Buchi Sepiatec SFC-250 User manual

Sepiatec SFC-250
Operation Manual
11594446 | A en

Imprint
Product Identification:
Operation Manual (Original) Sepiatec SFC-250
11594446
Publication date: 01.2023
Version A
BÜCHI Labortechnik AG
Meierseggstrasse 40
Postfach
CH-9230 Flawil 1
E-Mail: [email protected]
BUCHI reserves the right to make changes to the manual as deemed necessary in the light of experience,
especially with respect to structure, illustrations and technical details.
This manual is copyrighted. Information from it may neither be reproduced, distributed, or used for
competitive purposes, nor made available to third parties. The manufacture of any component with the
aid of this manual without prior written agreement is also prohibited.

Büchi Labortechnik AG Table of contents
Operation Manual Sepiatec SFC-250 iii
Table of contents
1 About this document...........................................................................................................6
1.1 Mark-ups and symbols ..........................................................................................................6
1.2 Trademarks ...........................................................................................................................6
1.3 Connected instruments..........................................................................................................6
2 Safety....................................................................................................................................7
2.1 Proper use .............................................................................................................................7
2.2 Use other than that intended .................................................................................................7
2.3 Staff qualification ...................................................................................................................7
2.4 Personal protective equipment ..............................................................................................8
2.5 Warning notices in this document..........................................................................................8
2.6 Warning symbols ...................................................................................................................8
2.7 Residual risks ........................................................................................................................8
2.7.1 Faults during operation.............................................................................................9
2.7.2 Hot surfaces .............................................................................................................9
2.7.3 Dangerous vapors ....................................................................................................9
2.7.4 Dangerous particles..................................................................................................9
2.7.5 Glass breakage ........................................................................................................9
2.7.6 Malfunction of a connected instrument (option)........................................................9
2.7.7 Malware infection due to connections with other devices or network.....................10
2.8 Modifications........................................................................................................................10
2.9 Warning signs......................................................................................................................10
3 Product description...........................................................................................................12
3.1 Description of function .........................................................................................................12
3.2 Configuration .......................................................................................................................14
3.2.1 Front view ...............................................................................................................14
3.2.2 Electric and electronic connections ........................................................................16
3.2.3 CO₂- and coolant connections ................................................................................17
3.2.4 Exhaust connections ..............................................................................................17
3.3 Scope of delivery .................................................................................................................18
3.4 Technical data .....................................................................................................................18
3.4.1 Sepiatec SFC-250 ..................................................................................................18
3.4.2 Ambient conditions .................................................................................................19
3.4.3 Solvents..................................................................................................................19
3.4.4 Detectors ................................................................................................................19
3.4.5 Pumps ....................................................................................................................19
4 Transport and storage ......................................................................................................21
4.1 Transport .............................................................................................................................21
4.2 Storage ................................................................................................................................21
4.3 Lifting the instrument ...........................................................................................................21
5 Installation..........................................................................................................................22
5.1 Installation site.....................................................................................................................22
5.2 Before installation ................................................................................................................24
5.3 Establishing electrical connections......................................................................................24

Table of contents Büchi Labortechnik AG
iv Operation Manual Sepiatec SFC-250
6 Software .............................................................................................................................25
6.1 Introduction..........................................................................................................................25
6.1.1 Starting the software...............................................................................................25
6.1.2 Entering values.......................................................................................................25
6.1.3 Exiting the software ................................................................................................26
6.2 Layout..................................................................................................................................26
6.3 MANUAL tab........................................................................................................................27
6.3.1 Rinsing the tubing and syringe ...............................................................................28
6.3.2 Starting the pump ...................................................................................................28
6.3.3 Setting detector signal to zero ................................................................................29
6.3.4 Changing CO₂ recycling module.............................................................................30
6.4 PARAMETER tab ................................................................................................................30
6.4.1 Setting new methods ..............................................................................................31
6.4.2 Setting gradient mode ............................................................................................32
6.4.3 Loading saved methods .........................................................................................33
6.4.4 Editing saved methods ...........................................................................................33
6.5 COLLECTION tab................................................................................................................34
6.5.1 Setting peak control................................................................................................38
6.5.2 Setting stack injection.............................................................................................38
6.6 SYSTEM tab........................................................................................................................39
6.6.1 Controlling the back pressure.................................................................................40
6.6.2 Controlling the CO₂ valve .......................................................................................40
6.6.3 Controlling the purge pump ....................................................................................41
6.7 RUN METHOD tab ..............................................................................................................42
6.7.1 Before starting a run ...............................................................................................43
6.8 Changing during a run .........................................................................................................44
6.8.1 Change No. of Injections ........................................................................................44
6.8.2 Change injection and run time parameter ..............................................................45
6.8.3 Change fraction parameter.....................................................................................46
6.9 CO₂ recycling module ..........................................................................................................47
7 Operation............................................................................................................................51
7.1 System start - up .................................................................................................................51
7.2 Positioning of the sample ....................................................................................................51
7.3 Positioning the fractionation bottles.....................................................................................52
7.4 Method selection .................................................................................................................52
7.5 Starting a run .......................................................................................................................53
7.6 Finishing a run .....................................................................................................................53
7.7 Shutting down the system ...................................................................................................54
7.8 Using the emergency switch................................................................................................54
8 Cleaning and servicing .....................................................................................................55
8.1 Pumps .................................................................................................................................55
8.2 Servicing the injection valves ..............................................................................................55
8.3 Cleaning the UV detector flow cell.......................................................................................55
8.4 Back pressure regulator valve .............................................................................................55
8.5 Fractionation valve ..............................................................................................................55
8.5.1 Cleaning the valve and replacing the rotor .............................................................55
8.5.2 Dismantling............................................................................................................. 56
8.5.3 Assembling .............................................................................................................56
8.6 Gas-liquid-separators (GLS)................................................................................................57

Büchi Labortechnik AG Table of contents
Operation Manual Sepiatec SFC-250 v
9 Help with faults ..................................................................................................................59
9.1 Troubleshooting...................................................................................................................59
10 Taking out of service and disposal..................................................................................60
10.1 Taking out of service ...........................................................................................................60
10.2 Disposal...............................................................................................................................60
10.3 Returning the instrument .....................................................................................................60
11 Appendix ............................................................................................................................61
11.1 Spare parts and accessories ...............................................................................................61

1 | About this document Büchi Labortechnik AG
6/62 Operation Manual Sepiatec SFC-250
1 About this document
This operation manual is applicable for all variants of the instrument.
Read this operation manual before operating the instrument and follow the
instructions to ensure safe and trouble-free operation.
Keep this operation manual for later use and pass it on to any subsequent user or
owner.
BÜCHI Labortechnik AG accepts no liability for damage, faults and malfunctions
resulting from not following this operation manual.
If you have any questions after reading this operation manual:
uContact BÜCHI Labortechnik AG Customer Service.
https://www.buchi.com/contact
1.1 Mark-ups and symbols
NOTE
This symbol draws attention to useful and important information.
RThis character draws attention to a requirement that must be met before the
instructions below are carried out.
uThis character indicates an instruction that must be carried out by the user.
ðThis character indicates the result of a correctly carried out instruction.
Mark-up Explanation
Window
Software Windows are marked-up like this.
Tab
Tabs are marked-up like this.
Dialog
Dialogs are marked-up like this.
[Button] Buttons are marked-up like this.
[Field names] Field names are marked-up like this.
[Menu / Menu item] Menus or menu items are marked-up like this.
Status Status is marked-up like this.
Signal Signals are marked-up like this.
1.2 Trademarks
Product names and registered or unregistered trademarks that are used in this
document are used only for identification and remain the property of the owner in
each case.
1.3 Connected instruments
In addition to this operation manual, follow the instructions and specifications in the
documentation for the connected instruments.

Büchi Labortechnik AG Safety | 2
Operation Manual Sepiatec SFC-250 7/62
2 Safety
2.1 Proper use
The instrument is designed for super fluid chromatography.
The instrument can be used in laboratories for the following tasks:
Purification
Separation of one or more compounds from a mixture
2.2 Use other than that intended
Use of any kind other than that described in Chapter2.1 "Proper use", page7 and
any application that does not comply with the technical specifications (See Technical
data) constitutes use other than intended. In particular, the following applications
are not permissible:
Use of the instrument in areas which require explosion-safe instruments.
Use of the instrument with solvents containing peroxides.
2.3 Staff qualification
Unqualified persons are unable to identify risks and are therefore exposed to greater
dangers.
The instrument may only be operated by suitably qualified laboratory staff.
These operating instructions are aimed at the following target groups:
Users
Users are persons that meet the following criteria:
They have been instructed in the use of the instrument.
They are familiar with the contents of these operating instructions and the
applicable safety regulations and apply them.
They are able on the basis of their training or professional experience to assess
the risks associated with the use of the instrument.
Operator
The operator (generally the laboratory manager) is responsible for the following
aspects:
The instrument must be correctly installed, commissioned, operated and serviced.
Only suitably qualified staff may be assigned the task of performing the
operations described in these operating instructions.
The staff must comply with the local applicable requirements and regulations for
safe and hazard-conscious working practices.
Safety-related incidents that occur while using the instrument should be reported
to the manufacturer ([email protected]).
BUCHI service technicians
Service technicians authorized by BUCHI have attended special training courses and
are authorized by BÜCHI Labortechnik AG to carry out special servicing and repair
measures.

2 | Safety Büchi Labortechnik AG
8/62 Operation Manual Sepiatec SFC-250
2.4 Personal protective equipment
Depending on the application, hazards due to heat and/or corrosive chemicals may
arise.
uAlways wear appropriate personal protective equipment such as safety goggles,
protective clothing and gloves.
uMake sure that the personal protective equipment meets the requirements of
the safety data sheets for all chemicals used.
2.5 Warning notices in this document
Warning notices warn you of dangers that can occur when handling the instrument.
There are four danger levels, each identifiable by the signal word used.
Signal word Meaning
DANGER Indicates a danger with a high level of risk which could result in
death or serious injury if not prevented.
WARNING Indicates a danger with a medium level of risk which could result in
death or serious injury if not prevented.
CAUTION Indicates a danger with a low level of risk which could result in mi-
nor or medium-severity injury if not prevented.
NOTICE Indicates a danger that could result in damage to property.
2.6 Warning symbols
The following warning symbols are displayed in this operation manual or on the
instrument.
Symbol Meaning
General warning
Instrument damage
Dangerous electrical voltage
Hot surface
2.7 Residual risks
The instrument has been developed and manufactured using the latest technological
advances. Nevertheless, risks to persons, property or the environment can arise if
the instrument is used incorrectly.
Appropriate warnings in this manual serve to alert the user to these residual
dangers.

Büchi Labortechnik AG Safety | 2
Operation Manual Sepiatec SFC-250 9/62
2.7.1 Faults during operation
If an instrument is damaged, sharp edges, glass splinters, moving parts or exposed
electrical wires can cause injuries.
uRegularly check instruments for visible damage.
uIf faults occur, switch off the instrument immediately, unplug the power cord and
inform the operator.
uDo not continue to use instruments that are damaged.
2.7.2 Hot surfaces
The column oven area and the heating elements of the device can become hot. If
touched they can cause skin burns.
uDo not touch hot surfaces or else wear suitable protective gloves.
2.7.3 Dangerous vapors
The use of the instrument can produce dangerous vapors that are capable of causing
life-threatening toxic effects.
uDo not inhale any vapors produced during processing.
uEnsure that vapors are removed by a suitable fume hood.
uOnly use the instrument in well ventilated areas.
uIf vapors escape from connections, check the ferrules an fittings concerned and
replace them if necessary.
uDo not process any unknown fluids.
uObserve the safety data sheets for all substances used.
2.7.4 Dangerous particles
The use of the instrument can produce dangerous particles that can cause life-
threatening toxic effects.
uDo not inhale any particles produced during processing.
uEnsure that particles are removed by a suitable fume hood.
uOnly use the instrument in well ventilated areas.
uIf particles escape from connections, check the ferrules an fittings concerned and
replace them if necessary.
uDo not process any unknown fluids or solvent mixtures.
uObserve the safety data sheets for all substances used.
2.7.5 Glass breakage
Broken glass can cause severe cuts.
Damaged glass components may implode if subjected to high pressure.
Minor damage to the ground joints impairs the sealing effect and may therefore
diminish performance.
uHandle the flask and other glass components carefully and do not drop them.
uAlways visually inspect glass components for damage every time they are to be
used.
uDo not continue to use glass components that are damaged.
uAlways wear protective gloves when disposing of broken glass.
2.7.6 Malfunction of a connected instrument (option)
A malfunction on a connected instrument can cause poisoning or death.

2 | Safety Büchi Labortechnik AG
10/62 Operation Manual Sepiatec SFC-250
uMake sure that the connected instrument is prepared and maintained according
to the user documentation.
2.7.7 Malware infection due to connections with other devices or network
Connections with other devices or a network can cause a malware infection to the
instrument.
uInstall antivirus software and firewall on the instrument before connecting it to
other devices or network.
2.8 Modifications
Unauthorized modifications can affect safety and lead to accidents.
uUse only genuine BUCHI accessories, spare parts and consumables.
uCarry out technical changes only with prior written approval from BUCHI.
uOnly allow changes to be made by BUCHI service technicians.
BUCHI accepts no liability for damage, faults and malfunctions resulting from
unauthorized modifications.
2.9 Warning signs
DANGER
Risk of electric shock
Risk of death by electrocution
uNever remove the housing.
uNever operate the instrument with removed housing or without the included
pumps.
uNever use any electric connections or cables that were not supplied with the
instrument.
DANGER
Risk of cold burns
Pressurized gases can generate low temperatures on depressurization.
uAlways wear appropriate personal protective equipment such as safety goggles,
protective clothing and gloves.

Büchi Labortechnik AG Safety | 2
Operation Manual Sepiatec SFC-250 11/62
DANGER
Hot surface
The column oven can reach temperatures of up to 70°C. The heating can heat up for
some time even after switching off power, due to internally stored energy.
uOpen door carefully so that the hot air can vent.
uOnly change columns after complete cool down.
uAlways wear appropriate personal protective equipment such as safety goggles,
protective clothing and gloves.
uMake sure the ventilation slits and the oven heater are not covered.
WARNING
Pressurized gas
The operating instrument is pressurized. Venting of pressurized gases and solvents is
possible. CO2 is suffocating in high concentrations.
uInstall gas detectors at site.
uCreate sufficient ventilation.
CAUTION
Risk due to falling objects or material
Personal injury or property damage
uNever place solvent bottles or other items on the top of the instrument.
NOTE
The pumps switch off automatically when reaching 400 bar.
The detector flow cell is stable up to a pressure of 300 bar. To prevent damage to
the flow cell, the back pressure regulator is programmed to switch off at 300 bar.
NOTE
The system my be pressurized after switching off.
Keep in mind, the escape of pressurized gases and solvents are possible.

3 | Product description Büchi Labortechnik AG
12/62 Operation Manual Sepiatec SFC-250
3 Product description
3.1 Description of function
The instrument is for preparative separations on small columns by means of
supercritical fluid chromatography (SFC).
Supercritical fluid chromatography has the ability to separate gram size samples.
The integrated gas-liquid-separators (GLS) allow fast and efficient removal of CO2
from separated components.
The instrument allows:
A mix of supercritical CO2 and one solvent.
With the optional solvent selection valve, the solvent can be chosen from four
different solvents.
Injection of liquid or solid sample
Separation on a column
Detection of the components by using a UV, ELSD or MS detector.
Collecting the desired fractions

Büchi Labortechnik AG Product description | 3
Operation Manual Sepiatec SFC-250 13/62
12 3 4 5 6 7
8
910
11
12
13
14
15
16
17
18
19
5 °C 70 °C 50 °C
1 Pumps 2 Column oven
3 Detector, flow cell 4 Back pressure regulator
5 Post-heater 6 Gas-liquid-separator
7 Fractions (8 pcs.) 8 Liquid CO2 60 - 75 bar
9 Pre-cooling 10 CO2 pump
11 Modifier pump 12 Sample
13 Modifier stream injection 14 Supercritical CO2 80 - 250 bar
15 BPR pressure value 80-250 bar 16 Add-on pump
17 Low pressure 18 CO2 exhaust
19 Depressurized fraction collection
The mobile phase in supercritical fluid chromatography consists of a supercritical
fluid and a liquid solvent. This fluid is a condensed gas above the critical temperature
and pressure. Its properties are those of a gas and liquid. Carbon dioxide (CO2) is
mostly used.
The CO2 is transported by the pump in a cooled state. The fluid gets heated above
the supercritical temperature in the column oven and mixed with the modifier. The
back pressure regulator maintains an elevated pressure to keep the mobile phase
above the critical point of CO2. The sample is then transported over the separation
column by the supercritical fluid. A modifier can be added to improve the elution of
organic substances.
After detection, the CO2 is separated from the eluted substances. The Detection and
Fraction valve position decides in which GLS the stream is going. The GLS separates
most of the CO2 and removes it from the system by an exhaust pipe and fed into the
ventilation system.

3 | Product description Büchi Labortechnik AG
14/62 Operation Manual Sepiatec SFC-250
The collected fractions contain the dissolved substances in the modifier and a small
amount of residual CO2 and optional in the organic from the add-on pump. The
fractions are available in highly concentrated form for further analysis.
3.2 Configuration
3.2.1 Front view
1
2
3
4
5
6
7
8
9
4
1 Touchscreen 2 Gas-liquid-separator
3 Fraction bottles 4 CO2 and modifier pump
5 Detector flap 6 Fraction collector valve
7 Column oven 8 Back pressure regulator cover
9 Add-on pump (optional)

Büchi Labortechnik AG Product description | 3
Operation Manual Sepiatec SFC-250 15/62
Column oven
1
2
3
4
5
6
7
7
8
1 Flow cell with optic fibre cable 2 Leak sensor
3 Injection valve 4 Column selector valve
5 Heating module 6 Filter
7 Check valves 8 Column holder

3 | Product description Büchi Labortechnik AG
16/62 Operation Manual Sepiatec SFC-250
3.2.2 Electric and electronic connections
1
2
3
4
5
6
7
1 Fuses and FI-circuit breaker 2 Main switch
3 USB connection 4 HDMI connection
5 LAN external connection 6 LAN internal connection
7 Emergency switch connection

Büchi Labortechnik AG Product description | 3
Operation Manual Sepiatec SFC-250 17/62
3.2.3 CO₂- and coolant connections
1123
1 Connection for cooling water supply 2 Outlet CO2 safety relief valve
3 Connection for CO2 supply
3.2.4 Exhaust connections
There are three exhaust connections on this instrument:
On the left side is the exhaust of the fractionation compartment.
On the right side is the outlet of the CO2 safety exhaust valve.
On the rear side is the GLS CO2 exhaust.

3 | Product description Büchi Labortechnik AG
18/62 Operation Manual Sepiatec SFC-250
3.3 Scope of delivery
NOTE
The scope of delivery depends on the configuration of the purchase order.
Accessories are delivered as per the purchase order, order confirmation, and
delivery note.
3.4 Technical data
3.4.1 Sepiatec SFC-250
Dimensions (W × D × H) 720 mm × 600 mm × 880 mm
Weight 130 kg
Power consumption Max. 1,300 W
Connection voltage Europe: 230 ± 10% VAC
Asia: 230 ± 10% VAC
USA: 120 ± 10% VAC
Frequency 50 / 60 Hz
Fuse 15 A
Over voltage category II
IP Code IP 20
Pollution degree 2
Minimum clearance on all sides 200 mm
Controller 15.6” touchscreen, 16:9 format
System Integrated Windows 10
Control software Prep SFC control software
Operating temperature range 15 − 25 °C
Operating pressure Max. 300 bar
Injection valve Electrically operated 6-port / 2-ways
valve
Column selection valve Electrically operated 11-port / 10-ways
valves
Back pressure regulator valve Electromagnetic check valve
Fractionation valve Electrically operated 11-port / 10-ways
valve
Tubing Stainless steel capillary tubes
OD: 1/16´´ ID: 1.0 mm or 0.5 mm or 0.25
mm
Pump head temperature control Chiller
Method development On analytical columns (ID: 4 − 4.6 mm)
Column dimension ID: 4 − 16 mm
Column length Max. 250 mm
Column oven temperature Ambient to 70 °C

Büchi Labortechnik AG Product description | 3
Operation Manual Sepiatec SFC-250 19/62
Standard column capacity 2
Max column capacity 8 (optional)
Fraction collection number 1 - 8 pcs.
Volumes unlimited
Pressure-less collection Standard
Syringe size 1.0 ml
Other volumes on request
Loop 0.5 ml
Stack injection Standard
Certificate CSA / CE
3.4.2 Ambient conditions
For indoor use only.
Max. altitude above sea level 2,000m
Ambient and storage temperature 15 − 25 °C
Maximum relative humidity 45 − 75% not condensing
3.4.3 Solvents
CO2Liquid
Organic modifier solvents 4 pcs.
Only with the optional solvent selector
valve.
3.4.4 Detectors
UV wavelengths range 190 − 500 nm
UV wavelengths selectable 1
DAD Standard
ELSD Optional
MS Optional
3.4.5 Pumps
CO2 Pump
Flow rate
150 ml/min
CO2 Pump
Pressure
Max. 400 bar
Modifier pump 2 pcs.
Modifier pump
Flow rate
30 ml/min
Modifier pump
Pressure
Max. 400 bar
Add-on pump Standard

3 | Product description Büchi Labortechnik AG
20/62 Operation Manual Sepiatec SFC-250
Flow rate accuracy ±2% at 2 − 50% of the flow range with
methanol (water 80/20 v/v)
Precision 0.25% from 1 ml/min to 20 ml/min at 20
°C