Evolve 70 Manual

70mm, 100mm and 140mm
Chromatography Columns

DELTA PRECISION LTD
Chromatography Column
Operating Manual and Installation Guide
70mm, 100mm and 140mm Columns

Revision History
Revision
Comments
Date
A
Initial release
21/11/2014
B
Photos updated
25/02/2015
C
Address Updated
21/06/2018
D
Pressure Equipment Directive updated
06/04/2021
E
Technical specification updated caster not available on 70-140mm columns
22/04/2021
F
Updated to reflect Evolve® Trademark status
12/07/2021
Evolve® is a registered trademark of Delta Precision Ltd. Evolve® is a registered trademark in the
EU, UK and US.
Manual Revision
Date
F
July 2021

Table of Contents
Column Hardware .......................................................................................................... 1
Mechanical Description ........................................................................................................................... 1
Technical Data. ...................................................................................................................................... 3
Weights and Dimensions ........................................................................................................................ 3
Pressure Drop ........................................................................................................................................ 4
Material Conformity ............................................................................................................................... 6
Chemical Compatibility .......................................................................................................................... 7
Safety Guidelines ............................................................................................................ 9
Pressure Equipment Directive (PED). ................................................................................................... 9
Over-pressurization ................................................................................................................................. 9
Handling ................................................................................................................................................ 9
Chemical compatibility .......................................................................................................................... 10
Installation and Column Set-up ................................................................................... 11
Installation. .......................................................................................................................................... 11
Column Operation ........................................................................................................ 15
Hydrostatic Leak Test ......................................................................................................................... 15
Media Preparation ............................................................................................................................... 16
Column Packing (Constant Flowrate Method) ...................................................................................... 16
Unpacking ........................................................................................................................................... 18
Cleaning and Storage ............................................................................................................................ 18
Maintenance ................................................................................................................. 20
Routine Maintenance Procedures-General guidelines .............................................................................. 20
Disassembly of 70mm, 100mm and 140mm Columns ......................................................................... 21
Reassembly of 70mm, 100mm and 140 column. ................................................................................... 24
Troubleshooting ........................................................................................................... 27
Leakage from adjuster seal .................................................................................................................... 27
Leakage from fixed bed support seal ..................................................................................................... 27
Leakage from inlet or outlet .................................................................................................................. 27
Excessive column back pressure ............................................................................................................ 28
Media Leakage .................................................................................................................................... 28
Kits, Spare parts and Accessories ................................................................................. 29
Spare parts and kits ............................................................................................................................. 31
Spare Parts and kits Cont. ................................................................................................................... 32
Accessories ............................................................................................................................................ 34
Contact Us .................................................................................................................... 35
Appendix 1 Chromatography Performance .................................................................. 36
Performance Measurement and Column Efficiency ................................................................................. 36
Performance Results From Evolve®Columns ........................................................................................ 38

Table of Figures
Figure 1.1 Evolve®Column Range 70mm, 100mm, 140mm, 200mm and 250mm columns .................... 2
Figure 1.2. Pressure Drop Evolve®70mm column ..................................................................................... 4
Figure 1.3 Pressure Drop Evolve®100mm column ................................................................................... 4
Figure 1.4 Pressure Drop Evolve®140mm column ................................................................................... 5
Figure 3.1 Diagram showing the location of the fixed bed support ............................................................. 11
Figure 3.2 Photograph showing the installation of the fixed bed support .................................................... 12
Figure 3.3 Diagram showing the location of the adjuster bed support ........................................................ 13
Figure 3.4 Photograph showing the location of the adjuster bed support ..................................................... 13
Figure 4.1 Schematic of the column with part identification ....................................................................... 17
Figure 5.1 Adjuster Assembly Evolve® 100 column .............................................................................. 22
Figure 5.2 Fixed Assembly Evolve® 100 column ................................................................................... 23
Figure 5.3 Diagram showing the location of the fixed bed support ............................................................. 24
Figure 5.4 Photograph showing the installation of the fixed bed support .................................................... 24
Figure 5.4 Photograph showing the location of the adjuster bed support ..................................................... 25
Figure 5.5 Diagram showing the location of the adjuster bed support ........................................................ 25
Figure 5.6 Photograph showing the final location of the adjuster bed support ............................................. 26
Figure 5.7 Photograph showing the alignment of marking on flowtube and adaptor to ensure component is
correctly tightened ....................................................................................................................... 26
Figure A1 Description of Column Efficiency Calculations ........................................................................ 37
Figure A2 Example of typical chromatogram. Evolve®250mm column. 2% Acetone sample, flowrate
100cm/hr, bed height 19.5cm. ................................................................................................... 38

1
Column Hardware
Mechanical Description
The Evolve®Column is designed specifically for use in the Biopharmaceutical Applications. The
column can be used in pilot or production scale Chromatographic processes.
A pressure rating of 4 bar, operating temperature range of 2-30OC and adjustable bed height
makes this column suitable for a wide range of chromatography media types and chromatographic
techniques.
The columns have the following design features.
The entire flowpath is non-metallic, which has the following advantages:
• Eliminates the corrosion risk and contamination of the process
• Allows the use of high concentration sodium chloride solutions and Guanidine
hydrochloride.
• Prevents any product inactivation through metal contact
Unlike other columns available on the market the Evolve®columns have an integral bed
support and seal assembly. The polypropylene mesh and seal are manufactured as a single
unit and fits directly on the flowcell without any fixings.
This has the following advantages.
• Ensure that maximum area is available to flow, promoting good chromatographic
performance and cleanability.
• Eliminates use of fixings to secure the bed support to the flowcell, removing the potential
for leak paths or dead flow zones.
• Reduces number of components in the column assembly, leading to easy operation and
maintenance.
Sealing of the adjuster flowcell to the tube wall is achieved by a moulded Santoprene seal.
Central activation of this seal ensures that compression is even. The mechanism to energise
and de-energise the seal is simple and easy to operate.
Positioning of the adjuster assemble within the tube is easy and simple using the central
adjustment mechanism.
Chapter
1

2
The column is composed of three main sub-assemblies:
• Adjuster assembly- consisting of the adjuster flowcell assembly, adjuster seal
components, adjuster flange and a central height adjust rod. This allows the
position of the adjuster to be varied using the height adjust rods over the range
20-520mm.
• Tube unit – consisting of the column tube, flanges and column stand or support
legs
• Fixed assembly – consisting of the bottom flowcell assembly and flowtube.
Figure 1.1 Evolve®Column Range 70mm, 100mm, 140mm, 200mm and 250mm columns

3
Technical Data.
70
100
140
200
250
Cat No.
AB070505
AB100505
AB140505
AB200505
AB250505
Tube id mm
70
100
140
200
250
Cross Sectional area cm2
38.5
78.5
154
314.2
490.9
Maximum Bed height mm
520
520
520
520
520
Minimum Bed height mm
20
20
20
20
20
Max Column Volume L
2
4
8
16.3
25.5
Adjustable Capacity L
1.9
3.9
7.7
15.7
24.5
Pressure rating bar
psi
4
58
4
58
4
58
4
58
4
58
Operating temp
2-30oC
Bed support pore size µm
8µm
Weights and Dimensions
70
100
140
200
250
Cat. No
AB070505
AB100505
AB140505
AB200505
AB250505
Adjuster Weight kg
4
5
7.5
12.7
15
Total Weight empty
kg
11.5
12.5
19
45.5
52
Total Weight Full*
kg
13
16
26
60
76
Maximum Diameter
ex casters mm
226
226
282
455
495
Maximum Diameter
inc casters mm 1
455
495
Maximum height mm
1523
1543
1545
1550
1550
Minimum
Clearance** mm
1620
1620
1620
1750
1750
1 casters are not available on 70-140mm columns
*based on bed height of 520mm and specific gravity 1.05,
**minimum height required to remove adjuster from tube unit.

4
Pressure Drop
The pipework and column contribute to the pressure drop of the chromatographic system.
Measuring the pressure drop from the column inlet to the column outlet shows the
contribution made by the column hardware. The tests were performed at ambient
temperature 15-20oC using DI water.
Figure 1.2. Pressure Drop Evolve®70mm column
Figure 1.3 Pressure Drop Evolve®100mm column
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0500 1000 1500 2000 2500
Linear Velocity cm/hr
Pressure bar
0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6
Volumetric Flowrate L/min
Evolve 70
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0200 400 600 800 1000 1200
Linear Velocity cm/hr
Pressure bar
0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6
Volumetric flowrate L/min
Evolve 100

5
Figure 1.4 Pressure Drop Evolve®140mm column
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
0500 1000 1500 2000 2500 3000
Linear Velocity cm/hr
Pressure bar
0 0.5 1 1.5 2 2.5 3 3.5 4
Volumetric flowrate L/min
Evolve 140

6
Material Conformity
All materials in contact with the process stream have been selected for their suitability for use
in equipment utilised in Biopharmaceutical applications and so either conform to relevant
sections of FDA code of Federal Regulations vol. 21 170-199 and/or passed USP class VI
test for in vivo toxicity.
Materials of construction – process-wetted parts
Column Component
Material
Column tube
Acrylic
Fixed flowtube
Polypropylene
Adjuster flowtube
Polypropylene
Sanitary clamp adapters
20% Glass filled polypropylene
Bed supports
Polypropylene
Fixed Bed Support seal
Santoprene®
Adjuster seal
Santoprene®
Flowtube seal (fixed and adjuster)
Santoprene®
Adjuster bed support seal
Santoprene®
Fixed flowcell
Polypropylene (black)
Adjuster flowcell
Polypropylene (black)
Antijet disc
20% Glass-filled polypropylene
Sanitary clamp seals
Santoprene® elastomer
Pipe spool
Polypropylene
Materials of construction – non process-wetted parts
Column Component
Material
Adjuster flange, tube flange and fixed flange, fixed flowcell back
plate (200 & 250 only).
Acetal Black
Seal adjust boss, seal adjust tube
Stainless Steel
Threaded central adjust rod
Acetal Black
Seal adjust handles (70mm,100mm), pipework clamp
Acetal blue
Seal adjust handles (140mm 200mm and 250mm)
Stainless Steel/ Acetal Blue
Lifting eye (200 and 250mm only)
Stainless Steel 316
Support Leg
Stainless steel 316
Foot (200 and 250mm only)
Stainless steel with rubber pad
Bolts, screws, washers
Stainless steel type 316

7
Chemical Compatibility
Acrylic columns are ideally suited for aqueous-based applications. See the table below for
detailed compatibility information.
Resistance
¨
Resistant; suitable for continuous use
¨
Not resistant; not recommended
¨
Limited resistance
¨
No information available
Notes
General
All solvents are at 100% concentration unless noted otherwise
Concentrations: % refer to w/w
All data is referenced to room temperature (15º – 25ºC)
Wetted Components
Non Wetted
Stainless Steel 316L
Chemical
1,2-dichloroethane
¨
¨
Acetic acid 25%
¨
¨
Acetone 5%
¨
¨
Acetonitrile
¨
¨
Ammonia aqueous<25%
¨
¨
Ammonium Sulphate 10-40%
¨
¨
Benzyl alcohol 1%
¨
¨
Benzyl alcohol 2%
¨
¨
Butanol
¨
¨
Calcium chloride 2
¨
¨
Calcium hydroxide 30%
¨
¨
Calcium hypochlorite
¨
¨
Chloracetic acid 50%
¨
¨
Chloroform
¨
¨
Chromic acid 10%
¨
¨
Citric acid
¨
¨
Copper sulphate
¨
¨
Dichloromethane
¨
¨
Dimethyl formamide
¨
¨
Dimethyl sulfoxide 10%
¨
¨
Disodium phosphate
¨
¨
Ethanol 20%
¨
¨
Ethanol 70%
¨
¨
Ethylene glycol (1,2-ethanediol)
¨
¨
Formaldehyde 50%
¨
¨
Glycerol (Glycerine)
¨
¨
Guanidine hydrochloride 6M
¨
¨
Hydrochloric acid 10%
¨
¨
Hydrofluoric acid
¨
¨

8
Chemical Compatibility cont.
Resistance
¨
Resistant; suitable for continuous use
¨
Not resistant; not recommended
¨
Limited resistance
¨
No information available
Notes
General
All solvents are at 100% concentration unless noted otherwise
Concentrations: % refer to w/w
All data is referenced to room temperature (15º – 25ºC)
Wetted
Non Wetted
Stainless Steel 316L
Chemical
Hydrogen peroxide
¨
¨
Industrial methylated spirit (IMS)
¨
¨
Methanol 20%
¨
¨
Methanol 50%
¨
¨
Methyl chloride (Chloromethane)
¨
¨
Methyl ethyl ketone (MEK)
¨
¨
Methylene chloride
¨
¨
Nitric acid 10%
¨
¨
Nitric acid 70%
¨
¨
Peracetic acid 300 ppm
¨
¨
Phosphoric acid
¨
¨< 55%
Potassium hydroxide 2M
¨
¨
Iso-propanol 20%
¨
¨
Iso-propanol 40%
¨
¨
n-propanol 20%
¨
¨
Sodium acetate
¨
¨
Sodium bicarbonate 20%
¨
¨
Sodium carbonate
¨
¨
Sodium chlorate
¨
¨
Sodium chloride 2M
¨
¨
Sodium chloride 6M
¨
¨
Sodium hydroxide 0.5M
¨
¨
Sodium hydroxide 2M
¨
¨
Sodium hypochlorite 200 ppm
¨
¨
Sodium nitrate
¨
¨
Sodium sulphate
¨
¨
Trichloroethene (Trichloroethylene)
¨
¨
Triton® X-100 surfactant
¨
¨
Urea 6M
¨
¨
Zinc chloride
¨
¨

9
Safety Guidelines
The following safety guidelines are presented.
Pressure Equipment Directive (PED).
European Pressure Directive 2014/68/EU relates to regulations that are mandatory and govern a
wide range of pressure containing vessels. The design and Manufacture of chromatography
column including the Evolve®column are affected by these regulations.
The regulations are divided into Categories and the Evolve® column has been assessed and found
that it falls within the Sound Engineering Practice (SEP) category. The foundation of the
assessment is based on the following;
• The range of Evolve® columns are liquid chromatography columns and when used in
accordance within their Operating Instructions there should be no gas e.g. air present in
the columns
• The substances that will be used in the column will remain in liquid form during the
entire operation at the maximum stated operating temperature.
In addition
• The maximum operating pressure is greater than 0.5 bar and less than 10 bar
• The product pressure x volume is equal to or less than 200 bar litres.
Over-pressurization
Users of these Chromatography Columns must ensure that the maximum operating pressure
is not exceeded. It is recommended that an over-pressurization protection device such as a
Pressure Relief Valve or equivalent is utilised.
Handling
Care should be taking when handling the Columns. Mechanical handling equipment or
manual handling aids should be utilised where personnel cannot safely lift or move the
Column or its subassemblies.
Chapter
2

10
Larger Columns and their subassemblies are particularly heavy and will necessitate a ‘2
Person’ lift and the subsequent use of mechanical handling equipment or manual handling
aids.
Provision is made for the use of a hoist. It is essential that ‘good practice’ is adopted
regarding the use of appropriate slings and associated equipment.
Chemical compatibility
This manual lists those substances for which the Columns are compatible. If substances not
listed are to be used the user must consider their effect as the integrity and safety of the
Column could be compromised.

11
Installation and Column Set-up
Installation.
The column is shipped preassembled. On receipt, it is recommended to check for damage or
loosening of components during transit.
Complete instructions for disassembly and reassembly can be found in the maintenance
section of the operating manual.
The following column preparation and installation procedures should be followed prior to
operation.
• Install bed supports and flowpath checks
• Set bed height.
• Level the column
Column Preparation
Installation of bed supports
The Evolve® range of columns features polypropylene, woven mesh bed supports,
integrated with a Santoprene seal. These do not require prewetting before use and are
required to be fitted directly onto the adjuster and fixed flowcell.
All handling of bed supports should be performed by a user wearing gloves.
Installation of Fixed Bed Support
1. Remove fixed flowcell from the column.
2. With the rough side facing towards the ribbed flowcell, press the edge of the seal into the
location groove on the side of the flowcell. See Figures 3.1 and 3.2 shown below
Figure 3.1 Diagram showing the location of the fixed bed support
Chapter
3
Smooth Side
Rough Side
Flow cell
Seal

12
Figure 3.2 Photograph showing the installation of the fixed bed support
3. Locate fixed flowcell into the tube unit and fix in place.
Leveling the 70mm, 100mm & 140mm Columns
After installation of the fixed flowcell and before installation of the adjuster unit, it is
recommended to level the column. This is achieved using the spirit level provided in the tool
kit and adjustment of the leveling studs located in the foot ring of the column.
1. Place the spirit level on the tube flange and adjuster the leveling studs to ensure
column is level before proceeding.
There is approximately 30mm free vertical movement available whilst still maintaining a safe
amount of thread engagement.

13
Installation of Adjuster Bed Support
1. The adjuster assembly should be removed from tube unit and placed on its side on a
clean work surface.
2. With the rough side facing towards the ribbed flowcell, press the edge of the seal into
the location groove on the side of the flowcell. See Figure 3.3.
Figure 3.3 Diagram showing the location of the adjuster bed support
3. Work around the circumference of the flowcell ensuring that seal is correctly fitted.
See Figure 3.4.
Figure 3.4 Photograph showing the location of the adjuster bed support
Seal Se tting R ing
Adjuster Seal
Bed Support
Seal
Smooth Side
Bed Support
Ro ug h Si d e
Flowcell
Seal
Adjuster Seal

14
Set Bed Height
1. Reinstate the adjuster unit in the tube, using the location pins and ensure the adjuster
seal is de-energized‘
2. Replace the flange fixing bolts underneath the tube flange and tighten.
3. Partially energized the seal and turn the height adjust handles clockwise to set the
required bed height.
4. Check the column is level by placing the spirit level on the adjuster flange.

15
Column Operation
This section provides general guidance on operating the Evolve® range of columns. It includes a
hydrostatic leak test to establish that the column is installed correctly and advice for the
preparation of the chromatography media.
Further advice specific to each media can be obtained from the media manufacturers; this section
only provides a general guidance.
Hydrostatic Leak Test
The purpose of this test is to check that there are no leaks from the column and the column is
ready to use. Due to the materials of fabrication, when the column is initially exposed to pressure
the column will flex. It is necessary to perform the hydrostatic leak test using a two step
procedure.
1. Ensure fixed end is correctly located and fixings tightened to the correct torque
setting. See table below.
Column Diameter
mm
Torque Setting
Nm
Torque Setting
Ft lbf
70mm
20
14.75
100mm
20
14.75
140mm
20
14.75
2. Fill the column with WFI or high quality water and position the adjuster at the
operating bed height.
3. Ensure the adjuster seal is wetted with buffer or other suitable liquid prior to
assembly. This helps ‘lubricate’ the seal to ensure correct seating.
4. Activate the adjuster seal
5. Ensure all air is removed from the column and any connections.
6. Attach a calibrated pressure gauge to the top inlet of the column
7. Raise the column pressure until 3bar (45 psi) and close bottom inlet valve. Wait
15mins for the pressure to stabilize.
8. Open the valve and increase the pressure to maximum working pressure 4bar (58psi).
9. Wait 15mins for the pressure to stabilize. Check the pressure and ensure pressure is 4
bar (58psi).
10. Wait for 15mins.
Due the nature of materials it is likely there will be a small pressure drop however; this
should not exceed 0.1 bar.
Chapter
4