 5. Pyridoxine,
6. Caffeine, 7. Riboflavin, 8. Retinol,
9.
δ
-Tocopherol, 10.
α
-Tocopherol,
A)
Columns: (A) TSKgel ODS-100V 3µm (4.6mmID x 15cm)
(B) TSKgel ODS-100Z 3µm (4.6mmID x 15cm)
Eluent: 20mmol/L Phosphate buffer (pH 7.0) /MeOH (20/80)
Flow rate : 1.0mL/min
Detection: UV @ 254nm
Temp: 40°C
Inj. volume: 10µL
Sample: 1. Uracil, 2. Toluene, 3. Ethyl benzene,
4. Quinizarin, 5. Amitriptyline
0
50
100
150
200
250
300
0
2
4
6
8
10
12
Retention Time (min)
mV
A)
B)
1
2.
3.
4.
5.
AF=0.99
AF=1.06
TSKgel ODS-100V
TSKgel ODS-100Z
figure 6
Analysis of vitamins
figure 5
Standard reference material SRM 879
Columns: (A) TSKgel ODS-100V 3 μm (4.6 mm ID x 15 cm L)
(B) TSKgel ODS-100Z 3 μm (4.6 mm ID x 15 cm L);
Eluent: 20 mmol/L Phosphate buffer (pH 7.0)/MeOH (20/80);
Flow rate: 1.0 mL/min; Detection: UV@254nm; Tep.: 40°C; Inj. volume: 10 μL;
Sample: 1. Uracil, 2. Toluene, 3. Ethyl benzene, 4. Quinizarin, 5. Amitriptyline
Columns: (A) TSKgel ODS-100V (4.6 mm ID x 15 cm L)
(B) TSKgel ODS-100Z (4.6 mm ID x 15 cm L);
Eluent: (A) 0.1% TFA in H
2
O; (B) 0.1 % TFA in ACN,
Gradient: 0 min (B: 0%) - 20 min (B: 40%) - 22 min (B: 100%) - 50 min (B: 100%);
Flow rate: 1.0 mL/min.; Temp.: 40°C; Detection: UV@280nm;
Inj. volume: 5 μL; Samples: 1. L-Ascorbic acid, 2. Nicotinic acid, 3. Thiamine,
4. Pyridoxal, 5. Pyridoxine, 6. Caffeine, 7. Riboflavin, 8. Retinol, 9.
δ
-Tocopherol,
10.
α
-Tocopherol, 11.
α
-Tocopherol acetate)
