Agilent Technologies 1260 Infinity II User manual

Agilent 1260 Infinity II
Multi-Angle Light Scattering Detector
User Manual

Notices
Manual Part Number
Document No: D0029607 Rev. B
Edition 07/2023
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© Agilent Technologies, Inc. 2023
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Safety Notices
A CAUTION notice denotes a hazard. It
calls attention to an operating
procedure, practice, or the like that, if
not correctly performed or adhered to,
could result in damage to the product
or loss of important data. Do not
proceed beyond a CAUTION notice
until the indicated conditions are fully
understood and met.
A WARNING notice denotes a hazard.
It calls attention to an operating
procedure, practice, or the like that, if
not correctly performed or adhered to,
could result in personal injury or
death. Do not proceed beyond a
WARNING notice until the indicated
conditions are fully understood and
met.
WARNING
CAUTION

Multi-Angle Light Scattering Detector User Manual 3
In this Guide….
This manual covers the Agilent 1260 Infinity II Multi-Angle Light Scattering Detector
(G7885A).
Please read this manual thoroughly before using the instrument!
Thank you for purchasing this detector from Agilent Technologies. You have
acquired a state-of-the-art instrument which enables you to perform unique
experiments and the characterization of the most complex particle, polymer and
protein samples.
This manual has been written for laboratory technicians who are responsible for
operation and maintenance of laboratory equipment. It is assumed that the user of
this manual is familiar with standard laboratory terminology. To ensure optimum
operation conditions, please read the manual completely and carefully before using
the detector. Use this product only in accordance with the guidelines and
instructions listed.
This manual describes in detail:
Characteristics
Operation and Maintenance
Important:
Do not use this instrument before completely understanding the full content of
this manual.
In the event that the instrument is loaned or transferred to a new owner please
ensure this manual is passed on with the device.
If this manual or any warning labels attached to the device become lost, get
damaged or are unreadable, promptly obtain replacement documents/ labels
from your nearest Agilent representative / distributor.
To ensure safe operation at any time, please read the safety instructions before
using the instrument.
In case of any uncertainty how to operate this detector, consult your Agilent
representative before starting to use this product.
NOTE Agilent offers basic, advanced and repeated trainings to support the user.

Multi-Angle Light Scattering Detector User Manual 4
Contents
1Introduction 6
After-Sales Service 7
Safety Instructions 8
Safety Symbols 16
Device Characteristics 17
2Operation 31
General Handling 32
Commissioning 35
User Interface 37
Changing the Solvent 39
Storage 40
Leak Detection 41
Vapor Detection 44
3Maintenance 46
Preventive Maintenance 47
Routine Maintenance 48
Detector Cleaning 49
4Troubleshooting 53
System Problems 54

Multi-Angle Light Scattering Detector User Manual 5
5Identified Cables 56
Cable and Connector Specifications 57
6Appendix 58
Waste Electrical and Electronic Equipment Directive 59
Agilent Technologies on Internet 60
Miscibility Chart of Solvents 61

Introduction
Multi-Angle Light Scattering Detector User Manual 6
1Introduction
After-Sales Service 7
Safety Instructions 8
Site Requirements 9
Power Considerations 10
Solvent Information 11
Using the Instrument 12
Maintaining the Instrument 13
Cleaning the Instrument 14
Handling Leak and Waste 14
Laser Warning 14
Static Electricity Warning 15
Safety Symbols 16
Device Characteristics 17
Delivery Content 17
General Purpose 17
Working Principle 18
Safety Functions 22
Physical Specifications 23
Performance Specifications 24
Detector Setup 26
Rear Side 28
Detection Angles 30
This chapter provides an overview of the principles and theory of Light Scattering
detection. Furthermore, the most important functions as well as specifications of
the detector are discussed.

Introduction
Multi-Angle Light Scattering Detector User Manual 7
After-Sales Service
If any problem occurs with this instrument, inspect it immediately and take
appropriate corrective action as described in chapter “Troubleshooting”on page 53.
If the problem persists or symptoms occur that are not described in this document,
disconnect the mains voltage and contact your Agilent representative.
Maintenance
Each component and the entire system should be checked periodically to ensure
that they function normally, otherwise the analysis data may not be precise and
reliable anymore. Typically, a yearly service is recommended.
Agilent offers special maintenance and service contracts to ensure maximum
lifetime and proper operation of the instrument during the entire life cycle. Before
shipment from the factory, this device was thoroughly inspected and tested.
The result of the inspection is documented in the attached protocol. Referring to
this protocol, please check the results of the hardware validation on a regular basis.

Introduction
Multi-Angle Light Scattering Detector User Manual 8
Safety Instructions
The user manual addresses persons who are qualified as chemical laboratory
technicians or have completed a comparable vocational training.
The following knowledge is required:
Fundamental understanding of liquid chromatography.
Familiarity with substances that are used for laboratory experiments.
Awareness regarding the health risks of chemicals.
Participation during a training by Agilent or an authorized person.
If you do not belong to this or a comparable professional group, you may not
perform the work described in this user manual under any circumstances. In this
case, please contact your superior.
The manufacturer does not warrant for any damage or injury resulting from
incorrect operation and maintenance, non-observance of the instructions in this
manual. Prior to delivery, this instrument has been tested for safe usage.
To ensure safe operation of the instrument, please read these safety
instructions carefully before use and follow them always while operating the
instrument.
Observe all of the Warning and Caution sections listed below. They are
extremely important for safety and to maintain a proper working device.
In this manual, Warning / Caution messages are always indicated using the
following conventions:
WARNING
A warning alerts you to situations that could cause physical injury or death.
Do not proceed beyond a warning until you have fully understood and met
the indicated conditions.
CAUTION A caution alerts you to situations that could cause loss of data, or damage of
equipment.
Do not proceed beyond a caution until you have fully understood and met the
indicated conditions.

Introduction
Multi-Angle Light Scattering Detector User Manual 9
Site Requirements
WARNING
•
Pay attention to the weight of the detectors. The detector weighs 16.5 kg
(36.4 lbs). Use proper lifting techniques to avoid potential injuries. Ensure
that you can handle the weight of the load.
•
Consider the entire weight of the combined system including all the
components. The lab table on which this detector is installed should be
sufficiently strong to support the total weight of the whole LC system. It
should be level, stable and have a depth of at least 600 mm. Otherwise, the
instrument could tip over or fall off the table.
•
Take measures to prevent the instrument from falling in the event of an
earthquake or other disaster. Strong vibrations could cause the detector to
fall over, resulting in injury and damage.
•
All other equipment connected to this instrument must be approved to an
appropriate safety standard and have reinforced insulation from the mains.
CAUTION
•
Avoid installation sites that are exposed to corrosive gases, strong vibrations, or
excessive dust. Otherwise, the performance and operability of the detector could
be impaired, and its lifetime may be shortened.
•Never place any liquid on top of the detector.
Bench Space
The module dimensions and weight (see Table “Physical Specifications”on page
23) allow you to place the module on almost any desk or laboratory bench. It needs
an additional 2.5 cm (1.0 inch) of space on either side and approximately 8 cm
(3.1 inches) in the rear for air circulation and electric connections.
If the bench shall carry a complete LC system, make sure that the bench is
designed to bear the weight of all modules.
NOTE The instrument is heavy and thus it is advisable to locate the system on a sturdy
bench.
NOTE Allow additional bench space for a PC if the instrument is to be computer-
controlled.

Introduction
Multi-Angle Light Scattering Detector User Manual 10
Power Considerations
WARNING
•
Ground the instrument.
To prevent electric shock in the event of an accident or electrical discharge,
the detector must be connected to protective earth. This is also important
to ensure stable operation and good data signals.
Ensure that all connected devices are grounded via the mains connector as
well.
•
Do not place heavy objects onto the power cord and keep any hot objects
away. This can damage the cord, resulting in fire, electrical shock, or
malfunction. If the cord is damaged, immediately unplug the mains power
cord and contact your Agilent representative.
•
Never modify the supplied power cord. Do not bend it excessively or pull on
it. Failure to do so could damage the cord, resulting in fire, electrical shock,
or malfunction.
•
Never replace the detachable power cord by an inadequately rated cord.
This can result in fire, electrical shock, or malfunction.
•
The power supply voltages and power consumption of this instrument are
listed in table “Physical Specifications”on page 23. Connect the detector
only to a power supply of the voltage indicated. Otherwise, fire or electric
shock may occur. Ensure that the power supply voltage is stable and has
sufficiently constant current capacity to operate all the components of the
system. If not, the detector will not operate at its rated performance levels.
•
If the power cord or its insulation gets damaged, unplug the cord and
contact your Agilent representative for a replacement cable immediately.
•
Only use mains connection cables admitted for use in your country. We
strongly recommend to only use cables supplied by Agilent.
•
In case of danger, immediately unplug the mains power cord either at the
wall power outlet, the power strip or on the rear of the devices.

Introduction
Multi-Angle Light Scattering Detector User Manual 11
Solvent Information
WARNING
•
Solvents used for Liquid Chromatography (LC) may be flammable and toxic.
Therefore, the room where the detector is installed, should be well
ventilated. Otherwise, solvent vapors could cause poisoning or may form an
explosive atmosphere, ignite, and cause fire.
•
LC can use large amounts of flammable organic solvents. Any open flame
in the surrounding of this instrument is strictly prohibited. Do not use the
detector in the same room with any other equipment that emits or could
potentially emit sparks. Sparks could cause fire. Provide fire extinguishers
for use in case of fire.
•
Provide protective equipment near the instrument. If solvent gets into the
eyes or on the skin, it must be flushed away immediately. Provide suitable
equipment such as eye wash stations / safety showers as close as possible
to the system. To avoid injuries due to spraying chemicals (e. g. by an
insufficiently connected tubing) it is advisable to always wear eye
protection when using the instrument.

Introduction
Multi-Angle Light Scattering Detector User Manual 12
Using the Instrument
WARNING
•
Use this instrument ONLY in the manner specified in this manual and for
the intended purpose. Using this detector in another manner or for any
other purpose may impair the protection provided by the instrument and
could cause accidents or unsafe conditions. The detector must not be used
for clinical or medical purposes.
•
When opening or closing the front cover, be careful not to pinch your fingers
to avoid any injuries.
•
Take thorough measures to prevent build-up of static electricity. Static
electricity could result in fire or explosions.
•
Always wear protective glasses and laboratory gloves when handling
solvents or samples.
Solvent getting into the eyes could cause blindness.
If solvent gets into the eyes, flush immediately with large amounts of water,
use eye wash stations, and seek medical treatment.
•
Always wear laboratory gloves when handling any toxic, biologically
infective samples or aggressive solvents.
•
Do not use flammable sprays (hair sprays, insecticide sprays, etc.) near the
instrument. They could ignite and cause fire.
CAUTION
•
Before the instrument is being powered off for a longer period, its flow lines
must be thoroughly flushed with filtered pure water to remove all remaining salt
from the fluid path. Failure to do this and damage caused by improper use and
precipitating buffer salts, or follow-up damages resulting from these actions,
are specifically excluded from the warranty.
•
Never let dry out the measuring cell. If the detector is disconnected from the
flow path, then close the “flow inlet” and “flow outlet” with the delivered plugs.
•
Never block the flow outlet of the detector when the inlet is connected to the
flow path of the LC system. An overpressure will cause internal leaks and can
damage the device.

Introduction
Multi-Angle Light Scattering Detector User Manual 13
NOTE Before flushing the detector with water, make sure that the eluent (organic eluent
with salt additives) used is miscible with water.
If necessary, first flush the detector with pure organic eluent and then switch to
water or use a transition solvent!
For more information concerning immiscible solvents, refer to section “Miscibility
Chart of Solvents”on page 61.
NOTE
For long time storage, flush the detector with a degassed mixture of pure water and
20 % methanol. Afterward, disconnect the detector from
the flow path and close the
inlet and outlet ports with the supplied plugs.
Maintaining the Instrument
WARNING
•
Disconnect the solvent flow and unplug the instrument before inspection,
maintenance, or replacement of any part. Otherwise, electrical shock, short-
circuit accidents, or other damages could occur.
•
Never remove the top cover of the detector while the unit is connected to
the mains power supply. This could cause injury or malfunction of the
instrument.
•
Do not remove screws and open the main cover of the instrument. This can
result in electrical shock, or malfunction of the instrument. The main cover
does not need to be removed for routine maintenance, inspection and / or
adjustment. Let your Agilent representative perform any repairs requiring
removal of the instrument's cover.
•
If the power cord plug gets dusty, remove the plug from the power outlet
and wipe the dust away with a dry cloth. Dust could cause fire.
•
Replace fuses only with fuses of proper type and capacity. Otherwise, the
safety regulations are not fulfilled.
If a fuse blows again after replacement, immediately unplug the mains
power cord and consult your local Agilent representative.
NOTE Replace parts only with original parts from Agilent. Any other parts may cause
damage and malfunction.

Introduction
Multi-Angle Light Scattering Detector User Manual 14
Cleaning the Instrument
WARNING
•
Never use pure alcohol or thinner solvent for cleaning the housing surface.
They could cause fire and discoloration.
To clean the surfaces of the closed housing, use a dry cloth and – if
necessary – a bit of glass cleaner.
•Do not allow flammable and / or toxic solvents to accumulate.
Handling Leak and Waste
WARNING
•
Liquids associated with this instrument may be classified as carcinogenic,
biohazardous, flammable, or radioactive. Should these liquids be used, it is
strongly recommended that this application be accomplished in an isolated
environment designed for these types of materials in accordance with
federal, state, and local regulatory laws, and in compliance with your
company’s chemical / hygiene plan in the event of a spill.
•
Dispose the waste liquid properly in accordance with the instruction by your
administrative department.
•
Never let hazardous liquids reach the public sewage system.
•
Before disposing of the equipment or sending it away for repair, you are
required to decontaminate the device in a technically correct manner.
All materials or fluids used for decontamination must be collected
separately and disposed of properly.
Laser Warning
WARNING
The laser inside the instrument is a class 3B laser device based on the norm
IEC 60825-1:2007. Do not open the detector. Hazardous laser radiation is
emitted when open. Avoid direct exposure to the beam. Never bypass the
safety guards to activate an opened device.

Introduction
Multi-Angle Light Scattering Detector User Manual 15
Static Electricity Warning
WARNING
•
Liquid Chromatography (LC) and other laboratory applications can use
flammable organic solvents as the mobile phase. Operators must be
constantly on guard against accidents involving fire or explosion. The major
cause of these accidents is static electricity. Devising preventive measures
for accidents involving static electricity is difficult as the causes can be
many, varied, individual, coincidental and / or simultaneous.
•
LC systems are often used in association with large quantities of
flammable solvents, thus the risk of large-scale damage is high, should an
accident occur.
•
The best way to prevent static electricity accidents is simply to prevent the
occurrence and accumulation of electrostatic charges. Therefore, the use of
antistatic shoes, tables, mats and floor covering is recommended.

Introduction
Multi-Angle Light Scattering Detector User Manual 16
Safety Symbols
The following safety symbols are attached to the cover on the back of the
instrument. They are intended to alert you to the following safety information.
Table 1 Safety Symbols
Symbol
Description
The apparatus is marked with this symbol when the user
should refer to the instruction manual in order to protect risk of
harm to the operator and to protect the apparatus against
damage.
Label indicates a CLASS 1 LASER PRODUCT.
Confirms that a manufactured product complies with all
applicable European Community directives. The European
Declaration of Conformity is available at:
http://regulations.corporate.agilent.com/DoC/search.htm

Introduction
Multi-Angle Light Scattering Detector User Manual 17
Device Characteristics
Delivery Content
The delivery of the detector includes the following components:
Agilent 1260 Infinity II Multi-Angle Light Scattering Detector:
with connection kit, Accessory Kit (tubing, cables, connectors, fittings, and
plugs)
User Manual (this document as PDF file on USB)
Declaration of Conformity to manufacturing specifications
The device is electronically and mechanically tested. The fluid system of the
detector is usually pre-filled with solvent (standard solvent is an 80 % water / 20 %
methanol mixture) as documented on a clearly visible adhesive label on the device,
if delivered with a system.
General Purpose
The Multi-Angle Light Scattering detector measures scattered laser radiation
reflected from sample material (nanoparticles / polymers) separated by an
LC system. This allows to determine molar mass, particle size and even branching
of particles or macromolecules (for more information, refer to section “Working
Principle”on page 18).
The detector uses high sensitive photon detectors with high-speed 24 bit A/D
converters as well as a thermally controlled and stabilized measurement bridge to
achieve best possible accuracy and performance. The integrated Ethernet LAN
interface allows to control the instrument and to transfer all measured data to a PC
running the Agilent WinGPC Software which can integrate further detectors like
multi-angle light scattering (one MALS detector per WinGPC Software license),
refractive index, and UV detectors.

Introduction
Multi-Angle Light Scattering Detector User Manual 18
Working Principle
For the determination of the mass and the size of particles, static light scattering
can be used. A laser beam illuminates the sample. Due to the electrical field of the
incident light, the electron shell of a molecule oscillates with the same frequency as
the incident light. Every charge that moves emits radiation, and so every molecule
emits light. Assuming that the sample has no conductivity, one can, to a first
approximation, treat this radiation as a dipole scatterer that emits radiation
isotropically.
Each atom in the sample thus acts as a light source. The sample consists of a large
amount of molecules that can have more than one scattering center if a molar
mass limit is reached (anisotropic scatterer). The more scattering centers there are,
the more light is emitted. This means that the total intensity of the scattered light
depends on the molar mass and sample concentration. The light from all these
scattering centers interferes, and this interference pattern can be used to obtain
information about the size of the sample.
Figure 1Schematic view of the light scattering setup

Introduction
Multi-Angle Light Scattering Detector User Manual 19
The scattering angle is defined as the angle between the passing laser beam and
the scattered ray of light, captured by a photon detector at the respective angular
position.
All incident light rays are in the same phase of maximum intensity. If they are
scattered at an angle of 0°, they still have the same phase. All partial rays have their
maximum and minimum at the same position. Due to the constructive interference,
this is where the highest signal intensity occurs.
If we go to larger scattering angles and consider two partial rays scattered at two
different centers, the partial rays have a phase shift due to the different positions of
the centers. The interference of the two rays is no longer purely constructive and
the intensity decreases. The greater the distance between the scattering centers,
the lower the intensity. This means that with increasing scattering angles an
intensity decrease is measured, which depends on the size of the particles. To
calculate the complete interference pattern, all scattering centers of the molecule
must be considered.
Thus, static light scattering takes advantage of two fundamental physical principles
when light interacts with matter:
The intensity of the scattered light is proportional to the product of sample
concentration times molar mass of the sample.
Small particles scatter light in all directions with nearly the same intensity
(isotropic scatterer) while large particles scatter light mainly in the forward
direction (anisotropic scatterer).

Introduction
Multi-Angle Light Scattering Detector User Manual 20
To utilize these facts for a determination of mass and size, the following
experimental setup is used:
Figure 2Scattering Geometry
Light is scattered by a particle and detected at different scattering angles. The
intensity of the scattered light is determined by several detectors mounted at
different angles
.
Several factors contribute to the measured intensity. The measuring cell contains a
collection of sample particles and solvent molecules. All these scatterer move and
produce a time-dependent interference pattern.
The time-dependent interference pattern due to particle motion can be neglected in
static light scattering, because the measuring time (about 0.1 sec) is much larger
than the typical time constants for particle motion (in the microsecond range,
depending on particle size). In dynamic light scattering, on the other hand, this time
dependence is measured, and from this the size of the particles is calculated.
The contribution of the solvent can be eliminated by subtraction. For batch
experiments, this means that the intensity distribution of the pure solvent is
measured first and subtracted from the result of the sample in the solvent. For
chromatographic runs, this means that the baseline is set to the solvent scattering
level and this baseline is subtracted from the peak. The remaining scattering
intensity is called the “Excess Rayleigh Ratio” in honor of Lord Rayleigh, who first
described the scattering of light mathematically in 1871.
In general, there are two contributions from the scattering of particles. The angular
dependence of the particles and how they sum up due to density fluctuations.