Tips and Tricks LC User

M i t
Maintenance

Edward Kim
Application Engineer
Agilent Technologies, Inc
May 20, 2009

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Agilent
g e t Technologies
ec o og es
1100/1200 Series HPLC
Maintenance

Pump
Modules

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Agilent HPLC System Flexibility

•Agilent LC systems are designed for flexible use with a wide

range of flow and pressure capabilities
– Many of the New 1200 Modules (i.e. 80 Hz DAD) are compatible with
older 1100 systems
– 1200 RRLC systems can run current, legacy and new high performance
methods
– Engineered Column Technology Allows Higher Performance Separations
With:
– Existing
g 1100 System
y
– Enhanced 1100 System
– Higher Pressure, Enhanced Flow Path 1200SL System

Separation needs will determine your choice!

Group/Presentation Title
Agilent Restricted
Month ##, 200X
The Agilent 1200 LC Systems
One Comprehensive Series of Solutions for your Applications

Application 1200 RRTM LC 1100/1200 1100/1200

Binary Quaternary

RRLC – Narrow Ultra-fast

Ultra fast YES NO NO
column length 15 – 50mm Delay volume too large Delay volume too large
(2.1)
High-resolution YES NO NO
column length 100 – 150mm Delay volume too large, Delay volume too large,
pressure limit too low
p pressure limit too low
p

RRLC – Std Ultra-fast YES YES YES

column length 15 – 50mm Modifications
(3.0, 4.6) recommended to reduce
delay volume

High-resolution YES NO NO
column length 100 – 150mm Pressure limit too low Pressure limit too low

HPLC – Narrow bore (2.1) YES YES NO

Modifications Delay volume too large
recommended to reduce
delay volume

HPLC – Standard bore (3.0, 4.6) YES YES YES

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Maximum Utilization of Entire Pressure Range
with Agilent 1100/1200 HPLC Systems
• Agilent 1100 and 1200 Instruments are Designed to Operate at Maximum
Pressure for Extended Periods
• Max Pressure Can be Maintained Across Full Flow Rate Range
– Flows of < 5mL/min Binary and < 10mL/min Quat are Easily Maintained
• Wide Flow Rate and Pressure Range Allows More Utilization
– One Instrument Configuration (1200SL) Can Run Current, Legacy and
Higher Performance Methods

• Attention to design details and strict adherence to manufacturing standards

permit routine operation at pressures up to the rated maximum.

• An Example is the piston/seal design specs and manufacturing techniques

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Quaternary Pump System
(G1354A)

Solvent Cabinet

Vacuum Degasser G1322A

Quaternary Pump G1311A

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Quaternary Pump
Principle of Operation

AIV
MCGV
Purge Valve
MCGV Proportioning Valve
AIV Active Inlet Valve
OBV Outlet Ball Valve
OBV Damper

Vacuum Chamber
Authoring Division Name File Name
Security Notice (if required)

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Binary Pump
with Solvent Selection Valve

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Binaryy Pumpp
Principle of Operation
Damper
Purge
valve Mixer

Waste Outlet Outlet

valve valve
Inlet
valve
Inlet
valve

A B

Group/Presentation Title
Agilent Restricted
Month ##, 200X
1100 and 1200 Pumps
Exploded
p View
Holding Screw

Purge
Outlet Valve Valve

P
Pump H
Housing
i
Plunger Housing Seals

Pistons Active Inlet Valve

Piston Support Rings

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Pump Check Valves

New style
G1312-60010
Cartridge

5062-8562 7

5062-8568 Old style

5
1 Outlet Ball Valve Purge
2 Iso/Quat Pump G1311-60012 6
Binary Pump G1312-60012
Valve
G1312-60009

Active Inlet Valve 1. Gold Washer 5001-3707 5. Gold Seal 5001-3707

(common to all)
2. Plastic cap 01018-21207 6. Cap(4pk) 5062-2485
3. Gold Seal 5001-3707 7. PTFE (5pk) 01018-22707
4 Cap(4pk)
4. 5062-2485
5062 2485

Group/Presentation Title
Agilent Restricted
Month ##, 200X
Pump Head with Seal Wash
Exploded View

1. Seal Wash Kit 01018-68722

2. Secondary Seal 0905-1175
3. Gasket, seal wash (6pk) 5062-2484

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Seal Wash
(Common to all 1100 pumps)

Washer
z Reduces seal wear when using mobile
phases
h with
ith salt
lt concentrations
t ti > 0.1
01M
Seal

z Optional for all LC 1100 Series Pumps

Support seal
assembly

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Agilent
g Technologies
g
1100 Series HPLC Maintenance

Autosampler

Group/Presentation Title
Agilent Restricted
Month ##, 200X
Autosampler Plumbing
From pump

To column compartment

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Autosampler
P i i l off Operation
Principle O i
Standard loop volume300µl
Total delay volume 300µl + Vinj
Minimal (bypass) delay volume 6.2ul

Vial gripper Metering device

Sampling
unitt
u

Rheodyne 7750
From pump
To column
z 4-port rotor seal

Widest dynamic injection range:

To waste
0 1 µl-1
0.1 µl-1.5
5 ml (w/addt
(w/addt'll hardware)

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Standard Injection Sequence
Metering
device

Sampling
unit

From pump
Prepare position
To column
6-port valve

To waste

Load position

Inject and run position

Impact of Needle Wash on Carry- Over with
Capped and Uncapped Wash Vials

Without needle wash

With needle wash, wash vial uncapped
With needle wash, wash vial capped

0.35 0.35

0.3 0.3

0.25 0.25
Primidone Phenacetine

Carry- over (%)

Carry -over (%)

0.2 0.2

0.15 0.15

0.1 0.1

0 05
0.05 0 05
0.05

0 0
0.1 0.5 1 5 0.1 0.5 1 5

Injection volume [µl] Injection volume [µl]

Group/Presentation Title
Agilent Restricted
Month ##, 200X
Autosampler
p Special
p Functions

z NEEDLE WASH
to reduce carry-over to absolute minimum

z MULTI- DRAW
for injection volumes greater than 100ul

z BYPASS MODE
for minimum delay volume

z INJECTOR PROGRAM
f iinjection
for j ti sequence flexibility
fl ibilit

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Autosampler
Rheodyne 7750 Injection Valve
z Motor-driven (36V motor)
z Switch <150 ms.
z End positions detected by microswitches.
z Ceramic stator.
z Vespel and Tefzel rotor seals available.
z No set screws required.
z Complete assembly as repair part
part.

Ceramic stator

4-port rotor seal

Test Functions-Autosampler
Functions Autosampler

z Maintenance Positions:

– Change needle
– Change needle seat
– Change metering seal
– Change gripper arm

z Step commands

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Agilent
g Technologies
g
1100 Series HPLC Maintenance

Thermostatted
Column
Compartment

Group/Presentation Title
Agilent Restricted
Month ##, 200X
Column Compartment
F t
Features
z Hold 3 30cm columns
z O
Operates
t from
f 10deg.
10d Cb below
l ambient
bi t ttemperature
t up tto 80
degrees C
z Peltier heating and cooling
z Two separate
p heating/cooling
g g zones
z Column ID module for column traceability
z Optional column switching valve

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Column ID Modules
Product number 79916OD-
Serial number 552 951001
Batch number 1675
Geometry [mm] 100x2.1
Stationary phase ODS
Particle size Hypersil
10 µm
Number of injections 999
Maximum pressure allowed [bar] 400
Maximum temperature recommended [°C] 70
Maximum pH recommended 12

ID ttag mustt b
be within
ithi 2mm
2 off the
th antenna,
t with
ith the
th HP logo
l up.

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Agilent
g Technologies
g
1100 Series HPLC Maintenance

Variable
Wavelength
g
Detector

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 V i bl Wavelength
Variable W l th Detector
D t t
Principle
p
Deuterium lamp
Source lens
Filter (holmium/cutoff)

Entrance slit
Don't disturb it's
holder!

SAMP photo diode Mirror #1

ADC board Flow cell
Grating

Beam splitter
Mirror #2

REF photo diode

ADC board

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Flow Cells
High Pressure
Standard and Micro

• Maximum pressure 40 Bar Maximum pressure 400 Bar

z Materials: SST, quartz, PTFE
Materials: SST, quartz, Kapton
z Standard cell: 10mm, 14ul
z Micro cell: 5mm,, 1ul 10mm, 14ul
z Gasket #2 has different aperture
High Pressure Cell

Standard and Micro

Cell

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Agilent
g Technologies
g
1100 Series HPLC Maintenance

Diode Array
Detector

Group/Presentation Title
Agilent Restricted
Month ##, 200X
Diode Array
y Detector
Features
E
Extended
d d wavelength
l h range up to 950nm
9 0
Micro-mechanical slit
Improved noise specification
Ch i off flow
Choice fl cells
ll

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Diode Array
y Detector
Principle of Operation
Support lens
Grating

Holmium oxide
filter

Spectro
lens
Deuterium lamp
Flow cell
Array
Tungsten lamp
Source lens
(Achromat) Micro mechanical Slit
Micro-mechanical

Coupling lens

Group/Presentation Title
Agilent Restricted
Month ##, 200X
 Flow Cells
Standard and Semi-micro Micro
Maximum pressure 400 Bar
z Maximum pressure 120 Bar
z M t i l SST,
Materials: SST quartz,Teflon
t T fl M t i l SST,
Materials: SST quartz,
t Kapton
K t
z Apertures of front and rear
6mm, 1.7ul
gaskets have different diameters
z Standard cell uses cell body for
additional
dditi l heat
h t exchange
h
z Semi-micro cell: 6mm, 5uL
z Standard: 10mm, 13uL

Mi
Micro C
Cellll
Different apertures

Standard and Semi-Micro

Semi Micro Cell

Group/Presentation Title
Agilent Restricted
Month ##, 200X
Chromatographic Results with Second Source
Lamp at 280 nm Wavelength - Small Peaks

Lamp from Agilent

Peak 1 S/N = 75
Technologies
Peak 2 S/N = 30
Peak 3 S/N = 45

Lamp from Second Peak 1 S/N = 10

Source Peak 2 S/N = 4
Peak 3 S/N = 6

Group/Presentation Title
Agilent Restricted
Month ##, 200X
Second Long-Life
g Lampp - 2000 Hours
PN: 2140-0813 (March 2003)

New coating material for the cathode

which improves life time to 2000 hours
Lamp ships with every new DAD since
Februaryy 2003 - CPL: Mayy 2003
A customer tested the lamp and reported 4000
hours life time

Group/Presentation Title
Agilent Restricted
Month ##, 200X
Summary

Over the past years we had been taken many efforts, invested money,
time and resources to make sure that we always have the best lamps for
our customers and made improvements for all our lamps.
Longer life time of our lamps reduces the cost of ownership of our
instruments substantially.y
We also make sure that we will continue our efforts to make the best
lamps available to our customers in the future.

Group/Presentation Title
Agilent Restricted
Month ##, 200X