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• Chapter 2 Software overview The Malvern Zetasizer software controls the system during a measurement and then processes the measurement data to produce either a size, zeta potential or molecular weight result
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• N o t e The Zetasizer Nano model, serial number, software and firmware version can be found by left-clicking the Nano icon in the right corner of the status bar
• Page 41: Navigating The Software C H A P T E R 3 Navigating the Software The Malvern Zetasizer software controls the system during a measurement and then processes the measurement data

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• As with any software, running other applications at the same time may affect the speed of the Zetasizer software
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Summary of Contents for Malvern Zetasizer Nano series

• Page 1 Zetasizer nano series user manual Malvern...
• Page 3 Zetasizer Nano User Manual MAN0485 Issue 1.1 April 2013 English...
• Page 4 United Kingdom. Tel + [44] (0)1684-892456 Fax + [44] (0)1684-892789 Zetasizer, Malvern and the 'hills' logo are registered trademarks in the UK and/or other countries, and are owned by Malvern Instruments Ltd. NIBS and M3-PALS are trademarks of Malvern Instruments.

• Page 5: Table Of Contents

  T able of Contents Introduction to this manual ......... 1-1 Introduction .
• Page 6 Table of Contents Zetasizer Nano Sample Preparation ......... 5-1 Introduction .
• Page 7 Zetasizer Nano Table of Contents Advanced features ......... 9-1 Introduction Solvent builder .
• Page 8 Table of Contents Zetasizer Nano Zeta potential theory ........13-1 Introduction .
• Page 9 Part 1 - Instrument guide...

• Page 11: Introduction To This Manual Introduction

  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.

• Page 12: 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.

• Page 13: Assumed Information

  Introduction to this manual Chapter 1 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”...
• Page 14 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.

• Page 15: Introducing The Zetasizer Nano

  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 ...
• Page 16 Chapter 2 Introducing the Zetasizer Nano 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 ...
• Page 17 Chapter 2 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.
• Page 18 Chapter 2 Introducing the Zetasizer Nano 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.
• Page 19 Introducing the Zetasizer Nano Chapter 2 Dip cell This is used to provide repeatable measurements of aque- (ZEN1002) 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 This is used to perform zeta potential measurements on a concentration...

• Page 20: Primary Measurement Techniques

  Chapter 2 Introducing the Zetasizer Nano 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-...
• Page 21 Introducing the Zetasizer Nano Chapter 2 Using this knowledge and the relationship between diffusion speed and size, the size can be determined. The above is a very simplistic explanation. A more detailed explanation can be found in zeta potential theory chapter. Why do we use it? A wide range of materials exist as molecules or particles that can be characterised by dynamic light scattering.
• Page 22 Chapter 2 Introducing the Zetasizer Nano A potential exists between the particle surface and the dispersing liquid which var- ies according to the distance from the particle surface – this potential at the slipping plane is called the zeta potential. Zeta potential is measured using a combination of the measurement techniques: Electrophoresis and Laser Doppler Velocimetry, sometimes called Laser Doppler Electrophoresis.
• Page 23 This is a calculation using a known molecular formula and applying the values from the periodic table. With the Zetasizer Nano series of instruments the molecular weight can now be determined by use of Static Light Scattering (SLS) measurement techniques.
• Page 24 Chapter 2 Introducing the Zetasizer Nano Note Malvern uses Daltons as the unit of molecular weight. Results are dis- played in kiloDaltons (kDa). Why do we want to know it? We need to know the molecular weight, so we can determine how many grams there are in 1 mole of a substance.

• Page 25: Hardware Components

  Introducing the Zetasizer Nano Chapter 2 Hardware components The following illustration shows a typical installation: ill 8803  Zetasizer Nano optical unit (as described in the next section) – measures the sample using red and green light wavelengths.  Cells – sample is presented in a selection of measurement cells so that it passes through the instrument’s laser path.
• Page 26 Chapter 2 Introducing the Zetasizer Nano Optical unit components This diagram shows the optical unit, the main component of the Zetasizer Nano system. ill 8804 The optical unit comprises the following:   Rear Panel Cell area   Status indicator Cuvette holder ...
• Page 27 Accessory connections – Two types of connection are available:  CAN Ports    Use these ports to connect any Malvern supplied accessory that requires a CAN connection (Controller Area Network). Consult the respective accessory manual for details. RS232 (I0I0I) Ports  ...
• Page 28 Malvern service personnel. Accessory output – A 12v output supply is provided on the rear panel to  connect to any Malvern supplied accessory that requires an external voltage source. Consult the respective accessory manual for details. Warning! Only connect Malvern approved accessories.
• Page 29 Introducing the Zetasizer Nano Chapter 2 The flow rate will be preset at 1.5ltr/min at 1 bar.  The instrument should be operated at a maximum of 30°C & 70% relative  humidity when the requested temperature is below the dew point. When operating at a humidity higher than 50%, the measurement should ...
• Page 30 Chapter 2 Introducing the Zetasizer Nano Cell access button Positioned in the middle of the Status indicator , pressing the button will open  the cell area lid. Cell area Warning! The system is capable of heating the cell to high temperatures. Care should be taken when removing the cells if a measurement has been performed at high temperatures.
• Page 31 Introducing the Zetasizer Nano Chapter 2 ill 8502 Cell area lid – Press the cell access button to open the lid, the lid will slowly  raise allowing access to the cell holder. On opening, two safety interlocks are activated. Laser safety interlock.
• Page 32 Chapter 2 Introducing the Zetasizer Nano The channel includes a pinch valve to hold the tubes in place and stop sample movement during the measurement. See the Inserting the cell section in Zetasizer Nano accessories guide for precise details on fitting the tubing. Cell basin –...
• Page 33 Introducing the Zetasizer Nano Chapter 2 Note When using glass or quartz cuvettes, first insert a disposable polystyrene cuvette into the cell holder. This ensures the cell clamp moves freely, and that no spilt sample has caused the mechanism to stick. Cuvette holder The cuvette holder is for storing the cells before and after use.
• Page 34 The cuvette holder includes a serial number, model number and option labels. These identify the instrument and should be quoted in any correspondence with Malvern Instruments. Refer to the identification table in the Introducing the Zetasizer Nano chapter. Cells and Cuvettes A range of cells and cuvettes are available to use with the Zetasizer instrument.

• Page 35: Software Overview

  Chapter 2 Software overview The Malvern Zetasizer software controls the system during a measurement and then processes the measurement data to produce either a size, zeta potential or molecular weight result. It displays the results and allows reports to be printed.
• Page 36 Chapter 2 Introducing the Zetasizer Nano Menu bar  The menu bar contains the main menu headings for all software functions. Items which end with a row of dots (...) will display additional windows. Similarly any items which end with an arrow ( ) will display a second menu.
• Page 37 Introducing the Zetasizer Nano Chapter 2 As a shortcut, a list of the most recently used measurement files are shown at the bottom of the menu for immediate opening. Exit will close down the software. Edit Menu This allows movement and manipulation of records in the Measurement file window(s).
• Page 38 Chapter 2 Introducing the Zetasizer Nano View Menu The View menu selects which reports will be shown in the measurement file window and which Toolbars are to be displayed. To see the Standard deviation, the Relative standard deviation, the minimum, the maximum or the mean of a selected number of records, a Statistics bar can be displayed.
• Page 39 Introducing the Zetasizer Nano Chapter 2 Refer to the Expert Advice section in Advanced features chapter for a description. Plate Navigator provides a graphical view of measurements made using a Zetasizer APS Instrument. Note A combination of the Edit and View menus are displayed when the right mouse button is clicked anywhere in the measurement file window.
• Page 40 The Macros submenu will show additional available programs. These are programs written by Malvern to enable the system to operate in a mode required for a specific application. These can be installed and selected using the Options feature.
• Page 41 Security Menu To prevent unauthorised changes, the Malvern software can be configured to limit each user’s access to various functions - e.g. modifying an SOP. Users are assigned operating permissions that allow, or restrict, access; refer to Security chapter for details.
• Page 42 Tutorials... provides a list of easy to follow tutorials describing various aspects of the system. Links to the Malvern website... including downloads and consumables. Tip of the day... will give hints on how to use the Zetasizer software; a different tip will be displayed each time this is selected.
• Page 43 Introducing the Zetasizer Nano Chapter 2 Toolbars  The toolbars contain a selection of tools that can be used to perform the most popular operations. Each tool will have its equivalent commands within the menu bar. For example, using the Open tool is equivalent to using the File-Open menu item.
• Page 44 Workspace windows. Selecting a report tab will display a predefined report as selected by the Report pages tab in the workspace windows. Malvern supplies several reports that give different views of the measurement settings and results, whilst custom reports can be generated using the Report Designer.
• Page 45 Introducing the Zetasizer Nano Chapter 2 Measurement display  When a measurement is being performed the measurement display is shown. The Measurement display shows the progress of the measurement. The screen display shown changes depending on the type of measurement being performed and the view tab selected.
• Page 46 Malvern logo ( ) or an (M) alongside. This identifies it as Malvern defined and cannot be overwritten. However, the Malvern defined parameters can be used as a template that can be altered and saved under a different name. Page 2-32...
• Page 47 Introducing the Zetasizer Nano Chapter 2 Units Conversion A number of parameters within the software have the option to change the units used to define the parameter, i.e. Temperature has the option to use either ºC (Celsius) or K (Kelvin). For all parameters where this option applies a scroll down list will be available alongside the parameter.
• Page 48 Chapter 2 Introducing the Zetasizer Nano Page 2-34 MAN 0317...

• Page 49: Making Measurements

  Making measurements Introduction After reading this chapter a user should be able to make simple measurements. The chapter goes through the basics from turning on the system to displaying the results of a measurement. The first section, “Quick guide to making a measurement” will run through the basic steps, giving an overview of the measurement process.

• Page 50: Quick Guide To Making A Measurement

  Chapter 3 Making measurements The sections that follow, Quick guide to making a measurement, etc, will focus on SOP measurements. Refer to the Using SOPs chapter for details on creating and managing a user’s SOPs. Quick guide to making a measurement This section will give a brief overview of the measurement process using an SOP.

• Page 51: Sample Preparation

  Selecting and preparing the measurement cell Malvern offers a range of cells and cuvettes for performing measurements with the Zetasizer system. The choice of cell or cuvette is dependent upon the type of measurement being performed and the sample that will be measured.

• Page 52: Making An Sop Measurement

  Chapter 3 Making measurements Filling the cell  Inserting the cell  Configuring the SOP parameters for the cell chosen.  Making an SOP measurement If a measurement is being made using an SOP, then all the hard work has already been done.
• Page 53 Making measurements Chapter 3 To create a new measurement file: Select File-New-Measurement File or A window is displayed allowing the new measurement file to be named and specify where it will be saved. Select Save. Note All measurement files have the extension .dts. This is added automatically to all new files.

• Page 54: Making A Manual Measurement

  Chapter 3 Making measurements Making a manual measurement Making a manual measurement is essentially the same as making an SOP measurement, except that, where as in an SOP measurement all the measurement options are pre-specified, here it will be necessary to set them immediately. Instead of starting an SOP, select Measure-Manual.
• Page 55 Making measurements Chapter 3 The measurement display for all measurement types is generally the same and shows a number of tabs representing the progress of the measurement sequence. The tabs displayed depend upon the measurement type selected. The diagram below shows the display for a size measurement. The features of the measurement display are: Button bar ...
• Page 56 Chapter 3 Making measurements Progress meter  The progress meter shows how far the measurement has progressed plus the number of measurements performed and the measurement runs completed. Also shown are the temperature, measurement position and attenuator settings. Tab views ...
• Page 57 Making measurements Chapter 3 Normal count rate Normal count rate display. ill 8451 If dust is present Dust present then sharp spikes will be observed. Measurement runs with dust present will be removed from the final measurement calculation by a dust filtration algorithm.
• Page 58 Chapter 3 Making measurements Contaminated or Noisy data bimodal sample = variable results Time Time ill 8495 Intensity PSD  Displays the result as an Intensity based particle size distribution. A summary table will displayed alongside the graph. Volume PSD ...
• Page 59 Making measurements Chapter 3 Intensity PSD  The result view will be updated after each of the individual concentration measurements. The result shown will be an evolving value from the data col- lected so far. Please refer to the description in the previous section. Count rate ...
• Page 60 Chapter 3 Making measurements Count rate  This displays the number of photons detected in kilo-counts per second. Phase Plot  This displays the phase shift that occurs during the zeta potential measure- ment. This is difference between the phase of the light scattered by the parti- cles and the phase of the reference beam.
• Page 61 Making measurements Chapter 3 Monitor enables the count rate signals to viewed before a measurement is run.  Multi-view Tab As the description in the previous section, except for Flow-mode specific view choices. Log sheet Tab Please refer to the description in the previous section. Expert Advice Tab Please refer to the description in the previous section.
• Page 62 Chapter 3 Making measurements Once these stages have been completed, the measurement proper will start; again the actual measurement sequence will depend upon the measurement being performed. The paragraphs that follow describe the three main measurement types the Zetasizer Nano performs: size, molecular weight and zeta potential. Size measurements The cell is inserted, Start is pressed and data collection begins.
• Page 63 Making measurements Chapter 3 Continue as above until all sample concentrations have been measured.  On completion of the last concentration the final result will be calculated.  The progress meter indicates the measurement progress during each stage. Zeta potential measurements The cell is inserted and Start is pressed.
• Page 64 Chapter 3 Making measurements Page 3-16 MAN 0317...

• Page 65: Records And Reports - Viewing The Results

  Records and Reports - Viewing the Results Introduction Once a measurement has finished the results will need to be reviewed. This chapter describes how to display the final measurement result. The final result is displayed in the measurement file window as either a measurement record or report.
• Page 66 Chapter 4 Records and Reports - Viewing the Results Aggregation and Titration records To view a sequence of Aggregation or Titration records, select the first record in the sequence; all subsequent records in that sequence will then automatically be selected. Outliners For both Aggregation and titration records, outliners are employed.
• Page 67 Records and Reports - Viewing the Results Chapter 4 The Outlined record view can be turned off and on by selecting View-View type- Sorted or Outlined or by right-clicking the mouse on the table, as shown above. Statistics bar Use this to display the Standard deviation, the Relative Standard deviation, the minimum, the maximum or the mean of a selected number of records.
• Page 68 Zetasizer, the filename and record number of the measurement and the date printed, plus the Malvern contact phone number. Each measurement type has a ‘standard’ report associated to it. The same result information is present in both the computer and printed versions, except for the diagnostic reports which contain extended information on the printed version.
• Page 69 Records and Reports - Viewing the Results Chapter 4 Units conversion As in the Records view, within the results section of each report, it is possible to change the units used to define some of the parameters displayed. If the option is available, right-clicking on a parameter will display a parameter icon enabling the units to be changed.
• Page 70 Chapter 4 Records and Reports - Viewing the Results Size measurements - standard report The standard report for size measurements is Intensity PSD (M). (PSD stands for Particle Size Distribution). ill 8444 The report is divided into four areas; these are described below. ...
• Page 71 Records and Reports - Viewing the Results Chapter 4 Duration used(s)  Duration of measurement, used in the analysis of the result. Cell type  This displays the cell type selected. Attenuator index  The laser power is automatically attenuated so that the count rate from the sample, especially high scattering samples, is within acceptable limits.
• Page 72 Chapter 4 Records and Reports - Viewing the Results  Results This section gives the results of the measurement. The values given here will be based either on intensity, volume or number, depending on which view tab is selected. The results section gives three pieces of information: Z-average size (also known as the “cumulants mean”) ...
• Page 73 Records and Reports - Viewing the Results Chapter 4 Lower values will be obtained for measurements on weaker scattering sample, or in turbid sample where background scattering is significant. Values >1 will only occur for samples with dust that makes the sample count rate fluctuate wildly.
• Page 74 Size quality report (M) This is a summary report detailing whether the result meets the Malvern Instruments quality criteria for a size measurement. The report indicates Result meets quality criteria, or Result does not meet quality criteria in which case...
• Page 75 Records and Reports - Viewing the Results Chapter 4 a list of parameters that do not meet the criteria, and probable reasons will be displayed. Some of these parameters will only be displayed if the research software has been used for the measurement. The parameters, followed by probable reasons displayed are: z Average is smaller than lower size display limit ...
• Page 76 Chapter 4 Records and Reports - Viewing the Results Multimodal fit error high  Data quality too poor for distribution analysis Sample too polydisperse for distribution analysis Inappropriate distribution analysis settings in Research Software Check cumulants and distribution errors  Sample too polydisperse for cumulant analysis - suggest rely on distribution analysis.
• Page 77 Records and Reports - Viewing the Results Chapter 4 Molecular weight measurements - standard report The standard report for molecular weight measurements is Molecular weight report (M). ill 7795 The report is divided into five areas; these are described below. ...
• Page 78 Chapter 4 Records and Reports - Viewing the Results  System details The System section gives details on settings configured during the measurement process. Please see the size description for details about Temperature, and Cell type.  Results The results section gives four pieces of information: Molecular weight ...
• Page 79 Records and Reports - Viewing the Results Chapter 4 Zeta potential measurements - standard report The standard report for zeta potential measurements is Zeta potential (M). The report is divided into four areas; these are described below.  Sample The Sample section gives details of the sample parameters. This includes the sample name, record number, measurement date and time, dispersant name, the SOP used and the measurement file name.
• Page 80 Please see the size description above for details on altering the graph. Other Zeta potential reports Other Malvern views available for zeta potential measurements include: Electrophoretic mobility (M) This report is effectively the same as the zeta potential report described, except the zeta potential result and graph are replaced by the electrophoretic mobility.
• Page 81 Sample concentration too high (sample looks turbid) Bubbles present in the path of the laser beam Cell not inserted correctly into the instrument A Technical note is available from Malvern that gives further details about this report. Zetasizer Nano Page 4-17...
• Page 82 Chapter 4 Records and Reports - Viewing the Results Flow-mode measurements - Standard report The Standard Flow-mode report, Chromatogram Summary (M), gives the same information as seen in a standard size report, plus additional information relating to the flow duration and rate used. The three main peaks in the measurement will also be shown, displaying the sample intensity, width, molecular weight and start and end flow volumes.
• Page 83 Records and Reports - Viewing the Results Chapter 4 Trend measurements (including Aggregation) Trend measurements can be viewed using the reports: Trend, Trend v Custom 1 and Trend v Custom 2 reports. Aggregationt measurements can be viewed using the Aggregation (M) report. A trend report gives the same information as seen in a standard size or zeta potential report, plus a graph showing the trend progression.
• Page 84 Chapter 4 Records and Reports - Viewing the Results Page 4-20 MAN 0317...

• Page 85: Sample Preparation

  Sample preparation Introduction Before filling the cell or cuvette that will be used in the measurement, the sample will need to be prepared. To ensure reliable and accurate measurements proper sample preparation is extremely important. Preparation of the sample for the different measurement types will involve specific preparation techniques.
• Page 86 Chapter 5 Sample Preparation An important factor in determining the maximum concentration the sample can be measured at, is the size of the particles. The table below can be used as an approximate guide to determine the maximum and minimum concentrations for different sizes of particles. The figures given are approximate values for samples with a density near to 1g/cm , and where the particles have a reasonable difference in refractive index to that of the dispersant,...
• Page 87 Sample Preparation Chapter 5 Maximum concentration For samples with small particle sizes, a maximum concentration does not really exist (in terms of performing Dynamic Light Scattering (DLS) measurements). However, in practice, the properties of the sample itself will set the maximum value.
• Page 88 Chapter 5 Sample Preparation Such fluctuations must be avoided and this determines the lower limit for the required concentration and for a lower limit in the number of particles. At least 500 particles should be present, however, a minimum of 1000 particles is recommended.

• Page 89: Preparing The Sample - Molecular Weight

  Sample Preparation Chapter 5 length of time ultrasonication is applied. Some materials can even be forced to aggregate using ultrasound. Emulsions and liposomes should not be ultrasonicated. Preparing the sample - Molecular weight Preparation of a molecular weight sample is similar to that followed for a size sample, though there are other aspects to consider.

• Page 90: Preparing The Sample - Zeta Potential

  Chapter 5 Sample Preparation Minimum concentration The minimum concentration of sample that should be used is defined by the excess scattering over the solvent which should be a minimum of 30%, e.g. If the solvent is toluene with a count of 150 kcps then the lowest sample concentration should be greater than 150 x 1.3 kcps (195kcps).
• Page 91 Sample Preparation Chapter 5 and the medium) and the particle size. The larger the particle size, the more scattered light it produces and hence the lower the concentration that can be measured. For example, consider a dispersion of a ceramic powder such as titania which has a particle refractive index of around 2.5.
• Page 92 Chapter 5 Sample Preparation Dilution Medium The continuous phase of most samples can arbitrarily be put into one of two categories: Polar dispersants are defined as those with a dielectric constant greater than 20  e.g. ethanol and water. Non-polar or low polarity dispersants are defined as those with a dielectric ...

• Page 93: Count Rate Meter

  Sample Preparation Chapter 5 Count rate meter After preparing the sample, the count rate can be checked by using the count rate meter. Selecting Tools-Count rate meter will display the following window. The Count rate is displayed in kcps (kilo counts per second), and enables the sample quality and concentration to be checked without having to start a measurement.
• Page 94 Chapter 5 Sample Preparation Drifting count rate  This may indicate that thermal gradients are present in the sample, and further time is required for temperature equilibration. Increasing/Decreasing count rate  A steadily increasing count rate will indicate an aggregating sample, while a decreasing count rate will indicate a sedimenting sample.
• Page 95 Sample Preparation Chapter 5 Zetasizer Nano Page 5-11...
• Page 96 Chapter 5 Sample Preparation Page 5-12 MAN 0317...

• Page 97: Introduction

  SOPs, deleting records and editing results can be reserved for specific users only. Within the Malvern Security system, one or more persons are set up as the admin- istrator. The administrator then controls access to the instrument by defining “User groups”...

• Page 98: Initial Start-up - Set Up The Administrator

  Chapter 6 Security Initial start-up - set up the administrator Note It is vital that at least one user has access to the configuration of the security system. Select Security-Configure Security. The first time the system is run, there will be just one user (Administrator) and one group (Administrators).

• Page 99: Enabling Security

  Security Chapter 6 Remember that after any changes to the security system - adding users, changing of permissions, etc. The new changes must be saved (User-Save). A window is dis- played on exiting to give a reminder on saving changes. Enabling security Within the Security configuration window select Options-Security settings and select the Enable security check box.
• Page 100 Chapter 6 Security Enter a Group name and a Description of the group’s purpose. Example names might be: Operators - General users of the system.  Supervisors - Skilled operators responsible for configuration and SOP crea-  tion. Administrators - Users authorised to configure the security system. ...

• Page 101: Users

  Security Chapter 6 Users Note Only users assigned to the administrator group can add or edit user prop- erties. Adding/editing a user Select Security-Configure security to enter the Security configuration win- dow. Select User-New User... to display the User properties window, alterna- tively double-click on an empty row.
• Page 102 Chapter 6 Security Password options User must change password at next logon  The change password window will be shown when the user next logs in, allow- ing the user to change their password. This allows the security administrator and a user to set up a user account without the administrator knowing the users final password.
• Page 103 Security Chapter 6 For a secure system, it is advisable for users to regularly change their passwords to prevent unauthorised access. The only possible exception is the security administrator’s account where forgetting the password to this account could prevent any further configuration of the security system. Zetasizer Nano Page 6-7...
• Page 104 Chapter 6 Security Page 6-8 MAN 0317...

• Page 105: Measurement File Window - Workspaces

  Measurement file window - Workspaces Introduction The Measurement File window is where all results (or records), and reports for each measurement file can be viewed, moved and analysed. Depending upon the Window options set, multiple measurement files can be viewed at the same time. Once a result has been calculated, the data can be displayed as either a record or report - by selecting the appropriate tab.

• Page 106: Measurement File Window

  Chapter 7 Measurement file window - Workspaces Malvern provides several default reports (identified with (M) in the name) that will be sufficient for most users. The user also has the ability to create their own custom reports to satisfy their individual needs. Details on creating and using custom reports are given in the Report Designer chapter.
• Page 107 Measurement file window - Workspaces Chapter 7 group of measurements, in the records view tab and then select the appropriate report tab. The report tabs will be greyed out if no measurement is selected. What measurement information is presented in the Report and Record tabs is controlled by the Workspace selection windows (below).
• Page 108 Use the Record view parameters to specify what parameters will be viewed as the column headings. Report pages allows selection of both Malvern created and customised reports. When selected, click OK. Once completed the new workspace can be selected using the View menu or Workspace toolbar.
• Page 109 Measurement file window - Workspaces Chapter 7 To Modify a Workspace: Select the workspace name from the toolbar list and the workspace configure icon , or from the Tools-Settings-Configure Workspace menu. This will display the Workspace Settings window for the selected workspace. Alter the settings as required and press OK;...
• Page 110 Chapter 7 Measurement file window - Workspaces Parameters can be added or removed from this list by using the Add and Remove keys Change the order in which the parameters are displayed by selecting a parameter, and pressing the Move up or Move Down buttons.
• Page 111 Measurement file window - Workspaces Chapter 7 Report tabs The report tabs show every report that has been selected for a workspace. With the Summary workspace selected the default reports are Intensity PSD (M), Zeta potential (M) and Molecular weight (M). These and other reports can be added or removed using the workspace windows.
• Page 112 Chapter 7 Measurement file window - Workspaces Moving, Copying and deleting records The are several ways to handle the information. One or more records can be copied from a Records view tab and “dragged and  dropped” directly into another open measurement file. Select a record or records, then move the cursor to within the selected records.

• Page 113: Using Sops

  Using SOPs Introduction An SOP measurement uses pre-set parameters, to ensure that measurements made on the same type of sample are made in a consistent way. SOPs are ideal if repeatedly measuring the same type of sample. Having to set the same parameters each time a measurement is made is tedious and also adds the risk of introducing errors into the settings - using SOPs avoids this.
• Page 114 Chapter 8 Using SOPs A further measurement type is Autotitration. Details on Titration SOPs are described in the Titrator accessory manual. Once a measurement has been completed it will be added to the list in the Records view tab of the Measurement file window. The SOP used can be viewed by right-clicking over the measurement record and selecting Extract SOP.

• Page 115: The Sop Editor

  Using SOPs Chapter 8 ing the icon. The measurement will now start without first showing the SOP editor. Alternatively if Browse for SOP... is selected and the icon is pressed. The SOP directory will be displayed where an SOP can be manually selected. The title of the SOP shown is that typed into the Sample name entry box in the Sample SOP window.
• Page 116 Chapter 8 Using SOPs Tree menu view On opening the SOP editor the window below will be displayed; the exact list displayed will depend upon the Measurement type selected. The SOP windows available for each measurement type will be displayed down the left hand side of the window, with the respective highlighted SOP window shown alongside.
• Page 117 Using SOPs Chapter 8 option applies a scroll down list will be available alongside the parameter. The Parameter unit chosen will be displayed in the final results and reports. The SOP windows available are: Measurement type Allows selection of the type of measurement that the SOP will perform, i.e. Size, Aggregation, Zeta potential, Molecular weight, Flow, Trend or Autotitration.
• Page 118 Chapter 8 Using SOPs Sample - Dispersant / Solvent This window allows the physical properties of the sample dispersant (or solvent) to be set. The refractive index and viscosity can be entered for the dispersant used. This is a common window for all SOPs with unique features dependent upon the measurement type.
• Page 119 Using SOPs Chapter 8 Measurement - Advanced Use the Advanced window to alter the cell and attenuator position settings. Measurement - Flow settings Enables the flow rate of the sample through the instrument and connecting tubing to be entered. This is a unique window for Flow SOPs. SZP measurement This option enables measurements specific to surface zeta potential measurements to be input.
• Page 120 Chapter 8 Using SOPs Trend - Sequence Enables the selection of the temperature trend parameters, i.e. start and end temperatures, and intervals between measurement steps. This is a common window for Size and Zeta trend measurements. Titration - Titrants The Titrants window enables the titrants used to be specified, and the pump and stirrer speed to be set.

• Page 121: Creating An Sop

  Using SOPs Chapter 8 SOP Editor Menu and Toolbar SOP file management, saving and associated software functions are handled using either the Menu bar or the Toolbar. The functions are described in the following table. Toolbar Menu item icon Description of function File-New Opens a new SOP File-Open...

• Page 122: Measurement Type Selection

  Chapter 8 Using SOPs Measurement type selection The measurement type must first be selected before entering the settings on the subsequent SOP windows. To Select the Measurement type Left click on  Measurement type and select the measurement type from the ...

• Page 123: Size Sops

  Using SOPs Chapter 8 Note The following Size SOP section describes the SOP windows shown when a Size Measurement type is selected. This includes all sizing SOP windows and all windows that are common to all SOPs. When requested in the Molecular Weight, Zeta potential and other SOP sections refer back to the Size SOP section for a description of the window.
• Page 124 Chapter 8 Using SOPs Note The Instrument configuration window is only displayed when the soft- ware is not connected to an instrument. When the instrument is con- nected, the software automatically detects the instrument type and sets the parameters accordingly. When creating an SOP, with no instrument connected, it is necessary to specify the Instrument configuration and indicate the options fitted to the instrument.
• Page 125 Using SOPs Chapter 8 Sample The components of the window are: Sample name: The name entered here should be a description of the sample being measured. If no name is entered the sample name entry in the Records view will be left blank. The name typed into the Sample name entry box will be displayed in the SOP toolbar browser.
• Page 126 Chapter 8 Using SOPs the concentration of an additive which is varied in subsequent measurements, then plotted as a function of the size of the material, for example to study its effect. Once in the record, the parameter will be available for selection for both the workspace and Records view, and for data export.
• Page 127 Using SOPs Chapter 8 Materials Manager From the list displayed, an available material can be selected from the list for inclusion in the SOP; alternatively a material can be added, modified or deleted from the list. To define a new material, press the Add... button to display the Material Properties window.
• Page 128 Chapter 8 Using SOPs Sample - Dispersant The Zetasizer software requires certain information about the physical properties of the dispersant that is used in the sample that is to be measured. Selecting the Sample - Dispersant window and pressing the button, displays the Dispersants properties manager where these properties can be defined.
• Page 129 Propanol 25C. Note Viscosity is temperature dependent. The Malvern defined dispersants include a built in viscosity calculation that determines the correct viscosity of the dispersant at any sample temperature.
• Page 130 Chapter 8 Using SOPs Sample - General Options Mark-Houwink parameters. Selecting the Define button displays the Mark-Houwink parameters window. This allows the Mark-Houwink parameters to be entered to enable a molecular weight to be calculated based on the dynamic light scattering data. a –...
• Page 131 Using SOPs Chapter 8 viscosity is the same as the dispersant viscosity is usually valid. If this is not the case, or at higher sample concentrations (over 0.1%) then the sample viscosity should be measured and this value used. If the viscosity of the sample is known only at certain temperatures select the Prompt operator to enter viscosity button.
• Page 132 Cell type Select the appropriate cell from the list. The choices match the range of cuvettes and cells available from Malvern Instruments. Once selected a description of the cells is given. To choose a cell, select the Cell type from the drop down list, then from the ...
• Page 133 Using SOPs Chapter 8 The software will automatically determine the measurement conditions for the cell type selected, i.e. cell position for size cells and applied voltage for the zeta potential cells. The Default selection shown will be appropriate to the measurement type selected - for a size measurement, the cell shown is the Disposable sizing cuvette.
• Page 134 Chapter 8 Using SOPs Choices are as indicated below: 175° Backscatter performs a “standard” measurement at an angle of 175°  12.8° Forward-scatter performs a measurement at an angle of 12.8°  Dual-angle performs a measurement at both angles - this measurement is ...
• Page 135 Using SOPs Chapter 8 Partial Results If it is likely that a measurement will not produce a correlation function that can be analysed - for example, measurement of a surfactant solution at lower than the critical micelle concentration - then the data collected can still be saved by selecting the Allow results to be saved containing correlation data only check box within the Partial Results area.
• Page 136 Chapter 8 Using SOPs Measurement - Advanced The Advanced window contains functions to alter the cell and attenuator position settings. For normal operations both will be at the default setting. Measurement position Positioning method Typically the measurement position that is automatically determined by the software will be appropriate for most sample measurements.
• Page 137 Using SOPs Chapter 8 Data Processing If characteristics are already known about the sample being measured, this window will allow an appropriate analysis model to be applied and so optimise the measurement calculation. Size ranges and measurement thresholds can be applied to the analysis to filter spurious peaks out prior to the analysis being performed.
• Page 138 Chapter 8 Using SOPs data processing. These are saved as .dass files. (The last s indicates the analysis setting has been created for a size or molecular weight measurement). Configure button The Configure button window enables various attributes of the analysis model to be altered.
• Page 139 Using SOPs Chapter 8 Note If no name has been given, the analysis model will be displayed as Cus- tomised analysis settings in the Analysis model entry box. Previously saved analysis models can be used, viewed and altered by pressing the Load button.
• Page 140 Chapter 8 Using SOPs and then the reports required. Any reports created using Report Designer will automatically be added to this list. If an Autotitration measurement is being performed, the print reports after each measurement option will be ‘greyed’ out and unavailable. This is to prevent a report being printed after each ‘individual’...

• Page 141: Molecular Weight Sops

  Using SOPs Chapter 8 Molecular weight SOPs Though the majority of the Sample and Measurement windows described below are unique to molecular weight measurements; when indicated please refer to the description in the Size SOP section. The Molecular weight SOP editor menu is: Sample Please refer to the Sample description in the Size SOP section.
• Page 142 Chapter 8 Using SOPs Materials Manager Selecting this window allows new material to be added, modified and deleted. To define a new material, press the Add... button to display the Material Properties window. Here, the material name, refractive index and absorption can all be specified.
• Page 143 Using SOPs Chapter 8 Solvents Manager Selecting this window enables solvents to be added, modified and deleted. To define a new Solvent, press the Add... button and choose from defining either a Simple dispersant or solvent or a Complex solvent. Please refer to the Sample - Dispersant description in the Size SOP section for details.
• Page 144 Chapter 8 Using SOPs Standards Manager Input and define the scattering “standard” properties of the liquid that will be used in the measurement: Temperature, Refractive index and Rayleigh ratio. This will nearly always be Toluene. The standard properties can be added, modified and deleted in the same way as detailed for the Solvent Manager above.
• Page 145 Using SOPs Chapter 8 many sample/solvent combinations this may be available in literature; while for novel combinations the dn/dc can be measured by use of a sensitive differential refractometer. Note This will require a refractometer capable of measuring to an accuracy of six decimal places.
• Page 146 Chapter 8 Using SOPs With Automatic selected, the software will automatically determine the measurement duration. With Manual selected the measurement will use the user defined settings Shape correction model This option allows selection of the shape correction model used for the measurement.
• Page 147 Using SOPs Chapter 8 These systematic variations can occur for a few reasons; possibly thermal drifting of the sample, poorly mixed sample, or instrument and laser stabilisation after initial switch on. (Wait 30 minutes after switch on for the laser to stabilise). If any variations are detected that are greater than the set values, then the measurement will be repeated until a result below the thresholds is achieved.
• Page 148 Chapter 8 Using SOPs Size Measurement With the Perform a size measurement check box selected a size measurement will be performed simultaneously with molecular weight. Measurement Duration The Measurement Duration setting can affect the accuracy and repeatability of the size results. Please refer to the Measurement description in the Size SOP sections, for details of the Measurement duration settings.
• Page 149 Using SOPs Chapter 8 Size Measurement - Advanced Measurement settings Please refer to the Measurements - Advanced description in the Size SOP section for a description of the Measurement Position and attenuator functions. Data Processing If the characteristics of the sample to be measured are known, this window will allow an appropriate analysis model to be applied and so optimise the measurement calculation.

• Page 150: Zeta Potential Sops

  Chapter 8 Using SOPs Zeta potential SOPs Though the majority of the Sample and Measurement windows described below are unique for zeta potential measurements, when indicated please refer to the description in the Size SOP section. The zeta potential SOP editor menu is: Sample Please refer to the Sample description in the Size SOP section.
• Page 151 Using SOPs Chapter 8 Materials Manager Selecting this window allows new material to be added, modified and deleted. To define a new material, press the Add... button to display the Material Properties window. Here, the material name, refractive index and absorption can all be specified.
• Page 152 Chapter 8 Using SOPs Dispersants Manager Selecting this window enables new dispersants to be added, modified and deleted. To define a new dispersant, press the Add... button and choose either a Simple dispersant or solvent or a Complex solvent. Please refer to the Sample - Dispersant description in the Size SOP section for details.
• Page 153 Using SOPs Chapter 8 For most particles in aqueous media select Smoluchowski: 1.5.  For non-aqueous measurements, or for very small particles in low conductiv-  ity media measurements, the Huckel approximation should be used: 1.0. Please refer to Zeta theory chapter for more information on the F(ka) model. Sample viscosity options This area is useful for customising the viscosity parameters.
• Page 154 Chapter 8 Using SOPs Measurement Duration The measurement duration setting may affect the accuracy and repeatability of the results. With Automatic selected, the software will automatically determine the measurement duration. This will be suitable for the majority of samples and can simply be left as the default.
• Page 155 Using SOPs Chapter 8 Measurement - Instructions Please refer to the Measurement - Instructions description in the Size SOP section. Measurement - Advanced In the Measurement - Advanced window there are two functions: Measurement settings and voltage. For normal operations both functions will be at the default settings. Measurement settings Fixing the attenuator setting can be used to enable the count rates of two samples to be compared, or to speed up the optimization stage prior to a measurement being...
• Page 156 Chapter 8 Using SOPs < 10mS/cm Aqueous Universal ‘Dip’ cell > 10mS/cm < 5mS/cm Aqueous High concentration cell > 5mS/cm > 30mS/cm 2.5V Non-aqueous Universal ‘Dip’ cell Negligible If the Automatic voltage selection is not appropriate for the sample being measured, double-click on the function (to change the settings from Yes to No).
• Page 157 Using SOPs Chapter 8 Analysis Models Auto mode This is the default setting. In this mode the software will determine the most suitable type of measurement scheme to perform after measuring the sample conductivity; this is useful if the sample to be measured is unknown. In Auto mode the result will reveal details about the number of modes and the shape of the zeta potential distribution.
• Page 158 Chapter 8 Using SOPs If it is known that all particles within the sample will fall within a certain zeta potential range, then the Zeta Display range can be set to improve the repeatability of the measurement result; similarly a lower threshold can be applied to eliminate undesired particles that are below a defined percentage of the particle population.

• Page 159: Flow-mode Sops

  Using SOPs Chapter 8 itself is not attached. This means that any subsequent changes to the Anal- ysis file will not change the SOP . The analysis settings file will have to be reloaded into the SOP for the changes to take place. Data Processing - Reports Please refer to the Data Processing - Reports description in the Size SOP section.

• Page 160: Trend Sops

  Chapter 8 Using SOPs Trend SOPs The Trend SOP Editor menu is: The Trend SOP allows size and zeta potential measurements to be performed and plotted as a function of temperature. Trend SOPs are similar to the standard size and zeta potential SOPs, the differences being the inclusion of a trend and changes in the measurement windows.
• Page 161 Using SOPs Chapter 8 Trend - Sequence The Trend - Sequence window enables the selection of the temperature trend parameters, i.e. start and end temperatures, and intervals between measurement steps. Start Temperature Input the temperature that the first measurement is to be made at. The instrument will initially cool, or heat, to this temperature before starting the measurement.

• Page 162: Titration Sops

  Chapter 8 Using SOPs Size Measurement The Trend and Aggregation measurement windows are generally the same as those used with either a size or zeta potential measurement SOP. The additional features are: At Each step gives the option to control the Equilibration time and Num- ...

• Page 163: Modifying An Sop

  Using SOPs Chapter 8 Modifying an SOP To modify an SOP, select File-Open-SOP. The SOP windows will be displayed, allowing any settings to be changed. Alter the SOP settings as required, and then save (select File-Save or File- Save as... as appropriate).
• Page 164 Chapter 8 Using SOPs Page 8-52 MAN 0317...

• Page 165: Advanced Features

  Advanced features Introduction This chapter contains the following advanced features and capabilities of the Zetasizer nano: Solvent builder  SOP player  Averaging results  Editing the result  Exporting the results  Options window  Expert advice  Zetasizer Nano Page 9-1...

• Page 166: Solvent Builder

  Chapter 9 Advanced features Solvent builder The Solvent builder is a database of components that can be added in different quantities for estimating the properties of solutions. The database is used for the calculation of Complex solvents; generally necessary when performing measurements involving proteins.
• Page 167 Advanced features Chapter 9 Note The Additive effect on both viscosity and refractive index is determined using 3rd order polynomial fits of data compiled in the “65th Edition of the CRC Handbook for Chemistry & Physics”. Building a Complex solvent To generate a Complex solvent, a few sample parameters have to be entered into the builder window.
• Page 168 A dispersant, or solvent, can be deleted by selecting the dispersant from the list and pressing the Delete... button. Note that it is not possible to modify or delete any dispersants or solvents that were specified by Malvern. Page 9-4 MAN 0317...

• Page 169: Sop Player

  Advanced features Chapter 9 SOP player The SOP player is a separate feature that enables a sequence of actions - SOPs, measurements, pauses or macros - to be constructed and then played. This sequence is called a playlist. This is useful for performing different measurement types consecutively without any user interaction;...
• Page 170 Chapter 9 Advanced features Note, that when first opened, the table on the left will normally be blank. The actions shown are only for reference. The SOP player controls are: Button Description Select this tab to add new items (or actions) to a playlist. Choices are SOP, Macro, Temperature, pH, Inject titrant and Pause.
• Page 171 Advanced features Chapter 9 Construct the playlist by selecting and configuring the Actions in the right pane; These are described in the following sections. The list can be arranged in order as required by dragging and dropping an action at the desired point. For each action, enter the action properties as required.
• Page 172 Chapter 9 Advanced features Selecting the SOP in the Playlist will display both the SOP properties and the sample details. Select each parameter to edit as required. The SOP used can be changed by selecting ; this will open the window above. To alter an SOP, use the SOP Editor.
• Page 173 The Macro used can be changed by selecting ; this will open the window above. Note The Macros available will be supplied by Malvern. Inject Titrant To add a titrant, select the Inject titrant icon. This will add a Titrant action into the playlist, with the properties displayed alongside.
• Page 174 Chapter 9 Advanced features A pause may be required to allow for thermal equilibration, or to allow a titrant to be delivered and dispersed into the sample before proceeding. Set temperature To add a temperature change or setting between each playlist action, select the Set temperature icon.
• Page 175 Advanced features Chapter 9 Go to pH To adjust the pH value between two playlist actions, select the Go to pH icon. This will add a Go to pH action into the playlist, with the properties displayed alongside. Alter the Properties as required. Enter the Titrant type used for each bottle - either acid, base or disabled.

• Page 176: Result Averaging

  Chapter 9 Advanced features Result averaging What is the average result of a selection of records? Highlight the required records and either select Edit-Create Average Result or right-click on the records and select Create Average Result from the displayed menu. An averaged result record will then be generated and added to the end of the measurement file.

• Page 177: Editing The Result

  Advanced features Chapter 9 Editing the result It is possible to re-analyse a measurement record using different measurement parameters. The re-analysed record will be added to end of the current file. Comments on the reasons for editing can be added and viewed in the report views. This option allows measurements to be reanalysed without the need for the instrument to be connected.
• Page 178 Chapter 9 Advanced features Note Each edit result window will be slightly different depending upon the measurement type originally performed. The picture above shows the size view. Multiple results can be edited by selecting the records required before selecting the edit result window.

• Page 179: Exporting Results

  Advanced features Chapter 9 Exporting results The Zetasizer software allows any of the parameters from one or more records, as well as graph and table data to be exported to another application such as Excel, Word or Wordpad. There are several ways to export the required information: One or more records can be selected in the Records view tab and moved via ...
• Page 180 Chapter 9 Advanced features Exporting using Menu controls To export a complete measurement file, or selection of records, first open the required file and select File-Export... Use the tabs - File, Parameter, Settings - and select the exporting options. File tab The data can be either Exported to file or Exported to Clipboard by ...
• Page 181 Advanced features Chapter 9 Parameters tab This enables exporting of selected parameters. Selecting Use records view parameters export parameters currently displayed in the Record view. User defined parameters can be exported by selecting Use export configuration parameters and choosing the required template. If necessary, a new template can be created or an existing one edited .
• Page 182 Chapter 9 Advanced features Settings tab The settings tab works in the same way as previously described. Parameters tab The list on the left displays all available parameters.  The list on the right displays all parameters that will be included in the export ...
• Page 183 Advanced features Chapter 9 A Dv(90) percentile can be viewed by adding Measurement-Size-D(v) and then typing 0.90 as the argument value. Column titles can be altered by double-clicking on them e.g. double-click on D(v) and alter to Dv(90) to reflect the particular percentile size viewed. Note The Export templates can also be selected and edited using the SOP crea- tion procedures - Export window.

• Page 184: Options Window

  Chapter 9 Advanced features Options window The Options window enables extra features and programs for the Zetasizer soft- ware to be installed, and to configure the default folder for saving of data. Page 9-20 MAN 0317...
• Page 185 Macros option. This window allows new macro programs (.zmac files) to be installed and selected. The Macro programs are written by Malvern to enable the system to operate in a mode required for a specific application. Highlighting the Macro in the left box will display the Macro details in the right box.
• Page 186 Chapter 9 Advanced features Press the Install new macro... button, select the macro from its location (CD,  USB memory stick, computer folder, etc.) and press Open. This will copy the macro to the Zetasizer/macros directory and add the macro to the list of available programs.
• Page 187 ER/ES Features (21 CFR Part 11) The Malvern security system can be upgraded to 21 CFR part 11 compliance by installing the Feature key. Once this is installed, 21 CFR part 11 security settings can be applied and audit trails can be viewed. If the feature key has been installed, the 21 CFR 11 icon on the status bar is displayed.
• Page 188 Chapter 9 Advanced features Measurement Window This option allows the appearance of the measurement display to be changed and selected. The appearance choices are shown below, for each one an anti-aliasing view is also available. Standard Style 1 Style 2 Style 3 Results This window contains a check box that will allow measurements results to be saved...
• Page 189 Advanced features Chapter 9 Instrument type This window provides the option to select which instrument type is currently in use. The software remembers the selection and uses the same mode the next time it is initialised. Theme Selection This option allows the appearance of the software to be changed. The appearance is based on various Windows colour schemes such as Windows XP, Office 2003, Office 2007, etc.

• Page 190: Expert Advice

  Chapter 9 Advanced features Expert advice Expert Advice enables a quality check to be performed on one or more completed measurement records. This will show how good the measurements are and whether they display any unwanted attributes, such as aggregation. To enable the Expert System, choose one or more records, right-click on the records and select Expert Advice, or select View-Expert Advice.
• Page 191 Note If the Microrheology feature key has been installed, a similar symbol will be displayed to indicate microrheology measurement advice. Note A Technical note is available from Malvern Instruments that further describes the Expert advice. Zetasizer Nano Page 9-27...
• Page 192 Chapter 9 Advanced features Page 9-28 MAN 0317...

• Page 193: Report Designer

  This chapter describes how to use the Report designer to design custom reports. These can display parameters which are not on the default reports. Note The Malvern Instruments default reports described will be sufficient for most users. These are identified with (M) appended to the name - e.g. Intensity PSD (M)

• Page 194: Overview

  Chapter 10 Report designer Overview This section introduces the Report Designer. Views - print and screen reports As the aspect ratio of a computer screen tends to be landscape, and a printed report tends to be portrait, each report has two versions - a print version and a screen version.
• Page 195 Report designer Chapter 10 Report contents The report can contain one or more of any of the following elements: Text – user-defined text.  Picture – graphics files can be added to customise the report.  Frame – used to separate parts of the report - inserts a border around an area. ...

• Page 196: Opening An Existing Report

  Chapter 10 Report designer These Edit menu commands are of general use: Edit, Copy and Paste – allow copying and pasting of text and graphics from  other programs. The toolbar buttons have the same function. Undo ( ) and Redo ( ) –...
• Page 197 Report designer Chapter 10 Button Use this to add: Button Use this to add: Pointer - select elements. Well-plate Text. Calculation. Graph – use the list button Picture. to select a Trend graph, or a Distribution graph. Table – use the list button Frame –...

• Page 198: Setting Up The Report Elements

  Chapter 10 Report designer Right-click on the element and use the menu this displays to configure its properties, as described below. Setting up the report elements Set the properties of any added element by selecting it and double-clicking to open the Properties window box.
• Page 199 Report designer Chapter 10 The parameter Properties window has the same tabs as the text window, plus a General tab to specify what the parameter is, as shown above: On the General tab click the Select button. In the Select a parameter window select the appropriate parameter (see below) and click OK.
• Page 200 Chapter 10 Report designer The Label selection area enables the parameter name to be added alongside the parameter value. Frame Use the Frame tool to split the report into relevant sections. With the element selected, use its Properties window to select the frame style – etched, raised, sunken, etc.
• Page 201 Report designer Chapter 10 Calculation Editor Use the Calculation Editor to load an existing calculation for editing or to create a new calculation. The language used, Sax Basic, is similar to Microsoft Visual Basic for Applications (VBA). Select the browse icon to choose functions from the library lists, which can be used to build up calculations.
• Page 202 Chapter 10 Report designer Setting Colours Choose the Color tab to set the high and low value colours - these are shown auto- matically on the plate as legend indicators. Using a report with a well-plate display Page 10-10 MAN 0317...
• Page 203 Report designer Chapter 10 When the report has been saved, it can be added to the workspace view. The report shows only those measurements that are currently selected - if no results are selected, the wells will be empty. An easy way to navigate through data that has been amassed through the measure- ment of a plate is to use the Plate Navigator, rather than the Records View.
• Page 204 Chapter 10 Report designer Size graphs The following size graphs can be inserted. Result This shows the size result of the measurement. It can be displayed in different ways using its Properties box. Correlogram Displays the correlation function, which is the output of the contents of each of the correlation channels.
• Page 205 Report designer Chapter 10 Zeta potential graphs The following zeta potential graphs can be inserted. Zeta Potential This shows the zeta potential distribution generated by the measurement. It can be displayed in different ways using its Properties box. Statistics The statistics graph will show the mean, standard deviation, maximum and mini- mum of a group of selected measurement records.
• Page 206 Chapter 10 Report designer The Trend plots selection area is used for selecting the y axes parameters. The y1 parameter column is used to select the axis on the left of the graph and y2 to select the axis on the right. To choose a parameter press the Add button, select the appropriate parameter and press OK.
• Page 207 Report designer Chapter 10 Titration tables This will insert a table of the titration parameter against the selected measurement type, i.e. zeta potential versus pH. The table, when used in the main Zetasizer application software, will automatically configure to the titration/measurement type result selected from the records list.

• Page 208: Selecting Elements

  Chapter 10 Report designer Any available parameter can be selected and inserted into the report. The parameter value displayed is taken from the selected measurement record. Selecting elements When selecting elements: To select one element, click on it.  To select multiple elements, hold down the Shift or Ctrl key and click the ...

• Page 209: Aligning And Sizing Elements

  Report designer Chapter 10 Aligning and sizing elements Select two or more elements and use the following options of the Layout toolbar or menu to align/size them: Button Function Button Function Layout-Align-Align left Layout-Make same width Layout-Align-Align right Layout-Make same height Layout-Align-Align top Layout-Make same size Layout-Align-Align bottom...

• Page 210: A Finished Report

  Remember if the custom report is to be printed then a Page version must also be created. Note New reports must be saved to the My DocumentsMalvern instru- mentsZetasizerReportsen-US (or ja-JP for Japanese) directory to be viewed in Records view.

• Page 211: Viewing The New Report

  The printed copy of a report shows the software version number and instrument serial number at the bottom of the page. This is important information if a call is made to the Malvern Instruments help desk. The name of the measurement file the report is based on is also shown.
• Page 212 Chapter 10 Report designer Page 10-20 MAN 0317...
• Page 213 Part 2 - Measurement theory...

• Page 215: Size Theory

  What is Dynamic light scattering? The Zetasizer Nano series performs size measurements using a process called Dynamic Light Scattering (DLS) Dynamic Light Scattering (also known as PCS - Photon Correlation Spectroscopy) measures Brownian motion and relates this to the size of the particles.
• Page 216 Chapter 11 Size theory The speckle pattern will consist of bright and dark areas. What causes these bright and dark areas? The diagram below shows the propagated waves from the light scattered by the particles. The bright areas of light are where the light scattered by the particles arrives at the screen with the same phase and interferes constructively to form a bright patch.
• Page 217 Size theory Chapter 11 Interpreting scattering intensity fluctuation data We know that the Zetasizer measures the fluctuation in scattering intensity and uses this to calculate the size of particles within the sample - but how does it do this? Within the instrument is a component called a digital correlator. A correlator basically measures the degree of similarity between two signals over a period of time.
• Page 218 Chapter 11 Size theory Using the correlation function How does the correlation function relate to the particle size? We mentioned earlier that the speed of particles that are being moved by Brownian motion is related to the size of the particles (Stokes-Einstein equation). Large particles move slowly, while smaller particles move quickly.
• Page 219 Size theory Chapter 11 The x-axis shows a distribution of size classes, while the y-axis shows the relative intensity of the scattered light. This is therefore known as an intensity distribution. Although the fundamental size distribution generated by DLS is an intensity distribution, this can be converted, using Mie theory, to a volume distribution.

• Page 220: Operation Of The Zetasizer Nano

  90° 175° ill 8683 With the Zetasizer Nano series, depending upon the particular model, the detector position will be at either 175° or 90°. Detection path Zetasizer...
• Page 221 Size theory Chapter 11 For samples that do not scatter much light, such as very small particles or sam-  ples of low concentration, the amount of scattered light must be increased. In this situation, the attenuator will allow more laser light through to the sample. For samples that scatter more light, such as large particles or samples of higher ...
• Page 222 Chapter 11 Size theory Contaminants such as dust particles within the dispersant are typically large  compared to the sample size. Large particles mainly scatter in the forward direction. Therefore, by measuring the backscatter, the effect of dust is greatly reduced.

• Page 223: Molecular Weight Theory

  What is Static light scattering? The Zetasizer Nano series performs molecular weight measurements using a process called Static Light Scattering (SLS). This is a non-invasive technique used to characterise the molecules in a solution.
• Page 224 Chapter 12 Molecular weight theory For samples where A >0, the particle ‘likes’ the solvent more than itself, and  will tend to stay as a stable solution. When A <0, the particle ‘likes’ itself more than the solvent, and therefore may ...
• Page 225 Molecular weight theory Chapter 12 The standard approach for molecular weight measurements is to first measure the scattering intensity of the analyte used relative to that of a well described ‘standard’ pure liquid with a known Rayleigh ratio. A common standard used in Static light scattering is Toluene, for the simple reason that the Rayleigh ratios of toluene are suitably high for precise measurements, are known over a range of wavelengths and temperatures and, maybe more importantly, toluene is relatively easy to obtain in a pure form.

• Page 226: The Debye Plot

  We can therefore stipulate that if the particle is small, Rayleigh scattering can be assumed and the Rayleigh approximation used. With the Zetasizer Nano series the applicable molecular measurement weight range is from a few hundred g/mol to 500,000 for linear polymers, and over 20,000,000 for near spherical polymers and proteins.
• Page 227 Molecular weight theory Chapter 12 The weight-averaged molecular weight (Mw) is determined from the intercept at zero concentration i.e. KC/R = 1/Mw (for c--> 0) where the Mw is expressed in  Daltons (or g/mol). The 2 Virial Coefficient (A ) is determined from the gradient of the Debye plot.
• Page 228 Chapter 12 Molecular weight theory Page 12-6 MAN 0317...

• Page 229: Zeta Potential Theory

  What is Zeta potential? The Zetasizer Nano series calculates the zeta potential by determining the Electrophoretic Mobility and then applying the Henry equation. The electrophoretic mobility is obtained by performing an electrophoresis experiment on the sample and measuring the velocity of the particles using Laser Doppler Velocimetry (LDV).
• Page 230 Chapter 13 Zeta potential theory ions within the boundary move with it, but any ions beyond the boundary do not travel with the particle. This boundary is called the surface of hydrodynamic shear or slipping plane. The potential that exists at this boundary is known as the Zeta potential. Electrical double layer Stern Diffuse...
• Page 231 Zeta potential theory Chapter 13 Imagine a particle in suspension with a negative zeta potential. If more alkali is added to this suspension then the particles will tend to acquire a more negative charge. If acid is then added to this suspension a point will be reached where the negative charge is neutralised.
• Page 232 Chapter 13 Zeta potential theory Sedimentation potential :  The electric field generated when charged particles move relative to a stationary liquid. Electrophoresis When an electric field is applied across an electrolyte, charged particles suspended in the electrolyte are attracted towards the electrode of opposite charge. Viscous forces acting on the particles tend to oppose this movement.

• Page 233: Laser Doppler Velocimetry

  Zeta potential theory Chapter 13 electrolytes containing more than 10 molar salt. The Smoluchowski approximation is used for the folded capillary cell and the universal dip cell when used with aqueous samples. For small particles in low dielectric constant media f(Ka) becomes 1.0 and allows an equally simple calculation.
• Page 234 Chapter 13 Zeta potential theory Detector Cell ° Intensity of scattered light Time ill 6777 The light scattered at an angle of 17° is combined with the reference beam. This produces a fluctuating intensity signal where the rate of fluctuation is proportional to the speed of the particles.

• Page 235: The M3-pals Technique

  With M3 the measurement can be performed anywhere in the cell, though with the Zetasizer Nano series it is performed in the centre of the cell. M3 consists of both Slow Field Reversal (SFR) and Fast Field Reversal (FFR) measurements, hence the name ‘Mixed Mode Measurement’.
• Page 236 Chapter 13 Zeta potential theory position in the cell, however there are a number of reasons for choosing to work at the centre. The measurement zone is further from the cell wall, so reduces the chance of  flare from the nearby surface. The alignment of the cell is less critical.
• Page 237 Zeta potential theory Chapter 13 the PALS technique is used to determine the particle mobility in this part of the sequence. Insignificant fluid flow Stationary Layer ill 7674 M3 measurement sequence An M3 measurement is performed in the following manner: A Fast field reversal measurement is performed at the cell centre.
• Page 238 Chapter 13 Zeta potential theory Overall, the application of PALS improves the accuracy of the measurement of low particle mobilities. This can give an increase in performance of greater than 100 times that associated with standard measurement techniques. This allows the measurement of high conductivity samples, plus the ability to accurately measure samples that have low mobilities.

• Page 239: Operation Of The Zetasizer Nano

  Zeta potential theory Chapter 13 Operation of the Zetasizer Nano - Zeta potential measurements In a similar way to the typical DLS system described in the size theory chapter, a zeta potential measurement system comprises six main components. First of all a laser is used to provide a light source to illuminate the particles within the ...
• Page 240 Chapter 13 Zeta potential theory detector will become overloaded. To overcome this an “attenuator” is used to  reduce the intensity of the laser and hence reduce the intensity of the scattering. For samples that do not scatter much light, such as very small particles or samples of low concentration, the amount of scattered light must be increased.
• Page 241 Index Append to file Symbols 8-28 Aqueous 5-8, 8-41, 8-44 .dass 8-26 Arguments 7-6, 9-18, 10-7 .dasz 8-45 Atomic weights .del 2-23, 7-8 Attenuation 8-24 .pag Attenuator 5-10, 8-37, 11-6, 13-12 .sop 8-51 Attenuator index .sopl 2-29 Audit trails Auto mode 8-45 Numerics Automatic attenuation...
• Page 242 Index Zetasizer Nano Large particles Drain channel 2-18 Small particles Drain port 2-18 Configure button Dry-air supply 8-26, 8-45 2-14 Connections Dust filtration algorithm 2-13 Control Palette (reports) Dynamic Light Scattering 10-4 2-6, 11-1 Cooling fans 2-13 Correlation function 11-4 Molecular weight Edit result 3-11...
• Page 243 2-26, 2-27 High temperature Macros (Options dialogue) 2-16 9-21 HT label Mains power input 2-12 Huckel approximation Making a measurement 8-41, 13-5 Malvern logo 2-32 Malvern personnel Icon Manual measurement 21 CFR Part 11 Mark-Houwink 2-32 8-18 Nano Marquee 2-31...
• Page 244 Index Zetasizer Nano Material properties manager Multi-view Tab 8-29 3-8, 3-10, 3-11, 3-12 Materials Manager Flow-mode 8-15, 8-30, 8-39 3-13 Maximum run 8-42 Measure menu 2-25 Measure-Manual Nano icon 2-31 Measurement Narrow band filter 2-27 Measurement - Advanced (SOP) Navigation 8-24, 8-34, 8-43 Measurement - Instructions (SOP) Navigation toolbar...
• Page 245 Zetasizer Nano Index Zeta Titration measurements 3-12 4-19 Photon Correlation Spectroscopy Report Designer 11-1 2-30 Pictures in reports Introduction 10-6 10-1 Plate Navigator Opening 2-25, 10-11 10-3 Plate Scheduler Report tabs 2-25 2-30, 4-4, 7-2, 7-7 Playlist Reports 8-2, 9-5 Polar Systems Reports (SOP) 8-7, 8-27, 8-37, 8-47...
• Page 246 Index Zetasizer Nano Serial number Data Processing - Reports 1-4, 2-2, 2-14, 2-20, 10-19 Service engineer Data Processing - Reports (Molecular 2-26 Settings sub-menu weight) 2-27 8-37 Data Processing - Reports (Size) 13-7, 13-8 8-27 Shape correction model Data Processing - Reports (Zeta potential) 8-34 8-47 Shape model...
• Page 247 Zetasizer Nano Index Sample - Temperature (Size) Systematic deviation 8-19 8-35 Sample - Temperature (Zeta potential) 8-41 Sample (Molecular weight) 8-29 Sample (Size) Tab views 8-13 Sample (Zeta potential) Tables in reports 8-38 10-14 Size Measurement - Advanced (Molecular Temperature (SOP) 8-6, 8-19, 8-33, 8-41 weight) Text in reports...
• Page 248 Index Zetasizer Nano Voltage 8-43 Volume distributions 11-5 Z-average Size Volume PSD Zeta potential 2-1, 2-7, 13-1 Molecular weight 3-11 Operation of... 13-11 Size 3-10 What is What is... 13-1 Zeta potential Distribution Well-plate 10-5, 10-10 Zeta 3-11 Window menu 2-28 Zeta potential measurements - standard report Workspace settings...
• Page 250 Printed in England Malvern Instruments Limited MRK0499-02 Enigma Business Park Grovewood Road, Malvern Worcs, WR14 1XZ, U.K. Tel: +44 (0) 1684 892456 Fax: +44 (0) 1684 892789...

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