No 900 Mixer AKTA PURIFIER Manual
User Manual: 900 Mixer
Open the PDF directly: View PDF 
.
Page Count: 43
amersham bioscience
amersham bioscience
amersham bioscience
amersham bioscience
ÄKTA system Training Guide
Purifier 100 / 10
ÄKTApurifier
Box
Monitor
Conductivity
pH
UV Detector
Pumps
Injector
Buffers
Pump A
Buffer switching
valve
Pump B
On-line
Filter
Outlet Valve
Mixer
ÄKTApurifier 10 / 100
A
System
pumps
Mixer
2
4
7
3
16
5
2
345
6
7
8
1
Fraction
collector
Frac-950
Conductivity
monitor
pH detector
In-line
filter UV
Cond.
pH
3 Wavelength
detector
Injection
valve (V1)
Outlet
valve (V4)
B
ÄKTApurifier + Sample Pump + Sample Valve
Fraction
collector
Frac-950
Conductivity
monitor
pH detector
Outlet
valve (V4)
3 Wavelength
detector
23456
7
8
1
UV
Cond
pH
Mixer
Injection
valve (V1)
23456
7
8
1
24
7
3
16
5
In-line
filter
System
pumps
AB
Sample
pump
Sample selection
valve (V5)
ÄKTApurifier 100 Air
Air
sensor
A
System
pumps
Mixer
2
4
7
3
16
5
2
345
6
7
8
1
Fraction
collector
Frac-950
Conductivity
monitor
pH detector
In-line
filter UV
Cond.
pH
3 Wavelength
detector
Injection
valve (V1)
Outlet
valve (V4)
B
23456
7
8
1Buffer
select
valve (V6)
A
System
pumps
Mixer
2
4
7
3
16
5
2
345
6
7
8
1
Fraction
collector
Frac-950
Conductivity
monitor
pH detector
In-line
filter UV
Cond.
pH
3 Wavelength
detector
Injection
valve (V1)
Outlet
valve (V4)
B
ÄKTApurifier + Buffer Valve
ÄKTApurifier 10 XT
A
System
pumps
Mixer
2
4
7
3
16
5
2
345
6
7
8
1
Fraction
collector
Frac-950
Conductivity
monitor
pH detector
In-line
filter UV
Cond.
pH
3 Wavelength
detector
Injection
valve (V1)
Outlet
valve (V4)
B
PUMP P-900
Mixer
Pump A Pump B
Proportion
Valve
A2 A1 B2 B1
Buffer
Acid /Alkali Salt Water
GRADIENT MIXER
ÄKTA 10 mls/min <6 mls/min
6 -14 mls/min
0.6 ml mixer
2 ml mixer
ÄKTA 100 mls/min <30 mls/min
30 - 50 mls/min
50 - 100 mls/min
2 mls mixer
5 mls mixer
12 mls mixer
PUMP P-900 Inlets
Water Salt
Buffer Select
Valve IV-908
Pump A Pump B
Mixer
A2 A1 B1 B2
Switching
Valves
1
2
3456
7
8
A11 A12 A13 A14 A15
A16 A17 A18
PUMP P-900 Manually Priming Buffer Inlet Lines
Open
Draw Air
out of
Buffer Line
Manual Flowpath
A1 or A2 BufferValveA
Manual Flowpath
B1 or B2 BufferValveB
Pump A Pump B
Common inlet A2 A1 B2 B1
Common inlet
PUMP P-900 Back - Wash Lines
20% Ethanol : Water
Pump Back-Wash
Lines
Wash OUT
Wash
IN
Buffer
IN
Buffer
OUT
High pressure
Piston seal
Ball outlet
check valve
Ball inlet
check valve
Piston
Backwash pump
diaphragm
IN-LINE
one-way
valve
Syringe fitting
ON-LINE FILTERS
P-901 ( 100 ml/min Pump )
High Flow Fibre Filter
Polymer Support
P-903 ( 10 ml/min Pump )
Low Flow HPLC Filter
LOAD (sample) INJECT (sample) LOAD (wash loop)
1
7
6
5
2
3
4
Pump
Column
Waste
PV-908
1
2
3
45
6
7
8
1
7
6
5
2
3
4
Pump
Column
Waste
PV-908
1
2
3
45
6
7
8
1
7
6
5
2
3
4
Pump
Column
Waste
PV-908
1
2
3
45
6
7
8
Sample
Sample
Sample
1
7
6
5
2
3
4
PUMP
LOAD
SAMPLE
Waste
COLUMN
Waste
LOAD
1
7
6
5
2
3
4
PUMP
Waste
COLUMN
Waste
INJECT
1
7
6
5
2
3
4
PUMP
Waste
COLUMN
Waste
WASTE
INJECTION VALVE INV- 907
SAMPLE LOADING METHOD
1
7
6
5
2
3
4
PUMP
LOAD
SAMPLE
Waste
COLUMN
Waste Superloop
Luer Lock
Adapter
1
7
6
5
2
3
4
PUMP
LOAD
SAMPLE
Waste
COLUMN
Waste
Needle Port
Always tighten by hand
with needle inserted
Do Not Overtighten
V
olume Max. allowed column
pressure
Code No.
10 ml 4 Mpa 18-1113-81
50 ml 4 Mpa 18-1113-82
150 ml 2 Mpa 18-1023-85
SUPERLOOPS
10 ml
150 ml
50 ml
BUFFER
FLUSH
SAMPLE
LOAD
SAMPLE
INJECT
SAMPLE
FLUSH
WASTE PUMP or
SYRINGE COLUMN COLUMN
Plunger
Buffer
Sample
Etched
Groove
Needle
Needle
Adapter
INV-907 Injection Valve - Injection Port
PEEK Valve Body
Valve Seal
Injection Needle Port
NeedleTube
Ferrule
INV-907 Injection Valve - Luer Adapter
PEEK Injection Valve Body
PEEK Injection Valve Body
PEEK Injection Luer Adaptor
PEEK Injection Luer Adaptor
Valve Seal
Valve Seal
Valve Channel Plate
Valve Channel Plate
Flow
End Beginning
PEEK Tubing
High Resistance
to Flow
Low Resistance
to Flow
Fluid Dynamics
Flow - Injecting a Loop
Fluid Dynamics
The way we imagine it Flows
The way it Actually Flows
Bubble Segmentation
•Partial Loading Loop = No more than 1/2 Loop Volume
•Total Loading Loop = Inject 2X Loop Volume
50ul Load
60ul Load
100ul Load
200ul Load
Waste
100ul Loop
LOADING A SAMPLE LOOP
1
7
6
5
2
3
4
Sample
Pump
Column
Inject
Load
Sample
pump
P-950
Injection valve
INV-907
Waste
LOADING A SUPER LOOP
1
7
6
5
2
3
4
Sample
Pump
Column
Inject
Load
Waste
Sample
Buffer
Injection valve
INV-907
MULTI-LOADING A SAMPLE LOOP
1
7
6
5
2
3
4
Inject
Load
PV-908
Samples
1
2
345 6
7
8
I/O
Injection valve
INV-907
Waste
Wash
Column
Pump
MULTI-LOADING A SUPER LOOP
Waste
1
7
6
5
2
3
4
Pump
Column
Inject
Load
PV-908
1
2
345 6
7
8
I/O
Wash
Samples
S3 S4 S5 S6 S8
Injection valve
INV-907
DIRECT LOADING ON TO COLUMN
Extra Tube
Inject
Load
1/16” or 1/8”
Tubing
7
6
5
2
3
4
Waste
1
Column Pump
Sample
REVERSE FLOW VALVE ( INV-907 )
Column
Pump
Downward Flow
Position 1
Extra Tube
MONITOR
Stop
Plug
1
7
6
5
2
3
4
1
Column Extra Tube
MONITOR
Stop
Plug
2
Upward Flow
Position 2
Pump
7
6
5
3
4
OUTLET VALVE PV-908
Default
To Fraction
Collector
DETECTOR
F1
F2
F3 F4 F5 F6
F7
F8
I/O
Waste
F3 F5 F6
F7
F8
F4
0
%B
100
50
Eluate
Fractionation
Peak
Fractionation
Flowthrough
Fractionation
0%B 100%B
Start
End
UNBOUND
BOUND
A1 A2 A3………………….
Inject
mAU
0
500
250
Waste
Fraction Collection
HiTrap Connections
M6 Male
1/16” Male
1/16” Female
M6 Male
M6 Female
1/16” Male
M6 male / 1/16” Female
18-1112-57
18-1112-55
Flow Cell Inlet
HiTrap Column
18-1112-58
M6 female / 1/16” male
1/16” Fingertight Fitting
Phenyl Sepharose
1/16” Female
Detector
Multiple HiTrap Arrangment ( Series )
Phenyl Sepharose
Detector
Phenyl Sepharose
Phenyl Sepharose
Detector
Phenyl Sepharose
Phenyl Sepharose
Phenyl Sepharose
Flow Cell
Traditional FPLC fitting to ÄKTA / HPLC
100 ul
Injector
1/16” Fingertight Fitting
18-1112-55
1/16” FEMALE / M6 FEMALE
18-1123-94
1/16” MALE / M6 FEMALE
18-1112-58
Flange M6 Fitting
Tubing Fittings
Flangless Inlet Connector
3/16” 18-1112-49
1/8” 18-1121-17
1/16” 18-1127-07
1/16” Fingertight Fitting
Luer Fitting Adapter
Fingertight Stop Plug
Flange Fitting Adapter
18-1112-55
18-1112-58
18-1112-51
18-1112-52
1/16”female X 1/16”female Union Ti
M6 male / 1/16” Female
18-1112-57
18-3955-01
18-1123-94
M6 female X 1/16”female Union
Flangless Buffer Inlet
Ferrule
3/16” 18-1112-48
1/8” 18-1121-18
1/16” 18-1127-06
1/16” PEEKTM Tubing
FPLC Flange Fitting
Luer Syringe
5/16” Stop Plug
18-1112-50
Union + 2 Fingertight Fittings
Tubing Connector
M6 male / O - ring
18-1017-98
FPLC Flange - 1/16” Fingertight
COMPRESSION FITTINGS
Untightened
Correctly tightened
Over tightened Tubing Restricted
Obstructed
PEEKTM Tubing
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
1/16” OD
18-1112-47
1/8” OD 18-1121-16
18-1119-74
G5 400mm
G6 160mm
G7 80mm
G8 130mm
G9 500mm
NB: Never use
the FR-904 with
the pH electrode
Tubing Kit Changes - ÄKTApurifier 10
GENERAL FLOW (at delivery)
0.25 mm i.d
Blue PEEK
Outlet Valve V4 (7ul)
FR-904
(0.2Mpa, 5ul)
pH Flow cell (100ul)
Accumulator
(40ul)
UV Flow cell
Conductivity
Flow cell (14ul)
100 ul
G6 G7
G8
G9
Injection valve V1
G5
Tubing Kit No Frac Frac 901 Frac 950
AKTApurifier 10 , 0.50 tubing kit 232 382
AKTApurifier 10 , 0.50 tubing kit + pH electrode 354 504
AKTApurifier 10 , 0.25 tubing kit 88 238
NB: Never use
the FR-904 with
the pH electrode
Tubing Kit Changes - ÄKTApurifier 10
HIGH FLOW
H6 160mm
H7 80mm
H8 130mm
H9 500mm
0.50 mm i.d
Orange PEEK
Outlet Valve V4 (7ul)
FR-904
(0.2Mpa, 5ul)
pH Flow cell (100ul)
Accumulator
(40ul)
UV Flow cell
Conductivity
Flow cell (14ul)
100 ul
H6 H7
H8
H9
Injection valve V1
G5
Tubing Kit No Frac Frac 901 Frac 950
AKTApurifier 10 , 0.50 tubing kit 232 382
AKTApurifier 10 , 0.50 tubing kit + pH electrode 354 504
AKTApurifier 10 , 0.25 tubing kit 88 238
G6 160mm
G7 80mm
G8 130mm
G9 500mm
NB: Never use
the FR-904 with
the pH electrode
Tubing Kit Changes - ÄKTApurifier 100
HIGH FLOW (at delivery)
0.75 mm i.d
Green PEEK
Outlet Valve V4 (7ul)
FR-904
(0.2Mpa, 5ul)
pH Flow cell (100ul)
Accumulator
(40ul)
UV Flow cell
Conductivity
Flow cell (14ul)
100 ul
G6 G7
G8
G9
Injection valve V1
G5
Tubing Kit No Frac Frac 901 Frac 950
AKTApurifier 100 , 0.50 tubing kit 232 382
AKTApurifier 100 , 0.75 tubing kit 470 620
AKTApurifier 100 , 1.00 tubing kit 769 809
AKTApurifier 100 , 0.50 tubing kit + pH electrode 354 504
AKTApurifier 100 , 0.75 tubing kit + pH electrode 672 862
AKTApurifier 100 , 1.00 tubing kit + pH electrode 955 995
AKTAbasic 100 , 0.75 tubing 290
L6 160mm
L7 80mm
L8 130mm
L9 500mm
NB: Never use
the FR-904 with
the pH electrode
Tubing Kit Changes - ÄKTApurifier 100
LOW FLOW
0.50 mm i.d
Orange PEEK
Outlet Valve V4 (7ul)
FR-904
(0.2Mpa, 5ul)
pH Flow cell (100ul)
Accumulator
(40ul)
UV Flow cell
Conductivity
Flow cell (14ul)
100 ul
L6 L7
L8
L9
Injection valve V1
L5
Tubing Kit No Frac Frac 901 Frac 950
AKTApurifier 100 , 0.50 tubing kit 232 382
AKTApurifier 100 , 0.75 tubing kit 470 620
AKTApurifier 100 , 1.00 tubing kit 769 809
AKTApurifier 100 , 0.50 tubing kit + pH electrode 354 504
AKTApurifier 100 , 0.75 tubing kit + pH electrode 672 862
AKTApurifier 100 , 1.00 tubing kit + pH electrode 955 995
A-900 Autosampler
COLUMN
1
7
6
5
2
3
4
PUMP
MANUAL
SAMPLE
LOAD
Waste
Waste
12
3
45
6
LOAD
Syringe
Sample
INV-907
A-900
COLUMN
1
7
6
5
2
3
4
PUMP
MANUAL
SAMPLE
LOAD
Waste
Waste
12
3
45
6
Syringe Sample
INV-907
A-900
LOAD
LOAD
INJECT
Needle Height
2mm
450ul
1mm
350ul
0mm
200ul
3mm
12ul
Standard Vial
Standard Vial
+
Insert
Needle Height
volume left
A-900 Sample Injection ( Flushed Loop Injection )
In INJECT, sample
flushes the valve
channel.
Min. ( 30ul ) with
Air Segment
Min. ( 35ul ) without
In INJECT, sample
flushes the valve
channel.
Min. ( 30ul ) with
Air Segment
Min. ( 35ul ) without
In LOAD The sample
is drawn up to
completely
fill the loop.
( no less than twice
loop volume
In LOAD The sample
is drawn up to
completely
fill the loop.
( no less than twice
loop volume
In INJECT sample
is washed onto
the column.
In INJECT sample
is washed onto
the column.
INJECT
INJECT LOAD
LOAD INJECT
INJECT INJECT
INJECT
Final internal
needle wash.
Final internal
needle wash.
waste
pump
column
sample
waste
sample
pump
Sample needle
waste
sample
waste
Wash
Reservoir
column
column
pump
waste
Air segment
Air segment recommended )
recommended )
INJECT
INJECT
Optional External
needle wash.
Optional External
needle wash.
pump
sample
transport
fluid
column
pump
column
sample
A-900 Sample Injection ( Partial Loop Injection )
The sample is drawn
up to partially fill the
loop with
programmed volume.
( no more than half
loop volume
The sample is drawn
up to partially fill the
loop with
programmed volume.
( no more than half
loop volume
In inject sample
is washed onto
the column.
In inject sample
is washed onto
the column.
INJECT
INJECT LOAD
LOAD INJECT
INJECT
column
pump
column
Sample needle
waste
sample sample
Sample needle
sample
pump
Sample needle
INJECT
INJECT
waste
Wash
Reservoir
Final internal
needle wash.
Final internal
needle wash.
waste
column
pump
waste waste
In INJECT, sample
flushes the valve
channel.
Min. ( 30ul ) with
Air Segment
Min. ( 35ul ) without
In INJECT, sample
flushes the valve
channel.
Min. ( 30ul ) with
Air Segment
Min. ( 35ul ) without
waste
recommended )
recommended )
pump
column
INJECT
INJECT
Optional External
needle wash.
Optional External
needle wash.
pump
sample
transport
fluid
column
sample
Air segment
Air segment
A-900 Sample Injection ( ul Pick-up Injection )
In inject, valve
flushes with
transport fluid
In inject, valve
flushes with
transport fluid
In inject sample
is washed onto the co
In inject sample
is washed onto th
The sample is
drawn up from
the sample vial
The sample is
drawn up from
the sample vial
The sample is
washed into
the loop with
transport fluid
The sample is
washed into
the loop with
transport fluid
pump
column
Sample nee
INJECT
INJECT
column
pump
Sample needle
LOAD
LOAD
column
pump
Sample needle
LOAD
LOAD
samp
sample sample
INJECT
INJECT
waste
sample
transport
fluid
waste
pump
INJECT
INJECT
waste
lumn
e column
pump
column
INJECT
INJECT
Optional needle
wash.
Optional needle
wash.
pump
dle
le
column
sample
transport
fluid
Wash
Reservoir
column
sample
Final internal
needle wash.
Final internal
needle wash.
B
A
0%
B20%
COLUMN TOP
Command
Effect
8 mls
System Compensation Volume
Equilibration Phase
COLUMN TOP
A
0%B
20%B
8mls
B
A
0%
COLUMN TOP
B100%
COLUMN TOP
Command 100%
Effect 16 mls
Gradient Delay Volume
100%B
Gradient Delay
Gradient Phase
A
Column Pressure ( compensation ) Flow Restrictor
Column Hardware Pressure Limits
Column Matrix Pressure Limits
Column Pressures Column Shatters DANGEROUS
Matrix Collapses EXPENSIVE
GEL Press Drop
0.3 MPa
(0.3 - 0.0)
GEL Press Drop
0.3 MPa
(0.3 - 0.0)
0.3 Reading
0.3 Reading
GEL Press Drop
0.1 MPa
(0.3 - 0.2)
GEL Press Drop
0.1 MPa
(0.3 - 0.2)
Pump
0.3 Mpa
Pump
0.3 Mpa
Atmos
Press
0.0 MPa
Atmos
Press
0.0 MPa
Column Press Drop
0.3 MPa
(0.3 + 0.0)
Column Press Drop
0.3 MPa
(0.3 + 0.0)
Colu
Colu
0.5 Reading
0.5
Pump
0.3 Mpa
Pump
0.3 Mpa
0.3 MPa 0.3 MPa
Gel Top Pressure
Pre-column pressure
= 0 MPa Pre-column pressure
= 0 MPa
mn Press Drop
0.5 MPa
(0.5 + 0.2)
mn Press Drop
0.5 MPa
(0.5 + 0.2)
Reading
0.2 MPa
Flow Restrictor
0.2 MPa
Flow Restrictor
0.0 MPa
0.0 MPa
0.2 MPa
0.0 MPa Gel Bottom Pressure
