Instruction Manual
For High Performance Liquid Chromatography
COSMOSIL PACKED COLUMN
| INTRODUCTION |
Thank
you for purchasing our
COSMOSIL Packed Column. Ensuring
maximum efficiency and long column life, we ask you to
read this manual carefully.
COSMOSIL Packed Columns are made of stainless steel and
packed with totally porous spherical silica-based materials.
Our COSMOSIL series covers most of the conventional and
special applications of normal phase chromatography, reversed
phase chromatography and available for both analytical
and preparative separation purposes. For the choice most
suitable for your task consult with our distributor or
contact directly Nacalai Tesque. |
| CARE
AND USE |
| |
1. |
Avoid
mechanical shock. |
| |
2. |
Connect
the column according to the flow direction indicated on
the label. |
| |
3 |
Let
through the column 20-30ml mobile phase before connecting
to the detector. |
| |
4. |
Use
scrupulously deaired mobile phase. Air bubbles generate
detection noise and accelerate column deterioration. |
| |
5. |
Use
only HPLC grade solvents. |
| |
6. |
Avoid
precipitation in the column. Check the solvent constitution
on the enclosed inspection record. |
| |
7. |
Keep
the pH of the mobile phase within the range of 2-7.5.
Buffer concentration is usually sufficient within the
range of 0.005-0.02M. Pass mobile phase through membrane
filter (less than 0.5?m in pore size) before use. When
using trifluoroacetic acid keep the concentration under
0.1%. |
| |
8. |
Keep
pressure under 200kgf/cm2. Take special care when using
highly viscous mobile phase. |
| |
9. |
After
performing reversed phase chromatography, first wash the
column with acid- and/or salt-free solvent, then with
acetonitrile or methanol. Store it tightly plugged. |
| |
10. |
After
normal phase chromatography, change solvent to halogen
free, non-polar (n-hexane or n-heptane). Keep the column
tightly plugged. |
| |
11. |
Filter
the sample before injection. Avoid precipitation at injection. |
| |
12. |
Removal
of the end filters or change of the end-fittings will
result in low performance of the column. |
| |
13. |
Do not tighten nuts more firmly than necessary. |
| |
14. |
Establish
perfect chromatography conditions by experimenting with
an analytical column of the same packing material before
employing a preparative column. Pay attention to impurities
without UV absorption or with longer retention time than
your sample. |
FOR
THE BEST PERFORMANCE OF COSMOSIL PACKED COLUMNS |
| |
1. |
In
order to maximize the column performance, minimize the
dead volume of mobile phase in the equipment by shortening
and/or narrowing the width of tubing. |
| |
2. |
Maintain
constant column and tubing temperature. |
| |
3. |
Avoid
injecting air, changing flow rate rapidly and changing
mobile phase at high flow rate. |
| |
4. |
Insoluble
matters from the pumping system, mobile phases, or samples
trapped in the filter (2?m) at the inlet of the column
may increase the pressure. Placing a 1?m prefilter between
the injector and the column is highly recommended to prevent
column clogging. |
| |
5. |
Use
guard column to protect both analytical and preparative
type columns from irreversible adsorption on the packing
material. Guard columns extend the lifetime of your valuable
analytical and preparative columns. |
| |
6. |
In
normal phase chromatography pay attention to the change
of retention time depending on the water content of the
mobile phase or at injection of large amount of polar
solvents. |
| |
7. |
We
recommend keeping the chromatography conditions constant,
since frequent changes of mobile phase shorten column
life. |
| TROUBLESHOOTING |
Trouble |
Cause |
Solution |
| increase
of pressure |
clogging
of the end filter
clogging of the packing material
precipitation in the column |
(1)
and/or (2)
(1)
(3) |
| poor
resolution |
contamination
of packing material
disorder of packing material |
(3)
unregenerable |
| split
peak |
void
in the column |
unregenerable |
unstable
baseline
(COSMOSIL PYE, NPE, Ph and PE type column) |
(4) |
|
| SOLUTION: |
| |
1. |
Disconnect
column from the detector. Let mobile phase through the
column in reverse direction at 0.5ml/min flow rate for
30 min. |
| |
2. |
Wash
the end filter or replace it with a new one. |
| |
3. |
Wash
the column with a solvent capable of dissolving the contaminants.
The column can be washed, if necessary, with water, acetonitrile,
methanol, ethanol, 1-propanol, 2-propanol, tetrahydrofuran,
chloroform, n-hexane and n-heptane. |
| |
4. |
Let
through the column approximately 10ml tetrahydrofuran. |
| PERFORMANCE
GUARANTEE |
The
strict quality control system of Nacalai Tesque supports
the customers with an individual "Inspection Report"
which accompanies each and every COSMOSIL and COSMOGEL
Packed Column (except guard columns) and an additional
"Certificate of Analysis" for the new COSMOSIL
5C18 MS-II and 5C18 AR-II (4.6mmID x150mm, 4.6mmID x
250mm). |
| INSPECTION
RECORD |
Contains
data of number of theoretical plates (N), peak asymmetry
(s), capacity factor (k') and separation factor (α).
| N: |
theoretical
plate number |
| TR: |
retention
time |
| W1/2: |
peak
half width |
| h: |
peak
height |
| s: |
peak
asymmetry |
| k': |
capacity
factor |
| α: |
separation
factor |
|
| CERTIFICATE
OF ANALYSIS |
| Validate
terms of the physical properties of the silica gel, the
carbon content, polar selectivity, hydrophobicity, silanol
capacity steric selectivity, inactive degree to basic
and chelating compounds. |
| I.
WARRANTY |
Nacalai
Tesque will change defective columns reported within
2 weeks of receipt. Nacalai Tesque approves return in
case of:
(1) Damage during the transportation caused by our incomplete
packing.
(2) Theoretical plate number measured according to the
test method specified in the Inspection Report is significantly
lower than guaranteed. (Please note that the plate number
decreases when using apparatus with large dead volume
or injecting big amount of sample.)
We cannot accept claims for deterioration of column
performance caused by taking off the end filters or
end-fittings, or long shelf life. Return shipment is
unacceptable unless we have given prior permission and
shipping instructions. |
Your
comments and questions are very welcome.
Email comments or question to gnow.info@sesres.com
|
