Modules Summaries and Reviews answers

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It covers the full functionality of the Agilent 6890/6850 ChemStation software with a focus on data analysis and reporting. 2. Course Outline with correlation to file names: Modules: out of the 12 H5926A modules ChemStation Configuration…….…………………………..…(mod03.ppt and mod03.doc) ChemStation Introduction .…………………….……….……(mod04.ppt and mod04.doc) Data Acquisition 6890 GC………………….…………..……(mod05.ppt and mod05.doc) Data Analysis………………..……………………………….(mod06.ppt and mod06.doc) Calibration ………………………………..…………….…….(mod10.ppt and mod10.doc) Sequences……………………………………………………..(mod11.ppt and mod11.doc) Reporting…………………………………………………..…(mod 12.ppt and mod12.doc) Introduction to the 6850 GC.(optional)……………………….(mod 13ppt and mod13.doc) Modules 2,7,8 and 9 of the H5926A course would be for reference and the lab exercises for those modules could be performed later by the customer. 3. For marketing purposes of this product see the Product Release Bulletin (if available): Product Release Bulletin available? No Product Release Bulletin location: 4. Instructor or Delivery Agent skills and training required to deliver the product: The following general skills are required in order to deliver customer education courses in the GC product line. Thorough technical knowledge in the use of GC as an analytical technique. Demonstrates technical knowledge in the operation of Agilent 6890 and 6850 GC- systems. Demonstrates technical knowledge in the operation of Microsoft Windows NT operating systems and software packages. Makes technical presentations which are professional and effective by: Pacing the presentation to match student comprehension based on observation and feedback. Demonstrating effective time management during course sessions in order to complete the required training material. Adapting the course material by citing specific examples appropriate for the students application. Relating personal experiences that reinforce the course material. Conducts quality training through proper preparation, organization, and use of materials. Accurately sets customer’s expectations while showing sensitivity to the customer’s needs. Provides feedback to SSBU about course materials where improvements can be made in the course organization or printed material. Demonstrates the ability to systematically solve problems during laboratory exercises in order to maximize the customer’s experience and minimize wasted time. Maintains classroom/equipment in working order and presentable condition. Ability to maintain a mature and professional attitude while dealing with problem students. Practices appropriate standards of business conduct as defined by Agilent. The following course specific skills are required in order to deliver H2606A Detailed knowledge of the principles of GC (theory and applications). Detailed knowledge and understanding of the A.04 to A.08 ChemStation software. Detailed knowledge of basic chemistry principles. Detailed knowledge of GC hardware and maintenance. Detailed knowledge of the original and the enhanced integrator Detailed knowledge of creating calibration tables, perform sequencing (with bracketing and cyclic recalibrations), and sequence summary reporting. Detailed knowledge of generating customized reports with custom report generator, and have familiarity with commands and macros Detailed knowledge of the security access included in Windows NT . Detailed knowledge of the Companion Software. Detailed knowledge of the Retention Time Locking Software. Detailed knowledge of the ChemStation Plus concepts. 5. Equipment (Instrumentation and Software) required for product delivery: The course is designed to accommodate Agilent 6890 optional 6850 GC customers with Rev. A.08.0X ChemStation Software. The classroom sessions with the offline laboratories are designed for laptop computers or ChemStation computers. The recommendation is a computer for each customer. A printer can be shared between no more than two ChemStations. Current Agilent 3D ChemStation Configuration and Software: G2070AA Rev.A.08 ChemStation Software CD-ROM with GC Companion Software and the Retention Time Locking software G2080AA The Microsoft Internet Explorer and Video drivers are necessary for laboratories. This course would be greatly enhanced when an Agilent 6890 or 6850 GC is available. Minimum Configuration PC Windows NT based system: HP Vectra Pentium 166 MHz Super VGA display (800 x 600) 1.2 GB hard disk CD-ROM dirve 48 MB RAM Laserjet family of printers Recommended PC Configuration HP Vectra 600 MHz Pentium Super VGA display 1024 x 768 4 GB hard disk CD-ROM drive 64 MB RAM Laserjet 5 with 4 MB RAM 3 ½ inch floppy drive Lecture material consists of electronic PowerPoint presentations. A high resolution (800x600) Proxima or NEC projector is required. The Student Manual is a Word97 document containing student notes for each PowerPoint slide and lab exercises for each module. Classroom Computer for slide presentation and software demonstration. Rev.A.08 Educational CE-ROM containing all files for the course delivery or a copy of this files downloaded from the EPI Warehouse. A floppy disk is provided with each book. It contains Data, Method, and Sequence Files that are used in the laboratory exercises. Each student should be given a blank floppy for saving additional files. Lab exercises include activities suitable for stand-alone PC’s without a GC attached. 6. Consumables required for product delivery: Module 3 requires each student to have a blank floppy disk. Modules 4, 5 require the use of Instrument configuration files loaded on a stand-alone PC in order to simulate the presence of an Agilent 6890 GC. Module 6 requires Data files and Method files that are provided on the Supplemental Floppy disk: Data files: CS_01, CS_02, CS_03, CS_04, CS_05, CS_06, CS_07, Unk_1, Unk_2 Method files: Std_Pack.m and Std_spli.m (these are methods using the Standard Integrator). Modules 10 , 11 and 12 require the use of files on the Supplemental Floppy. Module 13 is optional if 6850 users are among the students and requires the use of the Column Installation Video and the Controller Familarization Tutorial both on the ED- CD ROM. 7. Samples required for product delivery: no samples needed 8. Electronic Files required for product delivery: Rev.A.08 ChemStation Software CD-ROM with GC Companion Software and the Retention Time Locking software G2080AA. Rev.A.08 Educational CE-ROM containing all files for the course delivery or a copy of this files downloaded from the EPI Warehouse. A floppy disk is provided with each book. It contains Data, Method, and Sequence Files that are used in the laboratory exercises. Modules 4,5 requires the use of Instrument configuration files loaded on a stand-alone PC in order to simulate the presence of an Agilent 6890 GC (the same simulation will serve for the 6850 GC). 9. Tools required for product delivery: No tools required. 10. Activities required prior to product delivery: Ensure that the Agilent ChemStation are operational. The HP6890 Keyboard Tutorial should be loaded on each PC. Ensure the Instrument Configuration files are loaded in the Instrument subdirectory in order to simulate the presence of a 6890 GC. Instructor should arrive in enough time before the class starts to ensure proper and complete setup. 11. Activities required during product delivery: Supervision of the students during lab exercises. Instructor should refer to the “instructor notes” for specific information. 12. Activities required after product delivery: Ensure students complete required evaluation forms. Report any significant feedback from the customers or instructors to SSD via the Customer Feedback System. Shutdown equipment according to local facility requirements. User Task Analysis The user needs to know the basic chemistry to mechanisms. The user needs to know basics on Audience Analysis GC education levels range from lab technician chemist high school grad to Ph.D . Experience experience to several years. understand the separation Microsoft NT and LAN. being a college grad to levels range from no Modules Summaries and Reviews answers Module 2 Summary and Review Instructor’s Notes: Module 2 covers Basic GC concepts and definitions. Major components of a GC Considerations for Gases and Plumbing Explanation of the Split Inlet 6890 Keyboard Operation Lab Exercises Level 1 Lab Exercise: Lab 1: Hardware Familiarization and Inspection of the Split Inlet Lab 2: Inspection of the Purged Packed Inlet Lab 3: Column Installation Optional Lab Exercise: Lab 4: 6890 Keyboard Familiarization (Tutorial) The instructor should briefly familiarize the student with type of inlets and detectors, which are on the GC. Prior to the start of these labs: Initiate a ChemStation session and load the method "Def_GC.m and the sequence Def_GC.s. After the method and sequence are loaded, close out the instrument session. Thus, later when the students initiate the ChemStation, this default method and sequence will be brought up, the last method used prior to shutdown. Verify that the 6890 GC Keyboard Tutorial has been loaded on the PC. Prior to Lab 1: new septa and o-rings for the split inlet are required. Prior to Lab 3: The Column Installation segment from the Video: 6890/ALS/ChemStation Operation PN H5944-90006 should be shown prior to the start of Lab 3. Columns Needed for Lab 3: 30m x 0.32mm x 0.25um Phenyl Methyl Silicone (HP-5) PN 19091J413 30m x 0.53mm x 0.88um Methyl Silicone (HP-1) PN 19095J-023 Module 2 Review Answers List the primary components of a GC system. Gases, Inlets, Columns, Detectors, Data Acquisition and Data Analysis What are the differences between packed and capillary columns? Capillary columns are longer and much smaller i.d. as compared to packed columns. Cap columns are more efficient, produce sharper peaks. Cap column analyses are usually faster. Cap columns have limited sample capacity and work best with a split inlet or very small sample amounts. Why should traps be used in the gas plumbing? To prevent moisture and other contaminants from getting into the system. Why is it necessary to split away injected sample? Capillary columns have very limited sample capacity. If all of the sample injected is allowed on the column, the peaks would be very large and would overload the column. The exception is large diameter 0.53 mm (Megaboretm) or larger capillary columns. These columns can be used in a packed type inlet but will not provide as efficient separations as when used in a split inlet. Module 3 Summary and Review Instructor Notes: This module covers Introduction to Windows 95 and Windows NT Introduction to Lan capabilities Introduction to Configuration Editor Lab Exercises Level 1 Lab Exercises: Lab 1: Introduction to Windows NT Level 2 Lab Exercises: Lab 2: Working with Windows NT Lab 3: Configuring the ChemStation Lab 4: Using the ChemStation Scheduler Module 3 requires each student to have a blank floppy disk. Module 3 Review Answers What is the function of the ChemStation Configuration Editor? _ The ChemStation Configuration Editor detects the HP-IB interface in your PC and uses the instrument configuration information that you provide to enable the ChemStation to communicate with analytical hardware through the HP-IB bus. Where are chromatographic data files stored? In the path C:\HPChem\1\data Why should one keep the ChemStation CD-ROM readily available? In addition to the actual software, there are directories containing user information like installation manuals and user contributed libraries of macro programs. Also, if it is necessary to reload the software, the CD is required. 4. How does one view the contents of the ChemStation CD-ROM? Insert the CDROM and start Explorer. Double-click on the CD-ROM drive icon. How often should files be backed up? Files should be backed up on a regular basis. Lab personnel should determine the frequency and a procedure should become part of the standard operating procedures. Is it possible to back up the PC? _Yes, there are utilities that enable one to back up the entire hard drive and enable the PC to be restored in case of a major failure. What is the Emergency Recovery Kit? _An Emergency Recovery Kit is provided with the PC when it is shipped. The ChemStation comes with a CD-ROM and floppy disk that can be used to restore the hard drive in case of a major failure. ChemStation CD-ROM ChemStation CD-ROM ChemStation CD-ROM ChemStation CD-ROM ChemStation CDROM Module 4 Summary and Review Instructor Notes: This module can be treated as an interactive lecture/demo with the instructor performing the lab exercise with the students and giving pertinent explanations. If the computers are in a classroom environment, the demo is logistically straightforward. The Proxima or NEC projector system can be used to display the computer screen to the front of the classroom. If the PC’s are stand-alone, without a GC, the demo can be performed. It is necessary to copy the GC Instrument configuration and registration files from the Instrument Subdirectory from an active GC system to the Instrument Subdirectory of the stand-alone PC. The required files are: 6890gc1 Conf_off Config Hp68qk00 I7673.cfg Iq1.cfg New in Rev. A.06: GC Companion! In order to demonstrate the GC Companion View, the optional Companion software module must be loaded on the Instructor’s PC. There is an optional lab exercise in Module 5 which includes instructions for loading the software and demonstrates the use of the Companion. The Companion Lab can be performed on a Stand-alone PC. If the computers are in a lab environment, the instructor may need to be a little more resourceful in order to perform the demo/interactive lab. The Student Notes contain suggested activities as well as explanatory notes. An activity includes completing the “Tour the ChemStation” segment of the on-line tutorial. Module 4 Review Answers What is the difference between the Full Menu and Short Menu views? The Full Menu lists all the functions available for creating a method or sequence and performing an analaysis. The Short Menu lists functions appropriate for viewing parameters and performing an analysis. When the ChemStation software is first loaded, the default view is the Short Menu. What type of security features are available on your ChemStation? The log-on options for the ChemStation can be set so that one logs on as an operator or a manager. The operator only has access to the Short Menu view whereas the manager has access to the Full Menu. The user should maintain a record of the passwords used. For those using an NT operating system, additional security features are available within NT. What is Method Resolution? Method Resolution occurs when a new method is loaded and the method that was loaded did not match the GC’s current configuration. The ChemStation suggests reasonable revisions to resolve the configuration differences. Module 5 Notes and Review Instructor’s Notes: Module 5 covers Data Acquisition on the 6890 Creating a Method Setting up Acquisition Parameters The lab exercises address most of the screens which are reviewed in this module. Based on time constraints and the availability of PC’s in a classroom environment, the instructor may opt to simply point out the information contained in pages 2 through 61 and let the students proceed immediately to the lab exercises. The notes on pages 2 through 61 should be used as reference as the labs are completed. Lab 1: Creating a Method for the Split Inlet/FID Lab 2: Creating a Method for the Packed Inlet/TCD Lab 3: Making a Run with the Packed Method Lab 4: Making a Run with the Split Method Lab 5: GC Companion Verify that the method Dualcol has been loaded in the Method subdirectory of the PC’s. This method is for a 30m x 0.32 mm x 0.25 um HP-5 (PN 19091J-413) installed in a split inlet and FID and a 30 m x0.53 mm x .88um HP-1 (PN 19095J-023) installed in a packed inlet and TCD. If a stand-alone computer is used, this lab can be performed if the Instrument Configuration files have been loaded in the path: C:\hpchem\1. Make sure that the configuration editor has been set for a 6890. Initiate an On-line Session and ignore the error of a 6890 not being found. Obtain samples labeled BGC-1 and Hexane. BGC-1 contains 0.3% each of C14, C-15, and C-16 in C6 (Hexane). Information on ECD, NPD, and FPD is found on pages 53 through 61. New in Rev. A.06: Optional Lab 5 - GC Companion! Lab 5 requires the use of data files that are provided on the Supplemental Floppy disk. Module 5: Review Answers What are the primary parameters, which are required to be set before performing an analysis? Column information, inlet, autoinjector, detector, signal and reporting parameters. Name two ways to access the Instrument Parameters screen in order to set the Inlet, Column, and Detector parameters. In the Instrument menu, Edit Parameters or in the Method menu, Edit Entire Method. Also, the screen appears during Method Resolution. If you are new to the ChemStation, what is the best way to create a new method? Load the default method and select Edit Entire Method. parameters will be addressed. All of the key What is meant when the ChemStation asks if you would like to “Install a column as Col 1 or Col 2”? The ChemStation is asking you to identity the columns which you just installed or are already installed. Why is it important to know the column dimensions as accurately as possible? The column flow, linear velosity, and pressure are determined based upon calculations dependent upon the column dimensions. Does the ChemStation know acceptible ranges for various parameters? What does it mean if a parameter value shows up “red” in the Edit Parameters Screen? Yes, the flows, pressures, and temperature values must fall within acceptible ranges. If they do not, the parameter setting is show in “Red” and one is not able to leave the edit parameters screen until the value is adjusted within an acceptible range. Can the size of the peaks or the time scale of the signal plot be changed graphically during a run? Yes, select the arrows on the bottom corners of the plot window. See the notes on pages 39 and 40 of Module 4 for detailed explanation of the functions. Does changing the graphic picture of the peaks on the chromatogram change the amount of the peaks reported in the analysis report? No, it just changes the picture viewed. Module 6 Notes and Review Instructor Notes: Module 6 covers Data Analysis. Loading Signal files Setting Graphics Options Optimizing Integration Introduction to Manual Integration Level 1 Lab Lab 1: Introduction to Data Analysis Level 2 Lab Lab 2: Introduction to Manual Integration The labs in this module use Data files and Method files that are provided with this material. Data files: CS_01, CS_02, CS_03, CS_04, CS_05, CS_06, CS_07, Unk_1, Unk_2 Method files: Std_Pack.m and Std_spli.m (these are methods using the Standard Integrator). The labs in this module can be performed in an Off-line session and on Stand-alone PC’s. Lab 1 compares the results using the Standard versus the Enhanced Integrator. Though a very small difference in the integrated area, there is a difference of the computed area when using the standard versus the enhanced integrator. The instructor should note that for samples of smaller concentration, a more significant difference would be noted. Module 6 Review Answers What is the value of using an Off-Line Session? One can perform data analysis using an Off-line session while collecting data with an On-line session. How can peak names be automatically printed on the chromatogram report? What is the value of graphic annotation? If a calibration table is used, peak names are included in the table. In the Graphics, Signal Options screen, “checking on” peak names will allow them to be automatically printed on the chromatogram with each analysis. Annotations are manually added to a chromatogram in data analysis. The annotations will only occur for the data file selected for analysis. Annotations cannot be automatically added. When should one use the Standard Integrator versus using the Enhanced Integrator? The Enhanced Integrator is the new and more sensitive integrator. If new methods are created, this integrator should be used. The older, Standard Integrator has been available on earlier revisions of software. If methods have been created using the Standard Integrator and data collected is referenced to a historical data base, then that integrator should be used. When should one use the AutoIntegrate function? Before using timed integration events or manual integration, one should select AutoIntegrate. How and why are integration events used? If after performing Auto-Integration, the integration requires further optimization, then one should use the integration events available. The on-line Help offers definitions of the various integration events. It is recommended that if five or more events are used, one should reevaluate the chromatographic parameters and verify that they have been optimized. Integration events should not be used to overcome poor chromatography! What is the function of the “solvent peak off” integration event? Solvent Peak Detection On/Off: Peaks above a specific slope in units of mV/s are detected as solvent peaks, that lie outside the range of the analog-to-digital conversion. The trailing peaks are automatically tangent-skimmed; you don't need to switch on the tangent skim event. If solvent peak detection is off, droplines are drawn from the trailing peak instead of tangents. This function does not eliminate the solvent peak from being integrated, but defines how the peaks riding on the tail of a peak are handled. When should manual integration be used? Manual integration should only be used when an analyses results are an exception to the norm and the timed integration events did not achieve the needed results. Module 7 Notes and Review Instructor Notes: Module 7 covers Inlets. Fundamentals of Inlets Lab Exercises Lab 1: Split Ratio Determination Module 7 requires the instrument configuration and samples used in Module 5. Module 7 Review Answers In Lab 1, how did the manually calculated flow rates compare with the flow rates determined by the ChemStation? What might be the reason for the difference in the values? The manually calculated values were slightly different from those determined by the ChemStation. The 6890 has corrected values for atmospheric pressure conditions. In the manual formulas, we did not take that into consideration. Was there a significant change in the column flow rate when the Split Flow was changed? Why? No, changing the split vent flow should only change the area of peaks, not the retention times. the When the Split Flow was changed to 50 mL/min., what happened to the Split Ratio, area counts and retention time of C-16? The Split Ratio is reduced. The area counts increased since less sample was thrown away. The retention time should not have changed significantly. When the Split Flow was changed to 200 mL/min., what happened to the Split Ratio, area counts and retention time of C-16? The Split Ratio is increased. The area counts decreased since more sample was thrown away. The retention time should not have changed significantly When the Column flow was changed to 2.5 mL/min, did the split vent flow or split ratio change? Why? The split vent flow changed slightly since the total flow was not changed. Some of the split vent flow is now going to the column. The split ratio also changed since both column and split vent flows are in the formula for the split ratio calculation. Why would it be necessary to change the Split Ratio? To minimize the amount of sample allowed on the column in order to prevent column overload. Or to increase the amount of sample on the column in order to detect very small concentrations. Should one change the Split Ratio by changing the column flow rate? Why? No, one should only change the split vent flow. Changing the column flow also changes the retention times. Why is it necessary to save the method with a new name if parameters are changed? If one wants to go back to the old set of parameters, it is necessary to use a new name to distinguish the new set of parameters. Since heat is used to vaporize the sample in the injector, why not keep the inlet at 400 degrees? High inlet temperatures will degrade the septum and o-ring more rapidly. High temps may also decompose the sample. Module 8 Notes and Review Instructor Notes: Module 8 covers Columns. Fundamentals of Column Separation Lab Exercises Lab 1: Column Efficiency and Resolution Lab 2: Column Selectivity Lab 3: Oven Temperature and Column Compensation Lab 4: Retention Time Locking Module 8 requires each GC to have the following columns: 2 each PN 19095L-021 15 m x 0.53-mm x 1.0-um HP 50+ 1 vial of hydrocarbon mix labeled BGC-2 containing 0.3% each of C12, C-13, C-14, C-15, and C-16 and 0.15% each C-11OH and C-12OH in hexane. 1 vial of hexane Since Rev 6: Retention Time Locking Lab Exercise 4 Lab 4 requires the Retention Time Locking software and the use of data files that are provided on the Supplemental Floppy disk. Module 8 Review Answers 1.Why would one routinely monitor the number of theoretical plates or plates per meter for a column? Columns naturally loose plates over time. Monitoring the plates is a way of tracking the column’s deterioration over time. 2.What are the relationship between column diameter and efficiency? The more narrow the diameter, the better the efficiency. 3. In Lab 1, which column was more efficient? Which had better resolution? The 0.32 column was more efficient and had better resolution than the O.53 column. 4.In Lab 1, how would you classify the plate per meter values obtained: High? Medium? or Low? How might the efficiency of either column be improved? (Hint: what type of injection ports and flow rates were used? The values are much lower than one would expect for columns of those diameters. Most 0.53 columns would have at least 1000 plates per meters and 0.32 columns about 2000 plates per meters. The 0.53 column is installed in a packed inlet. Column overload is exhibited and negatively effects the efficiency. Performance factors for these columns with these analyses were not optimized, thus, the calculated plates per meter values do not represent the optimum. 5.What is the usefulness of the Van-Deemter Plot? The optimun linear velosity or column flowrate can be determined using the Van Deemter plot. 6.When would it be critical to carefully maintain the optimum linear velocity? When performing trace analyses and resolving very complex mixtures it is important to have the sharpest peaks, most optimum efficiency. 7 . G i v e n t h e f o l l o w i n g c o l u m n s , w h i c h w o u l d y o u e x p e c t m i g h t g i v e t h e b e s t e f f i c i e n c y a n d r e s o l u t i o n ? W h y ? 5 m x 0 . 5 3 m m x 2 . 6 5 mðm M e t h y l S i l i c o n e 2 5 m x 0 . 3 2 m m x 0 . 2 5 mðm M e t h y l S i l i c o n e 5 0 m x 0 . 2 m m x 0 . 3 3 mðm M e t h y l S i l i c o n e T h e l a s t one would theoretically achieve the best efficiency since it is the most narrow, and the best resolution since it is the longest. 8.Given the following columns, which would you use when analyzing a sample containing an unknown number or types of components? 25m x 0.2mm x 0.33 mðm M e t h y l S i l i c o n e ( H P - 1 ) 2 5 m x 0 . 2 m m x 0 . 1 7 mðm 5 0 % P h e n y l M e t h y l S i l i c o n e ( H P - 5 0 ) 2 5 m x 0 . 2 m m x 0 . 2 mðm C a r b o w a x 2 0 M ( H P 2 0 M ) O n e w o u l d u s e a t l e a s t t w o c o l u m n s o f d i f f e r e n t p o l a r i t y s t a t i o n a r y p h a s e t o e n s u r e s e p a r a t i o n o f a l l c o m p o n e n t s . T h i s p r o c ess is called dual column confirmation. Using three different stationary phases would provide even more confidence in separating all components. It would not matter which column was used first, since using all would be the best approach. 9.What is the likelihood that one or more of the above columns will provide the necessary analysis? Very likely. 10.Does the type of liquid phase effect the elution order of unsubstituted aliphatic hydrocarbons (i.e., C-12 through C-16)? No, straight chained hydrocarbons elute in order of carbon number. 11. In Lab 2, what happened to the elution order of the components when the type of stationary phase was changed from a non-polar to a medium polar stationary phase? The alcohols, which are polar, eluted later, relative to the straight chain hydrocarbons. They were retained longer on the more polar stationary phase. 12.In Lab 2 we had to edit the Instrument Parameters to update the column description when we changed columns. Why is it necessary to identify the correct column? The chemstation software and GC require accurate knowledge of the column dimensions in order to calculate the pressure- linear velocity-flow relationship. 13. If your column is not the length stated on the tag or is not identified in the list of column types, how do you update the method to include the correct column information? If the column length is within 5 meters of the original tag length, the new length can be edited in the “Calibrate currently installed column” option. If the column has changed more than 5 meters or is a column not listed in the list, one has to “Add new column to Inventory” and “Add new column model to catalog” typing in all the column parameters. 14.The corrected retention time of component X is 2.4 min on a 10m x 0.2mm x 0.33 s w r 0 A v t t a h e . b o h w m a t 1 o l e i e t e 7 u u f w n t mðm t m e s a c e mðm c o l u m n . W i t h t h e l o w r a t e a n d o v e n t e m p e r a t u r e , o u l d b e t h e e x p e c t e d c o r r e c t e d i o n t i m e o n a 2 0 m x 0 . 2 m m x c o l u m n ? t h e s a m e s i n c e t h e t o t a l o f s t a t i o n a r y p h a s e i s a b o u t m e i n b o t h c o l u m n s . O n e i s t h e l ength, but half the film thickness of phase. 15.What happens to the peak shape of later eluting peaks as the oven temperature is reduced? The peaks become broader. 16.What are the key advantages of temperature programming? Run times are usually shorter and the peaks are sharper thoughout the analyses. 17.What is the disadvantage of temperature programming? Baselines usually drift up as the temperature gradient becomes greater. 18.Will temperature programming improve the resolution of components that are difficult to separate? Maybe. It may be necessary to change the selectivity (type of stationary phase) if temperature programming does not work. What is the benefit of performing column compensation? Column compensation will electronically subtract the drifting background baseline from the chromatographic baseline, producing a flat baseline which yields more accruate quantitation. Module 9 Notes and Review Instructor Notes Module 9 covers Fundamentals of Detectors Thermal Conductivity Detector Flame Ionization Detector Lab Exercises Lab 1: Thermal Conductivity Detector Performance Lab 2: Flame Ionization Detector Performance The following samples are required for the lab exercises: BGC-3 containing 0.3% each of C-12, C-13, C-14, C-15, and C-16 in Hexane. Information on ECD, NPD, and FPD is found on pages 53 through 61 of Module 5. Module 9 Review Answers 1. Predict what would happen to the retention time and area counts if each of the problems below occurred. Use an "up" arrow to indicate an increase, a "down" arrow to indicate a decrease, "---" to indicate no change, and "NS" to indicate no signal. Problem Retention Time Area Counts FID Make-up Gas Off ------( Hydrogen Flow Increase ------( or ( Hydrogen Flow Decrease ------( or ( Air Flow Decrease ------( TCD Makeup Gas Off ------( TCD Make-up Gas Increase ------( TCD Reference Gas Off ------NS TCD Reference Gas Increased -----( TCD Reference Gas Decreased ------( Leak at TCD Column Nut ------( 2. Indicate which detector would be the best choice for the following analyses. TCD FID 200 ppm H2O ( 2% Butane ( ( 1 ppm Propane ( 150 ppm CO2 ( 1% Argon ( 12% Ethanol ( ( 3 ppm CO ((w/methanizer) 100 ppm Helium ( 20 ppm Helium ((optimized) 3. What is the function of the make-up or auxiliary gas? Capillary columns do not deliver as much carrier gas to the detector as packed columns do. Auxillary gas makes up the difference between what the column delivers and what the detector expexts to see. It affects the sensitivity of the detector response. 4. Based upon the responses in Lab Exercises 1 and 2, what is the significance of the response factors? How does the FID sensitivity compare to the TCD? The response factors are used to calculate the concentration of the components in an unknown mixture. The detector signal is related to a known concentration. In this case, the quantitative value is a very small number. 5. Why don't we normally use a TCD with narrow bore capillary columns? The sensitivity of the detector is quite low. 6. List the detectors in order of increasing sensitivity: FID, TCD, and ECD. TCD, FID, and ECD is the most sensitive. 7. What are the primary advantage and disadvantage of using the TCD? The TCD will respond to any component that is different from the carrier gas, but it is the least sensitive of the detectors. 8. What is the significance of the "clicking" sound of the TCD? The solenoid switching valve regulating the reference gas makes the clicking sound when the TCD electronics is turned on. 9. Why is it critical to properly set the air, hydrogen, and carrier gases for a FID? The ratio of flows affects the sensitivity response of the detector. 10. What is the significance of the noting the background signal output of the detectors? The background signal can be used to monitor the cleanliness of the detector and gases. If the system is contaminated, the background signal is elevated. It is a good idea to regularly note the signal. 11.What GC approach would you use to look for PCBs in transformer oil? The Electron Capture Detector is very sensitive to halogenated compounds. PCB’s are polychloro-bi-phenyls. 12.Why does the ECD require very pure carrier gas and why should it be maintained free of oxygen contamination? The ECD is very sensitive to oxygen. Module 10 Notes and Review Instructor Notes: Module 10 covers Calibration. Setting Up Single Level Calibrations Setting Up Multi-Level Calibrations Explanation of Calibration options Level 1 Labs Lab 1: Creating a Single Level Calibration in an On-Line Session Lab 2: Creating a Single Level Calibration in an Off-Line Session Lab 3: Creating a Multi-Level Calibration in an On-Line Session Lab 4: Creating a Multi-Level Calibration in an Off-Line Session Level 2 Labs Lab 5: Working with Calibration Settings Labs 2, 4, and 5 should be performed after completing the labs in Module 6. The methods created in Module 6, Lab 1: Dalab1s.m and Dalab1e.m will be used. They are available on the floppy disk, which accompanies this manual. Data files are also used in these labs: CS_01.d through CS_07.d and Unk_1.d and Unk_2.d. Labs 1 and 3 require samples to be analyzed using the Split Inlet and FID. These labs should be performed after Module 8 Labs have been completed. The 15m x 0.53 x 0.53 HP-50 should be column installed. Samples required: BGC-4 containing 0.3% of C-11 OH (undecanol) and 0.15% C-12 OH (dodecanol) in hexane. BGC-5 containing an unknown amount of C-11 OH with 0.15% C-12 OH added as the internal standard in hexane. Hexane solvent rinse. Lab 1 Discussion The Summary Table results on page 50 of Lab 1 should be compared for the class in order to determine the most accurate determination of the unknown amount. The most consistent results should be from the ISTD report. A Results table can be put on the white board and the various groups can record their values. Below is a suggested format for the table. Group 1 Group 2 Group 3 Group 4 Group 5 Area % Norm % ESTD ISTD Lab 2 Results Part 2: Calibration using the Enhanced Integrator Summary Report Method Unk_1.d C-11 OH Reported Amount Unk_2.d C-11 OH Reported Amount Callab2s (Stand Inte-ESTD-Hi Conc) 81.85590 -------------------- Calab2s2 (Stand Inte-ESTD -Lo Conc) ------------------0.53008 Callab2e (Enhan Inte-ESTD - Hi conc) 81.85411 --------------------- Calab2e2 (Enhan Inte-ESTD-Lo Conc) -------------------0.589134 Complete next section after Part 3 Calab2e3 (Enhan InteISTD- Lo Conc) ------------------- 0.50427 Calab2e4 (Enhan Inte-Norm%LoConc) ------------------8.266 % Lab 4: Part 2 Summary Table Method Unk_1.d C-11 OH Reported Amount Unk_2.d C-11 OH Reported Amount Calab2e 81.8541 ----------------------------------- Calab2e2 ------------------------------0.589134 Calab4e 84.16208 ------------------------------------Calab4e1 84.26995 1.43299 Lab 5 Summary Table Method Data File Amount of C-11 OH Calab4e1 (Linear, Force the Origin) Unk_2.d 1.43299 Calab4e1 (Linear, Force the Origin) CS_04.d 4.17733 Calab4e2 (Piecewise, Ignore the Origin) CS_04.d 1.50000 Calab4e2 (Piecewise, Ignore the Origin) Unk_2.d 0 Calab4e3 (Piecewise, Force the Origin) Unk_2.d 0.514558 Calab4e3 (Piecewise, Force the Origin) CS_04.d 1.50000 Module 10 Review Answers 1.Describe the steps needed to prepare a single level calibration table. Optimize the instrument and data analysis parameters (the method). Analyze the standard mixture. Create the Calibration Table. Save the method. 2. Describe the steps needed to prepare a multi level calibration table. Optimize the instrument and data analysis parameters (the method). Analyze the standard mixture with the lowest concentration. Create the Calibration Table with the first level amounts. Analyze the standard with the second concentration amounts. Add the Level to the Calibration table. Repeat the procedure until all levels are added to the table. Save the method. 3. Explain the following Calibration Setting parameters (Hint, you may use the ChemStation Help to find the definitions): Reference Window: The Retention Time windows are used by the ChemStation software to identify peaks in the integration results. By specifying a window rather than a specific retention time, the ChemStation can compensate for slight shifts of retention times from one run to the next. The used windows are a sum of an absolute part in minutes and a relative part given in percent of the expected retention time. The percentage part compensates for variations in retention time that are proportional to the expected retention time. The absolute part compensates for variations that are independent from retention time. The absolute part specifies the complete RT variation both negative and positive. Example a RT window of 5 specifies a deviation from -2.5 to +2.5. Multiplier A number by which all calculated results are multiplied before the report is printed. You can use the multiplier to change the scale of the results or correct for changes in sample composition during pre-analysis work. The multiplier can also be used for any other purposes, which requires the use of a constant factor. The multiplier is used for ALL report types. RF Uncal Peaks Allows you to define the response factor for unidentified compound peaks in each individual signal. Activate the For Signal drop-down list, select the signal to set up and define the response factor in one of the following ways: Using Compound Use this option to apply a response factor of a calibrated compound to unidentified peaks. If a calibration table is created, you can select the required compound from the combo box. With Rsp Factor Enables you to specify a response factor that is used in the calculation of uncalibrated peaks. This fixed response factor is not corrected during recalibration. With ISTD If you have specified a response factor and are doing an ISTD calculation with ISTDs set up in the calibration table, you can also select an ISTD compound from this combo box. This enables you to apply the ISTD correction to the response factor. No The unknown peaks are not reported. Note:To see any results of these calculations, ensure that Do Not Report is not selected in the Specify Report dialog box. Sample Defaults Allows you to enter data for off-line analysis. This default sample information is ignored if the "From Data File" option is selected in the Use Sample Data dialog box. Amount The amount of sample to be analyzed. The amount is necessary to calculate an ESTD% or ISTD% report. In these types of report the results are calculated as a percentage of the injected sample. Amount Units The units in which all amounts of the sample are measured, for example, ng/ul Multiplier A number by which all calculated results are multiplied before the report is printed. You can use the multiplier to change the scale of the results or correct for changes in sample composition during pre-analysis work. The multiplier can also be used for any other purposes, which requires the use of a constant factor. The multiplier is used for ALL report types. Dilution A number by which all calculated results are multiplied before the report is printed. You can use the dilution factor to change the scale of the results or correct for changes in sample composition during pre-analysis work. You can also use the dilution factor for any other purposes that require the use of a constant factor. The dilution factor is used for ALL report types. ISTD Amount The amount for each Internal Standard compound. The amount that is entered here is used as default if no sample information exits. Sample ISTD Information Specifies the known internal standard amounts in the unknown sample. Several ISTDs can be used. I# Identifies the internal standard compound. This I# corresponds to the I# in the Calibration Table. Default Calibration Curve Curve Type There are various possible curve-fit calculations for use with multiple- level calibration: Piecewise, Linear, Quadratic, Cubic, Exponential, Logarithmic, Power, Average Rsp/Amt Origin There are various ways to treat the origin when the response curve is plotted, you can ignore origin, include origin, force origin or connect origin. The default values are linear and include origin. The choices for the treatment of the origin are: Ignore Origin: The origin (0,0) is not used in the curve calculations. Include origin: The origin is used as one of the calibration points. Force Origin: The curve is forced through the origin: For example, y=mx y=ax^2+bx or y=ax^3+bx^2+cx Connect Origin: The linear segment is constructed between the origin and lowest calibration amount on the curve. Weight When the calibration curve is generated, you can specify the relative weighting (or importance), of the various calibration points. The weighting options are as follows: Equal: All calibration points have equal weight in the curve. Linear: A calibration point with amount x has the weighting 1/x normalized to the smallest amount. Quadratic: A calibration point with amount x has the weighting 1/x^2 normalized to the smallest amount. # Calibrations: A calibration point has weighting according to the number of calibrations for this point. For example, if level 1 has been calibrated twice, then the points on level 1 have weighting of two. No normalization is done. What concentration or amount values should your calibration standards have in reference to your unknown sample concentrations or amounts? The calibration standard amounts should bracket the range of concentrations expected in the unknowns. The lowest standard amounts should be lower than the lowest expected unknown amount and the highest standard amounts should be higher than the highest expected unknown amount. How important is curve linearity in reference to calculated accuracy? Very important, the more linear the curve, the more accurate the calculated value. How does one determine which set of curve fitting options is the best to use? Trial and error process selecting the various curve fitting options and determining which calculations match known standards analyzed as unknowns. How does one know that the calculated result is accurate? Analyze standard mixtures as unknowns to verify the accuracy of the method. Module 11 Notes and Review Instructor Notes Module 11 covers Sequences. Setting Up Sequences Setting Up Calibrated Sequences Understanding Batch Review Level 1 Labs Lab 1: Creating a Sequence for On-Line Analyses Lab 2: Creating Sequences with Reprocessing Lab 3: Creating a Dual Tower Sequence Level 2 Labs Lab 4: Creating a Calibrated Sequence Lab 5: Performing a Batch Review The method and data files used in this lab were created in previous modules and are also available on the floppy disk which accompanies this manual. Labs 1 requires samples to be analyzed using the Split Inlet and FID. This lab should be performed after Module 10 Labs 1 and 3 have been completed. The 15m x 0.53 x 0.53 HP-50 should be column installed. Samples required: BGC-4 containing 0.3% of C-11 OH (undecanol) and 0.15% C-12 OH (dodecanol) in hexane. BGC-5 containing an unknown amount of C-11 OH with 0.15% C-12 OH added as the internal standard in hexane. Hexane solvent rinse. Lab 4 Summary Table Data File Recal.s C-11 Amount Recalexp.s C-11 Amount Recalint.s C-11 Amount Recalbrk.s C-11 Amount Recal_01 150.7 150 Sample 1 84.1 83.7 84.1 84.1 Sample 2 0.59 0.59 0.59 0.59 Recal_02 28.3 150 Sample 3 4.2 22.1 4.2 4.2 Sample 4 0.59 3.12 0.59 0.59 Recal_03 15.0 150 Module 11 Review Answers When would one use a sequence for performing analyses? When automating analyses with multiple injections per vial or multiple methods. with What are the steps used to create a sequence? The method or methods used must be created and optimized. The sequence table is then created specifying the vials and methods to be used. If one updates a method, is it necessary to update the sequence? If the method has the same name as the one specified in the sequence, then it is not necessary to update the sequence table. When the sequence is started the method designated in the table will be loaded. If the method was saved with a different name, then the sequence table would require updating with the new method name designated. Why should one create subdirectories in the path: \\hpchem\1\sequence? Whenever the sequence is started, the data files will be saved in the subdirectory specified in the Sequence Parameters. There is the potential that data files can be written over when another sequence is begun. How would one designate the analysis of only vials 3,6,9, and 12 in a sequence that calls for the analysis of 25 vials? Use the Partial Sequence option to designate the vials to be run. What are three options for sequence recalibrations? Explicit recalibration Interval recalibration Bracketed recalibration Which option is probably the most accurate? The Bracketed recalibration option is theoretically the most accurate since the response factors are calculated using the results of the calibration standards run before and after the sample analyses. Module 12 Notes and Review Instructor Notes Module 12 covers Reporting. 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<xsd:enumeration value="Workstations"/> <xsd:enumeration value="X-ray Source"/> <xsd:enumeration value="ZORBAX"/> </xsd:restriction> </xsd:simpleType> </xsd:element> </xsd:sequence> </xsd:extension> </xsd:complexContent> </xsd:complexType> </xsd:element> <xsd:element name="Product" ma:index="26" nillable="true" ma:displayName="Product" ma:hidden="true" ma:internalName="Product"> <xsd:complexType> <xsd:complexContent> <xsd:extension base="dms:MultiChoice"> <xsd:sequence> <xsd:element name="Value" maxOccurs="unbounded" minOccurs="0" nillable="true"> <xsd:simpleType> <xsd:restriction base="dms:Choice"> <xsd:enumeration value="G2912A"/> <xsd:enumeration value="G2613A"/> <xsd:enumeration value="G2614A"/> <xsd:enumeration value="G2630A"/> <xsd:enumeration value="G2630B"/> <xsd:enumeration value="G2855B"/> <xsd:enumeration value="G2888A"/> <xsd:enumeration value="G2913A"/> <xsd:enumeration value="G2916A"/> </xsd:restriction> </xsd:simpleType> </xsd:element> </xsd:sequence> </xsd:extension> 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<xsd:extension base="dms:MultiChoice"> <xsd:sequence> <xsd:element name="Value" maxOccurs="unbounded" minOccurs="0" nillable="true"> <xsd:simpleType> <xsd:restriction base="dms:Choice"> <xsd:enumeration value="Air"/> <xsd:enumeration value="Analysis of Process Chemicals"/> <xsd:enumeration value="Antibacterial Drug Residues"/> <xsd:enumeration value="Bioanalysis"/> <xsd:enumeration value="Biological Warfare Agents (BWA)"/> <xsd:enumeration value="Chemical Warfare Agents (CWA)"/> <xsd:enumeration value="Components Analysis"/> <xsd:enumeration value="Contamination Control"/> <xsd:enumeration value="Environmental Monitoring"/> <xsd:enumeration value="Flavors"/> <xsd:enumeration value="Food"/> <xsd:enumeration value="Fragrances"/> <xsd:enumeration value="Natural Compounds & Additives"/> <xsd:enumeration value="Pesticides & Residues"/> <xsd:enumeration value="Silicon Wafer Analysis for Organic Contaminants"/> <xsd:enumeration value="Soil & Sediment"/> <xsd:enumeration value="Toxic 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<xsd:import namespace="http://purl.org/dc/elements/1.1/" schemaLocation="http://dublincore.org/schemas/xmls/qdc/2003/04/02/dc.xsd" /> <xsd:import namespace="http://purl.org/dc/terms/" schemaLocation="http://dublincore.org/schemas/xmls/qdc/2003/04/02/dcterms .xsd"/> <xsd:element name="coreProperties" type="CT_coreProperties"/> <xsd:complexType name="CT_coreProperties"> <xsd:all> <xsd:element ref="dc:creator" maxOccurs="1" ma:index="9" ma:displayName="Author"/> <xsd:element ref="dcterms:created" minOccurs="0" maxOccurs="1"/> <xsd:element ref="dc:identifier" minOccurs="0" maxOccurs="1"/> <xsd:element name="contentType" minOccurs="0" maxOccurs="1" type="xsd:string" ma:index="35" ma:displayName="Content Type"/> <xsd:element ref="dc:title" maxOccurs="1" ma:index="1" ma:displayName="Title"/> <xsd:element ref="dc:subject" minOccurs="0" maxOccurs="1"/> <xsd:element ref="dc:description" minOccurs="0" maxOccurs="1"/> <xsd:element name="keywords" maxOccurs="1" ma:index="7" 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