TR07A
An Innovative Size Exclusion Column Compatible with GFC/GPC
World's first silica-diol SEC column designed to work with LC and LC-MS compatible volatile salts

Compatible with hydrophilic polymers like proteins (GFC)
Compatible with hydrophobic polymers like polystyrene (GPC)
Uses volatile solvents as standard eluents
Extended lifetime under pH 1-8
Spherical porous silica / 3um particle / 30nm pore / Diol phase / pH 1-8 / MW up to 1MDa

Does not require NaCl for protein separation
Monomer 66 kDa
Dimer 132 kDa

BSA (Bovine Serum Albumin)monomer

UV 280nmELSD

monomer
Intrada SEC, 250x4.6mm
100mM ammonium formate
0.3mL/min (5MPa)
37 °C
10 µL (1mg/mL)

Protein separation
usingvolatile eluent

dimer

05Volatile eluent

dimer

10min

0510minIntrada SEC is a wide pore (300A), fully porous silica material with a bonded diol substitution stationaryphase. This innovative technology overcomes some traditional problems with diol-silica columns, such as
the need for use of high concentrations of non-volatile salts in the eluent and poor lifetime due to undesired
residual silanols effect. The next generation Intrada SEC column allows both aqueous durability like apolymer-based SEC column and the mechanical strength of a silica-based SEC column. This column can beused with volatile salts, such as 100mM ammonium formate, as the eluent for aqueous polymer analytes,
making it compatible with ELSD or MS detection.
Another benefit of the technology behind the Intrada SEC column is that it can be used for both aqueous
GFC (proteins, polysaccharides and nucleic acids etc.) and non-aqueous GPC (polystylene, PVAc etc.),
making it a real unified "SEC" column.
Imtakt USA (info@imtaktusa.com)
North America

Imtakt Corp./JAPAN (info@imtakt.com)
Other Countries

TR07AE-QH04

Ability to use volatile salt eluents may change SEC history

Traditionally, it has been required to use 100mM phosphate buffer + 300mM NaCl as an eluent for diolsilica SEC columns due to an undesired ionic effect, caused by exposed residual surface silanols. But use ofhigh concentrations of inorganic salt is too harsh for use with most LC systems and also their non-volatilenature makes them incompatible for use with an MS detector. The innovative Intrada SEC column with anovel diol substitution technology allows for the use of 100mM ammonium formate instead, which is avolatile eluent that is compatible with MS and ELSD detection. This truly revolutionary design may change
the history of SEC.
Intrada SEC may be the best SEC column
for use on MS detectors, due to its use ofvolatile eluents.
The figure to the left shows recombinanthuman serum albumin (rHSA) monomer
and dimer separation on LC-MS. The
molecular weight of the monomer wasdirectly analyzed as 66621Da, which ismore accurate than the traditional
caliburation curve method, which is basedon the hypothesis that all of the proteins areglobular shape.
We feel this shows that SEC-MS using
Intrada SEC column will likely be an
improved method for MW determination in
the future.

Determination of mAb purity is critical in
antibody drug science. To further support thiswork, Intrada SEC columns allow theseparation of not only monomers but also
oligomers and even fragments, as the figure tothe right shows.
For purification needs for products such as
biopolymers, antibodies or enzymes, full
preparative column dimensions are available(10 - 20 mm ID). Further adding to theconvenience of using Intrada SEC forpurification work, this unique material isdesigned to work conveniently with HPLC
instruments, due to its ability to utilize volatilebuffer eluents.

mAb, IgG
AntimAb- Fetoprotein

IgG
from human serum

Intrada SEC, 250x4.6mm
100mM ammonium formate
0.3 mL/min (5MPa)
37°C, 280nm
15 µL (0.5-1mg/mL)

Polymer quantification is also possible withIntrada SEC columns, as is shown below,
where accurate quantification of a 660 kDa
large protein is performed. Improved
separation performance and accuracy can also
be achieved by using our u-shaped multicolumn connections, which enable up to fourcolumns to be connected in series.

05min

101505min

10871

Product Information

Quantification
Thyroglobulin MW 660 kDa
ELSD
Intrada SEC
250x4.6mm

Separation Column

500

250 x 4.6 mm : WSE06
250 x 10 mm : WSEP6250 x 20 mm : WSEQ6200

100Height, log (mV)

100mM HCOONH40.3mL/min (5MPa)
37°C
ELSD
(spray cham. 50°C,drift tube 100°C)
5-50 µL (1mg/mL)

Guard Cartridge
504.6mmI.D. : GCWSES
10-20mmI.D. : GCWSEM
10Guard Holder
4.6mmI.D. : GCH11S
10-20mmI.D. : GCH12M

5051015 min

5151020Amount, log (µg)

250865

Versatile SEC column for peptides, proteins, bio- and synthetic polymers
Nearly all bio-polymers are compatible with the use of volatile

Peptides, Proteins / Bio-Polymers 100mM ammonium formate as the eluent solution, as shown in theexamples below.

Human SerumAmylase

Hemoglobin-Amylase (Bacillus sp.)

ELSD

ELSD

UV 280nm

UV 280nm

ELSD
UV 220nm
UV 280nm
0510100mM ammonium formate

15 min05872

Lactalbumin

min101505min

1015100mM ammonium formate

889

Casein0min

510155min

1015100mM ammonium formate

870

Ovalbumin

ELSD

ELSD
0510min

150510min

ELSD

150UV 280nm
5min

10100mM ammonium formate

150Myoglobinmin510015100mM ammonium formate

895

51015 min

51015 min

UV 280nm

UV 280nm

0100mM ammonium formate

894

Polyglutamic acid
HN

CHCH2CH2899

Lectin (Concanavalin A)
O

Concanavalin A(from Jack bean, 25.5 kDa)

CCHOO

ELSDn

poly-gamma-glutamic acid
(MW 1.5 - 2.5 MDa)051015 min

ELSD
UV 280nm

ELSD

00510100mM ammonium formate

51015 min05898

100mM ammonium formate

Proteoglycan(from nasal cartilage of salmon headca.450 kDa)

NH2O
NH

ELSD

CHCnSer

ELSD
0916

Proteoglycan
1 - 5 kDa

1015 min

100mM ammonium formate

901

Poly-L-Lysine

51015 minInsulin0051015 min

GclUA

GalNAc (6S)O-O
CH2O S OOO
HOO

15 min

O
O

OH

O

CH2OH
O
HOOO

OOOHXylGal

Gal

GclUA

HONH
O

CH2OH
OH
O OO

CH2CH
C OOH

OOOH
OH

OH
HNOH
O

UV 280nm
0UV 220nm
510100mM ammonium formate

15 min

8970n

ELSD
51015 min0100mM ammonium formate

Intrada SEC, 250x4.6mm, 0.3mL/min, UV or ELSD

3918

510100mM ammonium formate

15 min900

Pullulan

Mannan

1 000 000

CH2OH
O

CH2O

500 000

OH

CH2OH
O
OHOH

HOO

200 000

OH

OO

OH

Fucoidan

6Man 126Man 1Man 1226
Man 1Man 1Man 12263Man 1
Man 1
Man 1
Man 123633Man 1
Man 1
Man 1
Man 1
Man 122Man 1
Man 1

n100 000

6Man 1Mannanfrom Saccharomyces cerevisiae

HO50 000
SO3-MW
20 000

Fuc 110 000

4 Fuc 134 Fuc 1
33Fuc 1

SO3Fuc 1
42 Fuc133Fuc 1

Fucoidanfrom Fucus vesiculosus

5 000
2 000

RID

1 000
6ELSD
78910

ELSD

11 min0100mM ammonium formate

O
CH2O S OOOOHOO OO
COONH S OOHOOO S OOO-O S OOCOOOHOO-O S O

CH2OH
OOOO

O

O-O

OHnO

HN

O
O

OH

OHn10

COOO
O

CH2OH
OO

OH

Na+O

HN

OHnO

dermatan sulfate
chondroitinsulfate

909

HOO
O S OnOOH

15 minHyaluronic acid

CH2OO

O

nnheparin

O

5100mM ammonium formate

910

ELSD
O
-O S O
CH2OH
O
O

CH2O
O

015 min

Carrageenan
O-O S O
CH2OH
O
O

OO
OH

HN
OH

ELSD
CH2OH
O

10100mM ammonium formate

890

Glycosaminoglycan (GAGs)

5hyaluronic acid, sodium salt
from Rooster Comb-carageenan-carageenan

ELSD
510 min

510 min

5010 min

100mM ammonium formate

min1015 05min

10100mM ammonium formate

866

Lipopolysaccharide (LPS)

51505min1015100mM ammonium formate

907

908

20bp DNA Ladder

Dextran sulfate

500bp

Lipopolysaccharide
from E.coli O127

100bp
CH2

CH2

O
OSO3-O
OSO3-

OSO3-O
OSO3-

OSO3-

Na+200bp

O
OSO3.xNa+ n

400bp

dextran sulfate
sodium salt

20bp
180bp

300bp
160bp
140bp
120bp

ca.500 kDa

ELSD

UV 260nm

60bp 40bp
80bp

EDTAELSD

051015 min

100mM ammonium formate

0510917

10 min515 min

100mM ammonium formate

100mM ammonium formate

902867

Synthetic polymers Synthetic polymers have a wide range of polarities and solubilities, so it is necessary

to optimize both the sample solvent as well as the eluent, independently. Sample
solvent should have a high solubility for the polymer, and the eluent should have ahigher polarity than the sample, to ensure adequate elution.

Polylactide (PLA)

Polyvinylpyrrolidone (PVP)

poly DL-lactide(75 ~120 kDa)

Polyvinyl acetate (PVAc)
Vinyl Acetate, PolymerPolyvinylpyrrolidone K30
ca.40 kDa

H2CO

H2CNO
CH3CHCHOC OOnCH3nnn = 1400 - 1600

ELSD
0ELSD
510

acetonitrile / water = 80 / 2015 min

912

0ELSD
51015 min

100mM HCOONH4 / MeOH = 70 / 30 873

0510

THF (tetrahydrofuran)Intrada SEC, 250x4.6mm, 0.3mL/min, UV or ELSD
Imtakt USA (info@imtaktusa.com)
North America

Imtakt Corp./JAPAN (info@imtakt.com)
Other Countries

15 min874