ÄKTA system Training Guide
Purifier 100 / 10
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ÄKTApurifier
Buffers
Injector
Box
Monitor
On-line
Filter
Conductivity
pH
Outlet Valve
UV Detector
Mixer
Pumps
Pump A
Pump B
Buffer switching
valve
ÄKTApurifier 10 / 100
In-line
filter
1 7
6
2
3 4 5
Injection
valve (V1)
UV
3 Wavelength
detector
Cond.
Conductivity
monitor
pH
Mixer
pH detector
System
pumps
A
Fraction
collector
Frac-950
B
1
8
2
3
4
7
6
5
Outlet
valve (V4)
ÄKTApurifier + Sample Pump + Sample Valve
In-line
filter
Injection
valve (V1)
1 7
6
2
3 45
Mixer
UV
3 Wavelength
detector
Cond
Conductivity
monitor
System
pumps
A
pH
B
Sample
pump
pH detector
Fraction
collector
Frac-950
1 87
2
6
345
1 87
2
6
345
Sample selection
valve (V5)
Outlet
valve (V4)
ÄKTApurifier 100 Air
In-line
filter
1 7
6
2
3 4 5
Injection
valve (V1)
UV
3 Wavelength
detector
Cond.
Conductivity
monitor
pH
Mixer
pH detector
System
pumps
A
Fraction
collector
Frac-950
B
Air
sensor
8
1
2
3
4
7
6
5
Outlet
valve (V4)
ÄKTApurifier + Buffer Valve
In-line
filter
1 7
6
2
3 4 5
Injection
valve (V1)
UV
3 Wavelength
detector
Cond.
Conductivity
monitor
pH
Mixer
pH detector
System
pumps
A
Fraction
collector
Frac-950
B
1
8
2
3
1 87
2
6
345
Buffer
select
valve (V6)
4
7
6
5
Outlet
valve (V4)
ÄKTApurifier 10 XT
In-line
filter
1 7
6
2
3 4 5
Injection
valve (V1)
UV
3 Wavelength
detector
Cond.
Conductivity
monitor
pH
Mixer
pH detector
System
pumps
A
Fraction
collector
Frac-950
B
1
8
2
3
4
7
6
5
Outlet
valve (V4)
PUMP P-900
Mixer
Pump A
Pump B
Proportion
Valve
A2
Acid /Alkali
ÄKTA 10 mls/min
ÄKTA 100 mls/min
A1 B2
Buffer
B1
Salt
GRADIENT
<6 mls/min
6 -14 mls/min
<30 mls/min
30 - 50 mls/min
50 - 100 mls/min
Water
MIXER
0.6 ml mixer
2 ml mixer
2 mls mixer
5 mls mixer
12 mls mixer
PUMP P-900 Inlets
Mixer
Pump B
Pump A
Switching
Valves
Buffer Select
Valve IV-908
A2
A1
2
3
A11
A12
A13
1 8
4
A14
B1
Water
7
5
6
A16
A15
B2
A17
A18
Salt
PUMP P-900 Manually Priming Buffer Inlet Lines
Pump A
Common inlet
A2
Pump B
A1
Common inlet
Manual
Flowpath
Manual
A1 or A2
BufferValveA B1 or B2
B1
B2
Flowpath
BufferValveB
Open
Draw A
ir
out of
Buffer
Line
PUMP P-900 Back - Wash Lines
Pump Back-Wash
Lines
Syringe fitting
Wash OUT
IN-LINE
one-way
valve
Buffer
OUT
Ball outlet
check valve
High pressure
Piston seal
Piston
20% Ethanol : Water
Backwash pump
diaphragm
Ball inlet
check valve
Wash
IN
Buffer
IN
ON-LINE FILTERS
P-903 ( 10 ml/min Pump )
Low Flow HPLC Filter
P-901 ( 100 ml/min Pump )
High Flow Fibre Filter
Polymer Support
LOAD (sample)
INJECT (sample)
2
3
2
Pump
7
4
4
6
5
8
4
4
Waste
6
3
4
5
Sample
8
7
6
3
6
5
2
2
5
6
PV-908
1
8
7
7
2
Pump
7
3
PV-908
1
Sample
2
Pump
7
3
6
5
PV-908
4
1
1
1
3
Column
Column
Column
1
LOAD (wash loop)
Waste
5
Sample
Waste
INJECTION VALVE INV- 907
1
COLUMN
7
2
LOAD
SAMPLE
3
PUMP
6
4
5
Waste
Waste
LOAD
1
COLUMN
7
2
PUMP
3
4
6
5
Waste
Waste
INJECT
1
COLUMN
7
2
PUMP
3
4
Waste
6
5
Waste
WASTE
SAMPLE LOADING METHOD
1
COLUMN
2
LOAD
SAMPLE
7
PUMP
3
6
5
4
Needle Port
Waste
Waste
Always tighten by hand
with needle inserted
Do Not Overtighten
1
COLUMN
LOAD
SAMPLE
7
2
PUMP
3
6
4
5
Luer Lock
Adapter
Waste
Waste
Superloop
SUPERLOOPS
10 ml
50 ml
Buffer
150 ml
Plunger
BUFFER
FLUSH
SAMPLE
LOAD
SAMPLE
INJECT
SAMPLE
FLUSH
WASTE
PUMP or
SYRINGE
COLUMN
COLUMN
Sample
Etched
Groove
Volume
Max. allowed column
pressure
Code No.
10 ml
50 ml
150 ml
4 Mpa
4 Mpa
2 Mpa
18-1113-81
18-1113-82
18-1023-85
INV-907 Injection Valve - Injection Port
PEEK Valve Body
Injection Needle Port
Valve Seal
Ferrule
NeedleTube
Needle
Needle
Adapter
INV-907 Injection Valve - Luer Adapter
PEEK
PEEKInjection
InjectionValve
ValveBody
Body
PEEK
PEEKInjection
InjectionLuer
LuerAdaptor
Adaptor
Valve
ValveSeal
Seal
Valve
ValveChannel
ChannelPlate
Plate
Fluid Dynamics
Flow
PEEK Tubing
High Resistance
to Flow
Low Resistance
to Flow
End
Beginning
Fluid Dynamics
Flow - Injecting a Loop
The way we imagine it Flows
The way it Actually Flows
Bubble Segmentation
100ul Loop
Waste
50ul Load
60ul Load
100ul Load
200ul Load
• Partial Loading Loop = No more than 1/2 Loop Volume
• Total Loading Loop = Inject 2X Loop Volume
LOADING A SAMPLE LOOP
Injection valve
INV-907
1
Column
7
2
Load
3
Inject
Sample
4
Sample
pump
P-950
Pump
6
5
Waste
LOADING A SUPER LOOP
Injection valve
INV-907
1
Column
7
2
Load
Inject
3
Pump
6
4
5
Buffer
Sample
Waste
Sample
MULTI-LOADING A SAMPLE LOOP
Injection valve
INV-907
1
Column
7
2
Load
3
Inject
4
Pump
6
5
PV-908
1
8
2
3
I/O
4
5
7
6
Wash
Samples
Waste
MULTI-LOADING A SUPER LOOP
Column
1
2
Pump
Load
Inject
Injection valve
INV-907
7
3
4
Waste
6
5
PV-908
8
1
2
3
I/O
4
5
7
6
Samples
Wash
S3 S4 S5 S6
S8
DIRECT LOADING ON TO COLUMN
Column
1
Pump
7
2
3
Inject
Load
6
4
5
Extra Tube
1/16” or 1/8”
Tubing
Waste
Sample
REVERSE FLOW VALVE ( INV-907 )
1
7
2
Pump
3
5
4
Column
Extra Tube
6
Stop
Plug
MONITOR
Downward Flow
Position 1
1
7
2
Pump
3
4
Column
Extra Tube
5
6
Stop
Plug
MONITOR
Upward Flow
Position 2
OUTLET VALVE PV-908
DETECTOR
To Fraction
Collector
Default
F8
F1
F2
F3
I/O
F4
F5
F7
F6
F7
Waste
F8
F4
F3
F5
F6
Fraction Collection
Flowthrough
Fractionation
Peak
Fractionation
Eluate
Fractionation
100%B
0%B
mAU
100
50
Waste
End
D
N
U
O
B
N
U
D
N
U
O
B
Start
Inject
500
250
%B
0
A1 A2 A3………………….
0
HiTrap Connections
1/16” Female
1/16” Male
18-1112-55
1/16” Fingertight Fitting
1/16” Female
M6 Male
18-1112-57
M6 male / 1/16” Female
Phenyl Sepharose
HiTrap Column
M6 Male
M6 Female
18-1112-58
M6 female / 1/16” male
Flow Cell Inlet
1/16” Male
Multiple HiTrap Arrangment ( Series )
Phenyl Sepharose
Phenyl Sepharose
Phenyl Sepharose
Phenyl Sepharose
Phenyl Sepharose
Phenyl Sepharose
De
or
t
c
te
t or
c
e
t
De
or
t
c
te
De
Traditional FPLC fitting to ÄKTA / HPLC
Injector
1/16” Fingertight Fitting
18-1112-55
10
0u
1/16” FEMALE / M6 FEMALE
18-1123-94
l
Flange M6 Fitting
1/16” MALE / M6 FEMALE
18-1112-58
Flow Cell
Tubing Fittings
18-1112-55
1/16” Fingertight Fitting
1/16” PEEK
TM
Tubing
18-1112-58
Flange Fitting Adapter
FPLC Flange Fitting
18-1112-51
Luer Fitting Adapter
18-1112-52
Fingertight Stop Plug
Flangless Inlet Connector
3/16”
18-1112-49
1/8”
18-1121-17
1/16”
18-1127-07
18-1112-57
M6 male / 1/16” Female
Luer Syringe
18-1112-50
5/16” Stop Plug
Flangless Buffer Inlet
Ferrule
3/16”
18-1112-48
1/8”
18-1121-18
1/16”
18-1127-06
18-1017-98
Tubing Connector
M6 male / O - ring
18-3955-01
1/16”female X 1/16”female Union Ti
Union + 2 Fingertight Fittings
18-1123-94
M6 female X 1/16”female Union
FPLC Flange - 1/16” Fingertight
COMPRESSION FITTINGS
Untightened
Correctly tightened
Obstructed
Over tightened Tubing Restricted
PEEK Tubing
TM
Gray, 1/16”x 1.0 mm ( 7.85ul / cm )
Green, 1/16”x 0.75 mm ( 4.42ul / cm )
Orange, 1/16”x 0.5 mm ( 1.96ul / cm )
Blue, 1/16”x 0.25 mm ( 0.49ul / cm )
Teflon / Tefzel Tubing
3/16” OD
18-1112-47
1/8” OD
18-1121-16
1/16” OD
18-1119-74
Tubing Kit Changes - ÄKTApurifier 10
GENERAL FLOW (at delivery)
G5
10
0u
l
Injection valve V1
G9
Outlet Valve V4 (7ul)
Accumulator
(40ul)
UV Flow cell
G8
G6
0.25 mm i.d
Blue PEEK
G5 400mm
G6 160mm
G7 80mm
G8 130mm
G9 500mm
G7
Conductivity
Flow cell (14ul)
FR-904
(0.2Mpa, 5ul)
NB: Never use
the FR-904 with
the pH electrode
pH Flow cell (100ul)
Tubing Kit
AKTApurifier 10 , 0.50 tubing kit
AKTApurifier 10 , 0.50 tubing kit + pH electrode
AKTApurifier 10 , 0.25 tubing kit
No Frac
Frac 901
232
354
88
Frac 950
382
504
238
Tubing Kit Changes - ÄKTApurifier 10
HIGH FLOW
G5
10
0u
l
Injection valve V1
H9
Outlet Valve V4 (7ul)
Accumulator
(40ul)
UV Flow cell
H8
H6
H7
Conductivity
Flow cell (14ul)
0.50 mm i.d
Orange PEEK
H6 160mm
H7 80mm
H8 130mm
H9 500mm
FR-904
(0.2Mpa, 5ul)
NB: Never use
the FR-904 with
the pH electrode
pH Flow cell (100ul)
Tubing Kit
AKTApurifier 10 , 0.50 tubing kit
AKTApurifier 10 , 0.50 tubing kit + pH electrode
AKTApurifier 10 , 0.25 tubing kit
No Frac
Frac 901
232
354
88
Frac 950
382
504
238
Tubing Kit Changes - ÄKTApurifier 100
HIGH FLOW (at delivery)
G5
10
0u
l
Injection valve V1
G9
Outlet Valve V4 (7ul)
Accumulator
(40ul)
UV Flow cell
G8
G6
0.75 mm i.d
Green PEEK
G6
G7
G8
G9
G7
Conductivity
Flow cell (14ul)
FR-904
(0.2Mpa, 5ul)
160mm
80mm
130mm
500mm
NB: Never use
the FR-904 with
the pH electrode
pH Flow cell (100ul)
Tubing Kit
AKTApurifier 100 , 0.50 tubing kit
AKTApurifier 100 , 0.75 tubing kit
AKTApurifier 100 , 1.00 tubing kit
AKTApurifier 100 , 0.50 tubing kit + pH electrode
AKTApurifier 100 , 0.75 tubing kit + pH electrode
AKTApurifier 100 , 1.00 tubing kit + pH electrode
AKTAbasic 100 , 0.75 tubing
No Frac
290
Frac 901
232
470
769
354
672
955
Frac 950
382
620
809
504
862
995
Tubing Kit Changes - ÄKTApurifier 100
LOW FLOW
L5
10
0u
l
Injection valve V1
L9
Outlet Valve V4 (7ul)
Accumulator
(40ul)
UV Flow cell
L8
L6
0.50 mm i.d
Orange PEEK
L6
L7
L8
L9
L7
Conductivity
Flow cell (14ul)
160mm
80mm
130mm
500mm
FR-904
(0.2Mpa, 5ul)
NB: Never use
the FR-904 with
the pH electrode
pH Flow cell (100ul)
Tubing Kit
AKTApurifier
AKTApurifier
AKTApurifier
AKTApurifier
AKTApurifier
AKTApurifier
No Frac
100
100
100
100
100
100
,
,
,
,
,
,
0.50
0.75
1.00
0.50
0.75
1.00
tubing
tubing
tubing
tubing
tubing
tubing
kit
kit
kit
kit + pH electrode
kit + pH electrode
kit + pH electrode
Frac 901
232
470
769
354
672
955
Frac 950
382
620
809
504
862
995
A-900 Autosampler
INV-907
1
A-900
2
1
2
7
6
3
4
5
3
MANUAL
SAMPLE
LOAD
LOAD
6
5
4
Waste
COLUMN
Sample
PUMP
Waste
LOAD
Syringe
INV-907
1
A-900
2
1
6
3
4
5
3
MANUAL
SAMPLE
LOAD
INJECT
Syringe
Sample
7
2
4
6
5
Waste
Waste
COLUMN
PUMP
LOAD
Needle Height
Standard Vial
Standard Vial
+
Needle Height Insert
volume left
2mm
450ul
1mm
350ul
0mm
200ul
3mm
12ul
A-900
Sample Injection ( Flushed Loop Injection )
INJECT
INJECT
INJECT
INJECT
LOAD
LOAD
pump
waste
INJECT
INJECT
pump
waste
waste
waste
column
column
pump
pump
column
column
Sample needle
sample
INJECT,sample
sample
InInINJECT,
flushesthe
thevalve
valve
flushes
channel.
channel.
Min.( (30ul
30ul) )with
with
Min.
AirSegment
Segment
Air
Min.( (35ul
35ul) )without
without
Min.
Airsegment
segment
Air
sample
LOADThe
Thesample
sample
InInLOAD
drawnup
upto
to
isisdrawn
completely
completely
fillthe
theloop.
loop.
fill
noless
lessthan
thantwice
twice
( (no
loopvolume
volume
loop
recommended) )
recommended
Wash
Reservoir
sample
INJECTsample
sample
InInINJECT
washedonto
onto
isiswashed
thecolumn.
column.
the
sample
Finalinternal
internal
Final
needlewash.
wash.
needle
pump
waste
column
INJECT
INJECT
OptionalExternal
External
Optional
needlewash.
wash.
needle
sample
transport
fluid
A-900
Sample Injection ( Partial Loop Injection )
INJECT
INJECT
INJECT
INJECT
LOAD
LOAD
INJECT
INJECT
waste
pump
pump
waste
waste
sample
INJECT,sample
sample
InInINJECT,
flushesthe
thevalve
valve
flushes
channel.
channel.
Min.( (30ul
30ul) )with
with
Min.
AirSegment
Segment
Air
Min.( (35ul
35ul) )without
without
Min.
Airsegment
segment
Air
waste
waste
column
Sample needle
pump
pump
column
Sample needle
sample
Thesample
sampleisisdrawn
drawn
The
upto
topartially
partiallyfill
fillthe
the
up
loopwith
with
loop
programmedvolume.
volume.
programmed
nomore
morethan
thanhalf
half
( (no
loopvolume
volume
loop
recommended) )
recommended
column
column
Sample needle
sample
injectsample
sample
InIninject
washedonto
onto
isiswashed
thecolumn.
column.
the
Wash
Reservoir
sample
Finalinternal
internal
Final
needlewash.
wash.
needle
pump
waste
column
INJECT
INJECT
OptionalExternal
External
Optional
needlewash.
wash.
needle
sample
transport
fluid
A-900
Sample Injection ( ul Pick-up Injection )
INJECT
INJECT
LOAD
LOAD
pump
pump
waste
transport
fluid
sample
sample
Thesample
sampleisis
The
drawnup
upfrom
from
drawn
thesample
samplevial
vial
the
INJECT
INJECT
column
Sample needle
Sample needle
sample
sample
Thesample
sampleisis
The
washedinto
into
washed
theloop
loopwith
with
the
transportfluid
fluid
transport
pump
injectsample
sample
InIninject
washedonto
ontothe
theco
column
isiswashed
lumn
pump
waste
waste
Finalinternal
internal
Final
needlewash.
wash.
needle
Wash
Reservoir
pump
column
Sample needle
inject,valve
valve
InIninject,
flusheswith
with
flushes
transportfluid
fluid
transport
pump
column
column
INJECT
INJECT
LOAD
LOAD
column
sample
column
INJECT
INJECT
Optionalneedle
needle
Optional
wash.
wash.
sample
transport
fluid
System Compensation Volume
Equilibration Phase
A
B
COLUMN TOP
0%
A
B
COLUMN TOP
20%
Effect
8 mls
Command
20%B
0%B
8mls
Gradient Delay Volume
Gradient Phase
A
B
COLUMN TOP
0%
A
B
COLUMN TOP
100%
Effect
16 mls
100%B
Gradient Delay
Command 100%
Column Pressure ( compensation ) Flow Restrictor
Column Pressures
Column Hardware Pressure Limits
Column Shatters DANGEROUS
Column Matrix Pressure Limits
Matrix Collapses EXPENSIVE
0.3Reading
Reading
0.3
Atmos
Atmos
Press
Press
0.0MPa
MPa
0.0
Pre-column pressure
= 0 MPa
Pump
Pump
0.3Mpa
Mpa
0.3
0.3 MPa
GELPress
PressDrop
Drop
GEL
0.3MPa
MPa
0.3
(0.3- -0.0)
0.0)
(0.3
0.0 MPa
0.0MPa
MPa
0.0
0.5Reading
Reading
0.5
Pre-column pressure
= 0 MPa
Gel Top Pressure
Pump
Pump
0.3Mpa
Mpa
0.3
0.3 MPa
ColumnPress
PressDrop
Drop
Column
GELPress
PressDrop
Drop
GEL
Colu
mnPress
PressDrop
Drop
Colu
mn
(0.3++0.0)
0.0)
(0.3
(0.3- -0.2)
0.2)
(0.3
(0.5++0.2)
0.2)
(0.5
0.3MPa
MPa
0.3
Gel Bottom Pressure
0.1MPa
MPa
0.1
0.2 MPa
0.2MPa
MPa
0.2
FlowRestrictor
Restrictor
Flow
0.5MPa
MPa
0.5