KTA system Training Guide

Explorer 100 / 10

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KTAexplorer

KTAexplorer
Injector

Buffers

Box

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ham
amers

Column
Valves

Monitor
Conductivity
pH

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amers

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Outlet
Valve

UV Detector
3 Wavelengths
amersham bioscience

Pumps
am
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bios
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ence

KTAexplorer

Pump A

pH/cell
electrode

Buffer switching
valve

Sample
Valve

Sample
holder

KTAexplorer 100
In-line
filter
Injection
valve (V1)

1 7
6
2
3 45

Mixer
System
pumps

Column
selection
valve (V2)

UV

3 Wavelength
detector

Cond

Conductivity
monitor

pH

1 87
2
6
345

Sample
pump

1 87
2
6
345

1 7
6
2
3 45

Reverse
flow valve (V7)

pH detector

Fraction
collector
Frac-950

1 87
2
6
345
1 87
2
6
345

Buffer
select
valve (V6)

Outlet
valve (V4)

1 87
2
6
345

Sample selection
valve (V5)

Column
selection
valve (V3)

KTAexplorer 100 Air

In-line
filter
Injection
valve (V1)
1 7
6
2
3 45

Mixer
System
pumps

Reverse
flow valve (V7)
Column
selection
valve (V2)

UV

3 Wavelength
detector

Cond

Conductivity
monitor

pH

1 87
2
6
345

Air
sensor

Sample
pump

1 87
2
6
345

1 7
6
2
3 45

pH detector

Fraction
collector
Frac-950

1 87
2
6
345
1 87
2
6
345

Buffer
select
valve (V6)

1 87
2
6
345

Sample selection
valve (V5)

Column
selection
valve (V3)

Outlet
valve (V4)

KTAexplorer 10
In-line
filter
Injection
valve (V1)

1 7
6
2
3 45

Mixer
System
pumps

Column
selection
valve (V2)

UV

3 Wavelength
detector

Cond

Conductivity
monitor

pH

1 87
2
6
345

pH detector

Fraction
collector
Frac-950

1 87
2
6
345

1 87
2
6
345

Buffer
select
valve (V6)

Outlet
valve (V4)

1 87
2
6
345

Column
selection
valve (V3)

KTAexplorer 10 S
In-line
filter
Injection
valve (V1)
1 7
6
2
3 45

Mixer
System
pumps

3 Wavelength
detector

Cond

Conductivity
monitor

pH

1 87
2
6
345

Sample
pump

1 87
2
6
345

Column
selection
valve (V2)

UV

pH detector

Fraction
collector
Frac-950

1 87
2
6
345
1 87
2
6
345

Buffer
select
valve (V6)

1 87
2
6
345

Sample selection
valve (V5)

Column
selection
valve (V3)

Outlet
valve (V4)

KTAexplorer 10 XT
In-line
filter
Injection
valve (V1)

1 7
6
2
3 45

Mixer
System
pumps

Column
selection
valve (V2)

UV

3 Wavelength
detector

Cond

Conductivity
monitor

pH

1 87
2
6
345

pH detector

Fraction
collector
Frac-950

1 87
2
6
345

Buffer
select
valve (V6)

1 87
2
6
345

1 87
2
6
345

Column
selection
valve (V3)

Outlet
valve (V4)

PUMP P-900

Mixer
Pump A

Pump B

Proportion
Valve
A2

Acid /Alkali

KTA 10 mls/min
KTA 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

A14

B1

Water

7
5

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)

Column

Column

Column
2
3

Pump

Sample

Waste

Sample

PV-908
1

8
7

6
4

PV-908
1

Pump

7
3

6
4

PV-908

Pump

7
3

6
4

LOAD (wash loop)

Waste

Sample

Waste

INJECTION VALVE INV- 907

1

COLUMN

2
LOAD
SAMPLE

PUMP

Waste

Waste

LOAD
1

COLUMN

PUMP

3
4

6
5

Waste

Waste

INJECT
1

COLUMN

PUMP
3
4

Waste

6
5

Waste

WASTE

SAMPLE LOADING METHOD

1

COLUMN
2

LOAD
SAMPLE

PUMP

3
6
5

4
Needle Port

Waste

Waste

Always tighten by hand

with needle inserted

Do Not Overtighten

COLUMN
LOAD
SAMPLE

PUMP

3
6
4

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

General

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

Column

Load

Inject

Sample

Sample
pump
P-950

Pump

6
5

Waste

LOADING A SUPER LOOP

Injection valve
INV-907

Column

Load
Inject

Pump
6

Buffer

Sample
Waste

Sample

MULTI-LOADING A SAMPLE LOOP

Injection valve
INV-907

Column

Load

Inject

Pump
6

PV-908
1

2
3

I/O
4

7
6

Wash
Samples

Waste

MULTI-LOADING A SUPER LOOP

Column

1
2

Pump

Load
Inject

Injection valve
INV-907
7
3

Waste

6
5

PV-908
1
8
2
3

I/O
4

7
6

Samples
Wash
S3 S4 S5 S6

S8

DIRECT LOADING ON TO COLUMN

Column

Pump

2
3

Inject

6
4

Load

Extra Tube

1/16 or 1/8
Tubing

Waste
Sample

REVERSE FLOW VALVE ( INV-907 )

1
7

Pump

Column

Extra Tube

Stop
Plug

MONITOR

Downward Flow
Position 1
1
7

Pump

3
4

Column

Extra Tube

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

250

500

mAU

Flowthrough
Fractionation

Inject

A1 A2 A3.

Eluate
Fractionation
0%B

100%B

Peak
Fractionation

End

Fraction Collection

Waste

Start

50

100

%B

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

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t
De

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t
De

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