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Purifier 100 / 10
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ÄKTApurifier
Buffers
Injector
Box
Monitor On-line
Conductivity
pH
Filter
Pumps Mixer
Pump A
Buffer switching
Pump B valve
ÄKTApurifier 10 / 100
In-line UV 3 Wavelength
filter
detector
Injection
valve (V1) Cond. Conductivity
1 7 monitor
2 6
3 4 5
Mixer pH
pH detector
System
pumps
Fraction
A B collector
Frac-950
1 8
7
2 6
Outlet
3 5
4 valve (V4)
Injection UV 3 Wavelength
valve (V1) detector
1 7
2 6
3 45 Cond Conductivity
Mixer monitor
System
pumps
pH pH detector
A B
Sample
pump Fraction
1 87 collector
2 6
345 Frac-950
1 87
2 6 Outlet
345
valve (V4)
Sample selection
valve (V5)
ÄKTApurifier 100 Air
In-line UV 3 Wavelength
filter
detector
Injection
valve (V1) Cond. Conductivity
1 7 monitor
2 6
3 4 5
Mixer pH
pH detector
System
pumps
Fraction
A B collector
Frac-950
Air
sensor
1 8
7
2 6
Outlet
3 5
4 valve (V4)
Injection
valve (V1) Cond. Conductivity
1 7 monitor
2 6
3 4 5
Mixer pH
pH detector
System
pumps
Fraction
A B collector
Frac-950
1 8
7
2 6
Outlet
3 5
4 valve (V4)
1 87
2 6 Buffer
345 select
valve (V6)
ÄKTApurifier 10 XT
In-line UV 3 Wavelength
filter
detector
Injection
valve (V1) Cond. Conductivity
1 7 monitor
2 6
3 4 5
Mixer pH
pH detector
System
pumps
Fraction
A B collector
Frac-950
1 8
7
2 6
Outlet
3 5
4 valve (V4)
PUMP P-900
Mixer
Pump A Pump B
Proportion
Valve
A2 A1 B2 B1
GRADIENT MIXER
ÄKTA 10 mls/min <6 mls/min 0.6 ml mixer
6 -14 mls/min 2 ml mixer
ÄKTA 100 mls/min <30 mls/min 2 mls mixer
30 - 50 mls/min 5 mls mixer
50 - 100 mls/min 12 mls mixer
PUMP P-900 Inlets
Mixer
Pump A Pump B
Switching
Valves
Buffer Select A2 B1 B2
A1
Valve IV-908
1 8 Water Salt
2 7
6
3 5
4
Pump A Pump B
A1 or A2 BufferValveA B1 or B2 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 Buffer
valve OUT
Ball outlet
High pressure check valve
Piston seal
Piston
Backwash pump
diaphragm
Ball inlet
check valve
Wash Buffer
IN IN
ON-LINE FILTERS
Polymer Support
PV-908 PV-908
PV-908 1 8
1 8
1 8
7
7
7 2
2
2
3 6
3 6
3 6 4 5
4 5
4 5
Waste Waste
LOAD
1
COLUMN
2 7
PUMP
3
6
4 5
Waste Waste
INJECT
1
COLUMN
7
2 PUMP
3 6
4 5
Waste Waste
WASTE
SAMPLE LOADING METHOD
1
COLUMN
2 7 PUMP
LOAD
SAMPLE 3
6
4 5
Needle Port
Waste Waste
1
COLUMN
LOAD 2 7 PUMP
SAMPLE
3
6
4 5
Luer Lock
Adapter
10 ml
50 ml
Buffer
150 ml
Plunger BUFFER
FLUSH
SAMPLE
LOAD
SAMPLE
INJECT
SAMPLE
FLUSH
Sample
Etched
Groove
PUMP or
WASTE COLUMN COLUMN
SYRINGE
10 ml 4 Mpa 18-1113-81
50 ml 4 Mpa 18-1113-82
150 ml 2 Mpa 18-1023-85
INV-907 Injection Valve - Injection Port
Valve Seal
Ferrule
Needle
Needle
Adapter
NeedleTube
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
Bubble Segmentation
100ul Loop
Waste 50ul Load
60ul Load
100ul Load
200ul Load
Injection valve
INV-907
1
Column
2 7
Pump
Load 3 6
Inject 4 5
Sample Sample
pump
P-950 Waste
LOADING A SUPER LOOP
Injection valve
INV-907
1
Column
7
2 Pump
Load 3
6
Inject 4 5
Buffer
Sample
Waste
Sample
MULTI-LOADING A SAMPLE LOOP
Injection valve
INV-907
Column 1
7
2 Pump
Load 3 6
Inject
4 5
PV-908
1 8
2 7
I/O
3 6
4 5
Waste
Wash
Samples
MULTI-LOADING A SUPER LOOP
Injection valve
1 INV-907
Column
2 7
Pump
Load 3
6
Inject
4 5
PV-908
1 8
Waste 2
I/O 7
3 6
4 5
Samples
Wash
S3 S4 S5 S6 S8
DIRECT LOADING ON TO COLUMN
1
Column
2 7 Pump
3
6
Inject
4 5
Load
Extra Tube
1/16” or 1/8”
Tubing
Waste
Sample
REVERSE FLOW VALVE ( INV-907 )
1
2 7
Pump
3 6
5
4
Column Extra Tube Stop
Plug
MONITOR
Downward Flow
Position 1
1
2 7
Pump
3 6
5
4
Column Extra Tube
Stop
Plug
MONITOR
Upward Flow
Position 2
OUTLET VALVE PV-908
DETECTOR
To Fraction
Collector
Default
F1 F8
F7
F2 I/O
F3 F6
F5
F4
F7
Waste F4 F8
F5 F6
F3
Fraction Collection
Flowthrough Eluate Peak
Fractionation Fractionation Fractionation
500 100
ND
ND OU
OU B
250
NB 50
Inject
U
Start
End
Waste
0 0
A1 A2 A3………………….
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
ct or
te
De
Phenyl Sepharose Phenyl Sepharose Phenyl Sepharose
e ct or
t
De
Phenyl Sepharose Phenyl Sepharose
ct or
te
De
Phenyl Sepharose
Traditional FPLC fitting to ÄKTA / HPLC
Injector
Flange M6 Fitting
Flow Cell
Tubing Fittings
18-1112-55
TM
1/16” Fingertight Fitting 1/16” PEEK Tubing
18-1112-58
18-1112-51
18-1112-52 18-1112-50
Fingertight Stop Plug 5/16” Stop Plug
18-1112-57
18-1017-98
M6 male / 1/16” Female 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
Untightened
Correctly tightened
Obstructed
3/16” OD 18-1112-47
1/8” OD 18-1121-16
18-1119-74
1/16” OD
Tubing Kit Changes - ÄKTApurifier 10
G5
10
0u
l
Injection valve V1
G9
Accumulator
(40ul)
UV Flow cell
G8
G6 G7
Conductivity
Flow cell (14ul)
0.25 mm i.d
FR-904
Blue PEEK (0.2Mpa, 5ul)
G5 400mm NB: Never use
G6 160mm the FR-904 with
G7 80mm the pH electrode
G8 130mm
G9 500mm pH Flow cell (100ul)
HIGH FLOW
G5
10
0u
l
Injection valve V1
H9
Accumulator
(40ul)
UV Flow cell
H8
H6 H7
Conductivity
Flow cell (14ul)
G5
10
0u
l
Injection valve V1
G9
Accumulator
(40ul)
UV Flow cell
G8
G6 G7
Conductivity
Flow cell (14ul)
0.75 mm i.d
FR-904
Green PEEK (0.2Mpa, 5ul)
LOW FLOW
L5
10
0u
l
Injection valve V1
L9
Accumulator
(40ul)
UV Flow cell
L8
0.50 mm i.d L6 L7
Conductivity
Orange PEEK Flow cell (14ul)
L6 160mm FR-904
L7 80mm (0.2Mpa, 5ul)
NB: Never use
L8 130mm
the FR-904 with
L9 500mm the pH electrode
A-900 INV-907
1
2 1
2 7 PUMP
3 6
3 6
4 5 MANUAL 4 5
SAMPLE
LOAD
INJECT
Waste Waste
COLUMN LOAD
Syringe Sample
Needle Height
Standard Vial
Standard Vial
+
Needle Height Insert
volume left
2mm
450ul
1mm 3mm
350ul 12ul
0mm
200ul
A-900 Sample Injection ( Flushed Loop Injection )
INJECT LOAD INJECT INJECT
INJECT LOAD INJECT INJECT
Sample needle
InInINJECT,
INJECT,sample
sample InInLOAD
LOADThe Thesample
sample
InInINJECT
INJECTsample
sample Finalinternal
Final internal
flushesthe
flushes thevalve
valve isisdrawn
drawnup upto
to
isiswashed
washedonto
onto needlewash.
needle wash.
channel. completely
completely thecolumn.
column.
channel. the
Min.( (30ul
30ul) )with
with fillthe
fill theloop.
loop. pump
Min.
AirSegment
Air Segment ( (no
noless
lessthan
thantwice
twice waste
Min.( (35ul
35ul) )without
without loopvolume
loop volume
Min. column
Airsegment
segment recommended) )
recommended
Air INJECT
INJECT
sample
OptionalExternal
Optional External
transport
needlewash.
needle wash. fluid
A-900 Sample Injection ( Partial Loop Injection )
INJECT LOAD INJECT INJECT
INJECT LOAD INJECT INJECT
waste
pump pump pump pump
Thesample
sampleisisdrawn
drawn Finalinternal
internal
InInINJECT,
INJECT,sample
sample The InIninject
injectsample
sample Final
flushesthethevalve
valve uptotopartially
partiallyfill
fillthe
the needlewash.
wash.
flushes up isiswashed
washedonto
onto needle
channel.
channel. loopwith
loop with thecolumn.
the column.
Min.( (30ul
Min. 30ul) )with
with programmedvolume.
programmed volume. pump
AirSegment
Air Segment ( (no
nomore
morethan
thanhalf
half waste
Min.( (35ul
Min. 35ul) )without
without loopvolume
loop volume
Airsegment
Air segment recommended) )
recommended column
INJECT
INJECT
OptionalExternal
Optional External sample
needlewash.
needle wash. transport
fluid
A-900 Sample Injection ( ul Pick-up Injection )
INJECT LOAD LOAD INJECT
INJECT LOAD LOAD INJECT
waste
column column column
column
sample
sample sample sample
InIninject,
inject,valve
valve Thesample
The sampleisis Thesample
The sampleisis InIninject
injectsample
sample
flusheswith
flushes with drawnup
drawn upfrom
from washedinto
washed into isiswashed
washedonto
ontothe
theco
column
lumn
transportfluid
transport fluid thesample
the samplevial
vial theloop
the loopwith
with
transportfluid
transport fluid pump
Equilibration Phase
A
COLUMN TOP
B 0%
A COLUMN TOP
B 20% 8 mls
Effect Command
20%B
0%B
8mls
Gradient Delay Volume
Gradient Phase
A
COLUMN TOP
B 0%
A COLUMN TOP
B 100%
16 mls Command 100%
Effect
100%B
Gradient Delay
Column Pressure ( compensation ) Flow Restrictor
0.3Reading
0.3 Reading Atmos
Atmos
Press
Press
0.0MPa
0.0 MPa 0.5Reading
Reading
0.5
Pre-column pressure Pump Pre-column pressure Pump
Pump Pump
= 0 MPa 0.3Mpa
Mpa = 0 MPa 0.3Mpa
Mpa
0.3 0.3
GELPress
PressDrop
Drop ColumnPress
PressDrop
Drop GELPress
PressDrop
Drop ColumnPress
PressDrop
Drop
GEL Column GEL Column
0.3MPa
0.3 MPa 0.3MPa
MPa 0.1MPa
0.1 MPa 0.5MPa
0.5 MPa
0.3
(0.3- -0.0)
0.0) (0.3++0.0)
0.0) (0.3- -0.2)
0.2) (0.5++0.2)
0.2)
(0.3 (0.3 (0.3 (0.5
0.0MPa
0.0 MPa 0.2MPa
MPa
0.2
FlowRestrictor
Flow Restrictor