Selecting a Pre-Packed Column
1.
Choose
the particle size and pore size
HYDROCELL media are available in 5 µm, 10 µm, 20 µm, 30 µm and 50 µm
particle size of macroporous media for routine ion exchange, hydrophobic
interaction, reversed phase and nucleic acid analysis and purification.
Hydrocell media are also produced nonporous polymeric beads for
high-speed analysis and process control.
|
Particle Size |
Pore Size |
Applications |
|
5 µm
|
500Å, 1500Å
|
High
resolution, Reversed Phase media for protein, polypeptide and
oligonucleotide analysis, and small-scale separation. |
|
10 µm
|
500Å, 1500Å
|
Ion Exchange,
Hydrophobic interaction and Reversed Phase media for analytical to
small-scale preparative |
|
10 µm
|
nonporous
|
Same as 10 µm
porous particle except producing from nonporous beads for high
speed analysis and preparation. |
|
20 µm
|
500Å, 1500 Å |
Same as 10 µm
porous particle except for high pressure semi-preparative to preparative applications |
|
30 µm
|
500Å, 1500 Å |
Same as 10 µm
porous particle except for medium and low pressure preparative and
process applications |
|
50 µm
|
500Å, 1500 Å |
Same as 10 µm
particles except for medium and low pressure preparative and process
applications |
2.
Select
the column geometry
HYDROCELL columns are available in several sizes.
We recommend 4.6 mm ID column for routine lab scale use. For
semi-preparative and preparative work choose a larger diameter 7.8 mm,
10 mm or 21.2 mm column. For small scale work or more sensitive
detection, choose a narrow-bore 1.0 or 2.1 mm ID column. All standard Hydrocell
1500, 3000 and NP10 series of columns are constructed of PEEK or
Stainless steel materials with 2 µm Titanium frit.
|
Scale of Separation |
Column Dimension |
Recommended Flow |
|
|
Diameter (mm) |
Length (cm) |
Volume (mL) |
Volumetric (mL/min) |
Line velocity
(cm/hr) |
|
Micro (µg) |
2.1 |
15 |
0.52 |
0.20-1.20 |
350-2100 |
|
Lab (µg to mg) |
4.6
|
15 |
2.50 |
1.00-6.00 |
360-2200 |
|
|
7.8
|
15 |
7.16 |
2.50-15.0 |
310-1860 |
|
Semi-Preparative (mg) |
10 |
15 |
11.50 |
4.00-24.0 |
310-1860 |
|
Preparative (mg
to grams) |
21.2 |
15 |
50.60 |
15.0 -90 |
255-1530 |
3.
Match
the Column to the System
Each pre-packed HYDROCELL 1500, 3000 or NP10 series of columns are
produced from 10 µm and 20 µm particle size with average pore size of
500Å, 1500 Å or nonporous PS-DVB polymeric beads for high pressure
systems.
HYDROCELL 30 and 50 series of columns are
designed for medium and low pressure preparative systems. Choose the
column that is suitable for your system environment. This will ensure
that the back pressure generated on the column will not exceed the
pressure limit of your system while operating under higher flow rates.
Fig. 1 -
Pressure/Flow Relationship of NP10 series, 150x4.6mm columns
Fig. 2 - Pressure/Flow Relationship of 1500 series, 150x4.6mm columns
Fig.
3 - Pressure/Flow Relationship of Reversed Phase RP 10G, 150X4.6 mm
column
4.
Choose
the HYDROCELL Chemistry
|
Product |
Functional Group |
Particle Size (µm) |
Pore Size (A) |
Dynamic
Loading Capacity
(column volume/mL) |
|
|
|
DEAE 1500 |
Diethylaminoethyl |
10 |
500 |
50 mg |
|
DEAE 3000 |
Diethylaminoethyl |
10 |
1500 |
45 mg |
|
DEAENP10
|
Diethylaminoethyl |
10 |
Non-porous |
35mg |
|
QA 1500 |
Diethylmethylaminoethyl |
10 |
500 |
50 mg |
|
QA 3000 |
Diethylmethylaminoethyl |
10 |
1500 |
45 mg |
|
QA NP10 |
Diethylmethylaminoethyl |
10 |
non-porous |
35 mg |
|
CM 1500 |
Carboxymethyl |
10 |
500 |
45 mg |
|
CM 3000 |
Carboxymethyl |
10 |
1500 |
40 mg |
|
CM NP10 |
Carboxymethyl |
10 |
non-porous |
25 mg |
|
SP 1500 |
Sulfopropyl
|
10 |
500 |
45 mg |
|
SP 3000 |
Sulfopropyl
|
10 |
1500 |
40 mg |
|
SP NP10 |
Sulfopropyl
|
10 |
non-porous |
25 mg |
|
NS 1500 |
Quaternized
Amine |
10 |
500 |
60 mg |
|
|
|
C3 1500 |
Allyl
|
10 |
500 |
30 mg |
|
C3 3000 |
Allyl |
10 |
1500 |
25 mg |
|
C3 NP10 |
Allyl |
10 |
non-porous |
15 mg |
|
C4 1500 |
Butyl |
10 |
500 |
30 mg |
|
C4 3000 |
Butyl |
10 |
1500 |
25 mg |
|
C4 NP10
|
Butyl |
10 |
non-porous |
15 mg |
|
Phenyl 1500 |
Phenyl |
10 |
500 |
35 mg |
|
Phenyl 3000
|
Phenyl |
10 |
1500 |
30 mg |
|
Phenyl NP10
|
Phenyl |
10 |
non-porous |
20 mg |
|
|
|
RP 5T |
Base PS-DVB
|
5 |
140 |
60 mg |
|
RP 5G |
PS-DVB with
coating |
5 |
500 |
45 mg |
|
RP 10G |
PS-DVB with
coating |
10 |
500 |
40 mg |
|
RP 5S |
Base PS-DVB
|
5 |
500 |
50 mg |
|
RP 10S |
Base
PS-DVB |
10 |
500 |
45 mg |
|
RP 5D |
Base PS-DVB
|
5 |
1500 |
45 mg |
|
RP 10D |
Base PS-DVB
|
10 |
1500 |
40 mg |
| |
|
|
|
|
|
5. Recommended Uses
Ion Exchange Chromatography
|
|
DEAE 1500 |
Weak anion
exchanger-- for PI<7 proteins, peptides and oligonucleotides – use
with pH range 3-9 |
|
DEAE 3000 |
|
DEAE NP10 |
Same as DEAE
1500 and 3000, but producing from nonporous beads for fast separation and process
monitoring |
|
QA
1500 |
Strong anion
exchanger-- for PI <7 biomolecules such as proteins, oligonucleotides-use
with pH range of 3-11 |
|
QA 3000 |
|
QA NP10 |
Same as QA 1500
and 3000, but producing from nonporous beads for fast separation and process
monitoring |
|
CM 1500 |
Weak cation
exchanger-- for PI>7 biomolecules such as proteins, polypeptides- use
with pH range 5-12 |
|
CM 3000 |
|
CM NP10 |
Same as CM 1500
and 3000 , but produced from nonporous particle to speed up
the analysis and purification |
|
SP 1500 |
Strong cation
exchanger-- for PI>7 biomolecules such as proteins, Polynucleotides-
use with pH range 3-12
|
|
SP
3000 |
|
SP NP10 |
Same as SP 1500
and 3000, but produced from nonporous particles to
increase permeability and sensitivity |
|
NS 1500 |
Strong anion
exchanger for analysis and small scale preparation of
oligonucleotides with less than 1000 base pair |
Hydrophobic Interaction Chromatography
|
|
C3 1500 |
Strong
hydrophobic proteins and peptides |
|
C3 3000 |
|
C3 NP10 |
|
C4 1500 |
General
hydrophobic proteins and peptides |
|
C4 3000 |
|
C4 NP10 |
|
Phenyl 1500 |
Weak
hydrophobic proteins and peptides |
|
Phenyl 3000 |
|
Phenyl NP10 |
Reversed-Phase Chromatography
|
|
RP 5T |
General
Reversed Phase Chromatography (RPC) of small drugs and organics |
|
RP 5S
|
General RPC of
small peptides, polypeptides, nucleotides and oligonucleotides |
|
RP 10S
|
|
RP 5G |
General RPC of
small proteins, polypeptides and oligonucleotides |
|
RP 10G |
|
RP 5D |
General RPC of
large proteins, polypeptides, polynucleotides and DNA fragments |
|
RP 10D |
|
