YMC Pilot PLUS User manual

Original-
Manual
Low pressure chromatography column
YMC Pilot
PLUS
In order to avoid personal injuries and damage to equipment, please read this manual carefully and
act on the recommendations given. Keep this manual accessible for the user at all times.
Version 1.3
26.03.2018

YMC Europe GmbH
Schöttmannshof 19
46539 Dinslaken, Germany
[phone]
[email protected]
www.ymc.de
YMC Europe GmbH
YMC Pilot
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YMC Europe GmbH
Contents
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Contents
1 Information on manual ........................................................................................... 5
1.1 General................................................................................................................ 5
1.2 Explanation of warnings....................................................................................... 5
1.3 Structure of instructions....................................................................................... 6
1.4 Guidelines............................................................................................................ 6
1.5 Figures................................................................................................................. 6
2 Safety ....................................................................................................................... 7
2.1 Intended use........................................................................................................ 7
2.2 General safety instructions.................................................................................. 7
2.3 Personnel requirements....................................................................................... 7
2.4 Personnel protective equipment.......................................................................... 8
3 Specifications and overview.................................................................................. 9
3.1 Fields of application............................................................................................. 9
3.2 Specifications....................................................................................................... 9
3.3 Chemical Resistance.........................................................................................10
3.4 Overview............................................................................................................11
3.4.1 Adjustable end piece .................................................................................... 12
3.4.2 Column body.................................................................................................13
3.4.3 Column stand................................................................................................14
4 Delivery.................................................................................................................. 15
4.1 Delivery contents ............................................................................................... 15
4.2 Delivery inspection............................................................................................. 15
5 Requirements for Operation................................................................................ 16
5.1 Pilot column set up ............................................................................................16
5.2 Pressure test......................................................................................................16
6 Operation............................................................................................................... 18
6.1 Removal of adjustable end piece ......................................................................18
6.2 Pilot column packing.......................................................................................... 19
6.2.1 Slurry preparation......................................................................................... 19
6.2.2 Slurry column packing under flow.................................................................20
6.2.3 Slurry column packing with sedimentation ...................................................24
6.3 Column test and release criteria........................................................................27
6.4 Pilot column unpacking...................................................................................... 28
7 Cleaning and maintenance .................................................................................. 30
7.1 Notes for Cleaning-In-Place (CIP)..................................................................... 31
7.2 Cleaning and maintenance plan........................................................................32
7.3 Cleaning............................................................................................................. 33
7.4 Replacement of spare parts............................................................................... 36
7.4.1 Frit replacement in the adjustable end piece................................................36
7.4.2 Replacing the glass body .............................................................................37
7.4.3 Frit replacement column body......................................................................40
8 Troubleshooting.................................................................................................... 42
9 Storage, packaging and transport ...................................................................... 44
9.1 Storage .............................................................................................................. 44
9.2 Transport............................................................................................................44

YMC Europe GmbH
Contents
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10 Disposal................................................................................................................. 45
11 Appendix................................................................................................................ 46
11.1 Spare parts column stand.................................................................................. 46
11.2 Spare parts adjustable end piece......................................................................46
11.3 Spare parts column body 70 –100 mm ID........................................................ 47
11.4 Spare parts column body 140 –200 mm ID...................................................... 48
12 Index....................................................................................................................... 49

YMC Europe GmbH
Information on manual
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1 Information on manual
1.1 General
This manual includes important massages and information for the intended use of this
pilot column. The manual must be kept accessible for the user at all times.
In order to avoid injuries and damages, read this manual carefully prior to use and act
on the recommendations given.
1.2 Explanation of warnings
Warnings are highlighted by coloured signal word fields. Always read the full text of
the warning in order to effectively protect yourself from damage!
The following signal word fields marked by different colours 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.

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Information on manual
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1.3 Structure 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 levels:
Aim of the instruction
1. Action step
Impact of the action step to check whether the step has been performed
correctly.
2. Another action step
Figures within instructions:
A figure is only related to the action step which is listed directly below the figure.
1.4 Guidelines
The following guidelines are available in this manual:
•Contents at the beginning
•Index in the end
•Name of the main Chapter in the header
•Links to other Chapters, for example “Information in this manual“ ( Chapter 1, S.
5)
•Additional information and hints indicated with an i-symbol
Example for additional information
1.5 Figures
Figures can differ in the graphical presentation from the product.

YMC Europe GmbH
Safety
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2 Safety
This section gives an overview of all the important safety aspects for the protection of
personnel and for the safe and trouble-free handling of the YMC PilotPLUS column.
Other task-specific safety requirements are contained in the sections relating to the
respective operation. In addition, the national and local legal and other applicable
safety regulations must be observed.
2.1 Intended use
The pilot columns 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 specifications
Chapter 3.2, S. 9. The product is only for use with liquids that have a pressure <0.5
bar above the atmospheric pressure at the intended maximum temperature (40°C).
Every use which is outside these specifications is defined as not intended use. The
intended use includes the compliance with the advice and terms in the manual.
2.2 General safety instructions
In order to avoid accidents and injuries,
•operate the pilot column only within its intended use.
•follow all general and special safety instructions as well as all warnings in this
manual.
•follow danger signs and sign giving instructions.
•follow the rules and regulations for prevention of accidents at the site of operation.
•comply with all inspection and maintenance intervals.
2.3 Personnel requirements
The various tasks described in this manual make different demands on the
qualifications and training of the persons carrying them out.
The user must:
•have competent and proficient training.
•be trained in dealing with pressure devices.
•read and fully understand this manual before using this YMC PilotPLUS column.

YMC Europe GmbH
Safety
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2.4 Personnel protective equipment
In order to reduce the risks during working on the column, always use the
recommended personnel protective equipment.
The following protective equipment is required:
Symbol
PPE
Phase of life
Action
Protective gloves
Operation
•Column packing
•Column unpacking
Initiation
•Cleaning
Safety goggles
Operation
•All actions
Initiation
•Cleaning
•Pressure testing
Dust mask
Operation
•Slurry preparation
Protective clothing
Operation
•All actions
Initiation
•Cleaning
•Pressure testing

YMC Europe GmbH
Specifications and overview
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3 Specifications and overview
3.1 Fields of application
The YMC PilotPLUS pilot column is especially designed for pilot scale use. The column
is compatible with common BioLC separation modes (IEX, SEC, Affinity, IMAC, HIC,
MMC) and aqueous conditions. Due to the metal-free and the minimal dead volumes,
the column is perfectly suitable for the separation of biomolecules (e.g. proteins,
oligonucleotides or nucleic acids).
3.2 Specifications
PED* fluid group:
Group 1 referred to EU Guideline 2014/68/EU
PED* safety requirement:
Article 4.3 of EU Guideline 2014/68/EU
Column body:
Borosilicate glass 3.3
Calibrated precision glass (Duran KPG®)
Frits:
PE
Seals:
75 Fluoroprene XP41 blue
Connection:
FEP tubing ¼" OD
Wetted parts:
Borosilicate glass 3.3, FEP, PE, PTFE, PEEK, 75
Fluoroprene XP41 blue
Approved temperature range
4 –40°C
*Pressure Equipment Directive
Length Glass Body
[mm]
500
850
Inner Diameter
[mm]
70
100
140
200
70
100
140
200
Bed Length
[mm]
100 - 4501
250 - 8001
Volume [mL]
0.4
–
1.7
0.8
–
3.5
1.5
–
6.9
3.1
–
14.1
1.0
–
3.0
2.0
–
6.2
3.8
–
12.3
7.8
–
25.1
Pressure Limit [bar]
16
10
7
5
16
10
7
5
Total Weight Empty
Column [kg]
19
20
37
38
23
24
45
47
Footprint [mm]
620
750
620
750
Maximum Height [mm]
1,700
1,750
2,250
2,300
1Depending on packing material and packing method

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Specifications and overview
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3.3 Chemical Resistance
The chemical resistance depends on the wetted materials used Chapter 3.2, S. 9.
If the desired solvent is not listed, YMC will be pleased to advise you.
Material
Borosilicate
glass 3.3
FEP
75
Fluoro-
pren
XP41
PE
PTFE
PEEK
Lösungsmittel
Acetone
Good
Good
Unstable
Good
Good
Good
Acetonitrile
Good
Good
Limited
Good
Good
Good
Dihydrogen phosphate
Good
Good
Good
Good
Good
Good
Cyclohexane
Good
Good
Good
Limited
Good
Good
Dichloromethan
Good
Good
Limited
Unstable
Good
Unstable
0.1M EDTA (3%)
Good
Good
Good
Good
Good
Good
1M Acetic acid (6%)
Good
Good
Good
Good
Good
Good
Ethanol
Good
Good
Good
Good
Good
Good
Ethyl acetate
Good
Good
Unstable
Good
Good
Good
n-hexane
Good
Good
Good
Limited
Good
Good
Isopropanol
Good
Good
Good
Good
Good
Good
Methanol
Good
Good
Good
Good
Good
Good
2M Sodium hydroxide
(8%)
Good
Good
Good
Good
Good
Good
1M Hydrochloric acid
(4%)
Good
Good
Good
Good
Good
Good
8M Urea (36%)
Good
Good
Good
Good
Good
Good
1M NaCl
Good
Good
Good
Good
Good
Good
0,5M Na2SO4
Good
Good
Good
Good
Good
Good
YMC recommends avoiding the following chemicals:
•Strong oxidizing agents
•Halogenated compounds
•Chlorinated solvents
•Aromatic compounds (e.g. toluene)
•High concentrations of strong acids
•THF

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Specifications and overview
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3.4 Overview
A parts list with all wetted parts and non-wetted parts is in the appendix.
The YMC PilotPLUS column consists of the adjustable end piece, the column body and
the column stand. These parts can be assembled whilst operating the column without
any tools.
Adjustable end piece
Column body
Column stand

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Specifications and overview
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3.4.1 Adjustable end piece
1
Tubing
4
Star screw
2
Spindle
5
Closing disc (AEP)
3
Rotary Handle
6
Connecting flange
The adjustable end piece consists of a closing disc and the piston. The piston consists
of the spindle, the connecting flange, the plunger, the frit and the profile gasket. A
tubing goes through the spindle to the connecting flange. With the rotary handles in
the closing disc, the piston can be adjusted up and down. The closing disc is attached
with star screws to the column body.
Piston front
1
Spindle
4
Frit
2
Connecting flange
5
Profile gasket
3
Plunger (AEP)

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Specifications and overview
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3.4.2 Column body
1
Cylinder screw
6
Connector
2
Column disc
7
Glass body
3
Connector drainage tubing
8
Rod
4
Drainage tubing
9
Holding bar
5
Closing disc (CB)
The glass body is fixed between the column disc and the closing disc by the rods. The
glass body has a visible scale. Excessive solvent is removed during piston insertion
via the connector drainage tubing. The fixed end piece is assembled with the closing
disc (CB). The fixed end piece consists of the connector, the sealing washer, the
plunger (CB), the ring gasket B, the frit (CB) and the ring gasket A. The column body
can be assembled in the stand in two different positions.

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Specifications and overview
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Frit column body
1
Connector without cap nut
4
Frit (CB)
2
Plunger (CB)
5
Ring gasket A
3
Ring gasket B
3.4.3 Column stand
1
Levelling feet
3
Frame disc
2
Main pillar
4
Locking bolts
With the locking bolts in the frame disc the column body is attached to the column
stand. With the adjustable levelling feet, the pilot column can be aligned vertically.
Alternatively, the column stand is available with casters.

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Delivery
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4 Delivery
4.1 Delivery contents
•Pilot column
•Documentation folder:
–Brochure YMC Glass Columns
–Manual
–Overview drawings
–YMC Quality Control / Leakage Tests
–Certificate of conformity FDA/BSE/TSE/USP Class VI
4.2 Delivery inspection
YMC recommends retaining and storing the original packaging and the
foam cushion so that the column can always be shipped safely.
Delivery inspection
Delivery inspection is performed as follows:
1. Check the delivery for visible transport damage.
2. Check the delivery for the completeness of parts.
3. Compare the column description on the glass body with the data of the delivery
note.
Transport damages
Procedure, if you observe any visible transport damage:
1. Refuse or reserve 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 responsible supplying agent.

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Requirements for operation
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5 Requirements for Operation
5.1 Pilot column set up
Required devices:
•Drainage tubing
•Collecting vessel
The pilot column is set up as follows:
1. Make sure that the pilot column is standing on fixed ground.
2. Make sure that the pilot column is standing vertical in the final operating place.
3. Connect the drainage tubing to the drainage tubing connector.
4. Put the drainage tubing in the collecting vessel.
5.2 Pressure test
WARNING
Operation of pressure tests
Eye or skin injuries possible due to solvent leakage or incorrectly sealed connectors.
►Perform the pressure test behind a safety screen.
►During work on the pilot column, wear the prescribed personnel protective
equipment.
WARNING
Pilot column under high pressure
Eye or skin injuries possible due to remaining pressure in the tubing or in the pilot
column.
►Stop the pump during work on the pilot column.
►During work on the pilot column, wear the prescribed personnel protective
equipment.
A pressure test is used to check the tightness of both the seals in the pilot column and
also the tubing connectors. Before using the column for chromatographic applications,
it is recommended that a pressure test is performed to verify the correct assembly.
Prior to delivery of the column, the column has been tested according to the standard
procedure listed below. The relevant certificate is located in the documentation folder.

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Requirements for operation
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Standard procedure pressure test
1
Pilot column
4
Pressure gauge
2
Back pressure regulator
5
Gate valve
3
Gate valve
6
Pump
The pressure test should be performed as follows:
1. Assemble the system according to the schematic overview of a pressure test.
2. Turn the piston of the adjustable end piece in the DOWN direction as far as it
will go.
3. Fill the pilot column with isopropanol.
4. Increase the testing pressure in the pilot column by slow and stepwise
adjustment of the flow rate at the pump.
The testing pressure is the pressure limit of the column plus 10% (70
mm ID = 17.6 bar | 100 mm ID = 11 bar | 140 mm ID = 7.7 bar | 200
mm ID = 5.5 bar)
5. Keep the testing pressure constant for 10 minutes.
6. If leakages occur, check the connections. If required, replace tubing and/or
connectors.
If the testing pressure was constant for 10 minutes, the column is ready for use.
If a back pressure regulator is used, the maximum pressure is reached
easier.
Leakages at the piston of the adjustable end piece are often related to
impurities trapped in the seal.

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Operation
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6 Operation
WARNING
Risk of explosion
Severe injuries possible due to glass splinters.
►Never put the glass column under gas pressure!
In order to guarantee safe operation, the pilot column must be used for the intended
use only. ( Chapter 2.1, S. 7).
6.1 Removal of adjustable end piece
Required devices:
•Clean and soft surface
The adjustable end piece is removed as follows:
1. Turn the piston of the adjustable end piece in the UP direction as far as it will
go.
2. Remove the star screws from the closing disc (AEP).
3. Remove the adjustable end piece from the glass body and put it on a clean
and soft surface.

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Operation
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6.2 Pilot column packing
CAREFUL
Packing material
Eye and skin injuries possible through contact with packing material.
►Wear the prescribed personnel protective equipment.
Two different procedures are common for slurry packing the column:
•under flow,
•with sedimentation.
6.2.1 Slurry preparation
CAREFUL
Formation of dust with dry packing material
Lung irritation possible if packing material is inhaled.
►Read the material safety data sheet from the packing material manufacturer.
►Wear a dusk mask.
The use of a more dilute liquid slurry simplifies the column packing. The
total volume of the slurry should not exceed the maximum volume of the
column.
The removal of fines decreases the column back pressure and so
improves the performance of the packed column.
The slurry is prepared as follows:
1. Mix the solvent or buffer with the appropriate amount of packing material in a
suitable container (see manufacturer’s recommendations).
The total volume of the slurry must be less than the maximum column volume.
The amount of packing material determines the final bed length.
2. Mix the slurry with a stirring rod until the slurry is homogenous and avoid
introduction of air bubbles.
3. Check if the material is suitable for ultrasonication (see manufacturer’s
recommendations).
If the material is suitable for ultrasonication, degas the slurry in an ultrasonic
bath.
If the material is not suitable for ultrasonication, let the material settle.
4. Let the degassed slurry settle.
5. If fines are visible on the surface of the slurry, remove them.

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Operation
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6.2.2 Slurry column packing under flow
The column is packed with the slurry method under flow as follows:
1. Remove the adjustable end piece Chapter 6.1, S. 18.
2. Check the drainage tubing for correct connection.
3. Check the drainage tubing outlet for correct position in the collecting vessel.
4. Connect the pump with the tubing at the column body.
5. Start the pump.
6. Fill the glass body with solvent or buffer until 2-3 cm above the frit in the
column body (bottom-up).
This ensures that no air is trapped under or is inside the frit.
7. Close the valve in the tubing on the column body.
8. Connect the pump with the adjustable end piece.
9. Flush the piston of the adjustable end piece with slurry solvent until the frit of
the adjustable end piece is wetted completely.
10. Disconnect the pump from the adjustable end piece.
11. Pour the homogenous and/or degassed slurry into the glass body.
12. Rinse the inner wall of the column body with slurry solvent for removal of
particles.
By removing particles from the inner wall of the column body, a correct seal
with the profile gasket is guaranteed.
13. Fill the pilot column up to the edge of the drain function with slurry solvent.
14. Turn the closing disc of the adjustable end piece in the UP direction as far as it
will go.