AKTA Purifier Core 100

System Features
Flowrate: 0.01 100 ml/min System Pressure: 0 10 MPa Conductivity Range: 1uS/cm 999.9 mS/cm pH Range: 0 14 (+/-0.1 pH unit) UV Measurement: 190 700 nm in 1nm steps (3 Simultaneous)

KTA purifier Flowpath

In-line filter

UV
Injection valve (V1) 1 2 3 7 4

3 Wavelength detector

Cond
6 5

Conductivity monitor

Mixer System pumps

pH

pH detector

Fraction collector Frac-950

Sample

KTA System Practical Handling and Maintenance

Pumps
Purging, Backwash Filters Flow Restrictor and Pressure

Application of Sample
Injection Valve Loop Loading and Rules Superloop Direct Injection

Collecting Peaks
Flowthrough Fractionation Elution Fractionation Peak Fractionation Delay Volume

RECIPROCATING DISPLACEMENT PUMP

Wash OUT
IN-LINE one-way valve High pressure Piston seal Piston

Buffer OUT
Ball outlet check valve

Check for leakage. If there is a sign of liquid leaking between the pump head and the housing side panel or increased or decreased volume of rinsing solution, replace the piston Seals.

When changing eluent, it is important to remove trapped air. Backwash pump diaphragm Ball inlet check valve

Wash IN

Buffer IN

PUMP P-900 Back - Wash Lines

Pump Back-Wash Lines

The purge line is recycled back in the resevoir. Make sure that the resevoir lines are secured.

Syringe fitting

Wash OUT Buffer OUT

One-way valve

20% Ethanol : Water

Change MONTHLY Book says Weekly

Wash IN

Buffer IN

PUMP P-900 Mixer Pump A Pump B

Default Lines Pump A A1 Pump B B1

Any one Buffer passes through both pump heads. When purging a buffer line both pump heads must be purged.

Proportion Valve A2 A1 B2 B1 A B A B

The Mixer volume is best changed for different flow rates. When a command for a %B mix is received by the Pump it takes 3-4 X the mixer volume before it is achieved COLUMN TOP 0%
Mixer, 2mls

COLUMN TOP
KTA 10 mls/min KTA 100 mls/min GRADIENT <6 mls/min 6 -14 mls/min <30 mls/min 30 - 50 mls/min 50 - 100 mls/min MIXER 0.6 ml mixer 2 ml mixer 2 mls mixer 5 mls mixer 12 mls mixer

20%

8 mls* Effect Command 20% ( 3 to 4 x Mixer Volume )

This appears in the program as System compensation volume and Gradient delay volume

Seal
Buffer OUT

Only manually purge when pump contains large volume of air Unscrew valve and the seal moves out of the way. Buffer can then be sucked through the Pump. Note the check valves opening.

Air

Buffer IN

Pump A

Pump B

~Try This System Control. Manual. Flowpath. PumpA Inlet... A2.

Common inlet

A2

A1

Common inlet

B2

B1

Manual A1 or A2

Flowpath BufferValveA

Manual B1 or B2

Flowpath BufferValveB

Execute

Open

Draw A ir out of Buffer Line

ON-LINE FILTERS
Polymer Fibre Filter ( 5um pores ) Support Mesh ~ Stops fibres from entering the chromatography steam.

P-900 Pump High Flow Fibre Filter ( for Soft Gel Columns ) 18-1027-11 ( 10/pack)

18-1112-44 On-line Filter

Polymer Support mesh

Change MONTHLY / 3 MONTHLY Book says Weekly

Flow Restrictor and Pressure

10 0u

Injection valve V1

FR-904 (0.4Mpa, 4ul) FR-902 (0.2Mpa, 5ul)

Outlet Valve V4 (7ul)

Accumulator (40ul)

1. Suppresses Air Bubbles 2. Stops Siphoning Effect on Buffer 3. Closes Pump Check Valves efficiently

Conductivity Flow cell (14ul) FR-902 FR-904 (0.2Mpa, 5ul) (0.4MPa, 7ul)

NB: Never use the FR-904 with the pH electrode

pH Flow cell (100ul)

Column Pressure
TOP ~ Full resistance reflects the addition of the Flow Restrictor. BOTTOM ~ Resistance to flow, due to Flow Restrictor.

2 90 RF 0.5
0.3

MPa

0.3 Column Gel TOP = 0.5 MPa

TOP ~ Full resistance to flow, by gel bed. BOTTOM ~ No resistance to flow, open to atmosphere. This means you should ADD the 0.2MPa to the recommended Column Pressure, ie. HiTrap (0.3 +0.2)= 0.5MPa Pressure gradient down column (0.5 0.3) Pressure difference = (0.3 0.0) = 0.3 MPa

FR-902 0.2 MPa

Column Gel BOTTOM ==0.2MPa Column Gel BOTTOM 0 MPa

Column Pressure
WARNING ! However the set pressure must not EXCEED the recommended Column Hardware pressure.

Column Hardware Pressure Limits

Column Shatters DANGEROUS

Column Pressures

Column Matrix Pressure Limits

Matrix Collapses EXPENSIVE

pH Electrode and Flow cell ~ Never leave in line

pH electrode DUMMY electrode

Storage Reservoir
1:1 mixture Buffer pH 4 and 1M KNO3

Electrode Flow Cell

SAMPLE APPLICATION
Different ways of applying sample

INJECTION VALVE INV- 907

COLUMN LOAD SAMPLE 1 2 3 4 5 7 6 PUMP

Waste 1 2 4

Waste

LOAD
COLUMN 7 3 5 6 PUMP

Waste

Waste

COLUMN

INJEC T1
2 3 4 5

7 6

PUMP

Waste

Waste

WASTE

Loop Loading Rules

PEEK Tubing
High Resistance to Flow

Low Resistance to Flow

End

Beginning

Loop Loading Rules

Waste 100ul Loop
100ul Load 60ul Load 50ul Load
No Waste Exactly 100ul

200ul Load

Partial Loading Loop = No more than 1/2 Loop Volume Total Loading Loop = Inject 2X Loop Volume

Sample Partial Loading Instruction

1. Ensure that Sample loop is Twice the Sample Volume 2. Ensure that the Injection Valve (INV-907) is at LOAD position 3. Flush the Sample loop with at least Twice Sample loop volume, do not detach the Syringe 4. Switch (from UNICORN) the Injection Valve (INV-907) to INJECT position 5. Detach the Syringe containing Flush Buffer and attached the Syringe containing Sample 1. Switch (from UNICORN) the Injection Valve (INV-907) to LOAD position, then Inject your sample into the Sample loop 1. Leave the Sample syringe on the Injection Valve (Do not remove)

MANUALY LOADING SAMPLE LOOP

1 Sample 2

( sample volumes 25ul to 5.0 mls )

Injection Valve INV-907 IV-907

7 3 4 5 6

System Pump

LOAD
Syringe Needle Adaptor Sample Loop

Waste

While buffer is pumped through INJECTION VALVE to column Sample can be pushed ( LOADed ) into Loop

MANUALY LOADING SAMPLE LOOP

1 Sample 2

( sample volumes 25ul to 5.0 mls )

Injection Valve INV-907 IV-907

7 3 4 5 6

System Pump

Wash volume 2/3 loop vol. LOAD INJECT

Sample Loop

Waste

INJECTION VALVE changes from LOAD to INJECT Buffer pushes ( INJECTS ) sample onto column

MANUALLY LOADING SUPERLOOP ( sample volumes 1 ml to 150 mls )

Injection Valve INV-907 IV-907
1 Sample 2 7 3 4 5
50 40 30 20

INJECT LOAD
System Pump
6

SuperLoop

Leur Syringe Adaptor

10 0

Waste

INJECTION VALVE Changes from LOAD to INJECT to column While buffer is pumped through INJECTION VALVE System PumpsSuperloop Syringe Loads (injects) Sample from Superloop to column

LOADING SAMPLE LOOP with SAMPLE PUMP ( sample volumes >10 mls )
Injection Valve INV-907 IV-907
1 2 7 3 4 6
5

System Pump

LOAD

Sample Loop

Sample

NOTE: Sample Does not enter pump This means Minimal wastage
Sample Pump

Waste

LOADING SAMPLE LOOP with SAMPLE PUMP ( sample volumes >10 mls )
Injection Valve INV-907 IV-907
1 2 7 3 4 6 5

System Pump

Wash volume 2/3 loop vol. LOAD INJECT

Sample Loop

Sample

NOTE: Sample Does not enter pump This means Minimal wastage
Sample Pump

Waste

LOADING SUPERLOOP with SAMPLE PUMP ( sample volumes 5 ml to 150 mls )

Injection Valve INV-907 IV-907
1 2 7 3 4 5
50 40 30 20 10 0

INJECT LOAD
System Pump
6

SuperLoop

Sample

NOTE: Sample goes throughBuffer pushes NOTE: the pump and is onto the Samplepushed into superloop column

Waste

Sample Pump

DIRECT LOADING SAMPLE with SAMPLE PUMP ( sample volumes > 10 mls )
Injection Valve INV-907 IV-907
1 2 7 3 4 5 6

LOAD INJECT
System Pump

Extra Tubing

Sample

Waste

Sample Pump

DIRECT LOADING SAMPLE with SAMPLE PUMP / AIR SENSOR

Injection Valve INV-907 IV-907
1 2 ( sample volumes > 10 mls )

LOAD INJECT
7

Air Sensor
(watch)

System Pump
6

3 4 5

Sample

Less Wasted Sample No need for Accurate Volume

Waste

Air Stops Sample Pump

DIRECT LOADING SAMPLE with SYSTEM PUMP ( sample volumes > 10 mls )
1 2 7 3 4 5 6

Injection Valve INV-907

LOAD
System Pump

Inlet A1 Change To Buffer Inlet A1

Inlet A2

Buffer

Sample

DIRECT LOADING SAMPLE with SYSTEM PUMP and BUFFER VALVE

1 2 7 3 4 5 6

Injection Valve INV-907

( sample volumes > 10 mls )

LOAD
System Pump

8 7 6

Inlet A1

Change To Buffer
A12

2 3 4 A13 5

Buffer Selection Valve IV-908

BUFFER A11 INLET A18

Sample

Collecting Peaks
Different ways of collecting Fractions

The Fractionation Options

mAU 500
Outlet valve

0%B

100%B

%B 100

Tube

UNBOUND

Inject

UV Watch

250

Slope or Level
Start

50
End

X1
UV280 Conc %B

X2

X3

A1 A2 A3

Flowthrough Fractionation

Eluate Fractionation
Fixed volume

Peak Fractionation

Waste

FRACTION DELAY
Delay volume
START FRAC END FRAC

Waste

A1

A2

A3

A4

A5

Waste

Fraction Collector
vo lu m e De la y

A1 A2 A3 A4

Computer Screen Signal from UV monitor

Fraction Size = 1 ml Fraction delay = 1 ml

Delay volume continues collecting fractions after end of run until the delay volume is achieved

Optional Functionality