Académique Documents
Professionnel Documents
Culture Documents
Ad
d Topic:
T i Tips
Ti
and Tricks of LC User
Maintenance
Edward Kim
Application Engineer
February 19
19, 2010
2/18/2010
mA
U
Troubleshooting in HPLC
20
00
15
00
10
5
00
0
0
0
Page 2
1
0
1
5
Time
(min)
2
0
2
5
Pumping principle
Sampler principles
TCC principle
Detector principle
2/18/2010
2/18/2010
02274
DE
916
Where made?
Date code
JP = Japan
DE = Germany
US = USA
CN = China
FR = France
IIndividual
di id l module
d l counter
t
(how many have been made?)
2/18/2010
Degassers - Overview
G1322A Vacuum Degasser (aka Standard Degasser/Online Degasser)
G1379A Micro Vacuum Degasser 1100 Series (obsolete)
G1379B Micro Vacuum Degasser 1200 Series
2/18/2010
2/18/2010
2/18/2010
Comparison of degassers
G1322A:
better for applications using methanol
recommended for use with refractive index detector (RID)
better for high flow rates (>5 ml/min)
G1379A/B
better in all other cases
2/18/2010
Maintenance
Cleaning Flush the solvent tubes with
isopropanol.
When using buffers, flush with water.
May
M h
have tto exchange
h
th
the vacuum pump,
sensor, solenoid valve, or chamber.
Replace solvent inlet frits
Solvent
Pump
10
Pumps - Overview
G1310A IIsocratic
ti P
Pump: one solvent
l
t ((might
i ht b
be mixture)
i t )
G1311A Quaternary Pump: four solvents, low pressure mixing
G1312A Binary Pump: two (four) solvents, high pressure mixing
[G1312B: binary pump SL (RRLC)
Special pumps for other purposes: cap, nano, prep systems]
2/18/2010
Isocratic pump
2/18/2010
Quaternary/binary pump
Quaternary Pump
Binary Pump
high pressure
mixing
multi-channel
gradient valve (MCGV)
l
low
pressure mixing
i i
Agilent 1200 Series LC Agilent Restricted
Page 13
2/18/2010
Pump options
Solvent Selection Valve (P/N G1312-60000)
G1312 60000)
Normal Phase Seals
(P/N 0905-1420)
Seal Wash
(P/N 01018
01018-68722)
68722)
(P/N G1311-68711)
2/18/2010
2/18/2010
2/18/2010
2
1
problems at plateau 1
check OBV
OBV, plunger/seal 2
2,
purge valve, blanking nut
problems at plateau 2
check AIV, plunger/seal 1
also affected by chamber 2
Agilent 1200 Series LC Agilent Restricted
Page 17
2/18/2010
IF A2 & B2 failed:
everything after the both OBV could be
leaking
g including
g blank nut and p
purge
g valve
Agilent 1200 Series LC Agilent Restricted
Page 18
2/18/2010
Pump parameters
flow: volume delivered per time
time, range depends on pump type
composition: percentage for each solvent (bin./quat. pump)
stroke:
t k the
th volume
l
iin a range off 20 ll tto 100 ll off compressed
d
solvent delivered by one movement of piston 1
p
mixing
g but decreases lifetime of seals
a lower stroke improves
and valves
stroke
100 l
auto
settable stroke
20 l
0.1 ml/min
5 ml/min
flow
Agilent 1200 Series LC Agilent Restricted
Page 19
2/18/2010
2/18/2010
piston 2 delivers
pressure (ripple)
piston 1 delivers
piston 2 delivers
pressure drop
p
p
2/18/2010
compressibility
ibilit setting
tti ttoo hi
high
h
2/18/2010
2/18/2010
180 l to 480 l
2/18/2010
Autosamplers - overview
G1313A Autosampler (obsolete)
G1329A Thermostated Standard Autosampler (1100/1200)
[G1329B: ALS SL (RRLC) high pressure variant]
G1367A Well-Plate
Well Plate Autosampler (1100 Series)
G1367B High-Performance Autosampler (1200 Series),HiP ALS
[1367C: HiP ALS SL (RRLC)]
[1367D HiP ALS SL+ (RRLC,
[1367D:
(RRLC llow carry over als)]
l )]
2/18/2010
Mainpass
Pump
to Column
Draw Sample
(Bypass)
to Column
Inject
Pump
Pump
to Column
2/18/2010
2/18/2010
2/18/2010
Page 29
Seal
Pump
Column
Rotor Seal
2/18/2010
TCC principle
G1316A
2/18/2010
Options:
Column Switching Valve 6 or 10 Port
Specs:
Specs
10 degrees below ambient to 80 deg C
up to 80 deg C: max flow rate 5ml/min
2/18/2010
C l
Color
id
i.d.
Red
0.12 mm
Green
0.17 mm
Blue
0.25 mm
33
Fittings
g
Swagelok
Waters
0 130 iin.
0.130
0 090 iin.
0.090
Rheodyne
Parker
0.170 in.
0.090 in.
Valco
Uptight
0.080 in.
0.090 in.
Detectors
G1314A Variable Wavelength Detector (1100 Series) VWD obs.
G1314B Variable Wavelength Detector (1200 Series) VWD
[G1314C Variable Wavelength Detector SL (1200 Series) VWD SL]
[G1314D/E Variable Wavelength Detector SL+ (1200 Series) VWD SL+]
2/18/2010
Lamp Matrix
Detector
New
Original
DAD
G1315A/B/C
MWD
G1365A/B/C
DAD SL
G1315D/E
MWD SL
G1365D/E
1290 Infinity
G4212A (New)
VWD
Secondary
VIS Lamp
2140-0813
5181-1530
(2140-0820)
G1103-60001
2140-0820
5181-1530
(2140-0813)
G1103-60001
5190-0917
NA
G1103-60001
G1314A/B/C
G1314-60100
2140-0813
G1103-60001
VWD SL+
G1314D/E
G1314-60101
2140-0820
G1103-60001
UV VIS
8453A
2140-0605
2140-0813
G1103-60001
CE
G1600A
2140-0585
NA
NA
CE
G7100A (New)
5190-0917
NA
NA
Page 36
Group/Presentation Title
Agilent Restricted
Month ##, 200X
VWD principle
I0
I0
Agilent 1200 Series LC Agilent Restricted
Page 37
2/18/2010
Typical Column
Length
Typical Peak
Width
<= 5 cm
0.025
10 cm
0.05
20 cm
0.1
>= 40 cm
0.2
High-Pressure
Flow Cell
Standard Flow
Cell
For Pressure
Above 100 bar
0.05-0.2 mL/min
0.2-0.4 mL/min
0.4-0.8 mL/min
1-2 mL/min
1.0 mm
2.1 mm
3.0 mm
4.6 mm
0.05-5 mL/min
2/18/2010
Baseline stabilization
2/18/2010
2/18/2010
Wavelength calibration
Or: how to calibrate the rotation angle of the grating
0
1
1
656 nm
=>
>T
Two positions
iti
needed:
d d zero order
d and
d fifirstt order
d 656 nm emission
i i liline
of deuterium lamp
required after repair, part replacements or for PM
the 656 nm position is checked each time the lamp is switched on
Agilent 1200 Series LC Agilent Restricted
Page 41
2/18/2010
2/18/2010
lamp performance
cleanliness of flow cell
Agilent 1200 Series LC Agilent Restricted
Page 43
2/18/2010
2/18/2010
2/18/2010
VWD scan
During a method execution, a sample spectrum can be
acquired Therefore two scans are needed: a solvent/blank scan
acquired.
and a sample scan. The online spectrum is displayed as the
difference of both scans. The flow needs to be stopped
(flow = 0) during the sample scan.
2/18/2010
Flow Cells
Standard and Micro
High Pressure
Group/Presentation Title
Agilent Restricted
Month ##, 200X
I0
Measurement of I0?
Agilent 1200 Series LC Agilent Restricted
Page 48
2/18/2010
Page 49
2/18/2010
2/18/2010
2/18/2010
2/18/2010
Typical Column
Length
Typical Peak
Width
<= 5 cm
0 025
0.025
10 cm
0.05
20 cm
0.1
>= 40 cm
0.2
High-Pressure
Flow Cell
Standard Flow
Cell
0.01-0.2 mL/min
0.2-0.4 mL/min
0.4-0.8 mL/min
1-2 mL/min
0.3-1
0.3
1 mm
2.1 mm
3.0 mm
4.6 mm
0.05-5 mL/min
2/18/2010
Flow Cells
Standard and Semi-micro
Maximum pressure 120 Bar
M t i l SST,
Materials:
SST quartz,Teflon
t T fl
Apertures of front and rear
gaskets have different diameters
Standard cell uses cell body for
additional
dditi
l heat
h t exchange
h
Semi-micro cell: 6mm, 5uL
Standard: 10mm, 13uL
Micro
Maximum pressure 400 Bar
M t i l SST,
Materials:
SST quartz,
t Kapton
K t
6mm, 1.7ul
Mi
Micro
C
Cellll
Different apertures