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Part Number
N515-6071
Release
A
Publication Date
May 2001
Any comments about the documentation for this product should be addressed to:
User Assistance
PerkinElmer Instruments LLC
710 Bridgeport Avenue
Shelton, Connecticut 06484-4794
U.S.A
Or emailed to: AI.UserAssistance@perkinelmer.com
Notices
The information contained in this document is subject to change without notice. PerkinElmer makes no
warranty of any kind with regard to the material, including, but not limited to, the implied warranties of
merchantability and fitness for a particular purpose. PerkinElmer shall not be liable for errors contained
herein for incidental consequential damages in connection with furnishing, performance or use of this
material.
NOTE: TotalChrom is the updated version of the software previously marketed as Turbochrom. This
product is compatible with TotalChrom and Turbochrom version 6.1.x. The term TotalChrom has been
used throughout this document to denote either system.
Copyright Information
This document contains proprietary information that is protected by copyright.
All rights are reserved. No part of this publication may be reproduced in any form whatsoever or
translated into any language without the prior, written permission of PerkinElmer Instruments LLC.
Copyright 2001 PerkinElmer Instruments LLC.
Trademarks
Registered names, trademarks, etc. used in this document, even when not specifically marked as such,
are protected by law.
PerkinElmer is a registered trademark of PerkinElmer, Inc.
TotalChrom, AutoSystem, AutoSystem XL, and 600 Series Interface are trademarks of PerkinElmer
Instruments LLC.
Turbochrom is a trademark of Applera Corporation.
Table of Contents
Introduction
Using this Guide.......................................................................................................................... 1
Conventions Used in this Guide .................................................................................................. 1
The Help System ......................................................................................................................... 2
HS 40 System Connections
Hardware Connections ................................................................................................................ 3
HS 40 Headspace Analysis
Creating an HS 40 Control Method........................................................................................... 10
Creating a Sequence in TotalChrom.......................................................................................... 14
Setting up a Sequence and Displaying HS 40 Status................................................................. 14
Setting Up a Sequence from the HS 40 .............................................................................. 15
Displaying HS 40 Status .................................................................................................... 17
Index
Introduction
The HS 40 Control application is used on a TotalChrom system to set parameters and
obtain status reports from the HS 40 or HS 40XL Headspace Sampler. The HS 40
control application comprises the HS 40 Method Editor, HS 40 Status and Setup, and
HS MHE commands, which are on the TotalChrom Navigator Apps menu.
The Multiple Headspace Extraction (MHE) software application that is included with
the TotalChrom HS 40 Control software lets you analyze and report data and results
obtained by performing multiple extractions and injections from a headspace vial.
Terminology
The following terms are used to refer to program elements and the actions that you
perform to carry out tasks:
Choose The terms choose and select indicate actions that you perform. Choose
represents carrying out an action associated with a command or command button.
When you are likely to use the mouse to choose an action, mouse-specific terms
are used, such as click, right-click, and double-click. When you are likely
to use the keyboard to choose an action, the specific key is mentioned, such as
Enter or Tab.
Select The term select refers to highlighting an object or item or moving the cursor
focus. Selecting an item prepares it for an action: for example, when you select
text, it appears in reverse video. When you select a dialog box option, you
activate the option but the function is not carried out until you choose the OK
button, which closes the dialog box and sometimes completes another operation.
(QWHURUVHOHFW When you use the File Select or File Open dialog box to open a
file, the phrase enter or select is used to refer to the actions you can take to
open that file. When you enter a filename, you type it in the File Name text box
exactly as it exists. When you select a file, you browse for it on your computer or
your network. When you select a file, you will not introduce typographical errors.
Illustrations
Unless otherwise indicated, the values in the illustrations in this manual are examples
only. They are not intended to indicate the exact values you will see or to suggest the
values you should use.
Instrument Names
Throughout this manual, the name HS 40 refers to both the HS 40 and the HS 40XL
Headspace Samplers. Also, AutoSystem refers to both the AutoSystem and the
AutoSystem XL GC.
HS 40 System Connections
This section describes how to connect the HS 40 hardware to the TotalChrom system.
This section does not cover the installation and configuration of your computer.
Hardware Connections
The TotalChrom computer communicates with the HS 40 through the PerkinElmer
600 Series LINK Interface or the built-in integral LINK. The LINK acquires and
buffers digital data from the AutoSystem GC and controls the GC and HS 40.
The following procedure explains how to make the HS 40, AutoSystem, and LINK
interface connections for TotalChrom control. It assumes a certain level of knowledge
of laboratory instrumentation installation and of your TotalChrom system.
If you do not know how to perform some of the steps in this procedure, you will need
assistance in order to set up and install an HS 40 as part of your TotalChrom system.
Contact your local PerkinElmer representative to make an appointment for Service
Installation, or find a qualified person in your own organization.
This procedure assumes that you have done the following:
After installing the hardware, configure the LINK interface and the AutoSystem as
described in Chapter 3 of your TotalChrom Users Guide.
HS 40 System Connections
2.
Turn off and unplug all instruments, and disconnect the power cords from all
instrument rear panels.
3.
HS 40 System Connections
AutoSystem XL GC
Turbochrom
Serial/Host
or
IEEE
Connection
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HS 40XL
HS 40 System Connections
HS 40 System Connections
RS232
AutoSystem XL GC
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HS 40XL
Cable (P/N 6560-9010 or 6560-9025)
HS 40 System Connections
4.
5.
6.
Reconnect and plug in all power cords, and turn on all instruments.
HS 40 Headspace Analysis
When you run an HS 40 analysis, you do the following:
The AutoSystem GC and HS 40 methods must have the same filename and must
be stored in the same directory. For example, a sequence that references the
methods HS40.MTH and HS40.HSM, both stored the directory
C:\MYMETHODS\, is valid.
An AutoSystem sequence will not run if the two method filenames do not match.
For example, a sequence that references the methods HS40.MTH and
HS40METH.HSM will not run.
The Cycle Time value in the HS 40 method must be greater than or equal to the
time from one injection to the time at which the GC is ready for the next
injection. The Cycle Time value must include the oven cool-down and GC
equilibration times to allow the HS 40 to properly calculate and execute the
overlapping thermostatting in the HS 40.
The HS40TEST method and sequence files in the TotalChrom Examples subdirectory
contain values that can be used as a starting point when creating HS 40 methods.
10
HS 40 Headspace Analysis
The available text boxes reflect the options installed in your HS 40.
2.
4.
Select the appropriate thermostatting mode from the Sampling Mode list:
Constant Select this mode if you want to use the same thermostatting
time for all samples that are analyzed by this method. Constant mode is the
standard mode of operation.
Progressive Select this mode if you want to automatically increase the
thermostatting time for a series of samples. Sample 1 is thermostatted for the
value entered. Sample 2 is thermostatted for twice the entered value, Sample
3 for three times the entered value, and so on. This function is used for
initial method development to determine the thermostatting time required for
a specific application. The function can also be used for kinetic studies. Vial
position one must be filled. Subsequently, some vial positions can be empty
to increase the thermostatting time between some vials. For every vial
position after position one, the thermostatting time will be increased by the
length initial thermostatting time.
11
In the Injection group box, select whether you want to inject for a specific
length of time or inject a specific volume of sample.
If you select time injection, specify the desired injection time.
If you select volume injection, specify the column flow rate and the desired
injection volume. The HS 40XL will automatically calculate the
corresponding injection time.
6.
12
Specify the required temperatures for the sample vial, sampling needle, and
transfer line (sample transfer tubing) in the Temperatures group box.
HS 40 Headspace Analysis
For best reproducibility, set the needle and transfer temperatures to a value
5-10C higher than the sample temperature.
8.
9.
10. In the Options group box, select any accessories installed on your HS 40 that
you want to use during the run.
The Water Trap check box is available only when the CryoFocus check box
is selected. For more information on each HS 40 option, refer to the
appropriate accessory manual.
11. To calculate the period of time between injections (PII), choose Update PII.
TotalChrom queries the HS 40 for the PII time based on the method
parameters.
The PII value lets you see the effects of your time parameter changes on vial
throughput.
12. Complete the description on the Documentation tab, and choose OK.
13. Save the HS 40 method.
13
If you select One Per Row in the Method group box of the Global Parameters
dialog box, the Method column must contain the TotalChrom AutoSystem GC
method that has the same filename as the HS 40 method.
If you select Multiple Per Row in the Method group box, the Instrument method
column must contain the TotalChrom AutoSystem GC method that has the same
filename as the HS 40 method. The Processing and Calibration methods can be
different methods.
14
HS 40 Headspace Analysis
Choose HS 40 Status and Setup from the Navigator Apps menu to display
the HS 40 Status and Setup window.
2.
To display the Setup HS 40 dialog box, choose the Setup in the HS 40 Status
and Setup dialog box, or choose Set Up from the Instrument menu.
3.
If there is more than one GC, select the desired GC from the Instrument list.
4.
5.
In the Raw Data Path text, enter or select the path to the directory where you
want TotalChrom to store your HS 40 raw file data.
6.
In the Result Data Path text, enter or select the path to the directory where
you want TotalChrom to store your HS 40 result file data.
15
7.
In the Starting Row text box, you may change the row number with which
you want to begin the sequence operation.
8.
In the Ending Row text box, you may change the sequence row number with
which you want to end the sequence operation.
9.
10. Select Suppress Report/Plots if you do not want to print a report and plot.
11. To initiate the sequence setup and HS 40 run, choose OK.
TotalChrom verifies that the vials indicated in the sequence are in sequential
order with the correct number of injections indicated, downloads the
sequence to the LINK, and starts the HS 40 run.
12. To stop the run before the sequence is complete, choose Stop from the
HS 40 Status and Setup dialog box.
Do not use the Clear Setup command in TotalChrom to abort an HS 40 run.
If you want to abort (clear the TotalChrom setup) an HS 40 run after it has been set
up and is thermostatting, click the Stop button in the HS 40 Status and Setup window.
Choosing Clear setup from the TotalChrom Run menu will not stop the HS 40 from
thermostatting.
16
HS 40 Headspace Analysis
Displaying HS 40 Status
During a run the HS 40 Status and Setup dialog box displays the HS 40 status,
current parameter values, and current sequence.
Choose HS 40 Status and Setup from the Navigator Apps menu to display
the HS 40 Status and Setup window.
Extended Thermostatting The sample vial is undergoing a longer-thanprogrammed thermostatting while waiting for the GC to become ready for injection.
Pressurizing The sample vial is being pressurized to carrier gas pressure.
Injecting Sample gas is being introduced onto the column.
Withdrawing The sampling needle remains in the vial after injection has taken
place before being withdrawn or lowered into the vent position. The withdrawal time
period is designed to allow transfer of the injected sample into the separation system .
Venting The sample vial is being vented to atmosphere.
CryoFocus The sample is being focused cryogenically on the column over an
extended injection period to improve analytical sensitivity.
BackFlushing The HS 40 is backflushing the precolumn to remove higher boiling
analytes from the separation system and to shorten analysis time.
Analyzing GC analysis of a headspace sample is in progress. Depending upon the
method parameters used, the next sample in the sequence is most likely undergoing
thermostatting. The HS 40 is ready to pressurize and inject the next sample after the
thermostatting time has elapsed.
To print an HS 40 report or audit trail:
18
1.
2.
To print the audit trail for an HS 40 file, select Print from the Files menu in
the HS 40 Method Editor to open the Print Options dialog box, select Audit
Trail, and choose OK.
19
Component Menu Lets you select components for display in the Side-by-Side
Plots window and manually enter peak areas for extractions. The component selection
choices on this menu are available only when the Side-By-Side Plots window is
active.
Select HS MHE Application from the TotalChrom Apps menu to display the
MHE Results window.
2.
Choose New from the File menu to create a new MHE file.
Choose Open from the File menu and select an existing MHE file.
TotalChrom displays the Result Files and Side-by-Side Plots windows.
21
3.
To open one or more file(s) for the Calibration or Sample list, position the
cursor in the list in which you want to enter a result file, select Open Sample
Result (.RST) Files from the Samples menu, select the desired file(s), and
choose Open.
The files that you enter into a sample list should be consecutive runs
(extractions) from the same vial.
Once you have added at least two entries to a sample list, the MHE
application generates a regression analysis of those entries and continues to
update the analysis as you add more samples to the list.
4.
To save the regression results, choose Save or Save As MHE Results from
the File menu, and enter the MHE result file name <filename.mhe>.
To enter the peak areas manually, select Manual Area Entry from the
Component menu to display the Manual Area Entry dialog box.
2.
Enter the peak area for each extraction for which you want to calculate an
MHE curve.
After you have entered two peak areas, the MHE application calculates and
displays the MHE curve along with the total area (TA), the Y-intercept (A1),
and the slope of the regression curve. The total area is calculated as the total
area underneath the curve extrapolated to the X-intercept. TotalChrom
continues to update the curve as you enter new peak areas.
22
General Information
Component
6WXG\1DPH
Sample Name The name of the sample as entered in the Sequence Editor.
9LDO1XPEHU The number of the sample vial that contains the component
that is displayed.
6DPSOH$PRXQW
'LOXWLRQ)DFWRU
before injection.
With the Side-By-Sides window active, choose Select from the Component
menu to open the Select Component dialog box.
2.
To view the plot for a component in a sample list, select the desired list from
the options to the right of the component list, then select the desired
component, and choose OK.
TotalChrom displays the component data and plot in the Side-By-Sides Plot
window.
24
3.
To scroll for the selected component in another sample within the same list,
choose Next/Previous in the Side-by-Side Plots Component menu.
4.
To clear the MHE files, choose Abandon from the MHE Results File menu.
To print a report of the MHE analysis results, select Print from the Files
menu to open the Print Options dialog box.
2.
From the Print Option dialog box, choose the sample list(s), the desired
component(s), and the data you want to print.
Sample File List: Lets you print the list of TotalChrom result files for each
sample list selected in the Samples group box.
Component Table: Lets you print the table of statistical data on the
component that is currently on display, or for all components, for each
sample list that is selected in the Samples group box.
Component Plot(s): Lets you print the side-by-side plot(s) for the
component that is currently on display, or for all components for each
sample list selected in the Samples group box.
To change the paths to the Calibration and Sample Lists, choose Set File
Paths from the MHE Results Samples menu to open the Set File Paths dialog
box.
25
2.
Enter or select the path(s) for the TotalChrom result files, and choose OK.
Choose Erase File Lists from the File menu to open the Select Sample(s) To
Erase dialog box.
2.
Select the check box for each sample list that you want to erase, and choose
OK to clear all result files from the selected list.
Choose Sort Files from the File menu to open the Select Sample(s) To Sort
dialog box.
2.
Select the check box for each sample list that you want to sort, and choose
OK.
TotalChrom arranges the result files by injection date and time.
26
dC
= kC
dt
(1)
C = C0 e kt
(2)
27
Since the extractions are performed in steps, the time t may be replaced by the
number of extraction steps, n. The initial concentration C0 is replaced by the peak area
from the first extraction step, A1, which occurs at time t = 0 or n = 1. The constant k,
which now includes instrument parameters, becomes k*:
An = A1e k
( n 1)
(3)
ln An = k * ( n 1) + ln A1
(4)
A simple regression analysis by the MHE application arrives at a straight line plotted
through the values of the peak areas derived from three or four extraction steps. This
line gives the values for the slope (k) and y intercept (A1). If the MHE curve is not
linear, equilibrium is not established inside the headspace vial under the given
thermostatting conditions. In this case, neither MHE analysis nor a standard single
injection analysis with external or internal calibration will yield the correct results.
The total peak area of the volatile compound in the sample is then determined by
summing the partial peak areas from each extraction step. Using equation (3) as a
geometric progression, the total area An becomes
= A1 1 + e k * + e 2 k * + e 3k ...
(5)
Written in a more general form, the equation for the sum of the areas is
A1
1 ek
(6)
Applying the values determined by the regression analysis, the application arrives at
an extrapolated total peak area number that directly corresponds to the total amount of
the compound present in the sample.
Once the software has calculated the total area for a particular analyte in the sample
the equivalent concentration of the analyte in the sample is also calculated. Using
total peak areas derived from previous MHE analyses of a calibration standard, the
amount of analyte can be calculated and if the sample amount has been entered, the
concentration is subsequently displayed.
For more information about MHE theory and techniques, refer to the following:
28
Index
6
600 series LINK, HS 40 connections 6
A
audit trail 18
AutoSystem connections
HS 40 4
interfaces 4
ready/run signal 8
C
component information, MHE
about 23
viewing/printing 24
constant thermostatting 12
conventions, manual 1
cycle time 9, 12
D
data processing, delaying 16
E
equations, MHE 27
G
ground 8
H
hardware connections 3
HS 40 analyses
about 9
creating a method 11
creating a sequence 14
displaying status 17
printing audit trail 18
printing method parameters 18
setting up a sequence 15
starting 15
HS 40 connections
AutoSystem 4
interface 4, 6
ready/run signal 8
shield wire 8
HS 40 Method Editor 10
HS 40 methods
creating 11
printing audit trail 18
printing parameters 18
requirements 9
HS 40 MHE analyses See MHE analyses
HS 40 status 17
I
instrument configuration, updating 12
integral LINK, HS 40 connections 4
M
manual conventions 1
messages, HS 40 status 17
methods
GC 9
HS 40 11
HS 40 MHE 19
i
shield wire 8
side-by-side plots 22
starting a run 15
status, HS 40 17
stopping a run 16
suppress processing 16
MHE
about 27
files, managing 25
mode 12
regression curves 21
MHE analyses
about 19
creating a method 19
creating a result file 21
printing/viewing results 24
regression curves
displaying 22
entering peak areas 22
plotting 21
sequence 20
R
ready/run signal 8
regression curves, plotting 21
S
sample information
about 23
viewing/printing 24
sample lists
creating 21
managing 26
sampling modes 12
sequence
creating 14
HS 40 14
MHE 20
setting up 15
ii
theory, MHE 27
thermostatting
extended 17
mode 12
TotalChrom sequence See sequence