[ Care and Use ManUal ]
HsPgel Columns For HIgH-sPeed gPC analysIs
Contents
I. IntroduCtIon
II. HsPgel Column FamIlIes
III. ConneCtIng tHe Column to tHe gPC system
a. Column Connectors
b. Connecting an HSPgel Column
c. Solvents
IV. Column temPerature ranges
I. IntroduCtIon
Thank you for choosing a Waters HSPgel™ column. Every column
is individually tested and Certificates of Batch Analysis and a
Performance Chromatogram are provided with each column. It is not
recommended to use a guard column with the HSPgel columns, as it
will add significant volume and possible additional bandspreading
to the system. It is recommended, however, that a ~0.5 µm in-line
filter (pre-column) be installed to trap any particulates that may
potentially plug the column frit. The GPC analysis solvent must
be filtered prior to use through an appropriate ~0.5 µm filter. The
sample solutions should also be filtered prior to injection, unless the
sample molecular weight is very high and there may be a chance of
shear degradation.
HSPgel Columns for High Speed GPC Analysis 1
[ Care and Use ManUal ]
II. HsPgel Column FamIlIes
HSPgel columns are available in four offerings, three for organic GPC and
one for aqueous GPC. All columns have dimensions of 6.0 x 150 mm.
The descriptions of the columns are as follows:
• HSPgel HR series for room temperature, high resolution GPC
analysis of organic polymers. Shipped in THF and one solvent
changeover only.
• HSPgel RT series for routine room temperature GPC analysis
of organic polymers. Shipped in THF and multiple solvent
changeovers allowed.
• HSPgel HT series for GPC analysis of organic polymers from
room temperature to 180 °C. Shipped in either THF or ODCB and
may be transferred multiple times to other solvents (i.e., TCB at
0.15 m L /mi n) .
• HSPgel AQ series for room temperature analysis of aqueous
GPC analysis of water soluble polymers.
III. ConneCtIng tHe Column to tHe gPC system
Handle the column with care. Do not drop or hit the column on a hard surface
as it may disturb the bed and affect its performance.
1. Correct connection of 1/16-inch outer diameter stainless steel
tubing leading to and from the column is essential for high-quality
chromatographic results.
2. When using standard stainless-steel compression screw fittings, it is
important to ensure proper fit of the 1/16-inch outer diameter stain-
less steel tubing. When tightening or loosening the compression screw,
place a 5/16-inch wrench on the compression screw and a 3/8-inch
wrench on the hex head of the column endfitting.
Note: If one of the wrenches is placed on the column flat during this
process, the endfitting will be loosened and leak.
3. If a leak occurs between the stainless steel compression screw fitting
and the column endfitting, a new compression screw fitting, tubing and
ferrule must be assembled.
4. An arrow on the column identification label indicates correct direction
of solvent flow.
Table 1. Specifications for HSPgel Columns
Application Columns
Ultra-Hgh
Resolution
GPC
Room
Temperature
GPC
High
Temperature
GPC
Aqueous AQ 2.5 16,000 500- 2,000 4
* MW ranges base d on polystyrene c hain lengths for HR, RT and HT series, based on polyeth-
ylene oxide chain.
** Exclusion limits for AQ series extrapolated from highest MW polyethlene oxide standard
(~900,000 MW).
HR 1.0 18,000 100- 1,000 3
HR 2.0 18,000 500- 10,000 3
HR 2.5 18,000 1,000- 20,000 3
HR 3.0 18,000 2,000- 60,000 3
HR 4.0 18,000 10,000- 400,000 3
HR MB-L 18,000 500- 700,000 3
HR MB-M 16,000 1,000- 4,000,000 Mix of 3.5
HR MB-H 10,000 5,000- 10,000,000 10
RT 1.0 16,000 100- 1,000 3
RT 2.0 16,000 500- 10,000 3
RT 2.5 16,000 1,000- 20,000 3
RT 3.0 16,000 2,000- 60,000 3
RT 4.0 16,000 10,000- 400,000 3
RT 5.0 16,000 25,000- 4,000,000 3
RT 6.0 10,000 50,000- 10,000,000 3
RT 7.0 10,000 100,000- 15,000,000 3
RT MB-L 16,000 100- 1,000 3
RTMB-L/M 16,000 500- 400,000 3
RTMB-M 16,000 1,000- 4,000,000 3
RTMB-H 10,000 5,000- 10,000,000 Mix of 3.5
HT 1.0 12,000 100-1.000 5
HT 2.0 12,000 500- 10,000 5
HT 2.5 12,000 1,000- 20,000 5
HT 3.0 12,000 2,000- 60,000 5
HT 4.0 12,000 10,000- 400,000 5w
HT 5.0 12,000 25,000- 4,000,000 5
HT 6.0 8,000 50,000- 10,000,000 5
HT 7.0 8,000 100,000- 15,000,000 5
HT MB-L 12,000 100- 1,000 5
HTMB-L/M 12,000 500- 400,000 5
HTMB-M 12,000 1,000- 4,000,000 5
HTMB-H 8,000 5,000- 10,000,000 5
AQ 3.0 16,000 1,000- 60,000 4
AQ 4.0 10,000 10,000- 400,000 6
AQ 5.0 10,000 50,000- 4,000,000** 7
AQ 6.0 7,000 100,000-
AQ MB-H 7,000 500- 10,000,000** 9
Theoretical
Plates
MW Range*
10,000,000**
Particle
Size (µm)
9
HSPgel Columns for High Speed GPC Analysis 2
[ Care and Use ManUal ]
a. Column Connectors
Due to the absence of an industry standard, various column manufacturers use
different styles of chromatographic column connectors. The chromatographic
performance of your separation can be negatively affected if the style of your
column endfittings do not match the existing instrumentation tubing ferrule
setting. This section explains the difference between Waters style and Parker
style endfittings. HSPgel columns have Parker style endfittings.
The two styles of endfittings differ by the length of tubing that protrudes
from the ferrule. Parker style endfittings (e.g., HSPgel columns) require a
®
0.090-inch ferrule depth. Waters style endfittings (e.g., Styragel
0.130-inch ferrule depth. Examples of each endfitting are given below.
0.090 inches
Parker Ferrule Setting Waters Ferrule Setting
Figure 1. Ferrule settings.
) require a
b. Connecting an HSPgel Column
HSPgel columns have Parker style endfittings that require a 0.090-inch
ferrule depth. If tubing with a Waters style ferrule is connected to an HSPgel
column, the end of the tubing will bottom out before the ferrule reaches its
proper seating position and cause a leak. An example of this is given below.
Gap
Figure 3. HSPgel column connection.
As mentioned above, connecting a Waters style connector to an HSP-
gel column will leave a gap around the ferrule and create a leak. There
are two ways to correct this problem:
1. Tighten the screw a bit more. The ferrule will move forward, and
reach the sealing surface. Care must be taken not to overtighten
the connection as this may break the screw.
In a proper tubing/column connection the tubing touches the bottom of
the column endfitting with no void between them.
Figure 2. P roper tubing/column connection.
2. Cut the tubing, place a new ferrule on it and re-make the connection.
HSPgel Columns for High Speed GPC Analysis 3
[ Care and Use ManUal ]
Table 2. Flow-Rate Specifications for HSPgel Columns
Application Columns
Ultra-Hgh
Resolution
GPC
Room
Temperature
GPC
High
Temperatire
GPC
Aqueous AQ 2.5 0.30- 0.60 0.70 415
HR 1.0 0.25- 0.60 0.70 380
HR 2.0 0.25- 0.60 0.70 345
HR 2.5 0.25- 0.60 0.70 275
HR 3.0 0.25- 0.60 0.70 240
HR 4.0 0.25- 0.60 0.70 240
HR MB-L 0.25- 0.60 0.70 240
HR MB-M 0.25- 0.60 0.70 140
HR MB-H 0.25- 0.60 0.70 70
RT 1.0 0.30- 0.70 0.80 480
RT 2.0 0.30- 0.70 0.80 480
RT 2.5 0.30- 0.70 0.80 415
RT 3.0 0.30- 0.70 0.80 415
RT 4.0 0.30- 0.70 0.80 275
RT 5.0 0.30- 0.70 0.80 275
RT 6.0 0.30- 0.70 0.80 210
RT 7.0 0.30- 0.70 0.80 210
RT MB-L 0.30- 0.70 0.80 275
RTMB-L/M 0.30- 0.70 0.80 275
RTMB-M 0.30- 0.70 0.80 275
RTMB-H 0.30- 0.70 0.80 275
HT 1.0 0.30- 0.70 0.80 415
HT 2.0 0.30- 0.70 0.80 415
HT 2.5 0.30- 0.70 0.80 415
HT 3.0 0.30- 0.70 0.80 415
HT 4.0 0.30- 0.70 0.80 415
HT 5.0 0.30- 0.70 0.80 415
HT 6.0 0.30- 0.70 0.80 415
HT 7.0 0.30- 0.70 0.80 415
HT MB-L 0.30- 0.70 0.80 415
HTMB-L/M 0.30- 0.70 0.80 415
HTMB-M 0.30- 0.70 0.80 415
HTMB-H 0.30- 0.70 0.80 415
AQ 3.0 0.30- 0.60 0.70 415
AQ 4.0 0.30- 0.60 0.70 245
AQ 5.0 0.30- 0.60 0.70 210
AQ 6.0 0.30- 0.60 0.70 140
AQ MB-H 0.30- 0.60 0.70 140
Recommended
Flow Rate**
Maximum
Flow Rate
Maximum
Pressure
Dro p**
c. Solvents
To maintain maximum column performance, use high-quality chromatography
grade solvents. It is recommended to degas all solvents thoroughly before use
to prevent bubble formation in the pump and detector. The use of an on-line
degassing unit is also recommended. This will help to maintain a stable RI
detector baseline
Table 3. Availible Solvents for HSPgel Columns
Column
Type
HR* Tetrahydrofuran Toluene, xylene, chloroform dichlromethane,
RT** Tetrahydrofuran Toluene, xylene, chloroform, dichlromethane,
HT*** THF or ODCB Same as RT, and also: ODC B, TC B,
AQ**** Water Water with salts /buffe rs up to 0.80M,
* HR columns may be switched from T HF to one of the listed solvents one time only -
at 0.10 mL/minute.
** RT columns may be switched back and fort h to the listed solvents at the flow rates shown
in Table 4.
*** HT columns are shipped in either THF or ODC B. For work in TC B, the ODCB columns may
be directly switched over at 0.15 mL/min at >100 °C.
**** AQ columns may be used with up to 50% organic loading (methanol or acetonitrile).
Table 4. Recommended Flow Rates for Solvent Switchover
Solvent
Toluene 0.30 0.30 0.59 (20 °C)
Dichloromethane 0.30 0.30 0.43 (20 °C)
Chloroform 0.30 0.30 0.56 (20 °C)
DMF/DMAC 0.20 0.25 0.7 - 0.8 (25 °C)
DMSO 0.10 0.15 2.0 (25 °C)
NMP 0.10 0.15 1.65 (25 °C)
ODCB/TCB 0.10 0.15 1.3 - 1.5 (25 °C)
HFIP 0.05 0.05 1.32 (25 °C)
1-chloronapthalene – 0.05 2.94 (20 °C)
THF – – 0.55 (20 °C)
* (cp) - centiopise
.
Shipment Solvent Chargeable Solvent
(methylene chloride), dichloroethane
(methylene chloride), dichloroethane,
dimethylfomamide, dimethylacetamide,
n-methyl pyrrolidine hexafluoroisopropanol,
(HFIP), DMSO
o-chlorophenol, 1-chloronapthalene
pH 2 -12, 50% methanol or acetonitrile
RT Flow Rate
(mL /min)
HT Flow Rate
(mL /min)
Viscosity (cp) *
(at temperature
shown)
* Flow rates in mL/min.
** Pressure drops in PSI. Increasing column temperature can dramatically decrease.
HSPgel Columns for High Speed GPC Analysis 4
[ Care and Use ManUal ]
IV. Column t emPeratur e ranges
HR Series: Ambient – 45 °C
RT Series: Ambient – 85 °C
HT Series: Ambient – 180 °C
AQ Series: Ambient – 80 °C
Recommended maximum injection volume is 20 μL for each column type
– 1 μL typical.
Sales Offices:
Austria and European Export
(Central South Eastern Europe, CIS
and Middle East) 43 1 877 18 07
Australia 61 2 9933 1777
Belgium 32 2 726 1000
Brazil 55 11 4134 3788
Canada 1 800 252 4752
China 86 21 6156 2666
Czech Republic 420 2 617 11384
Denmark 45 46 59 8080
Finland 358 9 5659 6288
France 33 1 30 48 72 00
Germany 49 6196 400 600
Hong Kong 852 2964 1800
Hungary 36 1 350 5086
India 91 80 2837 1900
Ireland 353 1 448 1500
Italy 39 02 265 0983
Korea 82 2 6300 4800
Mexico 52 55 52 00 1860
The Netherlands 31 76 508 7200
Norway 47 6 384 6050
Poland 48 22 101 5900
Puerto Rico 1 787 747 8445
Russia/CIS 7 495 727 4490/ 290 9737
Singapore 65 6593 7100
Spain 34 93 600 9300
Sweden 46 8 555 115 00
Switzerland 41 56 676 7000
Taiwan 886 2 2501 9928
United Kingdom 44 208 238 6100
All other countries:
Waters Corporation U.S.A.
1 508 478 2000
1 800 252 4752
www.waters.com
©2012 Waters Corporation. Waters and Styragel are registered
trademarks of Waters Corporation. HSPgel and The Science of
What’s Possible are trademarks of Waters Corporation. All other
trademarks are the property of their respective owners.
June 2012 736000797 Rev D VW-P DF
HSPgel Columns for High Speed GPC Analysis 5
Japan 81 3 3471 7191
Waters Corporation
34 Maple Street
Milford, MA 01757 U.S.A.
T: 1 508 478 2000
F: 1 508 872 1990
www.waters.com
Loading...