761 Main Ave Chromatography Division
Norwalk, CT 06859 USA Product Service Information
_________________________________________________________________________________________
Name: TurboMass Sensitivity Test
Model: TurboMass Technical SDB: E640010B.STN
Engineering Department Date: May 4,2000
Page 1 of 39
For Perkin-Elmer Internal Distribution Only
Problem:
N/A
Purpose:
To provide the field with an updated procedure to determine TurboMass sensitivity in the EI
and CI modes. Earlier revision A of this document did not contain a procedure for CI mode
sensitivity, and a new 10pg OFN standard has been added for EI mode. This revision
supercedes previous versions.
Materials Required:
Part Number Description
Documentation: S270-1610 Software Guide
0993-6341 Hardware Guide
Test Equipment: 0023-0177 1µL Syringe or equivalent (qty.2)
N931-6282 PE5MS Column or equivalent
30m x 0.25mm i.d. x 0.25µm
Standards: E640-1017 GCMS Sensitivity Mix Test Std.
Lot # 301 or higher (for CI mode)
E640-1088 GCMS Sensitivity Mix Test Std.
10pg OFN in iso-octane (EI mode)
General:
This procedure gives all the parameters and procedures to be used in the signal to noise
determination of the TurboMass to verify proper performance of the instrument in both EI and
CI modes. This release replaces the earlier revision A document, which should be discarded.
Procedure:
Do not attempt this procedure with anything but the sensitivity test standards listed above.
Although it is possible to use other standards, this is not recommended. Follow the steps
given below to determine the sensitivity of the TurboMass system.
Although both EI and CI sensitivity procedures are provided, the CI mode is an option and
may not apply to some systems.
2
TurboMass Sensitivity Specifications
Sensitivity Test Conditions
The sensitivity specification of the TurboMass mass spectrometer is measured using the
following certified NIST traceable standards. Each of the first three test standards is in a solvent
of hexane, test standard part number is E640-1017. Also required is a 10pg OFN standard in
iso-octane: E640-xxxx
Test standard #1: This test standard is used for full scan sensitivity.
Consists of (1pg/µl octafluoronaphtalene CI-), (10pg/ul benzophenone CI+) and (2pg/µl
hexachlorobenzene not used for sensitivity calculation).
Test standard #2: This test standard is used for single ion recording sensitivity.
It consists of (0.1pg/µl octafluoronaphtalene used for EI+).
Test standard #3: This test standard is used for the identification of retention times and library
searching the NIST library (optional). It consists of (100pg/µl each of octafluoronaphtalene,
benzophenone, and hexachlorobenzene).
Test standard #4: This test standard is used for full scan sensitivity.
Consists of (10pg/µl octafluoronaphtalene used for EI+).
TurboMass parameters: The instrument scans from 50-300 amu at a rate of 0.60 seconds per
scan. Set by the user as 0.2 second scan time and a 0.2 second inter-scan delay.
Signal to noise: is defined as the ratio of the appropriate test standard peak height to the RMS
noise.
Sensitivity Summary - Electron Impact Ionization
(250 L/S Turbomolecular Pump)
Instrument setup time 2 hour (approx.)
Testing time 1 hour (approx.)
A 90 minute stabilization time is required prior to conducting the
Note:
sensitivity tests. This stabilization period is required after the
column is installed, and with Operate left on.
1. Install an unpacked 1mm splitless liner (PSS) or 2mm liner (CAP) and a Viton
o-ring in the injector.
2. Install the test column and pump the system down. The manifold pressure
should be <1.5X10-5 Torr after 20 minutes with 1ml/min helium flow.
3. Check air/water for accepactable limits.
4. Autotune and mass calibrate instrument.
5. Increase the multiplier to 600 volts.
3
6. Setup the GC method parameters (using pressure pulse method – PPC only):
Start temp 50°C
Oven hold time 1.5 min
Ramp rate 20°C
End temp 240°C
End hold time 1.5 min
Injector temp 250°C
Split flow 50mL
Timed event - .75 split = 0mL/min (splitless)
- .70 carrier = 4ml/min
1.5min split flow = 50ml/min
1.55 carrier = 1mL/min
Carrier flow 1 ml/min (PPC)
Carrier Time 1 999 min
Head pressure 18 psi (Std pneumatics)
Vacuum On (PPC)
compensation
7. Mass Spectrometer Method:
Transfer line temp 250°C
Source temp 150°C
Start Mass 50
End Mass 300
Scan time 0.2
Inter scan delay 0.2
Start time 4.0
End time 8.0
Solvent delay 4.0
8. Inject 1 microliter of test mix E640-1088.
9. Measure the 10pg/ul octafluoronaphthalene using the m/z 272 mass
chromatogram peak.
10. Elution time is at approx. 6.5 min.
11. Calculate the signal to noise using the signal to noise function.
Note: Use the 100pg OFN test mix#3 standard to verify spectral integrity
if necessary.
Use separate syringes for the 100pg and 10pg sensitivity
standards. Do not use the same syringe with the 10pg standard
after it has been used with any other standard.
4
Sensitivity Test Procedure (EI Mode):
A. Installation and Conditioning
1. Install the column into the split/splitless or programmable split/splitless
injector on the AutoSystem XL gas chromatograph equipped with standard
manual or programmable pressure control carrier gas pneumatics (PPC).
Install the column according to the procedure in the AutoSystem
Note:
XL GC User’s Manual, Chapter 6, beginning on page 6-8, using a
narrow-bore 2 mm I.D. quartz liner P/N N612-1002 (S/SL) or a 1mm
I.D. quartz liner P/N N612-1006 (PSS).
Install the liner with a Viton O-ring P/N N930-2783 (S/SL) or N610-
1747 (PSS). The ferrule used is a 0.04mm id P/N 0992-0104. The
column used in this test is a PE5MS, 30 meter, 0.25 mm ID,
0.25 um film thickness P/N N931-6282 or an equivalent.
Use gloves, not fingers. Cleanliness is essential!
2. In order to minimize column bleed, a new column must be conditioned for a
minimum of 1 hour at 310o C with 1 ml/min flow of helium in the constant flow
mode with PPC control or 20 psi head pressure with manual pneumatics. Set
the injector to 250o C. The split flow should be set to 50 ml/min, during the
column-conditioning period before installation onto the TurboMass.
3. During this time, the TurboMass zones should also be heated to remove any
water vapor and contamination from the vacuum system. Cap off the
TurboMass interface transfer line with a blank ferrule P/N 0992-0511 and
pump down to vacuum. Set the source temperature and the GC interface
transfer line temperature to 300°C.
4. At the end of 1-hour at 310oC, cool the column to ambient and cool the
TurboMass source and transfer line to under 100°C before venting for column
installation.
The TurboMass column installation procedure is explained in the
Note TurboMass Hardware Guide (P/N 0993-6341). Reference Chapter 3
pages 3-10 to 3-19.
To prevent crushing the column, do not over tighten the fitting.
5. The depth of the column is best set by first removing the inner source. Insert
a septum over the end of the column, followed by the nut and ferrule.
6. Cut the end of the column back by 1 cm and wipe with it with a methanol-
dampened Kim-wipe (or other lint-free wipe). Insert it through the transfer line
until it is visible inside the vacuum manifold (figure 1). To avoid
contamination, do not touch the column portion that is inserted into the
interface with your bare hands. Use protective gloves to avoid contact.
5
The end of column is half way from the
source block wall to the hole in Lens 1 plate
Figure 1 - View of the source block through the front of the TurboMass with inner source removed.
7. Gently tighten the nut and slide the septum up to it to mark the column
position. Loosen the nut and withdraw the column 2 cm. Insert the inner
source. Reposition the column and tighten the sealing nut.
Before removing the inner source, you must first withdraw the
Note column. This procedure is covered in more detail in the
TurboMass Service Manual.
8. Pump the instrument down. With 1.0mL/min of helium flowing into the mass
spectrometer, if a penning high vacuum gauge (optional) is fitted it should
come on in approximately 2.5 minutes. Wait until the pressure drops below
2.5 x 10-5 Torr in the vacuum manifold as read on the Penning gauge while
observing the tune page. This will take approximately 5 minutes (longer for
higher humidity).
9. The vacuum integrity of the combined gas chromatograph and the mass
spectrometer will be evaluated with the split vent open and a split flow rate
of 50 ml/min.
10. Heat the GC transferline to 250oC using the settings on the TurboMass Tune
page. Heat the injector in the gas chromatograph to 250oC. Heat the GC
oven to 50oC. Heat the mass spectrometer source to 150oC.
11. On the Tune page display masses m/z 4, m/z 18, m/z 28 and m/z 32. After 20
minutes of pumping the ratios should resemble the figure shown below. If a
leak free system exists continue the test, otherwise locate, and remedy the
source of the leak before continuing.
6
This is a typical air/water ratio. The complete leak checking
Note procedure and explanation is covered in the TurboMass Service
Manual. Mass m/z 4 is helium, 18 water, 28 nitrogen, 32 oxygen.
B. Setting Up the TurboMass Thresholding
The TurboMass Instrument Data Thresholding parameters allow the user to
control how the instrument collects data, specifying the amount of data to acquire
and save to the disk. Limiting the amount of data stored on disk is particularly
desirable when performing long GC runs.
Advanced Data Thresholding provides ion counting options.
7
1. To access the Acquisition Control Panel click on
The Acquisition Control Panel screen appears:
2. The instrument data threshold and advanced threshold (ion count) must be
set properly in order to meet the desired sensitivity for this test.
8
Select Set Instrument Threshold from the Instrument menu.
The Instrument Data Thresholding dialog box appears:
3. Set all Instrument Data Thresholding parameters as shown in the above
screen.
4. Click on Advanced.
9
The Advanced Instrument Data Thresholding screen appears:
5. Set all Advanced Instrument Data Thresholding control parameters as
shown in the above screen. (Note that normal customer operation is with the
Ion Counting Threshold set to 3, and should be reset after testing is
completed.) Click OK.
6. Click on the close box button x to close the Acquisition Control Panel and
redisplay the Sample List screen.
10
C. Setting Up the AutoTune and Calibration
1. Display the Tune page by clicking on the “sunglasses” icon.
The Tune page appears:
2. Ensure Operate is off, select Gas and check that the Pump Out Reference
Gas… option is shut off (use menu and not the icon).
11
3. Then activate Ref Gas from the Gas menu. This turns on the reference gas
solenoid allowing calibration compound FC-43 (heptacosa, PTA, etc.) to flow.
4. Wait one minute then turn Operate on. Modify all parameters including
multiplier voltage to keep m/z 69 on scale with a gain of 1.
5. Select AutoTune from the Other menu.
The AutoTune screen appears:
6. Click on Setup.
The AutoTune Setup dialog box appears:
12
7. Set all parameters as shown above. Click on OK.
The AutoTune screen re-appears:
8. Click on Start.
This initiates the AutoTune sequence.
After successful completion, calibrate the mass axis.
13
D. Calibrating the Mass Axis after an AutoTune has been Performed
1. Select Calibrate from the Instrument menu on the Acquisition Control
Panel (this will occur automatically if Prompt for Calibration was checked).
The Calibrate dialog box appears:
2. Ensure the Reference File is hepta.ref, as shown above.
Click on Start.
14
The Automatic Calibration screen appears:
3. Set Types and Process as shown above.
Click on Acquisition Parameters.
The Calibration Acquisition Setup dialog box appears:
15
4. Enter all parameters shown in the screen above.
Click on OK.
5. Select AutoCal Check Parameters from the Edit menu.
6. Step through each of the edit screens by clicking on AutoCal Check, Cal
Parameters, and Mass Measure Parameters.
Enter all parameters as shown below:
7. Click on OK.
8. Click on OK.
16
9. Click on OK.
Return to the Calibrate page:
10. Click on Start.
17
11. After the calibration ends the following dialog box appears:
12. Click on OK.
To Maximize the intensity of the m/z 272 region, use mass 269 and mass 281 of
the reference gas to manual tune for greater intensity.
Set the gain to 2 or 3 (keep peaks on scale) and multiplier to 600 volts. Display
mass 269 and 281. Maximize the peak heights while maintaining resolution,
using manual tuning controls; an example is shown below:
13. After Calibration and Tune are complete, use the Gas menu and turn off the
Reference Gas.
18
14. Select Pump Out Reference Gas from the Gas menu:
15. Pump out the reference gas for 25 minutes or until 69 m/z at a gain of 100 is
within x2 the height of the adjacent background contamination peaks. Leave the
pump out valve on and continue with the sensitivity test.
Save the tune file.
Next select the GC control Method Editor:
16. Display the GC parameters by selecting Control Options from the
Instrument menu:
19
The GC parameters are: Autosampler, Oven/Inlets, Carrier, Valves,
Detectors, and Instrument Timed Events.
17. Select Autosampler, then set Manual as the Injection Source.
18. Click on the Oven/Inlets tab and set the values shown in the following
screen:
20
Type in the parameters for the GC oven temperatures and the GC injector
temperatures as shown above. After typing the values, the screen visually
displays the oven program temperatures.
On the carrier gas control page, make the appropriate entries for the carrier
control being used.
21
If PPC control is available, set the column flow to control by flow and enter a flow
rate of 1ml/min. (For standard pneumatics, adjust the pressure regulator for 18psi
and enter this setpoint value into the method.)
19. Click on the Instrument Timed Events tab and set the values as shown in
the following screen.
These settings are for a splitless injection. In a GC fitted with PPC control,
they turn the split flow off just before the injection and set the carrier gas flow
to 4 mL/min. Then 1.5 minutes into the run the carrier is set back to 1 mL/min
and the split flow set to 50 mL/min.
20. Click on OK
21. Save the GC settings by selecting Save As.... from the File menu.
22. Type a File name ending in .mth for your GC method, then click on Save.
23. Click on Exit from the File menu.
24. For manual pneumatics system, adjust the split flow needle valve for a split
flow of 50ml/min. Set the initial split valve setting to “off’. Then enter a timed
event in the defined events list a 1.5 min to change the split valve to “on”.
22
E. MS Method Settings
1. Display the Acquisition Control Panel.
2. Select Mass Spectrometer from the Methods menu in the Acquisition
Control Panel.
The Scan Functions screen appears:
3. Click on Edit Function from the Functions menu or double click on the gray
area in line 1.
23
Enter the values shown in the following Function dialog box.
4. Click on OK.
The following screen appears:
Save the MS method with an appropriate file name.
24
F. Identifying the Components in the Test Mixture.
Using the test column and method parameters in the procedure above, the
following results are obtained with a1 ul injection of 10pg OFN test. Typical
retention time and ions are shown below for identification.
F. Calculating the Signal to Noise
1. From the Sample List page click Chromatogram from the View menu:
2. Select the file to be used to calculate the signal to noise.
The 10pg of octafluoronaphthalene (OFN) will not be able to be seen on the
total ion chromatogram. The peaks seen are low level contaminates from the
solvent that will not interfere with the test.
25
3. Select Mass from the Display menu:
26
4. Enter the mass of OFN (m/z 272). Click on OK.
It may also be necessary to select range 3 min to 10 min.
The following screen appears:
5. Expand the chromatogram using the left mouse button by click and hold and
drag the area. The elution time of OFN is approximately 6 - 7 minutes.
27
A screen similar to the following will be displayed:
6. Select Signal To Noise from the Process menu:
28
The following dialog box appears:
7. Select the options as shown above.
8. The Signal and Noise values will be obtained using the mouse, as described
in the following steps.
29
9. Click and hold the right mouse button and drag the cursor across the OFN
signal peak approximately 1/3 the distance up from the bottom of the peak.
Release the mouse button.
10. Left mouse click on the Noise box.
11. Click and hold the right mouse button and drag the cursor across the noise
and take approximately 0.5 to 1min of representative noise. (You should see
a noise band as shown above, not just occasional noise spikes.)
12. Click on OK. A screen similar to the following will be displayed, showing the
signal-to-noise value of the m/z 272 peak:
30
The signal to noise value must be greater than 100 to 1 RMS for EI+ full scan.
31
Sensitivity Test Procedure (CI Mode):
A. TurboMass Setup
1. Before attempting sensitivity testing in the CI mode, it is first necessary to
complete and pass the EI sensitivity test. Once this has been achieved, all
test conditions used for EI testing will be applied to CI (i.e.: column, carrier
flow rate, oven program, heated zones, etc).
2. Having completed the EI sensitivity test, cool the transfer line and source and
vent the TurboMass. When the manifold pressure reaches atmosphere, cool
the GC oven and follow the appropriate procedures to move back the
capillary column and remove the EI source.
3. Insert the CI source and tighten the holding screws. Reposition the capillary
column in the transfer line (for the proper column installation procedure refer
to Section A – Installation and Conditioning – of the EI Sensitivity Test
Procedure in this document).
4. With the CI source installed and the column in place, pump down the mass
spec. Wait for the pressure to drop below 2.5 x 10-5 Torr (approximately 5
minutes). Set the source temperature to 150°C and the transfer line to 200°C
and wait for stabilization.
5. Connect a methane gas source to the CI gas fitting on the back panel of the
TurboMass. Set the methane tank regulator to a delivery pressure of 15psi.
Methane gas must be used for the CI gas when performing CI
Note sensitivity tests. Minimum gas purity is 99.95%, and requires an
appropriate pressure regulator fitted with a delivery pressure
indicator gauge.
6. Use the Gas option on the Tune page to open the CI Gas solenoid valve and
the Pump Out CI Gas solenoid valve.
7. Slowly open the CI gas needle valve on the front panel of the TurboMass
while monitoring the vacuum pressure. Adjust the needle valve until the
vacuum is between 10-3 and 10-4 Torr.
8. Allow the CI gas line to pump out for 5 to 10 minutes and then close the
Pump Out CI Gas solenoid.
B. Setting Up the Tune for CI+
1. Use the Ion Mode option on the Tune page and select CI+.
2. Set the Electron Energy at 30eV, and the Multiplier voltage to 210V. Arrange
the spectral display on the Tune page to monitor the following masses:
m/z 17, 29, and 41. Turn Operate on and monitor peaks.
3. Slowly turn the CI gas needle valve and adjust the gas flow while monitoring
the mz/17 and 29 peaks. Optimum flow will produce maximum peak heights.
The objective is to make the peaks equal at 100%.
32
4. Leaving the Multiplier voltage set at 210V, adjust all the other source and MS
control sliders to achieve the best sensitivity and peak shape. When
optimized, the m/z 17, 29 and 41 peaks should look similar to those shown
below.
The source and MS control settings shown in these examples are
Note typical values – these will vary from instrument to instrument. The
objective is to achieve maximum sensitivity and good peak shape.
5. The following screen shows typical MS control settings for good tune:
6. Change the Multiplier voltage to 600V and return to the Sample List page.
33
C. Building a MS Method and Sample List
1. From the Sample List page open the Acquisition Control Panel, click on
Methods and create a MS method with the following parameters:
2. Click on OK and return to the Scan Functions screen. Enter a solvent delay of
4.5 minutes and save the method.
3. On the Sample List page, create a list with two sample lines, one for a 100pg
benzophenone standard (Mix #3) and one for a 10pg benzophenone standard
(Mix #1). Use the MS method created above and the GC method used in the
EI sensitivity test.
D. Identifying Components in the Test Mixture.
1. Start the sample sequence and inject 1uL of Mix #3 for the first run. Display
the TIC and add a second scan for m/z 183. Monitor the run for the
benzophenone peak (retention time approximately 10.5 minutes).
Note: Use a separate syringe for the Mix #3 and Mix #1 sensitivity
standards. Do not use a syringe in the 10pg standard after it has
been used with the 100pg standard.
34
2. Obtain a result similar to the following:
E. Calculating the Signal to Noise
1. Modify the MS method by going to the Instrument Data Threshold page and
making the following entries:
2. Click on Advanced and make the following entries on the Advanced
Instrument Data Threshold page:
35
3. Save the changes and make a second injection using the 10pg standard (Mix
#1) and identify the benzophenone peak on the 183 chromatogram.
4. Expand the plot around the benzophenone peak and initiate a signal to noise
calculation. The signal to noise value must be 10 to 1 RMS or greater for CI+
full scan.
If the minimum signal to noise level is not met, it may be
Note:
necessary to readjust the CI gas flow and then retune for
maximum sensitivity and peak shape. Then inject another 10pg
standard and calculate signal to noise.
F. Setting Up the Tune for CI-
1. On the tune page, click on the Ion Mode option and select CI-.
2. Click on the Gas option and activate the Reference Gas solenoid valve.
3. Set the Multiplier voltage to 300V, and set the spectral windows to display:
m/z 300, 452 and 633. Use the source and MS controls to tune for best
sensitivity and peak shape. Adjust the CI gas flow if necessary for maximum
sensitivity. Typical peak shapes should appear as follows:
36
4. Set the Multiplier voltage to 550V, click on the Gas option and turn off the
Reference Gas solenoid. Open the Pump Out Reference solenoid valve and
pump out the reference gas line for 10 minutes, then close the Pump Out
Reference Gas solenoid valve.
G. Identifying Components in the Test Mixture.
1. Using the same GC and MS methods created in the CI+ sensitivity
calculation, add a line to the Sample List page for a 1uL injection of Mix #1.
The CI- sensitivity is calculated using the 1pg octafluoronaphthalene
compound in the standard.
2. Start the run and inject 1uL of Mix #1. Monitor the TIC and Mass 272 for the
octafluoronaphthalene peak (approximately 6.5 minutes).
37
3. Expand the 272 chromatogram and initiate a signal to noise calculation on the
octafluoronaphthalene peak.
4. The signal to noise value must be greater than 20 to 1 RMS for CI- full scan.
If the minimum signal to noise level is not met, it may be
Note:
necessary to readjust the CI gas flow and then retune for
maximum sensitivity and peak shape. Then inject another 1pg
standard and calculate signal to noise.
38
Installation Test Results - TurboMass Mass Spectrometer
Company Date Tested
Address Call Number
Serial Number
User Name
Phone ( ) -
SPECIFICATION: ACTUAL VALUE:
(250 L/S Turbo Pump)
Sensitivity EI Mode:
EI+ Signal to noise full scan detection
1 pg Octafluoronaphthalene (m/z 272)
Signal to noise of 10:1 RMS.
Sensitivity CI+ Mode:
CI+ Signal to noise full scan detection
10 pg Benzophenone (m/z 183)
Signal to noise of 10:1 RMS.
Sensitivity CI- Mode:
CI- Signal to noise full scan detection
1 pg. Octafluoronaphthalene (m/z 272)
Signal to noise of 20:1 RMS.
CONFIGURATION TESTED:
MODEL NUMBER: SERIAL NUMBER:
Gas Chromatograph
Computer
Printer
TEST STANDARD: LOT NUMBER: EXPIRATION DATE:
GCMS Sensitivity Std. E640-1017
GASES:
Helium 99.999% Pure [ ] Reagent Gas: Purity:
This is to certify that the above tests have been performed on the TurboMass system.
[ ] meets [ ] does not meet the Perkin-Elmer Installation Specifications.
Authorized P-E Representative: Employee #:
COMMENTS:
39
