SEC
22
The best results are obtained when selecting a column with the sample’s
molecular weight in the linear portion of the calibration curve.
Column: TSKgel PW columns: A. G2000PW, B. G2500PW, C. G3000PW,
D. G4000PW, E. G5000PW, F. G6000PW, G. GMPW, all 7.5mm ID x 60 cm L
TSKgel PW
XL
columns: H. G2500PW
XL
, J. G3000PW
XL
, K. G4000PW
XL
,
L. G5000PW
XL
, M. G6000PW
XL
, N. GMPW
XL
, all 7.8 mm ID x 30 cm L; Elution:
distilled water; Flow rate: 1.0 m L/min; Detection: RI
Column: 1. TSKgel G3000PW
XL
, 2. G4000PW
XL
, 3. G5000PW
XL
, 4. G6000PW
XL
,
5. GMPW
XL
; Sample: a. thyroglobulin (660,000 Da), b.
g
-globulin (150,000 Da),
c. albumin (67,000 Da), d. ovalbumin (43,000 Da), e.
b
-lactoglobulin (36,000
Da), f. myoglobin (16,900 Da), g. cytochrome C (12,400 Da); Elution: 0.2 mol/L
phosphate buffer (pH 6.8); Flow rate: 1.0 mL/min; Detection: UV @ 280 nm
Columns: TSKgel G3000PW
XL
-CP, 7 μm, 7.8 mm ID x 30 cm L, TSKgel
G5000PW
XL
-CP, 10 μm, 7.8 mm ID x 30 cm L, TSKgel G6000PW
XL
-CP, 13 μm,
7.8 mm ID x 30 cm L
Mobile phase: 0.1 mol/L NaNO
3
; Flow rate: 1 mL/min; Detection: RI;
Temperature: 25°C; Samples: polyethylene oxides (PEO) standards,
polyethylene glycols (PEG) standards
Columns:TSKgelSuperOligoPW,SuperMultiporePW-N,SuperMultiporePW-M,
SuperMultiporePW-H (each 6.0 mm ID x 15 cm L);
Mobile phase: H
2
O; Flow rate: 0.60 mL/min; Detection: RI; Temperature: 25°C;
Samples: polyethylene oxides (PEO) standards, polyethylene glycols (PEG)
standards, ethylene glycol (EG) standards
Polyethylene glycol and oxide calibration curves
on TSK-GEL PW and TSK-GEL PW
XL
columns
Elution volume (mL)
Colu n:
Elution:
Flow Rate:
Detection:
TSK-GEL PW columns: A. G2000PW, B. G2500PW, C. G3000PW
D. G4000PW, E. G5000PW, F. 6000 , G. GMPW, all 7.5m ID x 60 cm
TSK-GEL PW
XL
columns: H. 25
XL
J 3
XL
K. G4000PW
XL
,
L. G5000PW
XL
, M. G6000PW
XL
, N. GMPW
XL
, all 7.8 m ID x 30 cm
distilled water
1.0 m L/min
RI
5
TSK-GEL PW Columns
TSK-GEL PW
XL
Columns
Molecular weight (Da)
10
15
10
20
15
10
2
10
3
10
4
10
5
10
6
H
J
N
K
L
M
10
2
10
3
10
4
10
5
10
6
G
E F
C
D
A B
Elution volume (mL)
Column:
1. G3000PW
XL
, 2. G4000PW
XL
, 3. G5000PW
XL
,
4. G6000PW
XL
, 5. GMPW
XL
Sample:
a. thyroglobulin (660,000 Da), b.
γ
-globulin (150,000 Da),
c. albumin (67,000 Da), d. ovalbumin (43,000 Da),
e.
β
-lactoglobulin (36,000 Da), f. my globin (16,900 Da),
g. cytochrome C (12,400 Da)
Elution:
0.2 mol/L phosphate buffer (pH 6.8)
Flow Rate: 1.0 mL/min
Detection: UV @ 280 nm
Molecular weight (Da)
10
4
10
3
10
5
10
6
10
7
5
1
3
2
4
a
b
c
d e
f
g
Protein calibration curves on TSK-GEL PW
XL
columns
4
6
8
10
12
Elution volume (mL)
figure 14
Polyethylene glycol and oxide calibration curves on TSKgel PW and
TSKgel PW
XL
columns
figure 15
Protein calibration curves on TSKgel PW
XL
columns
figure 16
Polyethylene glycol and oxide calibration curves for TSKgel PW
XL
-CP
columns
TSKgel G3
TSKgel G5
TSKgel G6
Mobile pha
Flow Rate:
Detection:
Temp:
Samples:
10
10
2
10
3
10
4
10
5
10
6
10
7
3
5
7
9
11
Elution time (minutes)
Log Molecular Weight (PEO)
TSKgel G3000PW
XL
-CP
TSKgel G5000PW
XL
-CP
TSKgel G6000PW
XL
-CP
figure 17
Polyethylene glycol, oxide and ethylene glycol calibration curves for
TSKgel SuperMultiporePW and SuperOligoPW
10
10
2
10
3
10
4
10
5
10
6
10
7
1.50
2.50
3.50
4.50
5.50
Elution time (minutes)
Log Molecular Weight
TSKgel SuperOligoPW
TSKgel SuperMultiporePW-N
TSKgel SuperMultiporePW-M
TSKgel SuperMultiporePW-H
TSKgel Su
TSKgel Su
TSKgel Su
TSKgel Su
Mobile pha
Flow rate:
Detection:
Temperatu
Samples:
Calibration curves for TSKgel PW / SuperMultiporePW Gel Filtration columns
