User Manual

Chromeleon 7 Extension Pack

ICH Templates - LOD LOQ (SN)

Thermo Scientific Dionex Chromeleon

Version 7.1 SR1
Document Revision: 1
October 2011
© Thermo Fisher Scientific
Table of Contents
1 Introduction 1
1.1 About this Document 1
1.2 Extension Pack 1
1.3 ICH 1
1.4 Parameters in Chromeleon 2

2 Requirements 3

3 Known Issues and Limitations 4

3.1 Limitations 4
3.2 Known Issues 4

4 Getting Started 5
4.1 Importing the ICH Templates 5
4.2 Description of the ICH LOD LOQ (SN) eWorkflow template 6
4.2.1 eWorkflow General 6
4.2.2 Sequence General 6
4.2.3 Sequence Layout 8
4.2.4 Resulting Sequence 9
4.3 Customizing the ICH LOD LOQ (SN) eWorkflow template 9
4.3.1 eWorkflow General 11
4.3.2 Sequence General 11
4.3.3 Sequence Layout 11

5 Step By Step Guide 13

5.1 Creating a Sequence 13
5.1.1 Launching the ICH LOD LOQ (SN) eWorkflow 13
5.1.2 Modifying the created sequence 14
5.2 Running the Sequence 15
5.3 Processing the Data 15
5.3.1 Processing Method 16
5.3.2 Updating the Component Table 16
5.3.3 Updating the Component Table - Advanced 17
5.4 Reviewing Results 18
5.5 Creating the ICH LOD LOQ (SN) report 19
5.5.1 Printing the report 20
5.5.2 Exporting the report 20

6 Description of the Report Sheets 22

6.1 Report Sheet “LOD LOQ (S/N)” 22
6.1.1 Limitations 22
6.2 Report Sheet “Overview” 22
6.2.1 Limitations 23
6.3 Other Report Sheet 23

7 Combined Validation Report 24

7.1 Importing Query 24
7.2 Modifying Query 24
7.3 Printing / Exporting the Query 24
1 Introduction

1.1 About this Document

The layout of this User Manual is designed to provide quick reference to sections of interest. However, we recommend that
you review the User Manual thoroughly before working with the Chromeleon 7 Extension Pack in order to obtain a full
understanding of the software.
This User Manual is provided "as is". Every effort has been made to supply complete and accurate information and all
technical specifications and programs have been developed with the utmost care. However, Thermo Fisher Scientific Inc.
assumes no responsibility and cannot be held liable for any errors, omissions, damage, or loss that might result from any
use of this manual or the information contained therein. We appreciate your help in eliminating any errors that may appear in
this document.
At various points throughout this document messages of particular importance are indicated by the following symbols:

Note: Indicates general information that points out specific items of interest.

Caution: Indicates areas where operations should be performed carefully.

The information contained in this document is subject to change without notice.

Chromeleon® and UltiMate® are registered trademarks of Thermo Fisher Scientific Inc.. Any other mentioned trade or company names are subject to the copyright and the
property and trademark rights of the respective companies.
All rights reserved including those for photomechanical reproduction and storage on electronic media. Without the written permission of Thermo Fisher Scientific Inc., no part
of this publication may be reproduced in any form (by means of photocopy, microfilm, or any other process) for any purpose or processed, copied, transmitted, or distributed
in any other form, independent from the means, electronic or mechanical, that is used.

1.2 Extension Pack

The Chromeleon Extension Pack contains templates that allow you to quickly and easily calculate results that require
advanced calculations, for example ICH LOD LOQ (Signal to Noise) determinations. All required items to perform the
calculation are combined in an eWorkflow:
• Sequence Templates

• Processing Methods

• Report Template

• Custom Variables

Furthermore the eWorkflow contains the following files:

• User Manual

• Example Sequence

1.3 ICH
The International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human
Use (ICH) provides recommendations on the validation of analytical procedures (Q2(R1)). Recommended tests include -

• Accuracy

• Detection Limit

• Linearity

• Precision

• Quantitation Limit

• Range

• Robustness
• Specificity

The calculation and reporting of all data requested by ICH can be very tedious and time consuming. The ICH method
validation templates provided in the Chromeleon 7 Extension Pack automate these steps and allow the quick and easy
creation of method validation reports.

1.4 Parameters in Chromeleon

To perform the ICH LOD LOQ (SN) calculations in the Chromeleon report template, additional information that cannot be
readily entered in Chromeleon is required. Table 2 gives an overview of the required information.

Parameter Description

TEST Identifies the ICH test. This is an injection custom variable that can be entered in the injection list.

Identifies the last injection of a sequence for a particular ICH test. This is an injection custom variable that can
Test_End
be entered in the injection list.

User definable option to specify if the noise measurement is taken from the blank or the actual chromatogram.
Measure_Noise
This is a component custom variable that can be entered in the component table in the processing method.

User defined ratio to be used for the LOD calculation. This is a component custom variable that can be entered
LOD_Ratio
in the component table in the processing method.

User defined ratio to be used for the LOQ calculation. This is a component custom variable that can be entered
LOQ_Ratio
in the component table in the processing method.

User defined start time for the noise measurement of the injection. This is a component custom variable that
Noise_Start
can be entered in the component table in the processing method.

User defined end time for the noise measurement of the injection. This is a component custom variable that can
Noise_End
be entered in the component table in the processing method.

Test Specifies if the test will be run for the corresponding component or not.
Table 1: ICH Parameters in Chromeleon
2 Requirements
The ICH templates can only be used with Chromeleon 7.1 SR1 or higher.
It is possible to export the ICH results in the Adobe® Acrobat® PDF format. Acrobat Reader version 5.0 or higher is needed
to read these reports.
3 Known Issues and Limitations

3.1 Limitations
• The template Processing Method file can handle 5 calibration standard levels by default, but additional levels can
be added.

3.2 Known Issues

• The “Print Preview” option does not work for graphical objects.
4 Getting Started

4.1 Importing the ICH Templates

All files required for a particular ICH test are contained in the relevant ICH Template eWorkflow. Each eWorkflow is named
according to the test e.g. ICH LOD LOQ (SN), and distributed as a Chromeleon backup file with .ewfx, extension. The files
are located on the Chromeleon 7 disc, in the folder \Extension Pack\ICH Templates.
1. In the Chromeleon Console go to the eWorkflows category.

2. In the Menu click File, Import From.

3. Select the relevant eWorkflow backup file and click Open.

4. Select in which Data Vault the eWorkflow must be imported and click Start (Figure 1).

Figure 1: eWorkflow import - Select Data Vault

The eWorkflow is now imported and visible in the list of eWorkflows (Figure 2A). A Chromeleon backup file of the demo data
(Figure 2B) and User Manual (Figure 2C) are directly accessible.

Figure 2: Imported eWorkflow

4.2 Description of the ICH LOD LOQ (SN) eWorkflow template
4.2.1 eWorkflow General
The eWorkflow General page is used to specify some general settings for the eWorkflow and to associate instruments,
methods, and documents. The eWorkflow General settings for the ICH LOD LOQ (SN) Template are displayed in Figure 3.

Figure 3: eWorkflow General page

Template settings are as follows:

Figure 3 Property Setting

eWorkflow State Approved for Use

eWorkflow Type HPLC

This eWorkflow contains all files for performing Limit of Detection and Limit of Quantitation
eWorkflow Description calculations based on the signal to noise ratio of the analyte peak. Please refer to the
attached user manual for information.

No instruments are associated with the ICH LOD LOQ (SN) Template eWorkflow. This
Instruments
eWorkflow can therefore be used for all instruments.

Processing Method ICH LOD LOQ (SN), containing the required custom variables for LOD LOQ
(Blank SD) calculations.
Report Template Method_Validation, containing all required calculations for LOD LOQ (Signal
Methods
to Noise).
View Settings ICH LOD LOQ to display chromatogram information in the Data Processing
Category.

Chromeleon backup file ICH LOD LOQ (SN) - Demo Data.cmbx, containing an ICH LOD LOQ
Attachments (SN) example
PDF document ICH LOD LOQ (SN) - User Manual.pdf
Table 2: Default eWorkflow General settings
Section 4.3 describes how these settings can be customized.

4.2.2 Sequence General

The Sequence General page is used to specify general settings for the sequence and select a default report and view
settings.
 Figure 4: Sequence General page
Template settings are as follows:

Figure 4 Property Setting

1
Default Report Template Method_Validation

2
Default View Setting ICH LOD LOQ

3 Defines the channel to be displayed as the default channel when opening an injection.
Default Channel
As this depends on the instrument and corresponding instrument method this is empty.

‘ICH LOD LOQ (SN)’ followed by current data/time (Example: ICH LOD LOQ (SN)
Sequence Name
20110511 161743)

Data Vault ChromeleonLocal

ICH/LOD_LOQ is the directory that the resulting sequence is stored in. If the folder in
Path this directory path does not exist, it will be automatically created when the first sequence
is created.

All signature steps to submit electronic signatures are checked, to enable a qualified
Electronic Signature
user to electronically sign the sequence and submit, review or approve it.

Comment This sequence is created using the ICH LOD LOQ (SN) eWorkflow.

Table 3: Default Sequence General settings

1
Report template to be used as the default template for presenting the data in a report.
2
View settings to be used as the default settings for presenting the data in the Chromatography Studio.
3
Channel to be displayed as the default channel when you open an injection.
Section 4.3 describes how these settings can be customized.

4.2.3 Sequence Layout

The sequence layout tab defines rules for the sequence layout, for example, the number of repetitions of LOD LOQ (SN)
injections. The sequence layout helps to create a sequence with a well-defined structure. A sequence layout comprises four
block types, each marked by a different color.

Figure 5: Sequence Layout page

Template settings are as follows:

Figure 5 Property Setting

By default the sequence is setup to make 3 LOD LOQ (SN) blank injections and 2 calibration
1 standard injections at 2 concentration levels (total of 5 injections). However, it is not mandatory to
Sequence Header
follow this approach. You can add or remove injections to meet your requirements. Injections can
also be set as multiple injections from the same vial in the same autosampler vial position.

2
Sample Block <Empty>

3
Bracket <Empty>

4
Sequence Footer <Empty>

Table 4: Default Sequence Layout settings

1
Sequence Header includes injections that always appear at the beginning of a sequence, such as blank injections.
2
Sample Block includes a line for each sample that needs to be analyzed. The sample block is repeated between brackets.
3
Bracket includes injections that will be performed between sample blocks (for example, calibration standards). A bracket
separates two sample blocks from each other.
4
Sequence Footer includes injections that always appear at the end of a sequence.
The settings in the sequence columns are as follows:

Sequence Column Setting

No. of inj. 1

Name Blank, Standard X

 Type Blank, Calibration Standard

Position <uneditable>

Volume 1µl

Instrument Method <empty>

Processing Method ICH LOD LOQ (SN)

Status <uneditable>

Inject Time <uneditable>

Weight 1.0000

Dilution 1.0000

IntStd 1.0000

ID <empty>

01, 02, 03, 04, 05 (depending on level of calibration standard i.e. 01 for first calibration standard and
Level
02, 03 and 04 for other calibration standards)

TEST LOD/LOQ (S/N)

Test_End <empty> except for last injection in list, which has LOD/LOQ (S/N) (End)

Comment <empty>
Table 5: Default sequence column settings

4.2.4 Resulting Sequence

When launching the ICH LOD LOQ (SN) eWorkflow template without making any changes, the resulting sequence will
look like Figure 6.

Figure 6: Resulting sequence

4.3 Customizing the ICH LOD LOQ (SN) eWorkflow template

The eWorkflow can be customized to fit any application. When doing so the number of steps to do before being able to run a
ICH LOD LOQ (SN) analysis is reduced to a minimum.
To start customization:
1. In the Chromeleon Console go to the eWorkflows category.

2. In the navigation pane select the ICH LOD LOQ (SN) eWorkflow template (Figure 7A).
 Figure 7: Start editing the ICH LOD LOQ (SN) eWorkflow templates
3. In the work area click Edit (Figure 7B). The eWorkflow editor now opens.

4. To save the eWorkflow with a new name click File, Save As.

5. In the Save eWorkflow dialog select the Data Vault (Figure 8A) to store the eWorkflow and enter a name (Figure 8B)
for the new eWorkflow.

Figure 8: Saving eWorkflow with new name

The eWorkflow can now be customized. The settings to modify are described in sections 4.3.1, 4.3.2, and 4.3.3.

Tip: For an extensive explanation of modifying eWorkflows and defining the sequence layout, refer to the Chromeleon 7
online help.
4.3.1 eWorkflow General

Figure 3 Property Customization

While customizing the eWorkflow the state can be set to In Development. This way it is clear to
eWorkflow State users the eWorkflow is still being worked on. When the customization is finalized the state can
be set back to Approved for Use.

eWorkflow Description Provide a descriptive text for the eWorkflow.

Add the instrument(s) that can run this ICH analysis. The eWorkflow can only be launched on
listed instrument(s).
Instruments
When no instruments are added, the eWorkflow can be used with all instruments accessible to
the user.

Add the instrument method(s) for the instrument(s).

Modify the processing method to:
Methods - have a suitable chromatogram integration
- include the required standard components and their concentration
- define the ICH calibration type

The current manual can be removed or other documents can be attached, for example a
Attachments
relevant SOP about running the ICH LOD LOQ (SN) application.

Table 6: Customization options for the eWorkflow General page

4.3.2 Sequence General

Figure 4 Property Customization

When instrument method(s) are added to eWorkflow General tab a list of corresponding
Default Channel
channels is available. Select a default channel.

If the default sequence name is not suitable, define a sequence name. Note that variables can
Sequence Name
be used for dynamic sequence names.

If the default ChromeleonLocal is not suitable, select the Data Vault where the sequences
Data Vault
should be stored.

If the default ICH/LOD_LOQ path is not suitable, select the folder location where the sequences
Path
should be stored. Note that variables can be used for dynamic folder names.

If electronic signatures are not used, uncheck the Submit electronic signatures option.
Electronic Signature
If electronic signatures are used, define the appropriate signature steps.

Comment Provide a comment for the created sequence.

Table 7: Customization options for the Sequence General page

4.3.3 Sequence Layout

Figure 5 Property Customization

Sequence Header Customize sequence header per your requirements.

Sample Block Customize sample block per your requirements.

Bracket Customize bracket per your requirements.

Sequence Footer Customize sequence footer per your requirements.

Table 8: Customization options for the Sequence Layout page

Sequence Column Setting

No. of inj. Enter the number of injections that should be injected from the same vial.

Name Enter relevant name

Type Must remain as Calibration Standard and Blank

Position This is a non editable field, which will be filled when creating the sequence.

Volume Enter desired injection volume

Select the correct instrument method. If the eWorkflow can be used on multiple instruments
Instrument Method
with different instrument methods, it’s better to leave this field empty.

When the processing method is updated and has a new name, select the correct processing
Processing Method
method.

Status <uneditable>

Inject Time <uneditable>

Weight 1.0000

Dilution 1.0000

IntStd 1.0000

01, 02, 03, 04, 05 (depending on level of calibration standard i.e. 01 for first calibration
Level
standard and 02, 03 and 04 for other calibration standards)

TEST LOD/LOQ (S/N)

Test_End Ensure the last injection has the entry LOD/LOQ (S/N) (End)

Comment Enter an appropriate comment for the injection

Table 9: Customization options for the sequence columns
5 Step By Step Guide
Using the ICH LOD LOQ (SN) eWorkflow template involves a simple process:
1. Creating a sequence, using the ICH LOD LOQ (SN) eWorkflow template. This creates a Sequence that checks the
expected result for the analyte peak against the actual result and expresses it as a percentage (e.g. % recovery).

2. Processing the data

3. Reviewing the results

4. Creating the ICH report

This section provides a step-by-step guide for using the ICH LOD LOQ (SN) template.

5.1 Creating a Sequence

5.1.1 Launching the ICH LOD LOQ (SN) eWorkflow
To create a sequence using the ICH LOD LOQ (SN) eWorkflow follow these steps:
1. In the Chromeleon Console go to the eWorkflows category.

2. Select the relevant eWorkflow (Figure 9A) and the instrument (Figure 9B) to use.

Figure 9: Launching customized eWorkflow

3. Click Launch (Figure 9C). The eWorkflow wizard is launched (Figure 10).
 Figure 10: eWorkflow wizard
4. Depending on the settings in the eWorkflow various settings have to be made. The minimum required entries are the
Number of samples and the starting position of the vials in the sampler. Optionally the setting Run sequence after
creation can be activated.

5. Click Finish. The sequence is now created (Figure 11).

Figure 11: Created sequence

5.1.2 Modifying the created sequence

Depending on the setup of the eWorkflow, some modifications to the sequence are required, e.g.:
• Modify the positions of the injections in the automated sample injector.

• Select the correct Instrument Method for all injections.

• Define the test level of each calibration standard by using the level field e.g. calibration standards at level 1 must have
01, etc.

• There is a column called “TEST” available. This is used to tell Chromeleon what test an injection is assigned to.
Injections must be assigned the test “LOD/LOQ (S/N)”.

• Finally, there is a column called “Test_End”. This is used to inform Chromeleon when the last injection for a specific
test has been made. You need to select the relevant value for the last injection i.e. for Standard 2 you need to use
“LOD/LOQ (S/N) (End)” (Figure 12).
 Figure 12: Setting Custom Variables

Note: All information in the sequence table can be updated after the sequence has run.

Note: If you do not want to use the data from a specific test, ensure that the sample type is set to “Unknown” and that the
injection is not assigned a “TEST” value.

5.2 Running the Sequence

Now that the sequence is created and modified, it can be started. Follow these steps:

Note: When the option Run Sequence after creation is used in the eWorkflow wizard (see section 5.1.1, Launching the
ICH LOD LOQ (SN) eWorkflow, page 14), the sequence is already running.

1. Select the sequence in the navigation pane.

2. Click Start (Figure 13A).

Figure 13: Starting the sequence

The sequence is now running (Figure 14).

Figure 14: Running sequence

5.3 Processing the Data

After the ICH LOD LOQ (SN) data has been acquired, it can be processed. All data processing steps are performed in the
Chromatography Studio. To open the Chromatography Studio from the Console, double click any object in a sequence.
5.3.1 Processing Method
The ICH LOD LOQ (SN) eWorkflow template contains a dedicated processing method. This processing method is designed
to show the processing parameters required by the test, and also to allow you to define your specifications for the test
(Figure 15).

Figure 15: ICH LOD LOQ (SN) processing method layout

The default Processing Method file for the LOD LOQ (SN) test shows 5 tabs
Detection: Allows you to define your peak detection parameters
Component Table: Allows you to name your peaks and define their specifications (e.g. minimum correlation coefficient)
Component Table - Advanced: Allows you to define the concentration of each peak at each calibration level
SST: Allows you to define System Suitability Tests
General: Allows you to define general settings
The major steps in using the Linearity Processing Method file are to update the Component Table (Figure 15A) and
Component Table – Advanced (Figure 15B).

Note: The processing method can have different layouts, consisting of different tabs and names. Therefore the
Processing Method tables and columns you see may look different from the ones in these pictures.

Tip: Choose the processing method layout appropriate for your application, or define your own layout by selecting
individual tabs via the Processing Method layout dialog ( ).

5.3.2 Updating the Component Table

If the processing method is not already updated, the major step is to update the component table (Figure 16).

Figure 16: Component table containing standard peaks only

In this table the calibration compound names (Figure 16A) are entered, including their corresponding retention times
(Figure 16B) and retention time window (Figure 16C) for peak identification. The evaluation type should remain as ‘Height’
(Figure 16D) and the calibration type can be set to either ‘Lin’ or ‘Lin, WithOffset’ (Figure 16E). ICH LOD LOQ (SN) specific
parameters (Figure 16F) are entered into six custom variables columns (Table 11).
 Component Table Column Description

Name Name of the peak.

Retention Time Expected retention time of the peak.

Window Allowed window for the expected retention time.

Eval. Type Allows you to choose the Y-axis for the linearity test.

Cal. Type Either “Lin” or “Lin, WithOffset” must be selected.

Test Specifies if the test will be run for this component or not.

Defines if the noise will be measured in the current chromatogram (Current) or in the blank
Measure_Noise
chromatograms (Blank).

LOD_Ratio Defines ratio for calculation LOD. Recommended value is 3.

LOQ_Ratio Defines ratio for calculation LOQ. Recommended value is 10.

Noise_Start The start time for the noise calculation. Noise is measured in the blank injections.

Noise_End The end time for the noise calculation. Noise is measured in the blank injections.
Table 10: ICH LOD LOQ (SN) specific custom variables in the component table

5.3.3 Updating the Component Table - Advanced

The Component Table - Advanced contains 5 predefined level columns numbered 01 to 05. Enter the concentration for
each calibration standard level in levels 01 to 05 for each component (Figure 17A). The units of concentration can also be
entered (Figure 17B).

Figure 17: Component table - Advanced

The steps involved in updating the component tables are as follows:

1. Open your sequence in the Studio, for example by double-clicking an injection in the Console.

2. Click the Data Processing category bar.

3. Show the Chromatogram Pane and the Processing Method Pane using the corresponding tools in the Data
Processing Home Ribbon tab (Figure 18).

Figure 18: Panes Group of the Data Processing Home Ribbon tab
In Component Table:
4. Ensure that your component is correctly identified by updating the retention time and retention time window.

5. Decide if you want to test the LOD LOQ (Signal to Noise) of each component or not by updating the “Test” field.

6. Define if the noise will be measured in the current chromatogram (Current) or in the blank chromatograms (Blank).
7. Define ratio for calculation LOD. Recommended value is 3.

8. Define ratio for calculation LOQ. Recommended value is 10.

9. Define if the start of the period you wish to determine noise for each component.

10. Define if the end of the period you wish to determine noise for each component.

Note: When measuring the noise in the blank, it is recommended to choose a section of the chromatogram that
corresponds to where the peak is expected to appear. For example, the EP recommends measuring a range of 20 times
the peak width at half height around the expected retention time of the peak.

When measuring the noise in the current chromatogram, it is recommended to choose a section of the chromatogram
that contains no peaks.

In Component Table - Advanced:

11. Enter the known amount of analyte in the Calibration Standard injections (e.g. concentration, weight, etc.), in the level
columns starting with 01.

12. Update the concentration units.

5.4 Reviewing Results

The ICH LOD LOQ (SN) eWorkflow template contains a report template. This report template is designed to report all ICH
test results in a convenient way including LOD LOQ (Signal to Noise). Once the sequence table and the processing method
are updated, Chromeleon immediately calculates the results.
To review the results switch to the Report Designer category (Figure 19).

Figure 19: Review results in the report designer category

In this view you will see that each ICH test has its own associated tab (e.g. Accuracy, LOD LOQ (Blank SD), Robustness,
…). Click on the “LOD LOQ (S/N)” tab to review your results. The ICH LOD LOQ (SN) report will now be visible (Figure 20).
For a detailed description of the report sheet, please refer to section 6.
 Figure 20: Reviewing ICH LOD LOQ (SN) results in the report template

5.5 Creating the ICH LOD LOQ (SN) report

The ICH report template is designed for a quick calculation of ICH results and creation of the report. The process to
generate the report is as follows:
1. Select the sequence containing the ICH results you want to report (Figure 21) and right click. Continue at section 5.5.1
to print the report or section 5.5.2 to export the report.

Note: When only a few of several samples need reporting, select those samples, right click, and select Print or Export.

Figure 21: Starting reporting the ICH results

5.5.1 Printing the report
To print the report, follow the steps below:
2. Right click and select Print Report (Figure 21A). The Print dialog now opens (Figure 22).

Figure 22: Print dialog when printing the ICH report

3. In the Report Template & Channel section, select the correct channel to print the data (Figure 22A).

4. In the Printer section, select the correct printer (Figure 22B).

Note: The automatic reporting options are configured to print the ICH LOD LOQ (S/N) test sheet. Please refer to section
7 for instructions how to print the results of all ICH tests together.

5. Click OK to start printing.

5.5.2 Exporting the report

The report can be exported as an electronic copy of the results in Adobe Acrobat PDF format. The electronic copy of the
report is automatically stored on the hard drive in the folder C:\Chromeleon_Export. The file name is
ICH_LOD_LOQ_(SN)_yyyymmddhhmmss.pdf, where:
• yyyy = calender year, for example 2011

• mm = month, for example 06

• dd = day, for example 19

• hh = hour, for example 11

• mm = minutes, for example 59

• ss = seconds, for example 23

This ensures that every file created has a unique name. To export the report follow the steps below:
2. Right click and select Export (Figure 21B). The Export dialog now opens (Figure 23).
 Figure 23: Export dialog when exporting the ICH report
3. In the Report Template & Channel section, select the correct channel to export the data (Figure 23A).

Note: The automatic reporting options are configured to export the LOD LOQ (S/N) sheet for injections with TEST
custom variable.

4. Click OK to start exporting.

6 Description of the Report Sheets

6.1 Report Sheet “LOD LOQ (S/N)”

The ICH guidelines state that “Determination of the signal-to-noise ratio is performed by comparing measured signals from
samples with known low concentrations of analyte with those of blank samples and establishing the minimum concentration
at which the analyte can be reliably detected. A signal-to-noise ratio between 3 or 2:1 is generally considered acceptable for
estimating the detection limit. For the quantitation limit a typical signal-to-noise ratio is 10:1.”. This sheet is designed to
provide an overview for all injections used in signal to noise determination. It shows the component information, details
injections used in determination and the calculated signal to noise results.

Figure 24: Report Sheet "LOD LOQ (S/N)"

6.1.1 Limitations
None.

6.2 Report Sheet “Overview”

The report sheet Overview provides a list of all injections that were used in determining the LOD LOQ Signal to Noise result
and the associated sequence of each.
 Figure 25: Report Sheet "Overview"

6.2.1 Limitations
None.

6.3 Other Report Sheet

The other report sheets are described in the respective manual of each individual test.
7 Combined Validation Report

7.1 Importing Query

The templates have been designed to allow you to quickly collate all your validation data and create a combined method
validation report. The process is as follows.
1. In the Chromeleon Console go to the Data category.

2. In the Menu click File, Import From.

3. Locate ‘Collate_Data.cmbx’ file on the Chromeleon 7 disc, in the folder \Extension Pack\ICH Templates. Click Open.

4. Select the correct Data Vault and directory that contains the method validation data.

7.2 Modifying Query

1. Open query.

2. In the Sequence search replace [ENTER_PATH] with the folder directory that contains the method validation (Figure
24A).

Figure 26: Query dialog for collating results

3. Save the query.

7.3 Printing / Exporting the Query

1. Click on Query Now. The query will automatically run and retrieve the relevant injections and show all of the
data in a virtual sequence (Figure 25).
 Figure 27: Virtual sequence containing all method validation data results.

2. To print the report, select Print, or expand and select Report on the Query tool bar (Figure 26A). This will open
the Print dialog.

Figure 28: Starting the reporting process

3. Select all relevant sheets of tests that you want to report.

 Figure 29: Printing collated results

4. To export the report, select the Studio button.

5. Then select the Report Designer category.
6. Click the Application button (Figure 28A) and select Export (Figure 28B).

Figure 30: Starting the exporting process

7. This will open the Export dialog (Figure 29). Ensure the option Apply to ‘Results of current injection query’ is
selected (Figure 29A).
Figure 31: Query Export dialog