Alltech® Model 3300 ELSD
Operation Manual
11593900A en
Imprint
Product Identification:
Operation Manual (Original) Alltech® Model 3300 ELSD
11593900A
Publication date: 04.2016
BÜCHI Labortechnik AG
Meierseggstrasse 40
Postfach
CH-9230 Flawil 1
E-Mail: quality@buchi.com
BUCHI reserves the right to make changes to the manual as deemed necessary in the light of experience, especially with respect to structure, illustrations and technical detail.
This manual is copyrighted. Information from it may not be reproduced, distributed, or used for competitive purposes, nor made available to third parties. The manufacture of any component with the aid of this
manual without prior written agreement is also prohibited.
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Safety Symbols used in this Manual:
The Danger symbol indicates a hazardous situation that, if not avoided, will result in death or serious injury.
The Warning symbol indicates a hazardous situation that, if not avoided, could result in death or serious injury.
The Caution symbol indicates a hazardous situation that, if not avoided, could result in minor or moderate injury.
The Notice symbol is used to highlight information that will optimize the use and reliability of the system.
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Important Safety Guidelines for the Model 3300 ELSD
Please read the following cautions and warning statements carefully before
using the Model 3300 ELSD:
• The equipment must be used as specified by the manufacturer otherwise
overall safety will be impaired.
• All service must be completed by qualified personnel only.
• Only use the power supply cord recommended by the manufacturer.
• Fire hazard, only use the same type and rated CERTIFIED fuse
(
• Shock hazard, disconnect power before replacing the fuse.
• Remove power cord from the power module to disconnect electrical power
from the unit.
• Check the MSDS forms and dispense and dispose of all reagents in
accordance with local and national regulations.
• Be sure to provide proper ventilation for all solvent vapors.
250V 5A F).
• Avoid open flames and sparks when using flammable solvents.
• If the unit is damaged and does not function properly, stop the unit safely
and contact the manufacturer immediately.
• Class 3B LASER radiation inside can cause severe eye damage. Do not
open or defeat interlocks. Avoid exposure to the beam.
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Table of Contents
1. Introduction ......................................................................................................................... 1
1.1 About the Model 3300 ELSD .................................................................................................................. 1
1.2 Principle of Operation ............................................................................................................................ 2
2. Installation ........................................................................................................................... 3
2.1 What You Will Need ............................................................................................................................... 3
2.2 Unpacking .............................................................................................................................................. 3
2.3 Controls and Features ........................................................................................................................... 4
2.3.1 Front Panel (Figure 2.1) ..................................................................................................................................... 4
.3.2 Back Panel (Figure 2.2) ..................................................................................................................................... 5
2
2.4 Making Electrical and Fluid Connections ............................................................................................. 6
3. Navigating the Software Interface ...................................................................................... 7
3.1 Main Screen ............................................................................................................................................ 7
3.2 Operation ................................................................................................................................................ 7
3.2.1 Mode ................................................................................................................................................................... 7
3.2.2 Start/Stop Run.................................................................................................................................................... 8
3.2.3 Lock/Unlock Detector ........................................................................................................................................ 8
3.2.4 Chart ................................................................................................................................................................... 8
3.2.5 Autozero ............................................................................................................................................................. 9
3.3 Method .................................................................................................................................................... 9
3.3.1 New ..................................................................................................................................................................... 9
.3.2 Open ................................................................................................................................................................. 10
3
3.3.3 Edit .................................................................................................................................................................... 10
3.3.4 Delete ................................................................................................................................................................ 10
3.3.5 Wizard ............................................................................................................................................................... 11
3.4 Configuration ....................................................................................................................................... 11
3.4.1 Alarm ................................................................................................................................................................ 11
3.4.2 Outputs ............................................................................................................................................................. 11
3.4.3 Display .............................................................................................................................................................. 12
3.4.4 Date and Time .................................................................................................................................................. 12
3.4.5 Language .......................................................................................................................................................... 12
3.4.6 Pressure Units ................................................................................................................................................. 12
3.4.7 Network ............................................................................................................................................................ 13
3.4.8 Timed Mode Changes ...................................................................................................................................... 13
3.5 Maintenance ......................................................................................................................................... 13
3.5.1 Manual Control................................................................................................................................................. 13
.5.2 Logs .................................................................................................................................................................. 14
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3.5.3 Tests ................................................................................................................................................................. 14
3.5.4 Files .................................................................................................................................................................. 14
3.5.5 Service Tech .................................................................................................................................................... 14
3.5.6 About ................................................................................................................................................................ 14
4. Routine Operation ............................................................................................................. 15
4.1 Safety .................................................................................................................................................... 15
4.2 Operating Notes ................................................................................................................................... 15
4.3 Selecting Initial Operating Conditions ................................................................................................ 15
4.4 Startup Sequence ................................................................................................................................ 15
4.5 Shutdown Sequence ............................................................................................................................ 16
4.6 Optimization Procedure ....................................................................................................................... 16
5. Maintenance ....................................................................................................................... 17
5.1 Cleaning Mode ..................................................................................................................................... 17
5.2 Nebulizer Cleaning Procedure ............................................................................................................. 17
5.3 Drift Tube Cleaning Procedure ............................................................................................................ 18
5.4 Optics Cleaning Procedure ................................................................................................................. 18
5.5 Fuse Replacement ............................................................................................................................... 19
6. Diagnostics and Troubleshooting ................................................................................... 20
6.1 Errors .................................................................................................................................................... 20
6.2 Performing Diagnostic Tests ............................................................................................................... 21
6.2.1 Nebulizer Gas Pressure Test .......................................................................................................................... 21
6.2.2 Gas Flow Test .................................................................................................................................................. 22
6.2.3 Optics Test ....................................................................................................................................................... 22
6.2.4 Optics Heating Test ......................................................................................................................................... 23
6.3 Diagnosing Baseline Noise ................................................................................................................. 25
6.4 Troubleshooting Charts ....................................................................................................................... 26
7. Appendix ............................................................................................................................ 28
7.1 Specifications ...................................................................................................................................... 28
7.2 Replacement Parts ............................................................................................................................... 28
7.3 Volatile Mobile Phase Modifiers .......................................................................................................... 29
Contact BUCHI .................................................................................................................... 30
8.
9. Useful References ............................................................................................................. 31
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1. INTRODUCTION
1.1 ABOUT THE MODEL 3300 ELSD
The Model 3300 ELSD is designed for use with High
Performance Liquid Chromatography (HPLC) systems to
analyze any sample compound that has sufficiently lower
volatility than the mobile phase. Some of its possible
application areas include carbohydrates, pharmaceuticals,
lipids, triglycerides, underivitized fatty and amino acids,
polymers, surfactants, nutraceuticals, and combinatorial
libraries.
Evaporative light scattering detection eliminates common
problems associated with other HPLC detectors. Refractive
Index (RI) detectors can be complicated by solvent front
interferences, baseline instability, and gradient
incompatibility. RI detectors can also have a less sensitive
response than ELSD. Low-wavelength UV can suffer
baseline drift with steep gradients and also requires that the
analyte contains a chromophore. ELSD does not have these
limitations. It can achieve stable baselines with multisolvent
gradients for improved resolution and faster separations.
Also, since ELSD response does not depend on the sample’s
optical characteristics, the sample does not require a
chromophore or fluorophore for detection.
The Model 3300 ELSD features the most advanced
evaporative light scattering detection technology available. It
provides sensitivity in the low nanogram range. An intuitive,
Windows-based software interface provides a nested menu
f options for detector operation, including Method Wizard for
o
simplified method development, multiple languages, and builtin Diagnostic tests for troubleshooting. The Model 3300 ELSD
also features compact, stackable dimensions, allowing it to fit
easily into limited laboratory space.
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1.2 PRINCIPLE OF OPERATION
The unique detection principle of evaporative light scattering
detection involves a three-step process: nebulization of the
column effluent to form an aerosol, solvent evaporation within
a heated drift tube, and detection of the non-volatile solute
particles in the light scattering cell.
NEBULIZATION
EVAPORATION
DETECTION
Lower mobile phase flow rates will require lower gas flow
rates for optimal nebulization. Substitution of a 2.1mm i.d.
column for your standard 4.6mm i.d. column will allow you to
greatly reduce the mobile phase flow rate while also
increasing the sensitivity of the analysis.
EVAPORATION
Evaporation of the volatile components in the aerosol occurs
in a heated, stainless steel drift tube. The optimum drift tube
temperature setting for an application will depend on mobile
phase composition, mobile phase flow rate, and on sample
volatility. Highly organic mobile phases require lower drift
tube temperatures for evaporation than highly aqueous
mobile phases. Lower mobile phase flow rates require lower
drift tube temperatures than higher mobile phase flow rates.
Semi-volatile analytes require the use of much lower drift
tube temperatures to obtain optimum sensitivity. The
optimum temperature should be determined by observing
the signal-to-noise ratio with respect to temperature.
Within the Model 3300 ELSD drift tube, a PTFE-coated,
stainless steel impactor plate is located three inches from
the nebulizer. The plate is perpendicular to the aerosol path
so as the aerosol contacts the plate, larger droplets exit
through the drain tube on the side panel. The remaining
droplets pass around the impactor and travel through the
drift tube to the optical cell for detection. The removal of the
larger droplets allows operation of the Model 3300 ELSD at
low temperatures, making it ideal for the analysis of semivolatile compounds.
Non-volatile impurities in the mobile phase or nebulizing gas
will produce noise. Using the highest quality gas, solvents,
and volatile buffers (preferably filtered) will greatly reduce
baseline noise. Noise will also increase if the mobile phase
has not been completely evaporated. Detector settings must
be carefully selected to ensure adequate mobile phase
evaporation.
NEBULIZATION
The column effluent from an HPLC separation enters the
nebulizer, where it is mixed with a steady stream of
nebulizing gas (usually nitrogen) to form an aerosol. The
aerosol consists of a uniform distribution of droplets whose
size is dependent on the gas flow rate used for the analysis.
The lower the gas flow rate used, the larger the resulting
droplets will be. Larger droplets scatter more light and
increase the sensitivity of the analysis, but they are also more
difficult to evaporate in the drift tube. There will be an
optimum gas flow rate for each method which will produce the
highest signal-to-noise ratio.
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DETECTION
The non-volatile sample particles emerge from the drift tube
in the mobile phase vapor and enter the light scattering cell.
In the optical cell, sample particles scatter light emitted by a
laser light source. The scattered light is detected by a silicon
photodiode, generating a signal proportional to sample
mass. This signal is sent to the analog output for data
collection. The advanced design of the Model 3300 ELSD
optical components1 provides excellent sensitivity for HPLC
nalysis.
a
1
US Patent #6229605; other patents pending
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2. INSTALLATION
2.1 WHAT YOU WILL NEED
In addition to the Model 3300 ELSD detector and its
accessories, the following will be needed for installation of a
complete chromatographic system:
Exhaust System:
A fume hood or other ventilation device located close
•
to the detector to remove the detector exhaust from
the laboratory.
Make sure the exhaust system provides adequate but not
excessive suction. Excessive suction can cause a noisy
baseline.
Gas Supply:
A supply of clean, dry nebulization gas, preferably
•
nitrogen, regulated from 65 to 80psig. 99.9% purity or
better is recommended. The gas source can be a highpressure gas cylinder, high-pressure liquid tank, or a
nitrogen generator.
HPLC System Components:
An HPLC pump, isocratic or gradient, capable of
•
low-pulsation solvent delivery at a flow rate ranging
from at least 0.1 to 1.5mL/min against pressures of
at least 3,000psig. Lower flow rate capabilities may
be necessary for smaller bore columns.
An autosampler or manual injection valve.
•
A column capable of separating the compounds of
•
interest.
A guard column or cartridge compatible with the
•
separation column is recommended to prolong
separation column lifetime.
A column heater, if needed.
•
A data system or integrator, capable of accepting
•
analog voltage data. 0-10mV or 0-1000mV systems
can be used.
Other:
HPLC-grade mobile phase solvents.
•
2.2 UNPACKING
The Model 3300 ELSD detector and its accessories are
shipped in the same container. Unpack components
carefully, making sure all items in the list below have been
included and are in good condition. Save the container and
packing material for future use.
The Model 3300 ELSD shipping container should contain the
following:
Model 3300 ELSD
•
Model 3300 ELSD Operating Manual
•
Model 3300 ELSD Performance Documents
•
Model 3300 ELSD Driver for Agilent ChemStation CD*
•
Power Cord
•
Signal Cable
•
PEEK Tubing: 1/16" o.d. x .005" i.d., 10’
•
SofGrip Fittings, 10/pk
•
Gas Tubing, 10’
•
Gas Fittings: 1/8” Brass Nut and Ferrule
•
Drain Tubing, 5’ (Reservoir not included)
•
Drain Tubing Clamp
•
Exhaust Adapter
•
Exhaust Tubing, 20’
•
14-Pin Connector
•
Open-End Wrench, 3/8" x 7/16"
•
Open-End Wrench, 1/4" x 5/16"
•
Hex Ball Driver, 3/32" x 5 1/4”
•
Hex Ball Driver, 7/64" x 4”
•
Drift Tube Cleaning Brush
•
Fuse, 5 Amp
•
*The Model 3300 ELSD Driver for Agilent ChemStation CD
can be used with Agilent ChemStation to control and collect
data from the 3300 ELSD. Agilent ChemStation must be
purchased separately.
The Model 3300 ELSD has been carefully shipped to ensure
that it is received in proper condition. Any damage to the
container or its contents should be reported immediately to
your local BUCHI Customer Service office which can be
found on the BUCHI website at: www.buchi.com. Please
refer to Section 7.5, Warranty, Returns, and Repairs, for
more information.
Refer to Section 7.2, Replacement Parts, for part numbers if
replacement parts are needed.
Only volatile buffers may be used in the mobile phase.
Refer to Section 7.4, Volatile Mobile Phase Modifiers, for a
list of suitable buffers.
Solvent reservoirs, tubing, inlet filters, paper, etc.
•
required for pump and data system operation. Consult
the appropriate manuals for requirements.
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2.3 CONTROLS AND FEATURES
2
3
9
5
4
6
8
Figure 2.1: Front Panel of 3300 ELSD; Door Removed
2.3.1 FRONT PANEL (FIGURE 2.1)
1. LCD (Liquid Crystal Display): The LCD displays the
software interface screens.
2. Soft Power Button: The Model 3300 ELSD can be
powered on/off from the front panel of the unit using the
soft power button. NOTE: The power button on the back
panel must be set to the On position in order for the front
panel button to be active.
3. Liquid Inlet: The column effluent tubing connects to the
LIQUID INLET with a 1/16" male fingertight fitting.
4. Nebulizer Liquid Tubing: The stainless steel tubing
carries the column effluent to the liquid inlet of the
nebulizer.
5. Impactor Cartridge: The cartridge can be removed in
order to access the drift tube for cleaning. Refer to
Section 5.3 for details.
6. Nebulizer: The column effluent and nitrogen gas are
combined within the nebulizer to form an aerosol. The
nebulizer is easily removed for cleaning by using a
counterclockwise twisting motion.
7. Nebulizer Gas Tubing: The gas tubing carries nitrogen
gas to the nebulizer gas inlet.
8. Drain Port: The DRAIN port is located on the right side
panel. The drain tubing and clamp included with the unit
are connected to the DRAIN port and then directed to a
waste reservoir (not included) located at bench level if the
unit is stacked or floor level. NOTE: The drain tubing
must not be submerged in the liquid inside the reservoir
during operation.
9. Keypad: The keypad is used to interact with the software
interface screens.
10. Removable Door (not shown): The front door can be
opened or removed to access the nebulizer and drift tube
for cleaning. To remove the door, gently pull the door
towards you.
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2.3.2 BACK PANEL (FIGURE 2.2)
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2
3
5
6
Figure 2.2: Model 3300 ELSD Back Panel
1. Signal Output: The signal cable included with the unit is
connected to the SIGNAL OUTPUT port on the back
panel of the unit and is used to send the analog signal to
the data collection device.
2. RS-232: This port is currently reserved for service tech
functions only.
3. USB Host: A pen drive (not included) can be inserted
into this port for file importing/exporting and software
upgrades. Also for possible future pc control capabilities.
4. USB Device: This port is currently reserved for service
tech functions only.
5. Ethernet: This port can be used with the 3300 ELSD
Driver for Agilent ChemStation. This port is also used for
service tech functions.
6. 14-Pin Connector: Outputs TTL/contact closure signals
or accepts signals from peripheral equipment.
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7. Exhaust Outlet: Nebulizer gas, mobile phase vapor, and
solute mist or particles produced during an analysis will
exit the detector through the EXHAUST outlet. The
exhaust outlet accepts the exhaust adapter and exhaust
tubing included with the unit, which must be then directed
to a fume hood.
8. Fan: Provides cooling airflow through the instrument. Do
not block.
9. Power Module: The power module contains a socket for
the incoming power cord. It also contains the main power
switch, which is used to turn the system power On/Off,
and the line fuse. The module is autoselecting for 85265V and contains a 5 Amp fuse.
10. Gas Inlet: The GAS INLET accepts the nebulizer gas
supply tubing. The inlet features a quick-connect port,
which requires no additional fittings. Gas pressure should
be regulated from 65 – 80psig.
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2.4 MAKING ELECTRICAL AND FLUID
CONNECTIONS
1. Unpacking the Unit: Remove the ELSD from its shipping
container and position it on a level surface near the
column outlet of your HPLC system and the fume hood.
Make sure there is free flow of air to the bottom of the
ELSD and to the cooling fan at the back panel of the
ELSD. Allow the detector to warm to ambient temperature
if necessary. Save the shipping container for future use.
2. Exhaust Adapter:
adapter provided in the accessory kit to the EXHAUST
port on the back panel of the unit.
3. Fill the Internal Drain Reservoir: Place a beaker under
the DRAIN port on the right side panel. Using a water
bottle or pipette, add water into the EXHAUST port on the
back panel of the unit until excess liquid is seen exiting
the DRAIN port. Filling the internal drain reservoir
prevents gas from escaping the drift tube.
4. Power Connection: Plug the power cord provided with
the unit into the power module on the back panel of the
detector. The module is autoselecting for 85-265V.
5. 14-Pin Connector: Insert the 14-pin connector into the
port on the back panel. Make the appropriate wiring
connections to the 14-pin connector depending on which
of the following functions are needed:
Start: Pin 1: Ground (-) Pin 2: Signal (+)
Pins 1 and 2 on the Model 3300 ELSD can accept a
TTL/contact closure signal to start a method run on the
detector.
Standby: Pin 3: Ground (-) Pin 4: Signal (+)
Pins 3 and 4 on the Model 3300 ELSD can accept a
TTL/contact closure signal to put the detector into
Standby mode.
Gas Shutoff: Pin 5: Ground (-) Pin 6: Signal (+)
Pins 5 and 6 on the Model 3300 ELSD can accept a
TTL/contact closure signal to turn off gas flow at the end
of a run. This signal is typically sent from an autosampler
or a data collection system. Consult the appropriate
manuals for wiring information.
Autozero: Pin 7: Ground (-) Pin 8: Signal (+)
ins 7 and 8 on the Model 3300 ELSD can accept a
P
TTL/contact closure signal from a start signal cable to
autozero the detector. This signal is typically sent from an
autosampler or a manual injection valve with a positionsensing switch. Consult the appropriate manuals for wiring
information.
Screw in the threaded exhaust
Only use the power supply cord
recommended by the
manufacturer.
External Output: Pin 9: Normally Open (NO) (+)
Pin 10: Common (-)
Pin 11: Normally Closed (NC) (+)
Pins 9, 10, and 11 on the Model 3300 ELSD can output a
TTL/contact closure signal to other instruments. Consult
the appropriate manuals for wiring details.
Fault Relay: Pin 12: Normally Open (NO) (+)
Pin 13: Common (-)
Pin 14: Normally Closed (NC) (+)
Pins 12, 13, and 14 on the Model 3300 ELSD can output
a TTL/contact closure signal to stop pump flow when an
error occurs on the detector. Consult the appropriate
manuals for wiring details.
6. Gas Connection: Connect the nitrogen gas supply
tubing to the GAS INLET on the back panel. The nitrogen
gas supply should be regulated from 65 – 80psig. A
stable gas flow and pressure are necessary for
reproducible results. The gas must be free of
contaminants, such as oil, water, particulates, or any other
non-volatile substances. A 0.1um gas filter is built into the
instrument.
7. Liquid Connection: Connect the column effluent line to
the LIQUID INLET with a 1/16" male fitting. The i.d. and
length of the tubing between the column and the detector
should be kept as small as possible to avoid band
broadening. 0.005" i.d. tubing is recommended for best
results. Tubing and fittings are included in the ELSD 3300
accessory kit.
8. Drain Setup: Attach the drain tubing and clamp included
with the detector to the DRAIN OUTLET on the right side
panel. Extend the tubing to a drain waste container (not
included) either at bench level if the detector is stacked or
at floor level. Make sure the container is sealed to prevent
solvent fumes from escaping. Monitor the liquid level in
the container during operation and decant excess liquid
when the level approaches the top of the container.
Do NOT allow the tubing to become
submerged in the liquid inside the
container. Remember that the waste
container will contain solvents from
your mobile phase and should be
disposed of properly.
9. Signal Output: Connect the signal cable provided with
the unit to the SIGNAL OUTPUT port on the back panel of
the detector and connect to your data collection device.
10. Exhaust Tubing: Connect the exhaust tubing included
with the unit to the EXHAUST outlet on the rear panel of
the unit. Extend the tubing to the hood/ventilation system.
There should be no low spots in the tubing where
condensate can collect.
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Make sure the exhaust system provides adequate but
not excessive suction. Excessive suction can cause a
noisy baseline.
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3. NAVIGATING THE SOFTWARE
INTERFACE
The Model 3300 software interface features a collapsible
menu located in the upper left corner of the LCD screen. The
following sections describe the software menu options in
detail. The Help Button, “?” can be pressed in many
software screens in order to access Help Screens which offer
assistance with using the software functions.
3.1 MAIN SCREEN
The Operation screen is the main screen displayed during
use of the instrument. This screen provides the following
information for the currently loaded method:
Method Name: Currently loaded method name.
•
Temperature: Set point and read value of the drift tube
•
temperature in °C. The temperature range is from 25.0 to
120.0oC. NOTE: There are two heated zones in the drift
tube, a lower zone and an upper zone. The value
displayed on the main screen is the average value for the
two zones.
Gas Flow: Set point and read value of the nebulizer gas
•
flow rate in L/min. The gas flow range is from 0.0 to
4.0L/min.
Gain: Current gain setting. Possible gain values are 1,
•
2, 4, 8, and 16. A gain setting of 1 produces an
unamplified signal, and each increase in gain setting
produces twofold signal amplification over the previous
setting.
Signal Output: The signal output in mV is displayed
•
when the instrument is in Run or Cleaning modes. The
output is not displayed in Standby or Heating modes. If
the output value exceeds 2.5V, the signal output will read
‘High’. If output drops below -500mV, the signal output
will read ‘Low’.
Method Run Time: The elapsed time for the method run
•
is displayed in the upper right corner if a method run has
been started. The time value will remain until the method
run has been stopped.
Chart: The chromatogram trace of up to 60 minutes will
•
be active for Run and Cleaning modes.
Mode: The current mode of the detector (Standby, Run,
•
Heating, or Cleaning).
Equilibrating Indicator: If the detector is in Run mode
•
and the heaters and gas flow have not reached their set
points, the screen will display “Equilibrating” next to the
Mode status until the equilibration requirements are met.
The equilibration requirements are: lower drift tube,
upper drift tube, and optics block must be within 1.5C of
their set points; gas flow must be within 0.3L/min of its
set point; the temperatures and gas flow must meet the
requirements for 60 seconds before the message
disappears.
Timed Mode Change Reminder (not shown): If a
•
timed mode change has been scheduled, a reminder for
the event will be displayed next to the Mode status.
Errors: Any errors that occur on the instrument will be
•
displayed in the lower right corner of the screen.
The software Menu is located in the upper left corner of the
screen. There are four main Menu headers: Operation,
Method, Configuration, and Maintenance. The following
sections describe the software functions within these
headers.
3.2 OPERATION
3.2.1 MODE
The Model 3300 ELSD has four operational modes: Standby,
Run, Heating, and Cleaning.
STANDBY MODE
Select Operation/Mode/Standby and press Enter to put the
detector into Standby mode. In Standby mode, the heaters,
gas flow, and laser are off. The signal output is not
displayed, and the chromatogram trace is not active. The
detector enters Standby mode after power-up with the last
saved method conditions.
RUN MODE
Select Operation/Mode/Run and press Enter to put the
detector into Run mode. In Run mode, the heaters, gas flow,
and laser are on. The signal output is displayed and the
chromatogram trace is active. Run mode is used for sample
analysis.
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