Malvern Zetasizer Nano Series User manual

Printed in England MRK0499-02
Malvern Instruments Limited
Enigma Business Park
Grovewood Road, Malvern
Worcs, WR14 1XZ, U.K.
Tel: [phone]
Fax: [phone]
www.malvern.com
user manual
user manual
ZetasizerZetasizer nano series
Zetasizer
Malvern
nano series

Zetasizer Nano
User Manual
MAN0485 Issue 1.1 April 2013
English

Copyright © 2007, 2009, 2012, 2013 Malvern Instruments Ltd.
Malvern Instruments pursues a policy of continual improvement due to technical development. We
therefore reserve the right to deviate from information, descriptions, and specifications in this
publication without notice. Malvern Instruments shall not be liable for errors contained herein or
for incidental or consequential damages in connection with the furnishing, performance or use of
this material. No reproduction or transmission of any part of this publication is allowed without the
express written permission of Malvern Instruments Ltd.
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Zetasizer Nano Page i
Introduction to this manual
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-1
About this manual. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-1
Access to the instrument. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-2
Assumed information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3
Where to get help . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3
Introducing the Zetasizer Nano
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-1
About the Zetasizer Nano . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-1
Primary measurement techniques . . . . . . . . . . . . . . . . . . . . . . . . . 2-6
Hardware components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-11
Software overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-21
Making measurements
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-1
Quick guide to making a measurement. . . . . . . . . . . . . . . . . . . . . . 3-2
Powering up the system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-2
Sample preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-3
Selecting and preparing the measurement cell . . . . . . . . . . . . . . . . 3-3
Making an SOP measurement . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4
Making a manual measurement . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-6
The measurement display . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-6
Records and Reports - Viewing the Results
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-1
Displaying the results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-1
Table of Contents

Table of Contents Zetasizer Nano
Page ii MAN 0485
Sample Preparation
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-1
Preparing the sample - Size. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-1
Preparing the sample - Molecular weight . . . . . . . . . . . . . . . . . . . . 5-5
Preparing the sample - Zeta potential . . . . . . . . . . . . . . . . . . . . . . . 5-6
Count rate meter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-9
Security
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-1
Initial start-up - set up the administrator . . . . . . . . . . . . . . . . . . . . . 6-2
Enabling security . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-3
User groups . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-3
Users. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-5
Measurement file window - Workspaces
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-1
Measurement file window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7-2
Using SOPs
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-1
The SOP editor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-3
Creating an SOP. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-9
Measurement type selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-10
Size SOPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-11
Molecular weight SOPs. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-29
Zeta potential SOPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-38
Flow-mode SOPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-47
Trend SOPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-48
Titration SOPs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-50
Extracting an SOP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-50
Modifying an SOP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-51
Distributing an SOP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8-51

Zetasizer Nano Table of Contents
Zetasizer Nano Page iii
Advanced features
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-1
Solvent builder . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-2
SOP player . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-5
Result averaging. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-12
Editing the result . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-13
Exporting results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-15
Options window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-20
Expert advice. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9-26
Report designer
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-1
Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-2
Opening an existing report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-4
Creating a new report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-4
Adding elements to the report . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-4
Setting up the report elements. . . . . . . . . . . . . . . . . . . . . . . . . . . 10-6
Selecting elements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-16
Aligning and sizing elements . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-17
Saving a report. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-17
A finished report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-18
Viewing the new report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-19
Other information shown on a report . . . . . . . . . . . . . . . . . . . . . 10-19
Protecting a report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10-19
Size theory
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11-1
What is Dynamic light scattering?. . . . . . . . . . . . . . . . . . . . . . . . 11-1
Operation of the Zetasizer Nano
- Size measurements. . . . . . . . . . . . . . . . . . . . . . 11-6
Molecular weight theory
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12-1
What is Static light scattering?. . . . . . . . . . . . . . . . . . . . . . . . . . . 12-1
The Debye plot. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12-4

Table of Contents Zetasizer Nano
Page iv MAN 0485
Zeta potential theory
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13-1
What is Zeta potential? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13-1
Laser doppler velocimetry. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13-5
The M3-PALS technique . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13-7
Operation of the Zetasizer Nano
- Zeta potential measurements . . . . . . . . . . . . . . . . . . . . 13-11

Part 1 -
Instrument guide

Zetasizer Nano Page 1-1
1
Introduction
to this manual
Introduction
This manual covers the operation of the Zetasizer Nano particle characterisation
system. The instrument measures the size of particles contained within a sample,
presenting data according to the user’s needs.
This user manual is a companion to the Zetasizer Nano Basic Guide, which
gives Health and Safety, maintenance, troubleshooting and other vital information
which all users must read.
About this manual
The manual contains the general information required by an operator; as well as
supervisor information that concentrates on administration and extension of the
system’s functionality.
Topics covered include:
An overview of the Zetasizer Nano hardware and software.
A guide to making measurements and how to read the results.
Sample preparation guidance.
Reporting - creating and editing reports.
System security.
Advanced features.
Additionally an Theory section which describes the basic measurement theories
involved with the measurements.

Chapter 1 Introduction to this manual
Page 1-2 MAN 0317
1
Warning!
The instrument and the samples to be measured may be hazardous if mis-
used. Users must read the Health and Safety information in the Basic
Guide before operating the system.
Access to the instrument
This manual refers to the various people who will have access to the instrument, as
follows.
Malvern Instruments personnel
Malvern Instruments personnel (service engineers, representatives, etc.) have full
access to the instrument and are the only people authorised to perform all service
procedures that may require the removal of the instrument covers.
Warning!
Unless advised within the content of this manual, removal of the instru-
ment covers by unauthorised personnel, even a supervisor, will invalidate
the warranty of the instrument.
Supervisor
The supervisor is the person responsible for the management and safety of the
instrument and its operation. The supervisor also trains the operators and can per-
form user maintenance routines.
Operator
An operator is a person trained in the use of the system. The operator can perform
some user maintenance routines identified.
Warning!
Failure to follow these guidelines could result in exposure to hazardous
voltages and laser radiation.

Introduction to this manual Chapter 1
Zetasizer Nano Page 1-3
Assumed information
Naming convention
Within this manual:
The Zetasizer Nano is referred to as “the optical unit” or “the instrument”.
The accessories or sample dispersion units are referred to as “MPT-2 Titrator”,
“the vacuum degasser”, or as “the accessory” or “the dispersion unit”.
The combination of the optical unit, one or more accessories or dispersion
units and the computer is referred to as “the system”.
Menu commands
Software menu commands are referred to in the form main menu-menu item.
As an example, the command Configure-New SOP refers to selecting the New
SOP item in the Configure menu. Menu commands are shown in bold text.
Where to get help
This section describes the available sources of information on the system. The pri-
mary sources of information on the system are instrument and accessory manuals
and the software’s help system. The manuals give an overview of the system as a
whole, while the online help system gives more detailed information on the Zeta-
sizer software. Some features are described in both the manuals and Help system.
Manuals
The following manuals are available for the Zetasizer Nano:
Basic Guide
A concise yet essential guide that provides an introduction to the system and its
usage covering Site requirements, Health and Safety and an instrument and
accessory overview. Finally a brief guide to making a measurement and view-
ing the results is included.
User Guide
The primary source of information about the Zetasizer Nano hardware and its use.
See About this manual earlier in the chapter.
Accessory and dispersion unit guides
Separate guides are supplied with the various Nano accessories and Titrator disper-
sion units. These provide essential information on the connection, operation and
maintenance of the dispersion units.

Chapter 1 Introduction to this manual
Page 1-4 MAN 0317
Help System
Each key software window has a Help button giving information specific to it.
Additional Info Tips are also available whenever is shown - simply move the
mouse pointer over this icon to see a relevant tip.
Technical support
Malvern Instruments offers several methods of technical support, which are
described in this section.
Email support
Note
This is the primary support method – only use the other methods if this is
not available at your location.
When contacting your local Malvern Instruments representative - details of all local
telephone numbers are available at www.malvern.com. Always quote the following
information:
Model and serial number of the instrument. The serial number is shown
on the Zetasizer software status bar - rollover the Nano icon to display.
The software version. To find this select Help-About in the software -
alternatively, move the mouse over the instrument status bar icon. The Appli-
cation Version is the main firmware version number to note.
Additional support
Contact the International Malvern Helpdesk if the local Malvern Instruments
representative is not available:
Telephone: [phone] or email: [email protected].
Note
This help line is primarily English speaking.
Remote support
Malvern Instruments offers a remote support service, delivered by an internet con-
nection. Benefits include fast and efficient fault diagnosis, as well as reducing
downtime and costs.

Zetasizer Nano Page 2-1
2
Introducing the
Zetasizer Nano
Introduction
This chapter provides information about the Zetasizer Nano hardware and soft-
ware, and identifies all major features of the system.
The following topics are covered:
A description of the purpose of the instrument, its accessories and simple
explanations of the primary measurement techniques
Detailed reference on the physical parts of the optical unit.
An overview of the primary software features.
Information about the parts of the cell, including how to insert and remove it.
The following topics are covered:
Note
Hardware maintenance - for details on how to maintain the hardware,
please refer to the Basic Guide and the Accessory manuals.
References will be made throughout the manual to the MPT-2 Titrator and other
accessories. Refer to the MPT-2 Titrator and Degasser, and the Zetasizer
Nano accessories user manuals where necessary.
About the Zetasizer Nano
The Zetasizer Nano range of instruments provides the ability to measure three
characteristics of particles or molecules in a liquid medium.
These three fundamental parameters are particle size, zeta potential and
molecular weight. Unique technology within the Zetasizer system allows these
parameters to be measured over a wide range of concentrations. Depending upon

Chapter 2 Introducing the Zetasizer Nano
Page 2-2 MAN 0317
2
the options and accessories used, the Zetasizer system also has the ability to per-
form:
Microrheology measurements
Autotitration measurements if an MPT-2 accessory is connected,
Trend measurements, including the determination of the Aggregation
point.
The Zetasizer range features pre-aligned optics and programmable measurement
position for the measurement of size and zeta potential over a wide concentration
range and precise temperature control necessary for reproducible, repeatable and
accurate measurements. In addition other key parameters such as conductivity and
with the MPT-2 Titrator, pH can be measured.
The Zetasizer Nano range has been designed so that a minimal amount of user
interaction is necessary to achieve excellent results. The use of Standard Operating
Procedures (SOPs) and features such as the Folded capillary cell alleviate the
need for constant attention.
The Zetasizer Nano range
This manual covers the operation and maintenance of the Zetasizer Nano particle
analyser series.
Note
The Zetasizer model, Serial number, software and firmware version can be
found by left-clicking the Nano icon in the right corner of the status bar.
Zetasizer Nano
instrument
Model
number Measurement types
Nano S (Red badge) ZEN1600 Particle size and Molecular weight
Nano S (Green badge) ZEN1500 Particle size and Molecular weight
Nano Z (Red badge) ZEN2600 Zeta potential
Nano ZS (Red badge) ZEN3600 Particle size, Molecular weight and
Zeta potential
Nano ZS (Green badge) ZEN3500 Particle size, Molecular weight and
Zeta potential
Nano S90 (Red badge) ZEN1690 Particle size - 90° optics
Nano ZS90 (Red badge) ZEN3690 Particle size and Zeta potential
- 90° optics
Nano ZSP (Red badge) ZEN5600 Particle size, Molecular weight and
Zeta potential, Microrheology

Introducing the Zetasizer Nano Chapter 2
Zetasizer Nano Page 2-3
Zetasizer Nano range options
The Zetasizer Nano series of particle analysers consists of a range of different
instrument models fitted with a choice of ‘red’ or ‘green’ lasers. The models and
their measurement specifications are described in the table below, with instrument
options following.
For complete measurement specifications refer to the Basic Guide.
Laser type
The laser fitted is identified by the colour on the oval badge on the cover.
Instruments with a black and red badge fitted to the instrument cover either
have a 4mW 632.8nm ‘red’ laser or a 10mW 632.8nm ‘red’ laser (Nano ZSP
only) fitted.
Instruments with a Black and green badge have a 532nm ‘green’ laser fitted.
Zetasizer Nano Size range maximum (diameter) Size range for Zeta potential (diameter) Molecular weight range (Daltons) Accessory option (see below)
S0.3nm to 10μm – 342Da to 2x107Da


Z– 3.8nm to 100μm – 

ZS 0.3nm to 10μm 3.8nm to 100μm 342Da to 2x107Da


S90 0.3nm to 5μm – 342Da to 2x107Da

*
ZS90 0.3nm to 5μm 3.8nm to 100μm 342Da to 2x107Da

*
Nano
ZSP 0.3nm to 10μm 3.8nm to 100μm 342Da to 2x107Da


(included
as standard)
ill * Compatible but not recommended because of low sensitivity.
Narrow band filter High temperature
Universal ‘Dip’ cell Flow-mode option
High concentration cell Microrheology
MPT-2 Titrator and Vacuum degasser Protein mobility

Chapter 2 Introducing the Zetasizer Nano
Page 2-4 MAN 0317
Generally red lasers are least suitable for blue samples, while green lasers are least
suitable for red samples.
90° optics
The S90 and ZS90 instruments use optics that have a 90° scattering detector angle
for size measurements.
Accessory options
A range of accessaries and options are also available for more advance measurement
strategies.
These accessories are fully documented in the Zetasizer Nano accessories
guide or MPT-2/Vacuum degasser user manual.
Narrow band
filter
This filter improves the signal for samples that fluoresce at
the wavelength of the laser fitted. If a filter is fitted, an option
label will be attached to the front of the cuvette holder
ZEN9051 - Narrow band filter for ‘Green’ badged Zetasizer
Nano S instruments
ZEN9052 - Narrow band filter for ‘Green’ badged Zetasizer
Nano ZS instruments
ZEN9061 - Narrow band filter for ‘Red’ badged Zetasizer
Nano S, Z and S90 instruments
ZEN9062 - Narrow band filter for ‘Red’ badged Zetasizer
Nano ZS, ZS90 and Nano Plus instruments
High
temperature
These instruments have an increased temperature range of
0to 120.
If fitted the Zetasizer Nano instrument label will include an
‘HT’ identification. An option label will also be attached to
the front of the cuvette holder.
(The high temperature option cannot be fitted to the Zeta-
sizer Nano ZSP)
ZEN9063 - Extends the upper temperature range of the
Nano series from 90to 120
MPT-2 Titrator
and Vacuum
degasser
The MPT-2 Titrator is used to perform pH, additive and dilu-
tion titrations. The degasser is used to remove any dissolved
gases from the titrants before dispensing by the titrator. This
prevents any gaseous escape during the titration, increasing
the accuracy and reproducibility of the measurement.
Cells and
Cuvettes
Overview of all cells available for zeta potential, size, molec-
ular weight, flow mode and microrheology measurements.
Folded capillary
cell (DTS1070)
Maintenance-free capillary cell primarily designed for zeta
potential measurements, but can also be used for size meas-
urements.

Introducing the Zetasizer Nano Chapter 2
Zetasizer Nano Page 2-5
Dip cell
(ZEN1002)
This is used to provide repeatable measurements of aque-
ous, and non-aqueous samples such as solvents. It is partic-
ularly suitable for measurements of valuable aqueous
samples where minimal sample quantity is important.
High
concentration
cell (ZEN1010)
This is used to perform zeta potential measurements on a
concentrated aqueous sample. The cell is suitable for a
broad range of conductivities.
Surface zeta
potential cell
(ZEN1020)
The surface zeta potential cell is intended for the measure-
ment of the zeta potential at the surface of a flat material in
an aqueous environment.
Flow-mode
option
Instruments fitted with the flow-mode option can be con-
nected to the output of, for example, a chromatography col-
umn, and be used as a light scattering detector.
The flow mode option is included as a standard fitment on
the Zetasizer Nano Plus.
ZEN1006 - Flow-mode option for Zetasizer Nano S and
Zetasizer Nano ZS.
Microrheology Microrheology is a new measurement type available to users
of the Zetasizer Nano ZS and ZSP. It allows the measure-
ment of the viscoelastic modulus of samples within the linear
viscoelastic region.
Microrheology measurements require a software key to
access the software features and functionality.
A microrheology starter kit containing a dip cell (ZEN1002)
and a selection of probe particles is available.
For use with the Zetasizer Nano ZS and ZSP.
Protein mobility Protein mobility is a measurement type available to users of
the Zetasizer ZSP.
The Zetasizer Nano ZSP has increased capability for the
measurement of small and weakly scattering molecules and
particles. This improved sensitivity improves the measure-
ment of zeta potential, with its primary purpose being the
ability to measure the zeta potential, or more appropriately,
the electrophoretic mobility of protein samples.
A dedicated measurement type is included for protein mobil-
ity and also a suite of new calculators for proteins.
For use with the Zetasizer Nano ZS and ZSP.

Chapter 2 Introducing the Zetasizer Nano
Page 2-6 MAN 0317
Primary measurement techniques
This section describes the basics of particle size, zeta potential and molecular
weight and why they are important. Greater detail on the measurement techniques
is given in Part 2 - Measurement theory.
Particle Size?
The particle size measured in a Dynamic Light Scattering (DLS) instrument is the
diameter of the sphere that diffuses at the same speed as the particle being meas-
ured.
The Zetasizer system determines the size by first measuring the Brownian
motion of the particles in a sample using DLS and then interpreting a size from
this using established theories.
Brownian motion is defined as:
“The random movement of particles in a liquid due to the bombardment
by the molecules that surround them”.
The particles in a liquid move about randomly and their speed of movement is
used to determine the size of the particle.
ill 6722
It is known that small particles move or diffuse more quickly in a liquid than larger
particles. This movement is carrying on all the time. So if we take two 'pictures' of
the sample separated by a short interval of time we can see how much the particles
have moved and therefore calculate their size.
If there has been a minimal particle movement between the two 'pictures' the parti-
cles in the sample can be inferred as large; similarly if there has been a large amount
of movement between the two 'pictures', then the particles in the sample can be
inferred as small.
1
Size nm
10 100
1000
Time
Small particles moving quickly
1
Size nm
10 100
1000
Time
Large particles moving slowly