II. Directions for Split Injections (General Use)

Injection port A [front port with black nut]
This port operates under split flow conditions (80:1) at 250 °C with a Rtx-5 (5% diphenyl 95% polydimethylsiloxane) Integra-guard capillary column and a head pressure of 14 PSIG. The guard column length is 10 m followed by an analytical column having the following dimensions: 30-m x 0.25-mm x 0.25-mm film thickness. Check the total flow of gas to injection port A by pressing FLOW followed by A on the GC keypad. If the meter reads something other than ~65, make the appropriate change in the flow rate by turning the TOTAL FLOW control knob, on Injection port A, until the meter reads ~65. The split ratio is now roughly 80:1. If you find that your sample is too dilute or too concentrated after injection, you may adjust the split ratio to compensate. See Appendix 1 for instructions. To use the mass selective detector with this column the internal valve inside the GC oven must be in the A position (valve knob set screw points left toward MSD). This is the default position of the valve! Do not change it. Sample injection should not exceed 5 mL in this injection port; additionally, the sample concentration should not exceed 1.0E-02 M (approximate concentration for NMR analysis). Too much sample will result in poorly shaped (fronted) peaks or cause damage to the MSD filament.

 

Injection Port A (black nut)                                   Total Flow Control Knob on Lower Left

Switching Valve in Position A

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MS ChemStation Software

The GC and MSD are controlled by a PC running MS ChemStation software. This package permits instrument control, data collection, and data processing.

Control-Data Collection. The control and data collection software utilities (GC/MS Top) are found under the 5970-Instrument-1 group in the Windows program manager. Once started you will see a window labeled Top. Load a method by pulling down the Methods menu and selecting Load.... Scroll through the method files and select either genlow or genhigh. Genlow is for compounds that are typical organics (bp < 220 °C and MW < 340), while genhigh is for compounds up to MW 500 and volatilization temperature of 270 °C. For instructions regarding creating your own method see Appendix 2.
 
 
 

 


To prepare the instrument for injection using the above method select Main Panel under the DataAcquisition pull-down menu. This will download all the parameters from the method to the GC and MSD; a new dialogue window "Acquisition" will appear. To inject a sample select Inject under the Data pull down menu. Enter the appropriate filename (8 characters maximum) and sample identification information in the boxes, then click OK. The software will check for system readiness. It is important to check that PURGE A is ON when using injection port A. If it is not, change it using the GC keypad. Follow the directions and inject your sample. Press START on the GC panel as soon as you have finished injecting your sample. This signals the software to begin the data collection protocols and a real time acquisition window is displayed. Never override the solvent delay unless an emergency situation arises. If all your compounds elute prior to the end of the run, select Stop Run in the real time acquisition window to conserve analysis time.

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Data Analysis. For data analysis use the StandAlone Data Analysis package. This way you can process data during analysis runs! Find the StandAlone Data Analysis icon in the 5970-Instrument-1 group of the Windows program manager. After the program loads you can process any data file you wish or take a ìsnap-shotî of the one currently running. To load a file for analysis pull down under File and select Load. Find the file in the dialog box and click OK. The total ion chromatogram (TIC) will be loaded into register R0 and displayed at the top of the window. Double click on a peak using the right mouse button to show the mass spectrum for a particular time point. If you wish to average the mass spectra over a certain time range, just hold the right mouse button down and drag over the appropriate time span in the TIC. You can expand any scale (in either the TIC or MS) by holding down the left mouse button and dragging the outline of a box in the window. To return to the original display just click the left mouse button once in the appropriate window. To take a ìsnap-shotî of a current run select Take Snapshot from the File pull-down menu. Process accordingly. To print the TIC, MS, or specify a window to print, select Print from the File pull-down menu. Select the appropriate response in the dialog box that appears. Remember that any zooming you do will be printed as such unless you reset prior to printing!
 
 



To obtain peak widths and areas for peaks in the TIC you have two options: A) Pull down Chromatogram and select Integrate. This will evaluate all the peaks above a given reference level established in your acquisition method. B) Type GET n on the command line, where n is an integer number. This will integrate the n peaks having the highest signal in the TIC and display the corresponding mass spectra. After either option select List Results under the Chromatogram pull-down menu. A new window with the integration results is displayed. Copy these values to the clipboard for pasting into other documents or print them.

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Advanced Data Analysis.

Library Matching - to see if a mass spectrum matches one currently in the NIST library, just display the mass spectrum and double click the RIGHT mouse button in the window containing the mass spectrum. The search results will be displayed and you have the option to print them. Scroll through the search results window to find the database compound that most closely matches your mass spectrum. Remember, not all the worldís compounds or their derivatives are in the database!

the white space above this line is the command line

Displaying multiple mass spectra - to display multiple mass spectra you must use a few macro commands. First, display two sequential mass spectra by clicking in the TIC with the right mouse button. Then type MERGE on the command line at the bottom of the window followed by a return. This will orient the two spectra in one window. Type DRAW on the command line and you will see them displayed. To add another mass spectrum, just display it as usual by selecting the spot in the TIC and clicking the RIGHT mouse button. Now repeat the MERGE and DRAW commands. Once you have decided upon the exact number of spectra you wish to display you have one more option. You can plot them all with the same Y-axis. To do this enter NORMALIZE 100 on the command line. This will normalize each spectrum to its base peak (ion fragment with the highest signal) and the 100 indicates the maximum value on the y-axis scale. Enter the DRAW command one last time to update the display.

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Appendix 1: Adjusting the Split Ratio

Use the graph below to assist you in setting the split ratio appropriately. With the 40 meter x 0.25 mm column and a head pressure of 14 PSIG, the column flow rate is roughly 0.86 mL/min. Set the total flow rate using the TOTAL FLOW control knob. Adjust the knob until the GC meter reads the flow value associated with the split ratio desired. Please note that the electronic flow meter DOES NOT display the split vent flow rate; it only approximates it! Thus, a meter reading of 40 does not mean the flow out the split vent is 40 mL/min.

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Appendix 2: Creating a Split Analysis Method

 


Method Creation. The control and data collection software utilities (GC/MS Top) are found under the 5970-Instrument-1 group in the Windows program manager. Once started you will see a window labeled Top. Load either the genlow or genhigh method depending on the sample type to analyze. Modify the existing method to create a new one by pulling down the Methods menu and selecting Edit Entire Method. Check all three boxes to edit. Make the appropriate changes in the ensuing dialogue boxes and save your file at the end with a new name (remember it must be 8 or fewer characters long).

A few important notes regarding the dialogue boxes are contained in this section. Enter comments regarding the method you are creating and your identity in the Method Information window. You should save a copy of the method with the data and only the Data Acquisition box should be checked as a method section to run. In the Normal Scan Acquisition window change only the solvent delay, mass range, and plot parameters. Keep in mind that the number of scans per unit time depends on the mass range scanned. As the scan range increases, the number of scans will decrease. Additionally, do not make the solvent delay too short!! The next window is the Temperature Information window. Injection port A temperature may be changed, but injection port B should stay at 200 °C and detector B at 280 °C. The real method creativity is associated with the oven program on the right hand side of the window. The maximum rate of heating or cooling is 70 °C/minute and the maximum temperature the oven should reach is 300 °C. A graphical display shows the run time and temperature program you enter. DO NOT change anything in the Injection Information dialog window; it is preset for split injections. The settings should read as follows: Injector port A: initial value ON, time on = 0.00 min, time off = 0.00 min; Injector port B: initial value OFF, time on = 0.00 min, time off = 0.00 min. If these are not set appropriately then the system will not function properly during injection and analysis. NOTE: when the purge time on and time off are set to zero the purge valve does not change from the initial conditions. The remainder of the dialog windows can be ignored. Remember to save the method with a new name; avoid overwriting the old method and reeking havoc with the system.

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