YMC Pilot Series User manual

Manual
Low pressure chromatography column
Pilot series
These instructions should be read prior to using the column

2Low pressure chromatography column - Pilot series
Introduction
Operating instructions
This guide enables the safe and efficient use of the low pressure Pilot chromato-
graphy columns. The manual is part of the product and must be kept accessible for
the user at all times.
The user must read this manual carefully and have understood its content before
attempting to use the column. A prerequisite for the safe operation is the compli-
ance with all specified safety and operating instructions in this manual. The safety
instructions are described in Section 1. In addition, the local occupational health
and safety and general safety regulations apply to the use of the product.
The following laws and standards were taken into account in the creation of this
manual:
• 2001/95/EG
• DIN EN 82 079-1
• VDI 4500 Blatt 1
The terms and conditions of YMC Europe GmbH apply.
CE certification
This product meets all requirements of applicable CE directives.
Copyright protection
The contents of this manual are protected by copyright. Their use is allowed only in
connection with the use of the product. Any further use is not permitted without the
written permission of YMC Europe GmbH.
How to contact YMC
YMC Europe GmbH
Schöttmannshof 19
46539 Dinslaken
Phone: [phone]
Fax: [phone]
Website: www.ymc.de
E-Mail: [email protected]
Translated operating instructions
Version: 2.0 (TAC_2014_06_03_V2.0_de)
Stand: 03.06.2014

3
Low pressure chromatography column - Pilot series
Contents
1. Safety............................................................................................................... 5
1.1. Explanation of warning messages............................................................ 5
1.2. Layout of instructions ............................................................................... 6
1.3. Fields of applications................................................................................ 6
1.4. Intended use............................................................................................. 7
1.5. Personnel requirements............................................................................ 7
1.6. Personal protective equipment ................................................................ 7
2. Delivery............................................................................................................. 8
2.1. Delivery contents...................................................................................... 8
2.2. Delivery inspection ................................................................................. 10
3. Starting operations ...................................................................................... 11
3.1. Safety instructions for use...................................................................... 11
3.2. Column construction .............................................................................. 12
3.3. Specifications ......................................................................................... 14
3.4. Chemical resistance ............................................................................... 15
3.5. General operations ................................................................................. 17
3.5.1. Tightening and loosening of the O-ring........................................ 17
3.5.2. Adjusting the stamp height........................................................... 17
3.5.3. Connecting the tubing.................................................................. 17
3.6. Dismantling............................................................................................. 18
3.6.1 Dismantling of the column............................................................. 18
3.6.2 Dismantling of the glass body ....................................................... 19
3.6.3 Dismantling of the plunger ............................................................ 20
3.6.4 Removing of the frit ....................................................................... 21
3.6.5 Dismantling of the fixed stamp...................................................... 21
3.7. Cleaning.................................................................................................. 22
3.8. Assembling............................................................................................. 23
3.8.1. Assembling the frit........................................................................ 23
3.8.2. Assembling the plunger................................................................ 23
3.8.3. Assembling the glass body .......................................................... 24
3.8.4. Assembling the column ................................................................ 26
3.9. Pressure test........................................................................................... 27
3.10. Packing adapter ..................................................................................... 29
Contents

4Low pressure chromatography column - Pilot series
Contents
Contents
4. Operation ...................................................................................................... 30
4.1. Notes on operation................................................................................. 30
4.2. Column packing ..................................................................................... 32
4.2.1. Column preparation...................................................................... 32
4.2.2. Slurry production.......................................................................... 33
4.2.3. Column packing with applied flow ............................................... 34
4.2.4. Column packing by sedimentation............................................... 37
4.3. Notes for Cleaning-In-Place (CIP) .......................................................... 39
4.4. Unpacking .............................................................................................. 40
4.4.1. Procedure when moving the upper piston ................................... 40
4.4.2. Procedure when using liquid flow ................................................ 40
5. Maintenance and care ................................................................................ 41
5.1. Maintenance plan .................................................................................. 41
5.2. Replacement of wearing parts ............................................................... 42
6. Troubleshooting............................................................................................ 42
6.1. Fault table............................................................................................... 43
7. Storage, packing and transport.................................................................. 44
7.1. Storage ................................................................................................... 44
7.2. Packing and transport ............................................................................ 44
8. Disposal......................................................................................................... 44
9. Other.............................................................................................................. 45
9.1. Available column versions ...................................................................... 45
10. You might also be interested in .................................................................. 46
11. Index .............................................................................................................. 48
12. Annex............................................................................................................. 49

5
Low pressure chromatography column - Pilot series
Safety
1. Safety
This section gives an overview of all the important safety aspects for the protec-
tion of persons and for the safe and trouble-free handling of the Pilot column.
Other task-specific safety requirements are contained in the sections on the res-
pective operator. In addition, the national legal and other applicable safety regula-
tions must be observed.
1.1. Explanation of warning messages
Warnings are highlighted by coloured signal word fields. Always read the full text of
the warning in order to effectively protect yourself from danger!
The following signal word fields marked by different colors and signal words distinct
security levels:
DANGER
Disregarding this warning will result in death or serious injuries.
WARNING
Disregarding this warning may result in death or serious injuries.
CAREFUL
Disregarding this warning may result in minor or moderate injuries.
ATTENTION
Disregarding this warning can result in damage to surrounding property.
Warnings are always in the same structure. They include signal word, type and
source of danger, consequences of neglect and security measures / avoidance.

6Low pressure chromatography column - Pilot series
1.2. Layout of instructions
The instructions relate directly to an activity. They are structured in a task oriented
manner. Always perform the individual steps in the order in which they are given.
Instructions are structured as follows and are marked by the corresponding symbols:
1.3. Fields of applications
Pilot columns are especially made for pilot scale applications. As they are made
from high quality, inert materials they can be used in normal phase and reversed
phase mode, depending on the actual specifications quoted. They are often used
for biochromatography separations with appropriate YMC stationary phases. This
ensures the best requirements for a high recovery of the analytes. The properties of
the biomolecules (for example proteins, oligosaccharides or nucleic acids) remain
unchanged. The columns are easy to handle and are flexible in the method of use.
Due to the wide portfolio of column dimensions, frit porosities and accessories
different applications may exceed the specification of standard Pilot columns. As
a consequence YMC offers custom made solutions with many possible options.
Pilot columns require the highest demands for the professional use in pilot scale.
The focus is good handling, mechanic stability and high flow rates for high efficien-
cy and economy.
Safety
► Aim of the instruction
1. Action step
üImpact of the action step to check whether the step has been
performed correctly.
2. Another action step
þResult of the entire instruction

7
Low pressure chromatography column - Pilot series
1.4. Intended use
Pilot columns are designed as low pressure chromatography columns. They are
only to be used for the separation of substances via chromatographic techniques.
The intended use of this product must be performed with regards to the specifica-
tions, ►chapter 3.3, page 14. At all time the product must be used within the
specifications. Every use which is outside this specifications is decelerated as not
intended use. The intended use includes the compliance to the advice and terms
in the manual.
The product is only for the use with liquids that have a pressure <0.5 bar above
the atmospheric pressure at the maximum temperature of 40 °C.
1.5. Personnel requirements
The various tasks described in this guide make different demands on the qualifica-
tions of the persons carrying out.
The user must
• have competent and proficient training
• be trained in dealing with pressure devices
• read and understand these instructions before use of Pilot columns.
1.6. Personal protective equipment
To reduce the risks of working on the column, always use the recommended per-
sonal protective equipment.
The following personal protective equipment is required.
Use protective gloves!
Wear safety goggles!
Safety

8Low pressure chromatography column - Pilot series
2. Delivery
2.1. Delivery contents
The pilot column is shipped in a wooden box.
Please check the column description on the
glass body with the delivery note. All of the de-
tails must match. Please contact YMC Europe
GmbH if it is not the case.
Picture 2.2: Example of a column description
Pilot columns are shipped as pre-assembled columns. The manual, the certificate
of pressure resistance, information material and if requested FDA certificates are
delivered with every Pilot column. Based on the label on the column it is always
possible to determine the column version via the product code and to identify the
column via serial number. Therefore you can get individual support if needed. The
product code includes information of inner diameter, length, frit porosity and the
solvent resistance. It also includes special specifications including custom made
versions, FDA certification or heating/cooling jacket. The product code is generated
via a combination of these parameters.
An overview of this information and also an example of generating the product code
is shown at the following table. The product code (PI100/500V0E-AB-FDA) includes
the information for a Pilot glass column: 100 mm inner diameter, 500 mm length, a
frit porosity of 10 µm, solvent resistance supplied with FDA certification.
Delivery
i
Co. No.: PI100/500V0E-AB-FDA
Ser.-No.: PI123456
ID: 100 x 500 mm
max. Pressure: 7 bar
Picture 2.1: Wooden box with the Pilot column

9
Low pressure chromatography column - Pilot series
Delivery
Table 1: column versions and part numbers
combinations
PI100/
PI140/
PI200/
PI300/
PI450/
500V
850V
0
(10 µm)
E
EPDM
K
Kalrez®
-AB
(aqueous
buffer)
-SR
(solvent
resistant)
-FDA
(FDA certified)
-T
(heating/cooling
jacket)
-SO
(custom made)
-SG
(rotatable column)
example
Co. No.
(part number)
PI100/ 500V 0 E -AB -FDA
ID 100 mm
column length
(glass body) 500 mm
frit porosity 10 µm
O-ring EPDM
type aqueous
buffer
feature FDA
certified
For custom made Pilot columns please contact YMC Europe GmbH.
Contact details:
YMC Europe GmbH
Schöttmannshof 19
46539 Dinslaken
Phone: [phone]
Fax: [phone]
E-Mail: [email protected]
Check list delivery contents:
• Pilot column
• Folder includes:
- Manual
- Spare parts lists
- Test certificate
- Certificate of quality control
- Material certificates (FDA*, TSE/BSE*)
*if ordered

10 Low pressure chromatography column - Pilot series
Delivery
2.2. Delivery inspection
Delivery inspection
uProcedure:
1. Comparison of the column description on the glass body with the data of
the delivery note.
2. Checking of the completeness of the delivery with the help of “check list
delivery contents” uchapter 2.1, page 9.
3. Verifying the delivery of visible transport damage.
Transport damages
uProcedure, if you recognise visible transport damage:
1. Refusal or reserved acceptance of the delivery.
2. Record the transport damage on the delivery note of the transporter or in
the transport documents.
3. Inform YMC Europe GmbH and/or the responsive customer consultant.
i
It is recommended to retain and store the original packing and the foam cushion so
that the column can always be shipped safely.

11
Low pressure chromatography column - Pilot series
Starting operations
3. Starting operations
3.1. Safety instructions for use
WARNING
Use correctly matched protective clothing when working with the
glass column.
WARNING
Do not put the glass column under gas pressure.
CAREFUL
Operate the column only with pressures within the stated specifica-
tions. Otherwise there is a risk of injury and material damage.
CAREFUL
The temperature range which is used may not exceed/fall below the
given specifications.
WARNING
Make sure that the wheel brakes are activated during the entire ope-
ration or storage time.
ATTENTION
Move the column with extreme caution in order to avoid the column
falling over.
ATTENTION
Make sure that the fluids used are always clean and free from particles,
as this can lead to blockages.
i
Use column only with chemicals that are proven applicable with the wetted materials.

12 Low pressure chromatography column - Pilot series
Starting operations
3.2. Column construction
Picture 3.1: Detailed drawing Pilot column
(Picture can differ from product supplied)
1: glassbody
2: rotatable rack
3: adjustment knob
4: internal spindel
5: external spindel
6: nut of internal spindel
7: nut of external spindel
8: column seal actuation nut
9: lock screw flex
10: lock screw fix
11: piston
12: force plug
13: Frit
42: Lifting eye bolt
43: Locking pin

13
Low pressure chromatography column - Pilot series
Starting operations
Pilot columns have a glass (or, on request, acrylic) body (1) which stands on a ro-
tatable rack (2). These columns can be rotated through 180 ° which ensures sim-
ple unpacking. There are pistons inside the column at the bottom and top of the
column. The construction of these pistons is similar to each other. The lower pis-
ton is shorter than the upper one and has no adjustment knob (3). The pistons
consist of an inner spindle (4) and an outer spindle (5) with inlet or outlet. Further-
more, here are the two terminal nuts (6/7) and the tension nut (8) located. Via the
end plate (9/10), the piston is connected to the column body. At the other end of
the piston is the centrepiece of the column: the distributor plate (25) with the
plunger (11). The plunger is attached to the the force plug (12) and the frit (13) but
is separated from the inner column.

14 Low pressure chromatography column - Pilot series
3.3. Specifications
AB-version for aqueous buffer
Temperature range: 4 - 40 °C
Column body: borosilicate glass 3.3, calibrated precision glass (Duran KPG®)
Frits: polyethylene 10 µm
Sealings: EPDM O-rings
Connection: FEP or polyethylene tubing AD ¼“
Wetted parts: borosilicate glass 3.3, EPDM, PVDF, PE, FEP
SR-version resistant against organic solvents
Temperature range: 4 - 40 °C
Column body: borosilicate glass 3.3, calibrated precision glass (Duran KPG®)
Frits: stainless steel 10 µm
Sealings: Kalrez®O-rings
Connection: stainless steel capillaries AD ¼“
Wetted parts: borosilicate glass 3.3, Kalrez®, stainless steel, PTFE
ID
[mm] Version
Glass
length
[mm]
Pressure
limit
[bar]
Bed length
[mm]
Volume
[l]
Total
weight
[kg]
Total
diameter
[mm]
Height
[mm]
300
AB
500 3 0 - 410 0 - 28 80 920 1430 - 1860
850 3 330 - 760 22,8 - 52 95 920 1780 - 2210
SR
500 3 70 - 430 4,8 - 29,5 129 920 1450 - 1810
850 3 420 - 780 29 - 53 135 920 1800 - 2160
450
AB
500 2 20 - 440 3,2 - 68 115 990 1450 - 1900
850 2 370 - 790 58 - 123 129 990 1800 - 2250
SR
500 — — — — — —
850 — — — — — —
Table 2: column specifications of different versions/dimensions
Starting operations

15
Low pressure chromatography column - Pilot series
Table 3: Used materials
3.4. Chemical resistance
The chemical resistance depends on the wetted materials used. From the resis-
tance of the respective materials the chemical resistance of the column can be
concluded. The materials used can be determined from the end of the part num-
ber and from tables 3 and 4.
It is generally recommended to avoid the following chemicals:
• Strong oxidizing agents
• Halogenated compounds
• Chlorinated solvents
• Aromatic compounds (e.g., toluene)
• High concentrations of strong acids
• THF
• SR version: salts in conjunction with extreme pH values, use of NaCl or NaBr
The wetted parts of the AB series Pilot column are completely free of metals. As a result,
most saline buffer solutions can be used without hesitation.
i
The aqueous buffer (AB) version has only limited resistance to organic solvents. Before
selecting organic solvents for use with AB series Pilot columns, please check the resis-
tance of the materials used to the corresponding solvent (see table 4, page 16).
i
When using solvent mixtures, the resistance may differ significantly from those of the
pure solvent.
i
Such statements are based on our current technical knowledge and experience as well
as on existing literature and the information from the commodity producers. These do
not exempt the users of our products from their own testing. Certain features or appli-
cations can be derived either expressly or by implication from our records.
i
Starting operations
PVDF PTFE Stainless
steel EPDM Kalrez PE FEP PA/PEEK
AB
(aqueous buffer) ✓ ✓ ✓ ✓ ✓ ✓
SR
(solvent resistance) ✓ ✓ ✓ ✓

16 Low pressure chromatography column - Pilot series
Table 4: Chemical resistance of the materials used at room temperature and 1 bar
If the desired solvent is not listed, we are pleased to help.
Contact details: YMC Europe GmbH, Schöttmannshof 19, 46539 Dinslaken
Phone: [phone], Fax: [phone], Website: www.ymc.de, E-Mail: [email protected]
i
Starting operations
Solvent PVDF PTFE Stainless
steel EPDM Kal- rez®PE FEP POM PEEK PA
Acetone limited good good good good good good limited good good
Acetonitrile limited good good unstable good good good unstable good good
Ammonium
dihydrogen
phosphate
good good good good good good good good good good
Cyclohexane good good good unstable good limited good good good good
Dichlormethane limited good good unstable good unstable good unstable limited unstable
EDTA good good good good good good good good good good
Acetic acid (5%) good good good good good good good limited good unstable
Ethanol (96%) good good good good good good good good good good
Ethyl acetate limited good good eing. good eing. good good good good
n-hexane good good good unstable good eing. good good good good
Isopropanol good good good good good good good good good good
Methanol good good good good good good good good good good
NaOH (1M) good good unstable limited good good good good good good
HCl (1M) good good unstable limited good good good unstable good unstable

17
Low pressure chromatography column - Pilot series
3.5. General operations
3.5.1. Tightening and loosening of the
O-ring
uTightening and loosening of the O-ring:
1. In order to tighten the O-ring (14), hold the in-
ternal spindle screw nut (6) firmly with one
hand and turn the column seal actuation nut
(8) clockwise towards the outer spindle (5),
until the O-ring completely touches the glass
body.
2. To loosen the O-ring (14), turn the column seal
actuation nut (8) counter-clockwise until the
O-ring (14) ) is completely free from contact
with the glass.
3.5.2. Adjusting the piston height
uAdjusting the piston height:
1. Loosen the O-ring (14) of the upper piston. To
adjust the piston hold the external spindle
screw nut (7) and turn the adjustment knob (3).
2. Once the height has been set, the O-ring (14)
must be tightened again. uchapter 3.5.1
3.5.3. Connecting the tubing
uConnecting the tubing:
1. Drive the screws, please consider torque (table below).
2. Cut pipe at right angles.
3. Slide union nut, cutting and sealing ring over the pipe end as shown.
4. Insert pipe end until it stops.
5. Screw on the union nut by hand, then tighten it gently with an appropriate
hex driver.
Synthetic
material torque
G 1/8 inch G 1/4 inch G 3/8 inch G 1/2 inch G 3/4 inch
PP 1.0 - 1.5 Nm 2.5 Nm 5.0 - 6.0 Nm 5.0 Nm 10.0 Nm
PVDF 1.0 - 2.0 Nm 5.0 Nm 5.0 - 7.0 Nm 10.0 Nm 15.0 Nm
PFA 0.3 Nm 0.5 Nm 1.0 Nm 2.5 Nm 5.0 Nm
PTFE 0.3 Nm 0.5 Nm 1.0 Nm 2.5 Nm 5.0 Nm
Starting operations
screw-in sealing ring
cutting ring
screw cap tubing
open
close
down
up
3: adjustment knob
5: external spindel
6: nut of internal spindel
7: nut of external spindel
8: column seal actuation nut
14: o-ring for piston

18 Low pressure chromatography column - Pilot series
3.6. Dismantling
i
3.6.1 Dismantling the column
uDismantling the column:
1. Remove the hexagon socket screws M6 (15) at the
upper end plate (9), holding the plunger in position.
2. Lift the plunger from the glass body (1) and put it with
the frit (13) down on a clean, padded surface.
In a column with rotating frame turn around the column
and remove the second plunger as described in points
1-2.
5. Now the O-ring (14), can be loosen carefully. Remove the plunger and put it with
the frit down on a clean, padded surface.
3. Tighten the O-ring (14) of the lower plunger uchapter 3.5.1, page 17.
4. Remove the screws (15) on the lower end plate (9), which keep the lower the plun-
ger in position.
i
Do not try to force the plunger out of the column
body. If the plunger does not move immediately, wet
the O-ring with a little isopropanol. This ensures less
friction and the plunger should be released easily
from the glass body.
i
With a column without removable lower plunger omit points 3 to. 5
i
Make sure that the piston will not fall out. This could damage and the assembled
tubing.
Starting operations
1: glassbody
15: plungerscrew

19
Low pressure chromatography column - Pilot series
3.6.2 Dismantling the glass body
uDismantling the glass body:
6. Follow the steps 1-5 uchapter 3.6.1 to remove the piston.
ATTENTION
Uneven loosening of the cap nuts may cause damage to the glass
body.
7. Loosen the cap nuts (16) each only half a turn.
Loosen the screw on the opposite side in rota-
tion.
8. Remove the cap nuts (16) and the washers (17)
at the upper column disc (18).
ATTENTION
After removal of the column disc, the
glass body is exposed. Make sure that
it will not fall over.
9. Lift the column disc (18) from the column
body. The O-ring (14) of the column disc can
now be also removed.
10. The glass body (1) is now exposed and can
be lifted from the lower column disc (34). (for
a Pilot column with a cooling jacket the outer
glass body is also exposed and can also be
removed.) The O-ring (14) on the lower co-
lumn disc (34) can now also be removed.
Starting operations
1: glassbody
16: cap nut
17: washer
18: column disc flex
19: counternut

20 Low pressure chromatography column - Pilot series
3.6.3 Dismantling of the plunger
11. Follow the steps 1-5 uchapter 3.6.1 to re-
move the piston.
12. Loosen the hexagon socket screws (20) of
both spindle screw nuts (6/7) and remove the
internal spindle screw nut (6), column seal
actuation nut (8), teflon disc (21) and the ex-
ternal spindle screw nut (7) in this order.
13. Pull the inner spindle (4) in the direction of
the piston out of the outer spindle (5).
14. The piston (11) is screwed onto the inner
spindle (6) and can be screwed off now (by
hand).
15. Loosen the tubing (22) with a suitable tool
from the piston (11).
i
The force plug is held in position only by the
piston. Make sure that the springs and the O-
ring (14) don’t fall off.
Starting operations
4: internal spindel
5: external spindel
6: nut of internal spindel
7: nut of external spindel
8: column seal actuation nut
11: piston
20: setscrew
21: teflon disc
22: tube