fineline™ 70-350 columns · 2020. 7. 13. · en-iso 14121-1, 14121-2 safety of machinery –...

GE Healthcare

FineLINE™ 70-350 columns

Operating InstructionsOriginal instructions

Table of Contents

Table of Contents

1 Introduction .............................................................................. 51.1 Important user information ................................................................................. 61.2 Regulatory information ......................................................................................... 71.3 FineLINE 70-350 columns description ............................................................ 9

2 Safety instructions................................................................. 112.1 Safety precautions ............................................................................................... 112.2 Labels .......................................................................................................................... 142.3 Recycling procedures .......................................................................................... 15

3 Installation .............................................................................. 173.1 Unpacking the delivery box .............................................................................. 173.2 Packing list ............................................................................................................... 183.3 Grounding the column ........................................................................................ 193.4 Connections ............................................................................................................. 19

4 Operation ................................................................................ 214.1 Leakage test ............................................................................................................ 224.2 Packing the column ............................................................................................. 264.3 Evaluating the packing ....................................................................................... 314.4 Recommended operating conditions .......................................................... 334.5 Unpacking the column ....................................................................................... 34

5 Maintenance ........................................................................... 355.1 General ....................................................................................................................... 355.2 User maintenance schedule ............................................................................ 365.3 Cleaning ..................................................................................................................... 375.4 Disassembly and assembly .............................................................................. 395.5 Component maintenance ................................................................................. 405.6 Storage ....................................................................................................................... 42

6 Troubleshooting ..................................................................... 436.1 General problems ................................................................................................. 436.2 Column packing related problems ................................................................ 45

7 Reference information .......................................................... 477.1 Specifications .......................................................................................................... 477.2 Chemical resistance ............................................................................................. 497.3 Service and training ............................................................................................. 507.4 Spareparts and accessories, ordering information .............................. 50

Appendix A Health and Safety Declaration/Liability Form .. 61

FineLINE 70-350 columns Operating Instructions 28-9603-78 AA 3

Table of Contents

4 FineLINE 70-350 columns Operating Instructions 28-9603-78 AA

Introduction 1

1 Introduction

Purpose of the Operating Instructions

The Operating Instructions provide you with the instructions needed to handle FineLINE 70-350 columns in a safe way.

PrerequisitesIn order to operate FineLINE 70-350 columns safely and according to the intended purpose the following prerequisites must be met:

• You should be acquainted with the use of general laboratory equipment and with handling of biological materials.

• You must read Chapter 2 Safety instructions‚ on page 11 of these Operating Instructions.

• Operators should have experience of working with pressure vessels as well as with equipment that provides pressurized media for axial compression packing control.

• The column should be installed according to the instructions in Chapter 3 of these Operating Instructions.

• The column must be used according to the instruction in Chapter 4 Operation‚ on page 21 of these Operating Instructions

In this chapterThis chapter contains important user information and a general description of FineLINE 70-350 columns and their intended use.


1 Introduction1.1 Important user information

1.1 Important user information

Read this before using FineLINE 70-350 columns

All users must read the Safety Instructions in Chapter 2 of these Operating Instructions before installing, using or maintaining the column.

Do not operate FineLINE 70-350 columns in any other way than described in the user documentation. If you do, you may be exposed to hazards that can lead to personal injury and you may cause damage to the equipment.

Intended useFineLINE 70-350 columns are intended for small scale production of biomolecules such as proteins, peptides and oligonucleotides.

FineLINE 70-350 columns must be used in accordance with the safety instructions and technical specifications given in this Operating Instructions.

FineLINE 70-350 columns must not be used for any other purpose.

Safety noticesThese Operating Instructions contain WARNINGS, CAUTIONS and NOTICES concerning the use of the product, with meanings as defined below.

WARNING WARNING indicates a hazardous situation which, if not avoided, could result in death or serious injury. It is important not to proceed until all stated conditions are met and clearly understood.

CAUTION CAUTION indicates a hazardous situation which, if not avoided, could result in minor or moderate injury. It is important not to proceed until all stated conditions are met and clearly understood.

NOTICE NOTICE indicates instructions that must be followed to avoid damage to the product or other equipment.


Introduction 1 Regulatory information 1.2

Notes and tipsNote: A Note is used to indicate information that is important for trouble-free and

optimal use of the product.

Tip: A tip contains useful information that can improve or optimize your procedures.

1.2 Regulatory informationThis section lists the directives and standards that are fulfilled by FineLINE 70-350 columns.

Manufacturing information

CE Conformity

International standards

Requirement Content

Name and address of manufacturer GE Healthcare Bio-Sciences AB, Björkgatan 30, SE 751 84 Uppsala Sweden

Place and date of declaration Uppsala, Sweden, May 2009

Identity of person authorized to sign DoC See EC Declaration of Conformity

Directive Title

2006/42/EC Machinery Directive (MD)

1997/23/EC Pressure Equipment Directive Applicable to FineLINE 200LP, 350P and 350LP columns only

94/9/EC Equipment and protective systems intended for use in potentially explosive atmospheres

Standard Description Notes

EN-ISO 12100-1, 12100-2

Safety of machinery – Basic concepts, general principles and design

Harmonized with 2006/42/EC

EN-ISO 14121-1, 14121-2

Safety of machinery – Principles of risk assessment

Harmonized with 2006/42/EC


1 Introduction1.2 Regulatory information

CE marking

The CE marking and the corresponding Declaration of Conformity is valid for the instrument when it is:

• used as a stand-alone unit, or

• connected to other CE-marked instruments, or

• connected to other products recommended or described in this Operating Instruction, and

• used in the same state as it was delivered from GE Healthcare, except for alterations described in the user documentation or explicitly authorized by GE Healthcare.

Regulatory compliance of connected equipment

Any equipment connected to FineLINE 70-350 columns should meet the safety requirements of relevant harmonized standards. Within the European Union, connected equipment must be CE-marked.


Introduction 1 FineLINE 70-350 columns description 1.3

1.3 FineLINE 70-350 columns description

FineLINE 70, 100 and 200

FineLINE 350

.

Part Description

1 Column top connection

2 Adapter

3 Hydraulic chamber connection

4 Top lid

5 Column tube

6 Bottom lid

7 Stand

8 Column bottom connection

9 Wheels with brakes (4 pcs)

1

2

3

5

78

9

4

6

200P 70 100LP

The layout and components of the 350 columns are identical to the 70/100/200 columns shown above.

350P350LP


1 Introduction1.3 FineLINE 70-350 columns description

Design overviewFineLINE 70-350 columns have been specially designed to meet the needs of a chromatography media for high resolution, process scale separations. They incorporate an axially adjusted adapter unit and special bed supports in the bottom lid and the adapter unit.

FineLINE 70-350 columns are packed using an axial dynamic compression packing technique.

The columns are intended for scale up and small scale production.

Design directivesFineLINE 70-350 columns can be considered a pressure vessel with variable volume.

All Standard FineLINE 70-350 columns are built and tested according to the PED standard for pressure vessels. They are individually inspected and pressure tested before delivery.

FineLINE 70-350 columns can also be produced according to the ASME design code. To order an ASME produced column contact your local GE Healthcare representative for ordering information.

MaterialsMaterials of construction used in FineLINE 70-350 columns are stainless steel and EPDM or PFR seals. The columns are easy to operate to the high standards of hygiene applied in the production of biopharmaceuticals.

Used materials are selected for their compatibility with the solvents most commonly used in chromatographic purification steps, in column maintenance and in cleaning processes, refer to Section 7.2 Chemical resistance‚ on page 49 for more information.

The stainless steel components are electropolished, which improves corrosion resistance and reduces friction and contamination. Electropolished surfaces reduce the risk of pyrogen contamination during the production of pharmaceuticals.

Polymeric materials used in FineLINE 70-350 columns have been tested for their biological reactivity according to USP XXIII. The materials have been tested in vitro according to the “Elution Test” and in vivo according to the “Systemic Injection Test” and the “Intracutaneous Test”.

ComponentsIn general, the components of the columns may be considered to contain “wet” parts, which come in contact with process liquids, and “dry” parts, which do not.

See Section 7.4 Spareparts and accessories, ordering information‚ on page 50 for detailed information regarding the components of FineLINE 70-350 columns.

Note: A special O-ring kit is recommended when eluents for RPC media are used (see Section 7.4 Spareparts and accessories, ordering information‚ on page 50 for details).


Safety instructions 2 Safety precautions 2.1

2 Safety instructions

This chapter describes safety precautions, safety labels and recycling of FineLINE 70-350 columns.

2.1 Safety precautions

IntroductionBefore installing, operating or maintaining the column, you must be aware of the hazards described in the user documentation. Follow the instructions provided to avoid personal injury or damage to the equipment.

The safety precautions in this section are grouped into the following categories:

• Explosive environment

• Flammable liquids

• Personal protection

• Installing and moving the column

• Column operation

• Maintenance

Explosive environment

WARNING Operators should have experience of working with pressure vessels as well as with equipment that provides pressurized media for axial compression packing control

WARNING In potentially explosive atmospheres only tools and protective equipment specially adapted to that environment should be used for operation and maintenance


2 Safety instructions2.1 Safety precautions

Flammable liquids

Personal protection

WARNING Always use suitable clothing when working in potentially explosive environments.

WARNING Explosion hazard. The chemicals listed in this Instructions for Use can be hazardous to use in explosive environments. Always consult the directives in force before using any chemicals for any type of use with the column.

WARNING Avoid discharge from static electricity by grounding the column. Use earthing wire connected to the plant earthing network. Refer to Section 3.3 Grounding the column‚ on page 19.

WARNING Explosive or flammable liquids. If potentially flammable or explosive liquids are used, consult local authorities regarding local rules and regulations before use.

WARNING Explosion hazard. To avoid building up an explosive atmosphere when using flammable liquids, make sure that the room ventilation is sufficient to remove any potentially explosive fumes.

WARNING Fire Hazard. Before starting the system, make sure that there is no leakage if potentially flammable or explosive liquids are used.

WARNING Hazardous substances. When using hazardous chemical and biological agents, take all suitable protective measures, such as wearing protective glasses and gloves resistant to the substances used. Follow local and/or national regulations for safe operation and maintenance of the column.


Safety instructions 2 Safety precautions 2.1

Installing and moving the column

Column operation

CAUTION After assembly, check that the column is tight and not leaking

CAUTION Risk for column tip over. The FineLINE 350L column must not be equipped with wheels.

CAUTION Check that the pressure relief valve is correctly adjusted. Otherwise, the weakest seal in the column will leak when its limit is exceeded.

CAUTION Do not adjust the feet during operation or when column is filled. If the feet are totally unscrewed, the column may loose stability or overturn.

WARNING Operators should have experience of working with pressure vessels as well as with equipment that provides pressurized media for axial compression packing control.

WARNING The design pressure must not be exceeded. See Section 7.1 Specifications‚ on page 47 for pressure limit.


2 Safety instructions2.2 Labels

Maintenance

2.2 LabelsThis section describes safety labels and labels concerning hazardous substances that are attached to the FineLINE 70-350 columns.

Labels on the column

EX marking

The EX marking code on the identification label defines the level of explosion protection of the equipment. The EX marking category of the equipment must be in conformity with the protection level of the room where it is used.

EX II 3 G

Equipment in the EX II 3 G category is designed to be capable of functioning in conformity with the operating parameters established by the manufacturer and ensuring a normal level of protection. Equipment of this category is intended for use in areas in which explosive atmospheres caused by gases, vapors or mists are unlikely to occur or, if they do occur, are likely to do so only infrequently and for a short period only.

CAUTION Use only spare parts delivered by GE Healthcare. This equipment is pressure classified with material specifications which must be preserved.

CAUTION After maintenance, make sure that the column is tight and not leaking.


Safety instructions 2 Recycling procedures 2.3

Column identification label

Symbols used in safety labels

2.3 Recycling proceduresThe equipment shall be decontaminated before decommissioning and all local regulations shall be followed with regard to scrapping of the equipment.

When taking FineLINE 70-350 columns out of service, the column should be emptied, cleaned and sterilized. The different materials must be separated and recycled according to national and local environmental regulations.

Identification label. (Any data shown are only example values.)

This label states the general specifications and utilities for this specific equipment.

The CE mark on the identification label shows that the column fulfills the requirements of the European pressure equipment directive (PED) 97/23/EC for FineLINE 200PL, 350P and 350PL columns and the directive for equipment in potentially explosive atmospheres (ATEX100) 94/9/EC.

Columns produced according to ASME design code have an ASME stamp and no CE-related marking.

Label Meaning

WARNING Read the user documentation before using the column. Do not open any covers or replace parts unless specifically stated in the user documentation.

The column complies with applicable european directives.


2 Safety instructions2.3 Recycling procedures


Installation 3 Unpacking the delivery box 3.1

3 Installation

3.1 Unpacking the delivery box

Unpack the equipment carefully and check the contents of the container against the packing list supplied with the column. Also check the equipment for signs of damage that may have occurred during transportation.

Report any damage immediately to your local GE Healthcare representative and to the transport company concerned. We recommend you keep the packaging material.

Note: Before delivery, the columns are pressure tested using 20% (v/v) EtOH. Some residual drops may remain in the column.

FineLINE 70-350 columns are supplied with the adapter unit taken out and placed upside down with the rod pushed through the center hole in the adapter lid down the column tube. We recommend you to leave the adapter unit in this position until you are ready to pack the column.

Standard wrenches are recommended in a non-explosive environment.

Refer to Section 5.4 Disassembly and assembly‚ on page 39, for further column assembly instructions.

Step Action

1 Unpack the column and check against the packing list (Section 3.2 Packing list‚ on page 18). Inspect items for obvious damage that may have occurred during transportation.

2 Remove the adapter unit from the adapter lid.

3 Remove the adapter lid from the column.


3 Installation3.2 Packing list

3.2 Packing listFineLINE 70-200 columns are delivered with 2 µm nets.

FineLINE 350 columns are also available with 10 µm nets, refer to Section Column assemblies‚ on page 52.

Table 3-1. Items included with FineLINE 70, 70L, 100P, 100LP, 200P and 200LP

Table 3-2. Items included with FineLINE 350 and 350L

Table 3-3. Items included with all FineLINE 70-350 columns

* Number of packages / number of items in each package. (multiply to get total number of items).

** Not part of column article, must be ordered separately (see Section 7.4 Spareparts and accessories, ordering information‚ on page 50).

Item FL 70 FL 70L FL 100P FL 100LP FL 200P FL 200LP Qty*Locking bar 18-1153-66 11-0034-15 11-0034-16 1/1Washer 13 x 24 - 18-1012-61 - 1/2Screw M10 x 160 mm 18-1106-83 - 2/2Screw M10 x 220 mm 18-1106-84 - 2/2Screw M10 x 375 mm - 18-1122-23 - 2/2Screw M12 x 195 mm - 11-0034-18 - 2/2Screw M12 x 460 mm - 11-0034-19 - 2/2Screw M16 x 338 mm - 11-0034-20 - 2/2Screw M16 x 516 mm - 11-0034-21 2/2Washer 17 x 30 - 18-1107-95 1/2Washer 10.5 x 30 18-1106-82 - 1/2Stand ** 18-1031-10 18-1031-20 1/1FineLINE column See Section Column assemblies‚ on page 52 for article numbers 1/1

Item FL 350P FL 350LP Qty*Screw M24 x 350 mm 18-0034-23 - 1/2Screw M24 x 495 mm - 18-0034-24 2/2Stand Included in column article 1/1FineLINE column See Section Column assemblies‚

on page 52 for article numbers1/1

Item Order no Qty*Ball valve 18-1105-37 1/1Grounding kit 18-1157-87 1/1Clamp 25 mm 18-1001-31 3/1TC gaskets 25 mm, i.d. 12 mm 18-0019-27 2/2Blind flange 25 mm 18-1001-25 2/1Operating Instructions 28-9603-78 1/1


Installation 3 Grounding the column 3.3

3.3 Grounding the column

For grounding purpose, a grounding accessory kit is available, see Section 7.4 Spareparts and accessories, ordering information‚ on page 50.

Connect one end of the accessory grounding cable to a ground terminal in the plant grounding network and the other end to the column ground terminal located on the stand, see Figure 3-1. The column ground terminal thread is M6.

Figure 3-1. Column ground terminals for FineLINE 350 (left() and 70/100/200 (right).

3.4 ConnectionsClamps and gaskets i.d. 6 mm, (FineLINE 70/100/200) or i.d. 12 mm (FineLINE 350) are required to connect the 25 mm sanitary flanged inlet/outlet to either valves or tubing of the same type.

Suitable preflanged tubing is available as an accessory.

Refer to Section Accessories‚ on page 58 for information regarding connection accessories.

WARNING When used in explosive environment, the column must be grounded to avoid discharge from static electricity.


3 Installation3.4 Connections


Operation 4

4 Operation

WARNING Operators should have experience of working with pressure vessels as well as with equipment that provides pressurized media for axial compression packing control.

WARNING The design pressure must not be exceeded. See Section 7.1 Specifications‚ on page 47 for pressure limit.

WARNING Hazardous substances. When using hazardous chemical and biological agents, take all suitable protective measures, such as wearing protective glasses and gloves resistant to the substances used. Follow local and/or national regulations for safe operation and maintenance of the column.

CAUTION Do not adjust the feet during operation or when column is filled. If the feet are totally unscrewed, the column may loose stability or overturn.

NOTICE There must be no air in the column during operation. Otherwise separation, cleaning and sanitization performance will deteriorate.

NOTICE When working with sealing, the nets and surfaces in contact with these components: - Never use sharp tools. - Do not scratch any such components or surfaces.


4 Operation4.1 Leakage test

4.1 Leakage test Leakage tests for the hydraulic chamber (H) and process chamber (P) shall be only be performed with liquid, for example water, in the column, i.e. no medium.

Figure 4-1 and Figure 4-2 shows the schematic set-up for the leakage test as required by the procedures described in this section

Equipment setup.

Figure 4-1. Test setup for hydraulic chamber leakage test.

Figure 4-2. Test setup for process chamber leakage test. Items are equal to Fig 4-1.

Part Description

1 Pressure gauge

2 Column top valve

3 Hydraulic outlet valve

4 Adapter unit

5 Column bottom valve

6 Hydraulic inlet valve

7 Pump

H Hydraulic chamber

P Process chamber

2

3

4

5

6

7

H H

P

1

12

3

4

5

6

7

H H

P


Operation 4 Leakage test 4.1

Hydraulic chamber leakage test

Note: See Section 7.4 Spareparts and accessories, ordering information‚ on page 50, regarding the components required by this procedure (see Fig 4-1).

Test procedure

Step Action

1 Close the column bottom valve (5).

2 Fill the column with water.

3 Mount the adapter, assembled to the top plate, onto the column.

4 Check that no air is trapped under the adapter net.

5 Check that the column top valve (2), the hydraulic inlet valve (6) and the hydraulic outlet valve (3) are open.

6 Set up the column according to Figure 4-1, the pump and pressure gauge are connected to the hydraulic chamber inlet (6).

7 Prime the hydraulic chamber (H) by pumping liquid through the hydraulic inlet valve (6), letting the air out through the outlet hydraulic valve (3).

8 Close the hydraulic outlet valve (3) when only liquid escapes the hydraulic inlet valve.Result: The adapter moves down while excessive water is removed through the adapter tube.

9 Stop the pump when the adapter is positioned at a suitable height, for example the same bed height that will be packed later.

10 Close the column top valve (2).

11 Pump water into the hydraulic chamber at a low flow rate while monitoring the pressure.

12 Stop the pump when the pressure reaches the maximum working or packing pressure.Note: If an ÄKTA system is used for this test set the flow to zero (flow=0)

instead of using Pause or Stop. Otherwise the pressure will not be monitored during the test.

13 Wait about 15 minutes until the pressure is stable.

14 Monitor the pressure on the pressure gauge. A recorder linked to a pressure controller gives more precise figures.


4 Operation4.1 Leakage test

Result

The pressure drop should not exceed 0.1 bar over a period of 5 minutes.

If the pressure drop exceeds this limit try to localize the source of leakage.

Begin with the valves and outlets for signs of leakage. If the leakage cannot be localized there this means that the adapter sealing system is most likely the cause. Unpack the column according to Section 4.5 Unpacking the column‚ on page 34.

Take necessary steps to find and correct the suspected problem.

Repeat the leakage test until tightness is confirmed.

Process chamber leakage test

Note: See Section 7.4 Spareparts and accessories, ordering information‚ on page 50, regarding the components required by this procedure (see Fig 4-1 and Fig 4-2).

Priming the hydraulic chamber and positioning the adapter

Step Action

1 Close the column bottom valve (5).

2 Fill the column with water.

3 Mount the adapter, assembled to the top plate, onto the column.

4 Check that no air is trapped under the adapter net.

5 Check that the column top valve (2), the hydraulic inlet valve (6) and the hydraulic outlet valve (3) are open.

6 Set up the column according to Figure 4-1, the pump and pressure gauge shall be connected to the hydraulic inlet (6).

7 Prime the hydraulic chamber by pumping liquid through the hydraulic inlet valve (6), letting the air out through the open outlet hydraulic valve (3).

8 Close the hydraulic outlet valve (3) when only liquid escapes the hydraulic inlet valve.Result: The adapter moves down and excessive water is removed through the adapter tube.

9 Stop the pump when the adapter is positioned at a suitable height, for example the same bed height that will be packed later.

10 Close the hydraulic inlet valve (6).

11 Mount the mechanical locking device.

12 Open the hydraulic outlet valve (3).


Operation 4 Leakage test 4.1

Testing the process chamber

Result

The pressure drop should not exceed 0.1 bar over a period of 5 minutes.

If the pressure drop exceeds this limit try to localize the source of leakage. Begin with the valves and outlets for signs of leakage.

If the leakage is seen in the open hydraulic outlet valve the most likely cause is that the o-ring that seals the adapter to the column wall is leaking.

Unpack the column according to Section 4.5 Unpacking the column‚ on page 34 and Take necessary steps to find and correct the suspected problem.

Repeat the leakage test until tightness is confirmed.

Step Action

1 Set up the column according to Figure 4-2, the pump and pressure gauge shall be connected to the column top valve (2).

2 While monitoring the pressure, pump water into the process chamber through the column top valve (2) at a low flow rate.

3 Stop the pump when the pressure reaches the maximum working or packing pressure.

Note: If an ÄKTA system is used for this test set the flow to zero (flow=0) instead of using Pause or Stop otherwise the pressure will not be monitored during the test.

4 Wait about 15 minutes until the pressure is stable.

5 Monitor the pressure on the pressure gauge. A recorder linked to a pressure controller gives more precise figures.


4 Operation4.2 Packing the column

4.2 Packing the column

General packing information

FineLINE 70-350 columns are designed specifically for use with separation media based on the SOURCE 15 and 30 matrix (see Section 7.4 Spareparts and accessories, ordering information‚ on page 50). The instructions supplied with these media also contain valuable information about column packing and operation.

The axial compression technique used causes the adapter to move down the column tube at a media specific preset pressure as the bed is packed. This simple and reproducible “one step” packing method results in well packed beds in as little as 10 minutes.

The requirement on pumps for packing FineLINE 70-350 columns with SOURCE is apart from the column diameter also to some extent dependent on the desired bed height and media to be packed. The pressure requirements listed below allow packing of all SOURCE media so far tested at bed heights down to 3 cm.

NOTICE Before packing the column for the first time, flush the column and tubings with CIP solution to check that all surfaces are clean.

NOTICE Some organic solvents may require changing to PFR O-rings, refer Section 7.2 Chemical resistance‚ on page 49.

NOTICE Pack the column at the temperature at which it will be used to prevent the hydraulic liquid expanding/contracting with temperature variations.


Operation 4 Packing the column 4.2

Pump characteristicsUse a high capacity pump (7) that meets the pressure and flow rate requirements as indicated in see Table 4-1 on page 28 with a minimum level of vibration.

The selected pump must fulfill the following requirements:

• Constant pulse-free flow.

• Flow independent of back pressure.

• Should withstand at least 10 bar back pressure.

• Liquid velocity of 10-20 cm/min is the recommended velocity to get optimal performance of the bed. However the column could be packed at 1 cm/min as long as the packing pressure can be reached.

Equipment setup

Figure 4-3. Setup for column packing

Note:

Note:

Note: Figure 4-3 resembles the FineLINE 350P but the schematics are valid for all columns covered by this instruction.

The column is used with tubing, TC clamps, gaskets and blind plates (stop plugs), dimension depending on column size.

Item Description

1 Pressure gauge

2 Pack eluent vessel return

3 Hydraulic chamber outlet

4 Adapter

5 Column bottom valve

6 Hydraulic chamber inlet valve,

7 Packing pump

8 Adjustable pressure relief valve, adjusted to packing pressure

9 Column top valve

H Hydraulic chamber

1

4

5

2

8

7

6

3

H H

9



Axial compression method

Unlike flow packing methods, axial compression does not require the operator to first determine a pressure/flow curve and calculate the optimal packing flow.

The critical parameter during packing is that the packing pressure is reached. The recommended packing velocity 10 - 20 cm/h (which corresponds to the volumetric flow given in Table 4-1) is used to get optimal performance of the bed as well as a fast packing methods.

The column could be packed at 1 cm/min as long as the packing pressure can be reached. In Table 4-1 packing parameters for SOURCE media is given.

Other media can be packed in these column with the same procedure. However, the packing pressure, packing solution, adapter velocity and the end point of the packing must be chosen based on the medium packed.

Table 4-1. Parameters for packing SOURCE 15 and SOURCE 30 in FineLINE 70-350 columns.

* These flow rates are used for optimal performance.

Note: See Section 7.4 Spareparts and accessories, ordering information‚ on page 50, regarding the components required by this procedure (see Fig 4-3).

NOTICE The amount of media needed will be more than the volume of the packed bed.

CAUTION Check that the pressure relief valve (8) is correctly adjusted to avoid overpressure. Failure to do so may cause column leakage or damage.

Column Recommended Packing flow rate [l/h] *

Packing pres-sure [bar]

FL 70/70L 23 to 45 6.5

FL 100P/100LP 50 to 100 6

FL 200P/200LP 190 to 380 5

FL 350P/350LP 580 to 1160 4


Operation 4 Packing the column 4.2

Preparations

Packing

Step Action

1 Align the column by adjusting the screws on the feet.

2 Ground the column If needed, see Section 3.3 Grounding the column‚ on page 19.

3 Connect the valve and pressure gauge needed for packing, see Fig 4-3:• Use ball valve (on/off) for hydraulic chamber connections (6) and (3)

• Use a 4 port (2 way) for the mobile phase outlets (5) and (9)

• Pressure gauge (1)

• Pressure-relief valve (8) which has been adjusted to the packing pressure, see Table 4-1

4 Connect the pump to the bottom valve (5).

5 Remove air from the bottom end-piece and nets by pumping packing solution upwards at 100-200 cm/h, use 20% ethanol for SOURCE. The bottom net should be covered by 2-4 cm packing solution.

6 Close the bottom valve (5) and disconnect the pump.

7 Connect the pump to the hydraulic inlet (6) of the column.

8 Prepare the desired amount of medium by diluting with packing solution.

NOTICE Before proceeding, it is important to read the steps 1 to 11 in section Packing thoroughly. Rehearse the packing procedure since it is important that these steps are done continuously and uninterrupted.

Step Action

1 Pour the slurry carefully into the column and if needed top with packing solution to approximately 0.5 cm below the rim.

2 Mount the adapter, which is assembled to the top plate, onto the column while tilting the adapter slightly to avoid air bubbles being trapped under the top bed support.

3 Tighten the adapter lid fixing screws using 10 Nm torque.

4 Prime the hydraulic chamber by pumping liquid through the hydraulic inlet valve (6), letting the air out through the open outlet hydraulic valve (3).



The column packing procedure is now complete.

When the column is packed and if the adapter is locked in packed position by the locking bar, it’s possible to exchange the hydraulic liquid.

For extended periods, we recommend 20% ethanol or 0.01 M NaOH in the hydraulic chamber.

Before the packed column is used, we recommend you to evaluate the packing result as described in Section 4.3 Evaluating the packing‚ on page 31.

5 Stop the pump.

6 Close the hydraulic outlet valve (3 when liquid only escapes the valve.

7 Open the column top valve (9).

8 Start the pump and lower the adapter a few millimeters until most of the air is removed.

9 Stop the pump and close the column top valve (9).

10 Simultaneously, open the bottom valve (5) and start the pump, adjusted to the desired packing flow.Result: The adapter moves down and the medium packs.

Note: Usually the pressure rises as the bed packs. Do not adjust the pressure during packing.

11 Continue until the adapter has stopped moving downwards and stabilized at its lowest position, then continue to run the pump for two more minutes.

12 Simultaneously, close the hydraulic inlet valve (6), close the bottom valve (5) and stop the pump.

13 Lock the adapter with the locking bar and locking screws.

14 Run the column upflow to remove residual air trapped in the upper net and adapter rod.

15 Measure the bed height.

Step Action


Operation 4 Evaluating the packing 4.3

4.3 Evaluating the packing

The efficiency of a column depends on, for example, how well it is packed. A poorly packed column gives rise to uneven flow, resulting in zone broadening and reduced resolution.

It is therefore important to have a method by which the column can be tested before it is put into operation. Such a method should be simple, quantitative and should not introduce contaminating materials.

It is also an advantage if the same method can be used to monitor column performance over its working life so that it is easy to determine when the medium should be repacked or replaced.

A widely used method for expressing the efficiency of a packed bed is in terms of the height equivalent to a theoretical plate (HETP) and the peak asymmetry factor (As). These values can be determined easily by applying a sample such as acetone or salt solution to the column.

Choice of test sample/eluentA sample like acetone or salt solution, for example NaCl, can be used to give a good indication of the column packing quality. The eluate is monitored by measuring the UV absorption at 280 nm or the conductivity. The resulting elution profile is used to calculate the HETP-value and asymmetry see below.

Both acetone and NaCl may interact with the matrix. If using NaCl on charged media such as ion exchangers, the bed must first be equilibrated with an eluent containing NaCl to a conductivity of about 35-40 mS/cm (corresponds to about 0.4 M NaCl) before applying a higher concentration NaCl as sample.

Acetone is not recommended as first choice for techniques such as HIC and RPC unless a solvent at a high enough concentration, to prevent interaction with matrix, is used as eluent. Consider the instructions for each specific media.

An alternative is to increase the running buffer concentration 3-fold and use as sample solution, provided that the running buffer contains salt to a concentration high enough that the sample does not interact with the matrix.

Test set-upOne general recommendation when choosing the test set-up, is to keep the external volume as small as possible. The external volume is defined as the volume upstream the column in between the point of sample application and the column inlet, and from the column outlet to the detector. The air trap and any filters should be bypassed during the sample application to avoid back mixing.

In Table 4-2 test recommendations for SOURCE media is shown. Other media may have other optimal test velocities. Running the HETP test at a higher velocity or using a larger sample volume will affect both HETP and AS values.


4 Operation4.3 Evaluating the packing

Table 4-2. Test set-up recommendation for SOURCE

When the test set-up has been decided and a column has been packed it is recommended to run three tests in a row to measure the repeatability of the method.

To get comparable results over time it is important to be consistent when the test set-up has been chosen. The equipment and conditions must be kept as constant as possible since changes in sample/eluent, velocity, sample volume, liquid pathway temperature etc. will influence the result.

HETP and asymmetry calculation

Calculate the height equivalent of a theoretical plate (HETP) and asymmetry factor (AS ) from the conductivity or UV curve (depending on sample used), as follows:

HETP calculation (see also Fig 4-4)

HETP = L/N N = 5.54 x (VR/wh)2

where, L = Bed height N = number of theoretical plate VR = Peak Retention volume wh = Peak width at half peak height VR and wh are in the same units

To facilitate comparison of column performance, the concept of reduced plate height (h) is often used. Reduced plate height is calculated as:

h = HETP/dp

where, dp = Particle diameter = Mean diameter of the beads (using same unit as for HETP).

As a guideline, a value of h<3 is normally well acceptable at the optimal test conditions.

Property Value

Sample volume 1% to 2% of bed volume

Sample/eluent Alt. 1) 0.8 M NaCl/0.4 M NaCl Alt. 2) 2% (v/v) acetone in ethanol/ethanol

Test velocity 60 cm/h

Detection NaCl, conductivity. Acetone, UV adsorption at 280 nm

NOTICE It is important that the column is properly equilibrated before the packing is evaluated.


Operation 4 Recommended operating conditions 4.4

Asymmetry calculation

The peak should be symmetrical and the asymmetry factor as close to 1 as possible (values between 0.8 and 1.8 are usually acceptable). A change in the shape of the peak is usually the first indication of bed deterioration due to use.

The asymmetry factor As describes the deviation from an ideal Gaussian peak shape and is calculated from the peak width at 10% of peak height:

AS = b/a

As a guideline, value of 0.8-1.8 is good.

Note: Note: If the results of the evaluation procedures described here prove unsatisfactory, refer to Chapter 6 Troubleshooting‚ on page 39, for resolution guidance.

4.4 Recommended operating conditionsFineLINE 70-350 columns require no special operating procedures. Apply generally recognized methods and routines for running process scale columns for the scale up or production of biopharmaceuticals.

When scaling up, remember to consider practical issues such as pressure limitations of related large scale equipment and the special aspects of liquid handling and process control at high volumetric flow rate.

Typically, a linear flow velocity for SOURCE media is in the range of 100 to 1000 cm/h.

Figure 4-4. Pressure/Flow curve in water for FineLINE 350, empty and packed with SOURCE 15Q and 30Q to 15 cm bed height.

Linear flow [cm/h]

Pres

sure

[kPa

]

0

200

400

600

800

1000

1200

100 200 300 400 5000

SOURCE 15Q SOURCE 30Q Empty column


4 Operation4.5 Unpacking the column

4.5 Unpacking the column

The item numbers in the procedure described below refer to Fig 4-3 on page 27.

NOTICE Before removing the locking bar, make sure the hydraulic chamber is filled

Step Action

1 Close the column top inlet (9) with the valve or a blind flange.

2 Attach a piece of tubing to the hydraulic chamber outlet valve (3).

3 Remove the locking bar completely.

4 Open the hydraulic chamber outlet valve (3). This will normally cause packing eluent to flow out through the valve as the pressure is released.

5 Unpack the column by applying a flow of approximately 100 to 200 cm/h upwards through the column bottom valve (5). This will cause the adapter (4) to rise.

6 To avoid loss of gel, stop the flow immediately when the adapter (4) reaches its highest position. Then unscrew the column lid and carefully remove the adapter (4) and the lid.


Maintenance 5 General 5.1

5 Maintenance

5.1 GeneralFineLINE 70-350 columns contain a minimum of components to facilitate cleaning and promote reliable operation. The columns require little regular maintenance.

Excluding the separation media, the main causes of column malfunction are blockage of the column net and worn O-rings.

We recommend filtering buffers and sample solutions before applying them to the column.

WARNING Hazardous substances. When using hazardous chemical and biological agents, take all suitable protective measures, such as wearing protective glasses and gloves resistant to the substances used. Follow local and/or national regulations for safe operation and maintenance of the system.


NOTICE When working with sealings, the nets or surfaces in contact with these components: - Never use sharp tools. - Do not scratch any such components or surfaces.


5 Maintenance5.2 User maintenance schedule

5.2 User maintenance schedule

Period Action

Every 5 cycles Clean-In-Place Refer to Section Cleaning-In-Place (CIP)‚ on page 37.

When repacking column

Nets Examine and wash in detergent solution or in an ultrasonic bath. Replace if required.

When O-rings loose flexibility

Replace O-rings Refer to Section 7.4 Spareparts and accessories, ordering information‚ on page 50.Note: Order solvent resistant seals, O-ring kit PFR (refer to

Section 7.4 Spareparts and accessories, ordering information‚ on page 50) if the EPDM O-rings supplied with the columns are not compatible with the solvent to be used.

As required Sanitization Refer to Section Sanitization‚ on page 38.Autoclaving Refer to Section Autoclaving‚ on page 38.


Maintenance 5 Cleaning 5.3

5.3 Cleaning

Recommended cleaning agents for column parts

All column parts can be cleaned with the most commonly used agents, such as detergents, ethanol, weak acids, sodium hydroxide and salt solutions.

Note: Note: Avoid sodium chloride solutions below pH 4.0. Rinse the column thoroughly immediately after it has been in contact with salt solutions.

Cleaning-In-Place (CIP)One of the most important aspects of production chromatography is the maintenance of the packed column by cleaning-in-place (CIP) procedures. CIP is the removal from the purification system of very tightly bound, precipitated or denatured substances generated in previous purification cycles.

If such contaminants accumulate on the column, they may affect the chromatographic properties of the column. If the fouling is severe, it may also block the column, increasing back pressure and reducing flow velocity.

Note: Reverse the flow direction through the column during CIP.

Suggested methods to remove strongly bound hydrophobic proteins, lipoproteins and lipids from SOURCE media packed in FineLINE columns are:

• Wash the column with 4-10 bed volumes of up to 70% (v/v) EtOH or 30% isopropanol followed by 3-4 bed volumes of water. Apply gradients to avoid air bubble formation when using high concentrations of organic solvents.

• Alternatively, wash the column with 1-4 bed volumes of 0.5% non-ionic detergent (e.g. in 1 M acetic acid) followed by 5 bed volumes of 70% (v/v) EtOH to remove the detergent, and 3-4 bed volumes of water.

WARNING Explosive or flammable liquids. If potentially flammable or explosive liquids are used, consult local authorities regarding local rules and regulations before use.

WARNING Fire Hazard. Before starting the system, make sure that there is no leakage if potentially flammable or explosive liquids are used.

WARNING Explosion hazard. To avoid building up an explosive atmosphere when using flammable liquids, make sure that the room ventilation is sufficient to remove any potentially explosive fumes.


5 Maintenance5.3 Cleaning

• To remove other contaminants, the following method can be used: Wash the column with 4 bed volumes of 0.5-1.0 M NaOH at 40 cm/h, followed by 2-3 bed volumes of start buffer.

These CIP protocols should be used as guidelines to formulate a cleaning protocol specific for the raw material to be applied.

Depending on the nature of the contaminants, different protocols may have to be used in combination. If fouling is severe, the protocols may have to be further optimized.

SanitizationSanitization reduces microbial contamination of the bed to a minimum.

Autoclaving

Step Action

1 Apply 2.5 column volumes of 1.0 M NaOH at a linear flow rate of 250 cm/h, using upward flow through the column.

2 Change flow direction and apply another 2.5 column volumes of 1.0 M NaOH with downward flow through the column.

3 Stop the pump and allow to stand for 60 minutes.

4 Re-equilibrate the column with start buffer.

NOTICE After autoclaving, a leakage tests is recommended, see Section 4.1 Leakage test‚ on page 22.

Step Action

1 Disassemble the adapter from the column

2 Remove the following parts from the column• Valves (contains elastomers that may shrink)

• Seals may shrink and cause leakage after repeated autoclaving)

• Wheels

3 Place the column and the adapter in the autoclave.

4 Autoclave at 121º C and 2.2 bar for 30 minutes


Maintenance 5 Disassembly and assembly 5.4

5.4 Disassembly and assembly

This section describes the assembly procedure. Disassembly is performed in reversed order.

Standard wrenches are recommended in a non-explosive environment (not supplied with the column).

During assembly and disassembly, refer to Figure 7-1 on page 54 or Figure 7-2 on page 55 depending on column model for clarification.

WARNING In potentially explosive atmospheres only tools and protective equipment specially adapted to that environment should be used for operation and maintenance

CAUTION Do not adjust feet during operation/when column is filled. The supporting feet can be totally unscrewed and can cause the column to overturn/loose stability

CAUTION After assembly, check that the column is tight and not leaking. Refer to Section 4.1 Leakage test‚ on page 22

NOTICE When working with sealings, the nets or surfaces in contact with these components: Never use sharp tools and Do not scratch any such components or surfaces.

NOTICE Clean the column with 20% ethanol before disassembly and assembly.

NOTICE Before removing the locking bar, make sure the hydraulic chamber is filled


5 Maintenance5.5 Component maintenance

Column tube assembly

Top lid assembly

Adapter assembly

5.5 Component maintenancePlease refer to Section 5.4 Disassembly and assembly‚ on page 39 regarding how to access the components covered by this section.

Care of filtersExcluding the media, the main cause of column malfunction is blockage of the column net. When columns are re-packed. Always check the nets and wash them in a detergent solution or in an ultrasonic batch or replace them.

Care of O-ringsUse a spatula to prise out worn out O-rings. O-rings need to be replaced at regular intervals since they lose their flexibility with use. Refer to Section Components for more details of O-rings available for FineLINE 70-350 columns.

Step Action

1 Mount the bottom lid (4) to the stand (18).

2 Place the O-ring (8) into the groove on the bottom lid (4).

3 Place the end-piece net (6) and the O-ring (7) onto the bottom lid.

4 Mount the column tube (3) on the end piece net. Make sure the O-ring is positioned in the right place. Tighten the fixing screws with 8 Nm torque.

Step Action

1 Mount the guide ring and piston seal(s) to the top lid.

2 Mount the O-ring (7) into the groove on the top lid.

Step Action

1 Fit the O-ring (8) into the groove on the adapter net (5).

2 Fit the O-ring (10) into the groove on the side of the adapter net.

3 Fit the O-ring (9) onto the adapter unit (2).

4 Mount the adapter net to the adapter unit.

5 Assemble the top lid onto the adapter rod. Lubricate with 20% ethanol if needed.


Maintenance 5 Component maintenance 5.5

Replacement of Teflon sealing washer in valves

The Teflon sealing washer is available as a spare part to the stainless steel 2 and 4 way valve.r

Figure 5-1. stainless steel 2 and 4 way valves exploded view.

Item Description

1 Body

2 Square of tapered plug

3 Teflon sealing washer

4 Positioning disc

5 Dished washer

6 Retaining nut

Step Action

1 Unscrew the retaining nut (6) at the bottom of the valve.

2 Insert the new Teflon sealing washer (3) in the positioning disc (4).

3 Insert the positioning disc (4) in the square of the tapered plug (the Teflon sealing washer has to touch the body).

4 Insert the dished washer (5) (the larger diameter has to touch the positioning disc).

5 Tighten the retaining nut (6) until the dished washer is flat.

6 Tighten the retaining nut up 1/4 turn.

7 Stabilize the valve for 24 hour minimum (as Teflon is a floating material).

8 Screw the retaining nut off 3/8 turn (the tightening torque of the retaining nut is 1.5 Nm).

2

34561


5 Maintenance5.6 Storage

5.6 Storage

Storage of a packed column for longer than 24 hours

Fill the medium process chamber with an appropriate antimicrobial agent, for example either 20% (v/v) ethanol or 0.01 M NaOH (check the instructions of the specific medium).

Lock the adapter mechanically to keep the axial compression.

Blind flanges or stainless steel valves can be used to seal off and prevent contamination of the packed bed.

Storage over extended periodsFor extended periods of storage it is recommended that the column is stored empty, dry and cleaned.

The top plate and adapter could be stored in the upper position in the column tube or as delivered, see Section 3.1 Unpacking the delivery box‚ on page 17.

NOTICE Low pH solutions containing chlorides can cause corrosion in stainless steel. Rinse thoroughly with clean water after each use with such solutions. Inspect the column regularly for signs of corrosive attacks, which will cause column damage if untreated.


Troubleshooting 6 General problems 6.1

6 Troubleshooting

6.1 General problems

Problem Check

High back pressure

Valves between pump and collection vessel not fully open.

Valves between pump and collection vessel blocked.

Column nets blocked, if so, clean or replace. Refer to Section 5.3 Cleaning‚ on page 37.

Air trapped in the nets. If so, wet and wash the nets with 20% (v/v) EtOH before use.

Prefilter blocked.

Tightly bound sample material on the medium. If found, refer to Section 5.3 Cleaning‚ on page 37.

Equipment before or after the column generates back pressure. For example: too small flow cell or sample/collection vessels positioned too low or high compared to the pump.

Connected tubing with too small diameter.

Pressure indicator faulty.

Gaskets misplaced or clamp connectors over tigthtened.

Column leakage Solvents used not compatible with O-rings and/or gaskets. Refer to Section 7.2 Chemical resistance‚ on page 49.

Note: Order solvent resistant seals, O-ring kit PFR (refer to Section 7.4 Spareparts and accessories, ordering information‚ on page 50) if the EPDM O-rings supplied with the columns are not compatible with the solvent to be used.

Tubing not compatible with all solvents? Refer to Section 7.2 Chemical resistance‚ on page 49.

Connections to and from the pump not mounted correct (loose or misplaced).

Air leaking into the column from the suction side of the pump.

Column not connected correctly.

Column pressure too high.

O-rings not positioned correctly.


6 Troubleshooting6.1 General problems

* In general, if air has entered the column, the column must be re-packed. However, if only a small amount of air has been trapped between the column net and the adapter head, it is possible to remove the air by pumping a solution with a temperature a few degrees centi-grade higher than the medium through the column in the opposite direction. After reverse pumping, test the column again prior to use (refer to Section 4.3 Evaluating the packing‚ on page 31).

Chromatograms / results not as expected

Liquid velocity not as intended.

Back-mixing has occurred.

Column incorrectly packed, refer to Section 4.2 Packing the column‚ on page 26, Section 4.3 Evaluating the packing‚ on page 31 and Section 6.2 Column packing related problems‚ on page 45

Column incorrectly equilibrated.

Procedure changes compared to previous batches. For example: Sample batch preparations, operation time intervals, precipitation, dilution or filtration.

Hardware changes compared to previous batches. For example: Filters or chromatographic system components.

Chemicals changes compared to previous batches. For example buffer constituents.

Buffers not prepared according to specifications.

Dead volumes no correctly filled with appropriate buffers and sample before running.

Infected and/or pyrogen contaminated column

Connections not secure.

Starting material contaminated.

In-going components (such as water, serum components in growth media, etc.) contaminated.

Column not cleaned correctly, refer to Section 5.3 Cleaning‚ on page 37.

Air trapped in column *

Buffers not prepared at the same temperature as the column.

Connections not fitted together correctly.

Closed valves not tightly shut.

Problem Check


Troubleshooting 6 Column packing related problems 6.2

6.2 Column packing related problems

If results are not as expected when evaluating the packing according to section X.X, first review the test set-up, then use this troubleshooting guide.

Problem Check Resolution

Leading peak Bed packed too hard. • Repack the column with decreased packing flow rate or pressure.

Tailing peak Media not packed hard enough.

• Repack the column with increased packing flow rate or pressure.

Double peaks

“Extra peak” leading the main peak.

Mainly vertical cracks in the bed caused by:– Pump pulsations. – Packed too hard. – Clogged nets.

• Repack the column with another type of pump or add an air trap in combination with a pressure relief valve.

• Decrease the packing flow rate or pressure.

• Repack with clean nets.

“Extra peak” tailing the main peak.

– Cracks in the bed or head space between medium and adapter. – Too loosely packed column. – Clogged nets.

• Repack the column. It may be possible to adjust only the adapter.

• Increase the packing flow rate or pressure.

• Repack with clean nets.

Poor stability

Increasing asymmetry factor:– Increased tailing – Decreased leading

– Too loosely packed bed leaving space between medium and inlet. – Column not properly equilibrated.

• Repack the column see comments on tailing, above.

• If the asymmetry factor changes from less than one to one, re-equilibrate the column and test again.

Decreasing asymmetry factor:– Increased leading – Decreased tailing

– Bed packed too hard. – Initial cracks getting worse. – Column not properly equilibrated.

• Repack the column, see comments on leading, above.

• If the asymmetry factor changes from greater than one to one, re-equilibrate the column and test again.

Decreasing number of plates.

Bed packed too loosely or too hard.

• Check if it is leading or tailing. See comments above.

• Repack the column.


6 Troubleshooting6.2 Column packing related problems


Reference information 7 Specifications 7.1

7 Reference information

7.1 Specifications

Table 7-1. Specifications FineLINE 70 and 100

Table 7-2. Specifications FineLINE 200 and 350

Property FL 70 FL 70L FL 100P FL 100LP

Weight Adapter unit 2 kg 2,3 kg 3,5 kg 4,3 kg

Column, complete 9 kg 12 kg 15 kg 20 kg

Height Adapter unit 450 mm 750 mm 450 mm 750 mm

Column, complete 1050 mm 1400 mm 1050 mm 1400 mm

Maximum inner length 350 mm 700 mm 350 mm 700 mm

Adapter movement 0 to 350 mm 50 to 700 mm 0 to 350 mm 50 to 700 mm

Inner diameter 70 mm 100 mm

Inlet and outlet diameter 4 mm

Footprint 480 x 480 mm

Property FL 200P FL200LP FL 350P FL 350LP

Weight Adapter unit 21,8 kg 24,2 kg 87 kg 91 kg

Column, complete 65 kg 80 kg 236 kg 260 kg

Height Adapter unit 500 mm 800 mm 575 mm 865 mm

Column, complete 1150 mm 1550 mm 1560 mm 2150 mm

Maximum inner length 350 mm 700 mm 375 mm 670 mm

Adapter movement 0 to 350 mm 50 to 700 mm 0 to 310 mm 50 to 600 mm

Inner diameter 200 mm 350 mm

Inlet and outlet diameter 6 mm 10 or 15 mm

Footprint 590 x 590 mm


7 Reference information7.1 Specifications

Table 7-3. Specifications common for all FineLINE 70-350 columns

Note: All used materials conform to US Pharmacopoeia requirements. EPDM = ethylene polypropylene diene PTFE = polytetrafluoroethene PFR = perfluorcarbon rubber

*) The design pressure must not be exceeded. A rough estimate with a security margin is to add the packing pressure to the operating pressure in the process chamber. Design pressure for this column corresponds to MAWP (Maximum Allowable Working Pressure) in ASME terminology.

Property Value

Recommended bedheight

SOURCE 15 30 to 150 mm

SOURCE 30 50 to 300 mm

Materials of construction

Column body Electropolished stainless steel 316L

Gaskets EPDM (or PFR and PTFE as accessories)

Nets Stainless steel 316L, 2 µm net or 10 µm net

Operating temperature 4ºC to 40ºC

Design pressure * 20 bar

Autoclavable Yes, refer to Section Autoclaving‚ on page 38


Reference information 7 Chemical resistance 7.2

7.2 Chemical resistance

WARNING Explosion hazard. The chemicals listed can be hazardous to use in explosive environments. Always consult the directives in force before using any chemicals for any type of use with the column.

No restriction Restricted short time use

Substance CAS no. Conc. by volume

Substance CAS no. Conc. by volume

Acetic acid 9035-69-2 ≤ 25% Hydrochloric acid 1 9004-54-0 0.1 M

Acetonitrile3 75-05-8 ≤ 70% NaCl2 8028-77-1 1 M

Acetone 8006-64-2 ≤ 10% Nitric acid 78989-43-2 0.5 M

Ethanol3 9003-99-0 100%

1) Not recommended. The stainless steel can be affected.

2) Can be used under normal running conditions. Do not use NaCl in storage solutions. Please note that NaCl can cause corrosion on stain-less steel in acid solutions (pH below 4.0)

3) If PFR-PTFE seals are used

Ethylene glycol 71767-64-1 50%

Formaldehyde 98615-67-9 1.5 M

Formic acid 992-98-3 ≤ 10%

Glycerol 8043-29-6 100%

Isopropyl alcohol3 8013-70-5 100%

Methanol3 67-56-1 100%

NaOH 95077-05-7 2 M

n-Propanol 71-23-8 100%

TFA 76-05-1 0.1%

Triton X-100 9081-99-6 100%

Tivan / TNBP - 1% / 0.3%

UREA 57-13-6 8 M

NOTICE Do not use the following chemicals in FineLINE 70-350 columns:

• Extreme oxidizers (such as peroxides).• Fluorine and halogenated compounds.• Chlorinated solvents (such as methylene chloride).• Esters (such as acetates).• Aromatic hydrocarbons (such as toluene).• High concentrations of strong acids.


7 Reference information7.3 Service and training

7.3 Service and trainingPlease contact your GE Healthcare representative regarding service and training for FineLINE 70-350 columns.

7.4 Spareparts and accessories, ordering information

Useful spare partsSee Section Components / spareparts‚ on page 54

• Bed supports FineLINE 70-200 columns are delivered with 2 µm nets. FineLINE 350 columns are delivered with 2 or 10 µm nets, refer to Section Column assemblies‚ on page 52.

• Seals Order solvent resistant seals (O-ring kit PFR, Section Accessories‚ on page 58) if the EPDM O-rings supplied with the columns are not compatible with the solvent to be used.

• Spare parts to keep on site A complete set of O-rings.

Useful accessoriesSee Section Accessories‚ on page 58

• Stand Applicable for 70, 100 and 200 columns (included in the 350 column). Enables stable positioning of the column during operation.

• PFR O-rings The PFR O-ring kit is recommended when eluents for RPC media are used.

• Connecting the column to your system Clamps and gaskets with applicable dimensions are required to connect the sanitary flanged inlet/outlet to either valves or tubing of the same type. Preflanged tubing is available from GE Healthcare.



Reference information 7 Spareparts and accessories, ordering information 7.4

• Assembly/disassembly of the column Standard wrenches are recommended in a non-explosive environment. In potentially explosive atmospheres only tools and protective equipment specially adapted to that environment should be used for operation and maintenance.

• Pressure gauge We recommend fitting a pressure gauge capable of measuring a negative pressure of -1 bar at the top mobile phase connection to indicate the pressure in the column. This monitors the operating pressure.

• Pressure relief valve Required for the packing procedure. It is connected between the pump and the hydraulic inlet to ensure flow delivery at constant pressure. A suitable pressure relief valve designated RL4 is available. Note:The valve is not supplied with the column and should therefore be ordered separately. As the pressure relief valve is just required when packing the column, only one valve will generally be needed irrespective of the number of columns in use.

• Grounding kit Required for grounding the column to avoid static electrical discharge. Must be fitted if the column is to be used in a potentially explosive atmosphere. See 4.2 Grounding the column for proper fitting of the grounding kit.

• Isolating the column after packing We recommend using stainless steel valves 2-or 4-way, to close off the top and bottom of the column and prevent contamination of the bed. For storage purposes the 25 mm blind flanges with clamps and gaskets can be used to seal off the column.


7 Reference information7.4 Spareparts and accessories, ordering information

Column assemblies

Note: For information regarding FineLINE 70/70L, 100P/100LP and 200P/200LP with other net mesh sizes than 2 µm or other sealings than EPDM, contact your GE Healthcare representative.

Note: The design directive for standard FineLINE 70-350 columns is PED. For information regarding columns designed according to ASME or other directives, contact your GE Healthcare representative.

Column Sealing inlet/outlet Net mesh Article numberFineLINE 70 EPDM 4 mm 2 µm 18-1152-98FineLINE 70L 18-1152-99FineLINE 100P 11-0027-98FineLINE 100LP 11-0027-99FineLINE 200P 6 mm 11-0031-14FineLINE 200LP 11-0031-15FineLINE 350P EPDM 10 mm 2 µm 11-0027-90

10 µm 11-0027-91PFR 10 mm 2 µm 11-0027-86

10 µm 11-0027-8715 mm 2 µm 11-0027-94

10 µm 11-0027-95FineLINE 350PL EPDM 10 mm 2 µm 11-0027-84

10 µm 11-0027-85PFR 10 mm 2 µm 11-0027-86

10 µm 11-0027-8715 mm 2 µm 11-0027-88

10 µm 11-0027-89



StandsFor FineLINE 70/70L, /100P/100LP/ and 200P/200LP, the stand is not included and must be ordered separately to obtain the complete setup, see Table 7-4.

The stand for FineLINE 70/70L and 100P/100LP has adjustable feet. Wheels with brakes are available as an accessory, see Section Accessories‚ on page 58.

The stand for FineLINE 200P/200LP has wheels with brakes as standard.

Table 7-4. Ordering information forFineLINE 70/70L, /100P/100LP/ and 200P/200LP stands

Item Article number

Stand for FineLINE 70/70L/100P/100LP 18-1031-10

Stand for FineLINE 200P/200LP 18-1031-20



Components / spareparts

Figure 7-1. Component parts of FineLINE 70/70L/100P/100LP (left) and 200P/200LP (right). Refer to Table 7-5 on page 56 for parts descriptions.



Figure 7-2. Component parts of FineLINE 350P and 350LP. Refer to Table 7-5 on page 56 for parts descriptions.



Table 7-5. Component descriptions and ordering information. Refer to Fig 7-1 on page 54 and Fig 7-2 on page 55. - : Not relevant. X: Item included

Item Description Material FL70/70L

OrderFL100P/100LP

noFL200P/200LP

FL350P/350LP Wet

1 Top lid 316L/EN1.4404 Included in column assembly article Yes2 Adapter unit 316L/EN1.4404 Yes3 Column tube 316L/EN1.4404 Yes4 Bottom lid 316L/EN1.4404 Yes5 Net, adapter

complete * **2 µm 316L/

EN1.440418-1153-61 11-0034-04 11-0034-06 10mm: 11-0034-08

15mm: 11-0034-12Yes

10 µm 18-1153-67 11-0034-72 11-0034-74 10mm: 11-0034-10 15mm: 11-0034-13

6 Net, end piece complete **

2 µm 316L/EN1.4404 18-1153-62 11-0034-05 11-0034-07 15mm: 18-1137-11 Yes10 µm 18-1153-68 11-0034-73 11-0034-75 15mm: 18-1126-00

7 O-ring 74.6 x 3.53 EPDM 18-1114-19 - - - YesPFR 18-1153-63 - - -

104.37 x 3.53 EPDM - 18-1103-89 - -PFR - 18-1105-50 - -

202.79 x 3.53 EPDM - - 18-8489-01 -PFR - - 18-1106-30 -

350 x 5 EPDM - - - 18-1153-72PFR - - - 18-1153-76

8 O-ring 5.3 x 2.4 EPDM 18-1103-92 - - YesPFR 18-1105-51 - -

7.3 x 2.4 EPDM - - 18-1103-90 -PFR - - 18-1106-31 -

15.2 x 3.5 EPDM - - - 18-1153-74PFR - - - 18-1153-78

9 O-ring 61.2 x 3 EPDM 18-1153-64 - - - Yes64.5 x 3 - 18-1105-48 -85.35 x 3.53 - - - 18-1153-73

PFR - - - 18-1153-7710 O-ring 61.9 x 3.53 PFR 18-1153-65 - - - Yes

91.67 x 3.53 EPDM - 18-1103-91 - -PFR - 18-1105-49 - -

187.3 x 6.99 EPDM - - 18-1106-26 -PFR - - 18-1106-29 -

329.5 x 6.99 EPDM - - - 18-1153-75PFR - - - 18-1153-79

11 Piston seal EPDM 18-1139-56 18-1106-28 18-1149-99 No12 Guide ring PTFE 18-1039-57 18-1106-27 18-1154-45 No



*) 350P/LP: the nets 11-0034-12, 11-0034-10 and 11-0034-13 are made of 316L.**) 350P/LP: the desgnation “10mm” and “15mm” refer to the column inlet/outlet diameter.

Note: It is recommended to keep a set of O-rings as spareparts on site.

13 Washer 316 X - No14, 15

Screw M8 x 35 316 according to SA194/EN10269

X - - NoM12 x 50 - - X -M8 x 45 - - - X

16 Screw M5 x 10 316 18-1107-23 - - - NoM6 x 12 - 18-1126-47 - -M6 x 16 - - X -M6 x 20 - - - 18-1154-46

17 Washer Seal EPDM 11-0034-14 - No6.4 x 20 316 - - - 11-0034-14

18 Stand 316 18-1031-10 18-1031-20 - No19 Screw M6 x 16 316 X - - No

M8 x 25 - - X -M12 x 30 - - - 18-1012-65

20 Washer 6.4 x 12 316 X - - No8.4 x 16 - - X -13 x 24 - - - 18-1012-61

21 Locking bar 316L/EN1.4404 18-1153-66 11-0034-15 11-0034-16 11-0034-17 No22 Washer 10.5 x 30 316 18-1106-82 - - - No

13 x 24 - 18-1012-61 - -17 x 30 - - 18-1107-95 -

23 Screw M10 x 160 316 18-1106-83 - - - NoM10 x 220 18-1106-84 - - -M10 x 375 18-1122-23 - - -M12 x 195 - 11-0034-18 - -M12 x 60 - 11-0034-19 - -M16 x 338 - - 11-0034-20 -M16 x 516 - - 11-0034-21 -M24 x 350 - - - 11-0034-22M24 x 350 - - - 11-0034-23M20 x 695 - - - 18-1155-46

24 Clamp Kit , including clamp, screw M4 x 12, washer 4.3 x 8

316 - - 11-0034-24 - No

O-ring 6.07 x 1.78 EPDM - - - 18-1154-48

Item Description Material FL70/70L

OrderFL100P/100LP

noFL200P/200LP

FL350P/350LP Wet



AccessoriesDescription For columns Material Qty per

packArticle no. Note

O-ring Kit PFR 70 PFR 4 18-1155-43 Includes: O-ring, item 7: 74.6 x 3.53 PFR 18-1153-63, O-ring, item 8. 5.3 x 2.4 PFR 18-1105-51, O-ring, item 10: 61.2 x 3.53 PFR 18-1153-65

100 4 18-1105-45 Includes: O-ring, item 7: 104.37 x 3.53 PFR 18-1105-50, O-ring, item 8: 5.3 x 2.4 PFR 18-1105-51, O-ring, item 10: 91.67 x 3.53 PFR 18-1105-49

200 4 18-1106-23 Includes: O-ring, item 7: 202.79 x 3.53 PFR 18-1106-30, O-ring, item 8: 7.3 x 2.4 PFR 18-18-1106-31, O-ring, item 10: 187.3 x 6.99 PFR 18-1106-29

350 1 18-1136-98 Includes: Two O-rings 350 x 5, Two O-rings 15.2 x 3.5, One O-ring 329.5 x 6.99, One O-ring 85.32 x 3.53

Grounding kit All - 1 18-1157-87Wheel set (4 pcs included)

70/100/200 - 1 18-1001-09350 - 1 18-1156-46

Valves - 25 mm TC4-port, 2-way, i.d. 6 mm

70/100/200 316L/PTFE 1 18-5757-01

4-port, 4-way, i.d. 6 mm

1 18-5758-01


350 1 18-1012-56


1 18-1012-57

Sealings washer Salvi valves

All PTFE 2 18-1128-69 Fits 18-5757-01, 18-5758-01, 18-1012-56 and 18-1012-57. For replacement, refer to Section Replacement of Teflon sealing washer in valves‚ on page 41

Ball valve, 12 mm 316L/PTFE 1 18-1105-37Pressure relief valve

70/100/200 (RL3, i.d. 6

mm)

1 18-1105-36

350(RL4, i.d. 10

mm)

1 18-1106-97

Gasket kit, pressure relief valve

70/100/200 (RL3)

316L/Viton 1 18-1105-52

350 (RL4) 18-1158-26Safety valve All 316 1 18-1156-37



SOURCE 15 and 30 mediaTable 7-6. Data files about SOURCE media details

Tubing with sanitary fitting, i.d. 6 mm, 25 mm TC

0.30 m 70/100/200 PVC 1 18-0005-420.75 m 1 18-0005-431.25 m 1 18-0005-441.50 m 1 18-0005-452.00 m 1 18-0005-47Tubing with sanitary fitting, i.d. 10 mm, 25 mm TC0.3 m All PVC 1 18-1012-850.9 m 1 18-1012-621,0 m 1 18-1156-381.4 m 1 18-1012-631.7 m 1 18-1012-642.0 m 1 18-1156-393.0 m 1 18-1156-40ConnectorsTC gasket 25/6 mm 70/100/200 PFR 5 44-5513-44

EPDM 2 18-0019-27TC gasket 25/8 mm PFR 5 44-5501-19

EPDM 10 18-1163-94TC gasket 25/12 mm

All PFR 5 44-5501-20EPDM 2 18-0200-00

TC gasket 50/22 mm

PFR 5 44-7133-01EPDM 1 44-5511-02

Clamp 25 mm 304 1 18-1001-3112 44-0568-01

Clamp 51 mm PFR 1 44-7134-01Blind flange 25 mm including gasket

304/EPDM 1 18-1001-25

Description For columns Material Qty per pack

Article no. Note

Media Data file article no.

SOURCE 15Q and SOURCE 15S 18-1123-65

SOURCE 30Q and SOURCE 30S 18-1107-12

SOURCE 15ETH, SOURCE 15ISO and SOURCE 15PHE 18-1128-86

SOURCE 15RPC 18-1123-50

SOURCE 30RPC 18-1129-73


Health and Safety Declaration/Liability Form Appendix A

Appendix A Health and Safety Declaration/Liability Form

GE Healthcare Biosciences

70-5009-65/AD Valid from: 2006-05-01

Health & Safety Declaration Form RA# __________________ and/orService Ticket/Request # _______________

1. Please note that equipment will not be accepted for servicing or return until this form is properly completed

2. Failure to complete the form completely accurately and truthfully can endanger GE Healthcare Bio-Sciences personnel and may lead to a delay in servicing the equipment.

3. Equipment not sufficiently cleaned prior to return to GE Healthcare Bio-Sciences may be subject to additional charges.

Liability Form (Servicing Only)

TO RECEIVE A RETURN AUTHORIZATION NUMBER OR SERVICE NUMBER, CALL LOCAL TECHNICAL SUPPORT or CUSTOMER SERVICE

Equipment type / Product No. _______________________________________ Serial No _______________________________________ I hereby confirm that the equipment specified above has been cleaned to remove any hazardous substances or sanitation solution with exception of the named liquids (see lower right on this form). I also agree to give GE Healthcare Bio-Sciences sole control of the system / equipment (specified above). Signed................................................................................ Name in Capital letters....................................................................... Position in Institution or company......................................................... Date (Year/month/date) 20........../........../..........

Specify if the equipment has been in contact with any of the following: Radioactivity (Must be decontaminated prior to service/return) yes no Radiation Safety Officer Signature (required if yes) _________________________________________________

Infectious or hazardous biological substances yes no (please specify) ____________________________________________________________ (Must be decontaminated prior to service/return)

Other Hazardous Chemicals (please specify) yes no _____________________________________________________________ (Must be completely removed prior to service/return)

Phone number where GE Healthcare Bio-Sciences can contact you for additional information concerning the system / equipment Phone No........................................................................... Liquid in equipment is:

Water Ethanol None, empty

I hereby confirm that I have taken full responsibility and control over the system / equipment: Engineer Signed................................................................................ Name in capital letters....................................................................... Date (Year/month/date) 20........../........../..........

Changes / Improvements on system or equipment See PM document See Service report None The owner of the system / equipment confirms by signing in the service report that he has accepted to take the full responsibility and control.


Appendix A Health and Safety Declaration/Liability Form


For local office contact information, visit www.gelifesciences.com/contact

GE Healthcare Bio-Sciences ABBjörkgatan 30751 84 UppsalaSweden

www.gelifesciences.com

GE, imagination at work and GE monogram are trademarks of General Electric Company.

BioProcess, FineLINE, ÄKTA and SOURCE are trademarks of GE Healthcare companies.

All third party trademarks are the property of their respective owners.

© 2009 General Electric Company—All rights reserved.First published Nov. 2009.

All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them. A copy of these terms and conditions is available on request. Contact your local GE Healthcare representative for the most current information.

GE Healthcare UK LtdAmersham Place, Little Chalfont, Buckinghamshire, HP7 9NA, UK

GE Healthcare Bio-Sciences Corp800 Centennial Avenue, P.O. Box 1327, Piscataway, NJ 08855-1327, USA

GE Healthcare Europe GmbHMunzinger Strasse 5, D-79111 Freiburg, Germany

GE Healthcare Japan CorporationSanken Bldg. 3-25-1, Hyakunincho, Shinjuku-ku, Tokyo 169-0073, Japan

28-9603-78 AA 11/2009

imagination at work

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