SWIFS Controlled Substances Resources Documents v2.0 - reformatted 61 pages.pdf

8/6/2019 SWIFS Controlled Substances Resources Documents v2.0 - reformatted 61 pages.pdf

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Controlled Substances Resource Documents


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Under development


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Texas Controlled Substances and Related Statutes

Click the following link and refer to Sections:481 Texas Controlled Substances Act482 Simulated Controlled Substances

483 Dangerous Drugs485 Abusable Volatile Chemicals

http://tlo2.tlc.state.tx.us/statutes/hs.toc.htm
http://tlo2.tlc.state.tx.us/statutes/hs.toc.htmhttp://tlo2.tlc.state.tx.us/statutes/hs.toc.htm


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GAS CHROMATOGRAPHY AND

GAS CHROMATOGRAPHY/MASS SPECTROMETRY

Gas Chromatography (GC)

Chromatography is a physical method of separation that involves the differential distribution of asubstance between two phases: one phase is a stationary bed of large surface area and the otherphase is a mobile phase, which percolates through the stationary bed. In GC, the mobile phase isa gas, and in order for a substance to be amenable to GC analysis, it must be capable of beingvolatilized in the GC inlet.

The chromatographic separation process occurs as a result of repeated sorption-desorption of asubstance between the stationary and mobile phase as the substance travels along the stationarybed. Separation of two or more substances occurs when the distribution coefficients between themobile and stationary phases are different for the substances. As a substance elutes from the end

of a GC column, it is detected by a flame ionization detector, mass spectrometer, or otheranalytical detector.

In a flame ionization detector, the gas exiting the column is mixed with hydrogen and air andburned. Ions and electrons form in the flame and decrease resistance in the electrode gap. Thisallows current to flow in the detector circuit. Current flow is monitored and charted as achromatogram. The amount of current generated is dependent upon the type and number of ionsformed in the flame. Under consistent instrument conditions, a particular substance will burn inthe same manner; therefore, the amount of current generated is proportional to the amount of substance burned allowing quantitation of the material when compared to known standards. Toimprove accuracy and consistency of quantitation, an internal standard is often added to a sampleand the relative response between sample and internal standard is used for quantitation.

GC can also give a very good indication of the identity of a substance; although it does notusually provide conclusive identification of a substance. Under consistent instrument conditions,movement of a specific material through a GC column will take a constant period of time. Thus,the time it takes a substance to travel through a GC column (retention time) is consistent for aparticular substance. Therefore, retention time on the GC column provides a very goodindication of substance identity when compared to a standard material.

Gas Chromatography/Mass Spectrometry (GC/MS)

A mass spectrometer may be the detector for a GC; in this case, the instrumentation is calledGC/MS. Mass spectrometry provides a unique identification of the materials tested; it can alsobe used for quantitation. As a substance elutes from the GC and enters the MS, it is bombardedwith an electron beam. The molecule fragments into characteristic ions depending upon therelative strength of its chemical bonds and chemical structure. Positively charged fragments of the molecule are directed down the analyzer and impinge upon the detector of the MS.

Dallas County Institute of Forensic Sciences 1 GC/MS OverviewControlled Substances Laboratory Version 2.0


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Directions for Operation of the HP 6890 GC withG2070AA Chemstation

Users are referred to the appropriate instrument manuals for additional information.

There are two configurations for each instrument. One is ONLINE and one is OFFLINE . Theonline must be selected to run a sequence or to operate the instrument. The offline mode is tolook at data or reprint reports while the instrument is in operation. The offline mode looksexactly like the online mode; however, it will not run samples or control the instrument in anyway.

FID Detector: FID detectors are used on the instrument. The output should be constant andstable. This may change with instrument usage or with maintenance of the detector. If the flameis out, the instrument will automatically light the flame when a method is loaded. At this time,there is no need to manually light the flame. If the flame will not light, seek assistance from a

supervisor.

Pausing the sequence: If the sequence must be paused during an analytical run to change an airtank or for another purpose, simply choose RUN CONTROL from the menu bar and selectPAUSE SEQUENCE. When finished, resume the sequence by choosing RUN CONTROL fromthe menu bar and RESUME SEQUENCE. It will begin at the next line from where it stopped.The sequence does not stop immediately. It will not stop until the current sample run iscomplete.

Dallas County Institute of Forensic Sciences Operation of the HP 6890 GCControlled Substances Laboratory 1 Version 2.0


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Sequence Parameters Screen: This screen can be accessed through the menu bar underSEQUENCE.

The subdirectory is the name of the data path in which the computer will store the dataaccumulated from the sequence.

Enter the subdirectory information in the following format: month (3 letters), day (2 numbers),and letter (a, b, c); for example, SEP23A

Currently initials are not entered in the operator box. Everything else in this screen is a defaultvalue and should not be change or updated at this time. When finished simply hit the OK buttonto exit.



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Sequence Table Screen: This screen is where the user enters the sequence to be run and sampleinformation. This screen may be accessed by a single click of the left mouse button on thecarousel picture or by selecting it under the SEQUENCE item on the menu bar. The criticalinformation that must be present is the vial location, sample name, method name, inj/vial , andsample type . All other cells should be left blank. Any numbers in these blank cells could causethe sample to be injected improperly, calculated wrong or any number of other errors.

To select a line move the cursor to the line number cell on the screen. This will produce anarrow. Click the left mouse button on this arrow and it will select the entire line (darkening theline). Drag the mouse down any number of lines to select the entire group of lines. Once a lineor group of lines has been selected, the user may cut, copy, insert, insert vial range, or appendline.

The Insert button will insert a blank line above or in front of the line that you have selected.



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The Insert Vial Range button will prompt you to enter information. Type in the appropriateinformation and hit the OK button . DO NOT put any number in the injection volume box. This will override the method and inject that amount, in microliters, onto the column. If leftblank, it will default to the method for that information. The information that is inputted will beinserted above or in front of the line that has been selected.

The Append Line will put a blank line after the line that has been selected.

The Cut button will cut the selected line. It is stored until another action takes place, i.e.:another cut or a copy. It may be pasted into another place in the sequence once it has been cut orsimply discarded.

The Copy button will copy the selected line but leave the line in place. It may then be pastedinto another place in the sequence.

The Paste button will paste the selected cut or copied line above or in front of the newly selectedline.

At this point, if the sequence has been saved, select the Run Sequence button to start a run. If thesequence has not been saved, select the OK button to exit this screen and save the sequencebefore returning. If the sequence is already running and samples are added to the sequence thensimply select the OK button after the samples have been added. Then save the sequence (noneed for the save as function).



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Reports are printed as a sample run is completed using macros written into the method

parameters. If a report must be reprinted or reprocessed, use the following instructions:

Reprinting a report: If the instrument is running a sequence, use the offline mode of theinstrument. From the Data Analysis screen select:

Select File from the menu bar and select Load Signal1. In the Folders: box, selected the correct data path where the signal is

stored. For example, C:\hpchem\1\data\aug05c.2. In the File name: box, selected the correct data signal to be analyzed, by

clicking on the name with the left mouse button, or retype the filename of the signal desired to print. Select OK.

3. If the Integrate and print report after load button is selected, the report

will be automatically generated. If it is not selected, the chromatographcan be manually integrated utilizing the Chemstation software.

This will print out a formatted report. Another way to reprint a report is go into the offline modeand load the sequence for the report that you want to reprint. Select the partial sequence. At thispoint simply mark the line or lines that you want to regenerate and it will print the report in theproper format. This is the preferred way to reprocess a batch or reprocess more than a singlesample.



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GC Maintenance Schedule

Daily/with use Change wash vial solventsCheck gases

Weekly Change septumChange liner and o-ring

Monthly Backup data

As needed Change gold sealTrim columnChange gas tanksReplace columnClean split side arm

Clean FID and jetMaintenance activities should be consolidated to minimize instrument down time. For example,if it is time to change the gold seal and cut the column, it may be more prudent to change theliner and septum at that time or hold off changing the gold seal until the liner and septum arechanged.

The maintenance interval is not a rigid schedule and should be based upon workflow. Unlessthere is an immediate problem, a chromatographic run should continue to completion, and themaintenance performed at the end of a run. Where duplicate instrumentation exists, only oneinstrument at a time should be scheduled for preventive maintenance to avoid multipleinstruments being out of service at the same time.

Ultraclean Technique:

Liners, gold seals, and certain other parts must be handled using ultraclean techniques to avoidcontamination of the part with oils from the skin, plasticizers from plastic laboratory bench-coats, etc. Do not handle an ultraclean part with your hands; wear gloves or use a freshKimwipe. Lay the ultraclean part on a clean Kimwipe.

Changing the Wash Vial Solvent:

Discard waste solvent into the hazardous waste. Replace spent solvent in wash vials with freshsolvent(s) and replace the vials in the appropriate location on the instrument autosampler.

Changing the Liner and Septum:

Tools needed: inlet wrench and tweezers.

Dallas County Institute of Forensic Sciences GC Maintenance HP6890Controlled Substances Laboratory 2 Version 2.0


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NOTE: Turn off carrier gas; if the septum nut is removed without turning off the carrier gas, theglass wool plug may be blown out.

Use ultraclean technique at all times when handling the new liner.

1.

Cool the inlet to < 80o

C to minimize oxidation and prevent burns.2. Turn off the carrier gas on the GC by using the keypad on the front panel of the GC: Fromthe front panel on the GC, push the FRONT INLET button, scroll down using the button tothe Total flow line. Turn off the flow by pressing the OFF button. Once maintenance iscomplete, turn the flow back on by pressing the ON button.

3. Once the inlet is cool and the gas is off, remove the septum nut. Remove and replace theseptum. Reinstall and tighten the septum nut. These are not ultraclean parts but care shouldbe taken to avoid any unnecessary contamination.

4. Remove the nut covering the liner. Remove the old liner and discard.5. Using ultraclean technique, carefully slide an o-ring over the end of the new liner,

approximately from the top of the liner, and slide into the inlet. The glasswool plug

should be nearest the bottom of the liner.6. Replace and tighten the nut and turn the inlet temperature and carrier gas flow on.7. A blank should be run to bake out the new liner before analyzing samples.

Changing the Gold Seal:

Tools needed: appropriate screw driver (Phillips/Torx/etc.), 9/16 wrench, wrench

Note: The gold seal is located at the bottom of the inlet. It is accessed through the nut in theoven where the column comes out of the inlet.

Use ultraclean technique at all times when handling the gold seal.1. Cool the inlet to < 80 oC to minimize oxidation and prevent burns.2. Turn off the carrier gas flow (follow the procedure listed above for Changing the Liner and

Septum).3. Remove the column from the inlet. (Once the column is removed, the ferrule will need to be

replaced and the column cut.)4. Remove the insulator cup (if present) to reveal the nut housing the gold seal.5. Remove the nut and discard the used gold seal and washer.6. Insert the new washer and gold seal into the nut:

a. Insert the washer into the nut; the washer goes between the gold seal and the nut.b. Insert the gold seal into the nut with the grooves visible on top. (These grooves are theexits for the split gas during split injection and for the purge flow after a splitless injection.)

7. Reinstall the nut containing the gold seal and washer, and tighten.8. Replace the insulator cup over the nut (if present).9. Replace the ferrule and trim the column as described below.10. Reinstall the column and check for leaks with a leak detector. (If you turned off the carrier

gas, turn it back on and let it flow for a few minutes before you check for leaks.)11. Turn on the inlet temperature.



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12. A blank should be run to bake out the system before analyzing samples.

Trimming the Column and Installing a New Column (Inlet/Detector):

Tools needed: column cutting wafer, inch wrench

NOTE: Hydrogen and airflow to the detector must be turned off any time the detector end of thecolumn is removed from the FID unit.

Any time a column is removed from the inlet/detector, the ferrule should be replaced and thecolumn should be trimmed. The inlet end of the column may be cut several inches to removeactive sites and restore separation capacity.

1. Cool the inlet and detector to < 80 oC. This will minimize oxidation and prevent burns.2. Turn off the carrier flow following the procedures in Changing the Liner and Septum.3. Remove the column from the inlet and detector by loosening the column nuts.

4.

Remove the column nut(s) and ferrule(s). The ferrule, which is made of graphite, may bestuck to the nut; remove all ferrule particles.5. Slide a septum, column nut, and ferrule onto the free ends of the column (tapered end of the

ferrule should point away from the column nut, the flat side toward the nut).6. Cut two or three inches off the end of the column:

a. The column should be cut by scoring one side with a wafer and then snapping the columnat the score.b. Inspect this cut with a magnifying glass. This cut must be clean and contain no roughedges. If there are rough edges, repeat until a clean cut is achieved or seek guidance.c. Wipe the end of the column with a Kimwipe and methanol, or hexane.d. Place the column back into the inlet or detector. When installed, the column shouldprotrude 5 mm (4 mm to 6 mm) beyond the ferrule into the inlet. Use the septum as a guidefor this measurement.e. Tighten the column nut so that the column does not slide with a gentle tug.

7. The detector end of the column should be prepared in the same manner.8. Insert the column into the detector until it comes to a stop, finger tighten the nut, pull the

column back approximately 1 mm or as noted by the manufacturer. Use the septum as aguide for this measurement. Tighten the column nut.

9. Turn on the carrier gas, and check for leaks with a leak detector.

Conditioning the Column:

If conditioning is not done properly the column may be ruined.1. Allow the carrier gas to flow through the column for approximately one hour with the GC

oven at room temperature.2. Ramp the oven temperature at 10-15 degrees per minute to the final conditioning

temperature. The final conditioning temperature should be 10 degrees higher than themaximum oven temperature to be used in the method but should not exceed 10 degreesbelow the maximum operating temperature of the column as recommended by themanufacturer.



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3. Condition the column several hours or overnight.

Cleaning FID Jets or Replacement:

Refer to HP 6890 Series Gas Chromatograph Maintenance and Troubleshooting Manual.

Cleaning the Split Side Arm:

The split vent side arm is the exhaust for split gasses and compounds that are purged off duringan injection. This can become very dirty and without maintenance can cause deleterious results.

1. Cool the injector temperature and turn off the gasses.2. Remove the liner from the inlet as specified in the Changing the Liner and Septum section.3. Remove the autosampler tray, tower, fan cover, and the top rear instrument cover.4. Remove the split side arm from the inlet to the filter located at the rear of the instrument.5. Inspect the ends of the side arm to make sure they are not clogged. If they are, use an old

syringe to unclog the ends.6. Using vacuum, pull a solvent such as chloroform through the side arm and into waste.7. Repeat step 4 using a solvent such as methanol and then air to dry.8. Using a small brush or Q-tip dipped in chloroform clean out the inlet arm where the side arm

attaches.9. Reconnect the side arm to the inlet and the filter.10. Replace the covers and autosampler tray and tower.11. Change the liner, o-ring and septum.12. Turn on the temperatures and gases.13. Check for leaks.


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QUICK REFERENCEROUTINE CALIBRATION AND OPERATION

AGILENT AND HP 5973 MASS SELECTIVE DETECTOR

Standard Spectra Autotune:

For each day the MS is used a Standard Spectra Autotune shall be performed prior to routineinstrument operation and after instrument maintenance:

From the Instrument Control Screen select Instrument , select Perform MS Autotune ,select Standard Spectra Tune from the Select Tune Type menu (see attached). Tune isautomatically saved in the Stune.u directory. Review the Tune Report and file for futurereference. Criteria for the Standard Spectra Tune are as follows:

1. Low background: less than 150 peaks2. Low water and air: less than 10%

3.

Correct mass assignments 0.2 amu (69, 219, 502)4. Symmetrical, smooth mass peak shapes5. Consistent mass peak widths (0.55 0.1) (Default value of Drug MS is 0.55)6. Isotope mass assignments should be 1 amu greater than parent peaks7. Appropriate EM voltage: 1000-2800 electron volts. If the voltage is not within these

limits, review the history of the electron multiplier or consult a supervisor. The EMvoltage will increase over time as the source becomes dirty with use. Cleaning thesource should return the EM voltage to a normal operating level.

8. Mass 69 abundance should be 150,000-400,0009. Typical relative abundance:

- Mass 69 = 100 %

-

Mass 219 = 35-99%- Mass 502 = 2-5 %10. Proper isotope ratios:

- Mass 70/69 = 0.5-1.6 %- Mass 220/219 = 3.2-6.4 %- Mass 503/502 = 7.9-12.3 %

Dallas County Institute of Forensic Sciences GC/MS Quick ReferenceControlled Substances Laboratory 1/08

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Target percent relative abundances are set for certain other PFTBA masses. The system willcome as close as possible to the values shown:

- Mass 50 1 %- Mass 131 55 %

-

Mass 414 3.5 %

If the tune meets the criteria listed, the chemist who performed the tune will initial and circle theinitials. All other chemists using the instrument that day will review the tune for acceptance of data and initial.

Autotune:

For each week the MS is used an Autotune shall be performed to provide a baseline for instrument troubleshooting (a Standard Spectra Autotune should be performed after the Autotuneand before routine instrument operation):

From the Instrument Control Screen select Instrument , select Perform MS Autotune ,select Autotune from the Select Tune Type menu (see attached). Tune is automatically savedin the Atune.u directory. Review the Tune Report and file for future reference. Acceptablecriteria for the Autotune are as follows:

1. Low background: less than 150 peaks2. Low water and air: less than 10 %3. Correct mass assignments 0.2 amu (69, 219, 502)4. Symmetrical, smooth mass peak shapes5. Consistent mass peak widths (0.55 0.1) (Default value of Drug MS is 0.55)

6.

Isotope mass assignments should be 1 amu greater than parent peaks7. Appropriate EM voltage: 1000-2800 electron volts. If the voltage is not within theselimits, review the history of the electron multiplier or consult a supervisor. The EMvoltage will increase over time as the source becomes dirty with use. Cleaning thesource should return the EM voltage to a normal operating level.

8. Mass 69 abundance should be 150,000-400,0009. It is normal at times to have a base peak of 219 instead of 6910. Relative abundance:

- Mass 69 = 100 %- Mass 219 = 70-150%- Mass 502 = 3 %

11. Isotope ratios:- Mass 70/69 = 0.5-1.6 %- Mass 220/219 = 3.2-6.4 %- Mass 503502 = 7.9-12.3 %


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If the tune meets the criteria listed, the chemist who performed the tune will initial and circle theinitials. All other chemists using the instrument that day will review the tune for acceptance of data and initial.

Failure of Tune To Meet Acceptable Criteria:

If the weekly Autotune or daily Standard Spectra Autotune fails, the operating chemist will take

note of any error messages generated by the Chemstation Software, check all sources of leaks for tightness, and inform a supervisor. The supervisor will evaluate instrument operation andattempt to correct any problem. If the problem is resolved, an Autotune and Standard SpectraAutotune shall be performed, and applicable technical, maintenance, and repair informationdocumented in the GC/MS Maintenance Logbook. If the problem cannot be resolved, theinstrument will be marked out of service and a supervisor will arrange for instrument repair.

GC/MS Maintenance Schedule:

GC/MS maintenance is documented in the GC/MS Maintenance Logbook located near theinstruments.

Daily/with use Tune (Standard Spectra Autotune)Change wash vial solventsCheck gases

Weekly Tune (Autotune)Change liner and o-ring

Monthly Change gold sealBack up data and sequences to CD or disc

6 months Clean SourceCheck autocal vial

Yearly Replace rough pump oil

As needed Replace septum (minimum-weekly)Trim column


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Change gas tanks


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Replace columnChange filamentClean split side arm

To Set Up and Run a Sequence on the GC/MS System:

From the MS Top Screen select Sequence , select Edit Sample Log Table Click on blank line in the table. The box labeled as Type should read Blank for method

blanks and Sample for unknown samples. Use the tab key or mouse to move to the Vial box and enter the corresponding vial number to

the unknown sample in the autosample tray. Move to Method and enter the name of the method to be used for the current sample. (For a

list of methods, hold the shift and ? keys in this field.) Supply the Sample Name and any Miscellaneous Information (if any), and the Expected

Barcode . Use the Repeat, Cut, Copy, and Paste buttons as appropriate to add samples to the table.

Repeat copies the highlighted line, increments the vial number, places the new lineimmediately after the highlighted one. Copy copies the highlighted line without change.Use Paste to position the copied line where you wish.

When finished entering unknown samples into the sample log table, click OK. From the MS Top Screen select Sequence , and select Run . Enter correct data file name

(i.e. C:\HPCHEM\1\DATA\ or C:\MSDCHEM\1\DATA\), insure that Full Method ischecked under Method Sections to Run, that Inject Anyway is checked under OnBarcode Mismatch, and click OK . Select Sequence and Save from the menu task bar.Enter correct sequence path name and click OK (ie. C:\HPCHEM\1\SEQUENCE\ or C:\MSDCHEM\1\SEQUENCE\). From the MS Top menu task bar select Sequence and Print Sequence . Print Full format.

From the MS Top Screen select Sequence , and select Run . Start sequence table byclicking on Run Sequence.

To Analyze MS Data:

To load a data file: From Data Analysis Screen menu task bar, select File , and select LoadData File . Select a data file name and click OK , or double click the left mouse button on thedata file name. The total ion chromatogram (TIC) for the data file is loaded and displayed inwindow [2].

A data file must be loaded to perform any of the following tasks.

To integrate a chromatogram: From Data Analysis menu task bar, select either Chromatogram/Integrate or Chromatogram/AutoIntegrate.

To select a spectrum: Double-click the right mouse button on the time point of interest inthe chromatogram. The mass spectrum appears in window [1].


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To print a spectrum: From Data Analysis task bar menu, select File , and select Print .Choose either Select Window or TIC & Spectrum from one of the options and the correctwindow desired.

To zoom in: Position the pointer at one corner of the area you wish to expand in achromatogram or spectrum. Press and hold the left mouse button while dragging the mouseto select the area you wish to expand. Release the mouse button. The selected area expands

to fill the existing window. To zoom out: Position the pointer anywhere in the zoomed window. Double-click the left

mouse button. To average spectra: Position the pointer in the chromatogram at the starting time for the

range you want to average. Press the right mouse button while dragging the mouse to theend of the range you want to average. Release the mouse button. The spectra in the selectedrange are averaged and the averaged spectrum is displayed in window [1].

To subtract two spectra: Select a spectrum (double-click the right mouse button in thechromatogram). Select the spectrum to be subtracted (double-click the right mouse button inthe chromatogram). Select Subtract from the Spectrum menu. The spectrum selectedsecond is subtracted from the first spectrum and the resulting spectrum is displayed inwindow [1].

To Use Spectral Libraries:

To search a single spectrum: In Data Analysis , load a data file. The TIC is displayed.Select a spectrum. The selected spectrum appears in a window below the chromatogram.Initiate the library search by double-clicking the right mouse button in the windowcontaining the spectrum. When the search is complete, the search results appear on thescreen. The spectrum for the unknown, the reference spectrum you select from the list of hits, and, if available, the chemical structure of the reference compound is displayed.

To print library results: Click the Print button to print a copy of the displayed spectra.Click Done button to clear the library search results from the screen.

References:

HP5973 MSD/HP6890 Series GC Quick Reference Calibration and Maintenance procedure in the GC/MS Operation Procedure notebook


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GC/MSD Chemstation and Instrument Operation Student Manual Vol #1 Course#H4043A


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Gas Chromatography/Mass Spectrometry

CALIBRATION AND MAINTENANCE

Agilent/HP 6890 GC with 5973 Series Mass Spectrometer

All chemists are responsible for ensuring the proper operation of the gas chromatograph/massspectrometers (GC/MS). A chemist is assigned primary responsibility for overseeingmaintenance and troubleshooting of the GC/MS. All chemists are responsible for running tunes,routine operation, and general housekeeping of the instrumentation as needed.

I. Calibration

A. Standard Spectra Tune

The Standard Spectra Tune ensures standard response over the full mass range. This tune

is recommended to optimize mass spectral library searches. Therefore, calibration of theGC/MS is performed each day of use by using the Standard Spectra Tune. Tuning should be performed after instrument maintenance and before operation.

For the 5973 instruments tune as follows:1. Select the instrument control screen.2. Select the Instrument menu item.3. Select Perform MS Autotune menu item.4. Select tune to perform Standard Spectra Tune.5. Review the tune using criteria listed below.

Acceptable criteria for the Standard Spectra Tune are as follows:

1. Low background: less than 150 peaks2. Low water and air: less than 10%3. Correct mass assignments 0.2 amu (69, 219, 502)4. Symmetrical, smooth mass peak shapes5. Consistent mass peak widths (0.55 0.1)6. Isotope mass assignments should be 1 amu greater than parent peaks7. Appropriate EM voltage 1000-2800 electron volts. If the voltage is not within these

limits, review the history of the electron multiplier or consult a supervisor. The EMvoltage will increase over time as the source becomes dirty with use. Cleaning thesource should return the EM voltage to a normal operating level. (If it does not, theEM may be going bad.)

8. Mass 69 abundance should be 150,000 - 400,0009. Typical relative abundance:

- Mass 69 = 100 %- Mass 219 = 35-99%- Mass 502 = 2-5 %

10. Proper isotope ratios:

Dallas County Institute of Forensic Sciences GC/MS Maintenance HP6890Controlled Substances Laboratory 1 Version 2.0


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- Mass 70/69 = 0.5-1.6 %- Mass 220/219 = 3.2-6.4 %- Mass 503/502 = 7.9-12.3 %

In addition, the standard auto tune sets targets for the percent relative abundances for

certain other PFTBA masses. The system will come as close as possible to the valuesshown:- Mass 50 1 %- Mass 131 55 %- Mass 414 3.5 %-

If the tune meets the criteria listed, the chemist who performed the tune will initial andcircle the initials. All other chemists using the instrument that day will review the tunefor acceptance of data and initial.

B. Autotune

Autotune maximizes instrument sensitivity over the mass range, using PFTBA masses69, 219, and 502. This tune is used in troubleshooting instrument operation. Therefore,an autotune is performed once a week and the tune filed for future reference in evaluatinginstrument operation. A Standard Spectra Tune is performed after the Autotune and prior to routine operation.

Acceptable criteria for the Autotune is as follows:

1. Low background: less than 150 peaks2. Low water and air: less than 10%3. Correct mass assignments 0.2 amu (69, 219, 502)4. Symmetrical, smooth mass peak shapes5. Consistent mass peak widths (0.55 0.1)6. Isotope mass assignments should be 1 amu greater than parent peaks7. Appropriate EM voltage: 1000-2800 electron volts. If the voltage is not within these

limits, review the history of the electron multiplier or consult a supervisor. The EMvoltage will increase over time as the source becomes dirty with use. Cleaning thesource should return the EM voltage to a normal operating level.

8. Mass 69 abundance should be 150,000 - 400,0009. It is normal at times to have a base peak of 219 instead of 6910. Relative abundance:

- Mass 69 = 100 %- Mass 219 = 70-150%- Mass 502 = 3 %

11. Isotope ratios:- Mass 70/69 = 0.5-1.6 %- Mass 220/219 = 3.2-6.4 %- Mass 503/502 = 7.9-12.3 %



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If the tune meets the criteria listed, the chemist who performed the tune will initial andcircle the initials. All other chemists using the instrument that day will review the tunefor acceptance of data and initial.

C. Quick Tune

Quick Tune provides re-tuning for optimum response and resolution, and for accuratemass assignment. Only the mass axis, peak widths, and EM voltage are adjusted; thelenses are unaffected. This tune may be used to rapidly check tuning after maintenance.The Standard Spectra Tune should be run prior to routine instrument operation.

D. Failure of a Tune to Meet Acceptable Criteria:

If the Autotune or Standard Spectra Autotune fails, the operating chemist will take noteof any error messages generated by the Chemstation Software, check all sources of leaksfor tightness, and inform a supervisor. The supervisor will evaluate instrument operation

and try to correct the problem. If the problem is corrected, an Autotune and StandardSpectra Autotune shall be performed, and applicable technical, maintenance, and/or repair information documented in the GC/MS Maintenance Logbook. If the problemcannot be resolved, the instrument will be marked as out of service, and the supervisor will arrange for instrument repair.

II. Maintenance

GC/MS maintenance is documented in the GC/MS Maintenance Logbook located near the instruments.

GC/MS Maintenance Schedule

Daily/with use Tune: Standard Spectra AutotuneChange wash vial solventsCheck gases

Weekly Tune: Autotune followed by Standard Spectra TuneChange liner and o-ring

Monthly Change gold sealBack up data and sequences to CD or disc

6 months Clean SourceCheck autocal vial

Yearly Replace rough pump oil

As needed Replace septumTrim column



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Change gas tanksReplace columnChange filamentClean split side arm

Maintenance activities should be consolidated to minimize instrument down time. For example,if it is time to change the gold seal and cut the column, it may be more prudent to change theliner and septum at that time or hold off changing the gold seal until the liner and septum arechanged.

The maintenance interval is not a rigid schedule and should be based upon workflow. Unlessthere is an immediate problem, a chromatographic run should continue to completion, and themaintenance performed at the end of a run. Where duplicate instrumentation exists, only oneinstrument at a time should be scheduled for preventive maintenance to avoid multipleinstruments being out of service at the same time.

Ultraclean Technique: Liners, gold seals, and certain other parts must be handled using ultraclean techniques to avoidcontamination of the part with oils from the skin, plasticizers from plastic laboratory bench-coats, etc. Do not handle an ultraclean part with your hands; wear cotton gloves or use a freshKimwipe. Lay the ultraclean part on a cloth wrapping or on a clean Kimwipe.

Changing the Wash Vial Solvent:

Discard waste solvent into the hazardous waste. Replace spent solvent in wash vials with freshmethanol, and replace the vials in the appropriate location on the instrument autosampler.

Changing the Liner and Septum:

Tools needed: inlet wrench and tweezers.

NOTE: Turn off carrier gas; if the septum nut is removed without turning off the carrier gas, theglass wool plug may be blown out.

Use ultraclean technique at all times when handling the new liner.

1. Cool the inlet to < 80 oC to minimize oxidation and prevent burns.2. Turn off the carrier gas on the GC by using the keypad on the front panel of the GC: From

the front panel on the GC, push the FRONT INLET button, scroll down using the button tothe Total Flow line. Turn off the flow by pressing the OFF button. Once maintenance iscomplete, turn the flow back on by pressing the ON button.

3. Once the inlet is cool and the gas is off, remove the septum nut. Remove and replace theseptum. Reinstall and tighten the septum nut. These are not ultraclean parts but care should

be taken to avoid any unnecessary contamination.4. Remove the nut covering the liner. Remove the old liner and discard.



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5. Using ultraclean technique, carefully slide an o-ring over the end of the new liner,approximately from the top of the liner, and slide into the inlet. The glasswool plugshould be nearest the bottom of the liner.

6. Replace and tighten the nut and turn the inlet temperature and carrier gas flow on.7. A blank should be run to bake out the new liner before analyzing samples.

Changing the Gold Seal:

Tools needed: appropriate screw driver (Phillips/Torx/etc.), 9/16 wrench, wrench

Note: The gold seal is located at the bottom of the inlet. It is accessed through the nut in theoven where the column comes out of the inlet.

Use ultraclean technique at all times when handling the gold seal.

1. Cool the inlet to < 80 oC to minimize oxidation and prevent burns.

2.

Turn off the carrier gas flow (follow the procedure listed above for Changing the Liner andSeptum).3. Remove the column from the inlet. (Once the column is removed, the ferrule will need to be

replaced and the column cut.)4. Remove the insulator cup (if present) to reveal the nut housing the gold seal.5. Remove the nut and discard the used gold seal and washer.6. Insert the new washer and gold seal into the nut:

a. Insert the washer into the nut; the washer goes between the gold seal and the nut. b. Insert the gold seal into the nut with the grooves visible on top. (These grooves are theexits for the split gas during split injection and for the purge flow after a splitless injection.)

7. Reinstall the nut containing the gold seal and washer, and tighten.8. Replace the insulator cup over the nut (if present).9. Replace the ferrule and trim the column as described below.10. Reinstall the column and check for leaks with a leak detector. (If you turned off the carrier

gas, turn it back on and let it flow for a few minutes before you check for leaks.)11. Turn on the inlet temperature.12. A blank should be run to bake out the system before analyzing samples.

Trimming the Column (Inlet):

Tools needed: column cutting wafer, inch wrench

Any time a column is removed from the inlet, the ferrule should be replaced and the columnshould be trimmed. The inlet end of the column may be cut several inches to remove active sitesand restore separation capacity.

1. Cool the inlet to < 80 oC. This will minimize oxidation and prevent burns.2. Turn off the carrier flow following the procedures in Changing the Liner and Septum.3. Remove the column from the inlet.



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4. Remove the column nut and ferrule. The ferrule, which is made of graphite or graphite/vespel, may be stuck to the nut; remove all ferrule particles.

5. Slide a septum, column nut, and ferrule onto the free end of the column (tapered end of theferrule should point away from the column nut, the flat side toward the nut).

6. Cut two or three inches off the end of the column:

a. The column should be cut by scoring one side with a wafer and then snapping the columnat the score. b. Inspect this cut with a magnifying glass. This cut must be clean and contain no roughedges. If there are rough edges, repeat until a clean cut is achieved or seek guidance.c. Wipe the end of the column with a Kimwipe and methanol, or hexane.d. Place the column back into the inlet. When installed, the column should protrude 5 mm(4 mm to 6 mm) beyond the ferrule into the inlet. Use the septum as a guide for thismeasurement.e. Tighten the column so that the column does not slide with a gentle tug.

7. Turn on the carrier gas, and check for leaks with a leak detector.

Installing a New Column:

Tools needed: column cutting wafer, inch wrench, MSD installation tool

1. Cool the injector and vent the detector. After the MS has vented, open the vacuum manifold.2. Turn off the carrier flow following the procedure in Changing the Liner and Septum.3. Loosen the column nuts from the injector and the transfer line from the detector, and remove

the column.4. Place a nut and ferrule on each end of the column. (The flat side of the ferrule goes toward

the inlet nut and tapered side out, and the tapered end slides into the MS transfer line nut,with the flat side out.) NOTE: These are different types of ferrules. See supervisor for instruction.

5. Cut about 2-3 inches from the inlet end of the column using techniques described in Cuttingthe Column.

6. Install the new column into the inlet.7. An MS column, which has been developed for use in a mass selective detector has

extremely low column bleed and can be conditioned while in the detector. MS columns canalso be conditioned prior to installation into the GC/MSD interface.

8. The column can be installed in the mass spec detector either with an installation tool or without.a. Without the tool: Slide the column into the GC/MSD interface until it can be pulled

through the vacuum manifold, trim 1-2 cm off the end of the column, clean the outside of the column with methanol or hexane, adjust the column so it protrudes 1-2 mm past theend of the interface, hand tighten the nut, readjust the column as necessary and tightenthe nut to turn. (Check tightness after one or two vent cycles.

b. With the tool: Slide the column into column installation tool, trim 1-2 cm off the end of the column, clean the outside of the column with methanol or hexane, adjust the columnso it protrudes 1-2 mm past the end of the tool, hand tighten the nut, slide the septum totouch the end of the nut, use two wrenches to tighten the nut turn, remove the columnand nut from the installation tool (total length from the septum to the end of the column is



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176 mm), clean the outside of the column with methanol or hexane, insert the columninto the interface, and tighten the nut to turn. (Check tightness after one or two ventcycles.

9. Close the vacuum chamber, and pump down the mass spectrometer.

Conditioning the Column:

Tools: Carrier gas, wrench

If conditioning is not done properly the column may be ruined. Do not use hydrogen tocondition a column.

1. Allow the carrier gas to flow through the column for approximately 5-15 minutes with theGC oven at room temperature

2. Ramp the oven temperature at 5-15 degrees per minute to the final conditioning temperature.The final conditioning temperature should be at least 10 degrees higher than the maximum

oven temperature to be used in a method. Do not exceed the manufacturers recommendedmaximum operating temperature.3. Hold this temperature and allow carrier gas to flow for several hours, or overnight.4. Return the GC oven temperature to a low standby temperature.

Cleaning the Split Side Arm:

The split vent side arm is the exhaust for split gasses and compounds that are purged off duringan injection. This can become very dirty and without maintenance can cause deleterious results.

1. Cool the injector temperature and turn off the gasses.2. Remove the liner from the inlet as specified in the Changing the Liner and Septum section.3. Remove the autosampler tray, tower, fan cover, and the top rear instrument cover.4. Remove the split side arm from the inlet and the filter located at the rear of the instrument.5. Inspect the ends of the side arm to make sure they are not clogged. If they are, use an old

syringe to unclog the ends.6. Using vacuum, pull a solvent such as chloroform through the side arm and into waste.7. Repeat step 4 using a solvent such as methanol and then air to dry.8. Using a small brush or Q-tip dipped in chloroform clean out the inlet arm where the side arm

attaches.9. Reconnect the side arm to the inlet and the filter.10. Replace the covers and autosampler tray and tower.11. Change the liner, o-ring and septum.12. Turn on the temperatures and gasses.13. Check for leaks.

Cleaning the Source for Mass Spectrometers/Changing the Filament/Fill Autocal Vial/ Change Rough Pump Oil:

Follow instructions provided in the Agilent/HP 5973 Hardware Manual.


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Operation of the 5973 Series Mass SpectrometerUtilizing G1701BA and G1701DA, Versions B.01.00 and D00.00

Mass Spectrometer Chemstation Software

Overview

The Chemstation software is a windows based application and usable on Windows95, 98, NT and 2000. The following instructions are intended as a general guideand do not represent all of the ways that the software may be used. In depth literature ispublished by the manufacturer and should be used as the key reference andtroubleshooting guide. Online help is also available at www.agilent.com . Users maybegin using the Chemstation software by clicking on the Top icon located on the desktop.There are several ways to move around in the Chemstation software. The primarymethod of movement from one screen to another in the software is through the View menu item. If in the Top Level , Instrument Control or Data Analysis , the View menuitem will allow the user to move to any of the other screens.

Users are referred to the appropriate instrument manuals for additional information.

Top Level Screen

From the Top Level screen the user has the ability to edit, print, load, and save methodsand sequences. This screen enables the user to begin instrument operation.

Instrument Control Screen

From the instrument control screen the user has the ability to modify instrument

parameters such as injector, inlet, column and oven parameters, mass spec temperatures,and electron multiplier voltages. The user can also load, edit, save and print methodsfrom this screen. This screen enables the user to monitor instrument parameters during arun. Instrument tuning is also done from this screen.

Data Analysis Screen

From the data analysis screen the user can edit the data analysis portion of a method, editand select libraries, review total ion chromatograms, review individual spectra, compareunknown spectra to known libraries, and print reports. Data analysis is also the locationof the quantitation databases used in quantitative methods.

Storage of Data files, Methods and Sequences

The hard drive is partitioned into two drives, drive C:\ and drive D:\. The C:\ drive is thelocation of the Chemstation and the operating system (Windows NT 4.0 or Windows2000). The D:\ drive is designated for storage of other installed software. For theoperation of the Mass Spectrometer Chemstation there are three key areas of storage.

Dallas County Institute of Forensic Sciences Operation 5973 GC/MSControlled Substances Laboratory 1 Version 2.0
http://www.agilent.com/http://www.agilent.com/


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Data is stored in C:\hpchem\1\data\ or C:\msdchem\1\data\ . Data consists of data.ms files containing total ion chromatograms and their spectra, pre_post.ini files containinginformation on the status of the instrument during the run of the indicated file, and logfiles containing information regarding the sequence that was run.

Methods are stored in C:\hpchem\1\methods\ or C:\msdchem\1\methods\ . Methodscontain all of the information required to execute a run. Methods contain the requiredmacros and instrument parameters such as oven ramping, pressure and flow parameters,injection temperatures and specifications, and instrument temperatures, as well as otherinformation.

Sequences are stored in C:\hpchem\1\sequence\ or C:\msdchem\1\sequence\ .Sequences contain the information required to complete a series of injections utilizing theautosampler tower, tray and barcode reader. The information consists of vial number,sequence line number, sample name, method name, and operator information.

I. Sequences

Loading a sequence or creating a new sequence

From the top level:

1. Select Sequence menu item.2. Select Load menu item.3. Select the default.s or a previous sequence

and OK.

This sequence of events will load the selectedsequence in the instruments memory. Proceed toEdit sequence section to edit the sequence or Runninga sequence section to run the loaded sequence.

Edit sequence

Once the desired sequence has been loaded you may want to edit the sequence. From thetop level:

1. Select Sequence menu item.2. Select Edit Sample Log Table menu item.

The edit screen will appear at this time with the sequence that the user had previouslyloaded. The user may edit, cut, copy, paste, or repeat lines in the sequence. Informationthat is required is Type, Vial, Method, and Expected Barcode. Line and Data File will becompleted automatically by the Chemstation software. The user may wish to include aSample Name and/or Miscellaneous Information.



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Edit a sequence while it is running

The user may also edit the sequence while a sequence is running to add specimen to therunning sequence. If the sequence is running the menu items will not be available andthe user must select the Edit Samp Log Tbl button from the center screen. At this time

the edit sequence screen will appear.

Saving a sequence

After the user has loaded and edited a sequence, thesequence must be saved. From the top level:

1. Select the Sequence menu item.2. Select the Save menu item.

The default location to save sequences isC:\hpchem\1\sequence\ or C:\msdchem\1\sequence\. If the location varies from the default location, locate thedefault and then save. The format in which to save thesequence is: month, day, and letter designation (ie:Jul29A).

Running a sequence


There are several ways to begin a sequence. The user may select Run , Load andRun , or Position and Run . The Run menu item will begin the current sequence.The Load and Run sequence menu item will allow the user to load a new sequence


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and then run it. The Position and Run menu item will allow the user to begin thecurrent sequence at a user specified location within the current sequence. To begin asequence from the top level:

1. Select the Sequence menu item.

2.

Select the Run, Load and Run, or Position and Run menu item.3. The user will be prompted to enter the data path for the storage of the datacollected during the sequence run. The following screen will appear:

4. Make sure that Full Method and Inject Anyway are selected. The user may

or may not want to select the Overwrite Existing Data Files option.5. Type the correct path for the storage of data. The default path isC:\hpchem\1\data\ or C:\msdchem\1\data\. The default location is where theChemstation looks for data from the data analysis screen. If the user does notstore the data in the default location be aware of the location that was selected.

6. Select Run Sequence button.

If the sequence does not begin at this time, check the above screen and the selections tomake sure that the proper selections have been made. If Load and Run or Positionand Run are selected the user will be prompted to load a sequence or select a positionwithin a sequence before the above screen appears.

Using Keywords

The Chemstation allows the use of keywords within a sequence. These keywords allowthe user to perform certain functions within the sequence such as running a command ormacro, tuning the instrument, or pausing the sequence. The most common keyword usedis Pause . The Pause command will allow the user to pause the sequence after aninjection or series of injections are made, or a method is complete. To use a keyword:



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1. From the Sample Log Table: 2. Select the next

line to be run,copy and paste so

there are twoidentical lines.3. Select the

Keyword command in theType box.

4. This will promptthe user to typethe Keyword thatis to be used.Select the

keyword desiredin the keywordbox.

5. Select OK.

When the current method is complete the keyword command will be executed.

II. Methods

The method in the Chemstation software is the location of all of the instrumentparameters and data analysis parameters that will take place during the execution of amethod in a sequence. Methods should only be created or modified by those withknowledge of the required parameters or experience with instrument parameters.

Creating a new method

To create a new method the user may select menuitems from either the Top Level screen or theinstrument control screen. From either screen:

1. Select the Method menu item.2. Select the Load menu item.3. Load the default method or load an

existing method and modify to createyour new method.

4. Select the Save menu item to save themethod with an appropriate name.



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Editing an existing methodIf the user needs to edit a method or check the contents of a method the method may beaccessed from the Top Level screen or the instrument control screen. From eitherscreen:

1.

Select the Method menu item.2. Select the Edit Entire Method menu item.3. Edit or review the method.

Save the method ONLY if aware of the changes that have been made. DO NOT save themethod if unaware of the changes made.

III. Data Analysis

The user may enter data analysis in several different ways. The user may open it fromthe icon on the desktop, from the Top Level screen during the execution of a sequence,or from the View menu item from anyscreen.

Loading a chromatogram (data file)


The data file contains the total ion chromatogram and the spectra for the identifiedcompounds in the chromatogram. To load a chromatogram:

1. Select the File menu item.2. Select the Load Date File menu item.3. Select the appropriate data file from the correct

data path and select OK . (The data file will be inthe location that was specified when the sequencewas started. The default location isC:\hpchem\1\data\ or C:\msdchem\1\data\.)


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Reviewing Data

Once the chromatogram has been loaded it will appear in window [2]. It will appear asthe total ion chromatogram (TIC) with peaks representing detector response tocompounds and their retention times. There are many items that the user may want to use

to review data. These include, but are not limited to, integrating, searching for specificions, comparing unknown spectra to known libraries, generating reports, and printingspectra.

The TIC will be normalized on the largest peak. This means that the largest peak will becompletely visible from the top of the peak to the baseline. Large peaks in a TIC maymake smaller peaks not visible when looking at the normalized chromatogram. If theuser wishes to look at the smaller peaks the zoom function may be used. To do this leftclick and hold the mouse, drag a box around the area to be analyzed and release. Thiswill zoom in on the area that the box was drawn around.

Generating, comparing and printing spectra

To generate a spectra at a specific TIC retention time double click the right mouse buttonat the point of interest. This will generate a mass spectrum of the area that was clickedon and it will appear below the TIC in window [1].

The method that is loaded will determine what library or libraries have been selected foruse (see searching libraries for instructions on changing). To compare the generatedspectrum with the library that is specified in the method:

1. Double click the right mouse button while the cursor is in window [1].



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The resulting spectrum will be the difference of the second obtained spectrum removedfrom the first obtained spectrum.

Printing reports

There are many ways to print reports and a number of reports are available in theChemstation software. The most common way to print a report for a designated methodis to:

1. Load the appropriate TIC.2. Select the Method menu item.3. Select the Run Method menu item.4. This will run the data analysis portion of the method that has been selected in

data analysis. It will print the report as if the sample had been run in asequence.

The user may also want to use the options available under the quantitate and tools menu

items.

Using the Quantitate and Tools menu items the user will be able to use the Chemstationreport format and to reprocess quantitative and qualitative reports linked to customreports.

Searching Libraries

There are many different libraries that are available in the Chemstation ranging from in-house libraries to commercially purchased libraries. The libraries are located on the C:\ drive of the computer in C:\database. To change the libraries that are automaticallysearched in a method:



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1. Select the Spectrum menu item.2. Select the Select Library menu item.3. This will prompt the user to type in the name of the

library to be used.

This will affect the library that is used when the user iscomparing an unknown spectrum to a library spectrum.

Note: Mass Spectral libraries are used only as a tool inidentifying an unknown. Chemists must consider a variety of factors before reaching a conclusion.

Addition of New Mass Spectra to Libraries

If a new compound or standard is obtained, it is important to have this compounds massspectra in a spectral library. The spectral library to place the new compound must beselected, usually it will be dcflab.l. (See instructions above to select the appropriatelibrary.) To add a mass spectrum to a library:

1. Select the Spectrum menu item.2. Select the Edit Library menu item.3. Select the Add New Entry menu item.

4. Enter information into appropriate cells: Name, Mol. Formula, Mol. Weight,etc.

5. Select the Include in search box.Dallas County Institute of Forensic Sciences Operation 5973 GC/MSControlled Substances Laboratory 12 Version 2.0


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6. After entering the information, select OK , and the Chemstation software willadd the spectra into the library in the C:\database\ file path.

7. The Chemstation software will also assign the new spectra a number.

Parametric Retrieval is located under the View menu item in Data Analysis screenonly. Parametric retrieval can be used to search a user designated library for compoundsbased upon name, molecular weight, CAS number, library entry number, etc. However,the information must be in the library before the search parameters will work.Occasionally all of the information may not be available with in-house libraries.



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uning the 5973

d in the maintenance log and thereference collection found near each instrument.

T The tuning instructions and parameters are locate


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PROCEDURE: ANALYTICAL BALANCE

1) Monthly Calibrationa) At the beginning of each month, analytical balances should be calibrated and their

linearity checked using certified weights that are located in the Drug Lab.

b)

Using the Balance Maintenance Log Calibration Check logsheet, record the date,actual weight, and the difference in the last decimal place measured.c) This difference must be within +/- 3 counts in the last place measured.d) If the difference is out of control, then the balance must be evaluated and recalibrated.

i) Ensure that the balance is level and clean.ii) Refer to the balance manual for calibration procedure or seek supervisory assistance.

e) Once the balance is recalibrated, the linearity should be checked using three certifiedweights (lower, middle and upper range), and the results recorded on the BalanceMaintenance Log - Calibration Check. Also, include the chemists initials, the date, thecertified weights used, actual weight measured, and the difference between the lastdecimal place measured and the target weight.

2)

Daily Calibration Check a) Each day before the balance is used, the calibration must be checked.b) On the Balance Maintenance Log sheet record whether the balance is level and clean.c) Also record the difference (in counts) between the measured secondary weight and its

target weight.d) The difference must be within +/- 3 counts in the last place measured.e) If the difference is out of control, then the balance must be recalibrated.

i) Refer to the balance manual for the calibration procedure or seek supervisoryassistance as needed.

f) Once the balance is recalibrated, the linearity should be checked using three certifiedweights (lower, middle and upper range), and the results recorded on the BalanceMaintenance Log Calibration Check logsheet. Also, include the chemists initials, thedate, certified weights used, actual weights measured, and the difference in the lastdecimal place measured.

3) For balances with flashing recalibration messages:a) If the secondary weight is within +/- 3 counts after recalibration, no further action is

necessary other than recording results in Balance Maintenance Log.4) For balances with automatic recalibration:

a) No additional action is needed other than daily calibration checks.5) Seek supervisory assistance in any situation in which a balance cannot be calibrated properly

or when balance operation appears unusual.

Dallas County Institute of Forensic Sciences 1 Analytical BalanceControlled Substances Laboratory Version 2.0


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The top entries and the Environmental factors (Temp., Barom. P., Rel. Hum.) can be editedbefore Starting a Run. Once started, you are locked out of these areas. After the last weight isreceived, the Environmental factors can be revised again, if necessary. If these are changed afterthe last weight is received, the program will also recalculate the calibration results and statistics.

After confirming the correct Input Mode (Balance), shown in the Balance ID field, ensure thatthe device and method are the ones you want before running the calibration. The followingdescribes some of the components (others found in the manual) of the Run Calibration screen: Temp. -- This field contains the liquids temperature in degrees Centigrade. This

information can be read from the meter found on the wall behind the balance. As notedearlier, this can only be edited at the start of a Calibration Run.

Barom. P. -- This field contains the barometric pressure in mm of mercury ( inner most scaletimes 10). This information can be read from the barometer found on the wall behind thebalance. As noted earlier, this can only be edited at the start of a Calibration Run.

Rel. Hum. -- This field contains the value for the relative humidity, with the units as %.This information can be read from meter found on the wall behind the balance. As noted

earlier, this can only be edited at the start of a Calibration Run. Abort Run -- Deletes all current weight, time and volume information for this run.

Environmental factors are not changed. Device selection, operator name and method are notchanged.

Redo Sample -- This button becomes active once you have entered the Tare weight and thefirst calibration sample. Clicking on it cancels the results entered for the most recent sampleonly, and resets the Run Line to the previous sample so you can re-send the weight fromthe balance. Please note the Redo Sample button does not allow you to re-dispense thesample, only to correct an error in entry. If you have a problem with dispensing the sample,consider using the Void button at the end of the Run (see details below), or abort thecalibration and start again.

Void Sample -- This button becomes active only at the end of the Calibration Run, andbefore you click on Accept Run . If you experience problems dispensing one or moresamples, you may choose to exclude these from the calculations for the Run, rather thanabort the Run and start over. In effect, the calculations are performed with fewer sample(s).When the Run is finished, you will be able to move the cursor up and down through thecumulative weights shown on the Calibration Run display grid. Position the cursor overthe sample to be voided, then click on the Void button. The calculated Volume for thissample will be replaced with the word VOID. You are able to Unvoid the sample bypositioning your cursor over the voided cumulative weight and clicking on the Void buttonagain. This reverses all effects of clicking Void .

Accept Run -- This button only becomes active when the run is complete. Clicking on itaccepts the Calibration Run, stores the results, then inactivates the button and the report willbe automatically printed.

Done -- Leaves the Calibration Run screen and goes back to the WorkList or Inventory List.

Dallas County Institute of Forensic Sciences 2 Pipette CalibrationControlled Substances Laboratory Version 2.0


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Run Calibration Automatically (Through the Balance) First, ensure that the balance is calibrated properly by following the QA Procedure for theAnalytical Balance, then proceed as follows from the Run Calibration screen: Making sure that the correct device (pipette) and method are listed, edit the Environment

factors (Temp., Barom. P., Rel. Hum.) Start the calibration by clicking on Start Run button. Place your container with liquid on the balance. Tare the balance and click on Tare Wgt, which sends the weight over to the Calibration

Run Table. Important: Tab over to Sample Wgt. Pipette a volume into the container. Once a stable weight has been obtained, press Enter, which sends the weight over to the

Calibration Run Table. Continue until all volumes have been recorded. Using Abort, Void Sample and Redo

as necessary. At the completion of the calibration , either click on Accept Run or Abort Run. Print a hard copy of the report, sign and file next to the balance or with the Quality Manager. Another device may be chosen by pulling down the device list. Once the calibration(s) are completed, click on Done. Log off by clicking on Password and choosing LogOff.



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Calibration of PipettersProcedure Summary

There may be three reasons for checking calibration of a pipetter: Receiving a new pipetter into the laboratory. See the Quality Manager prior to calibration. Regularly scheduled calibration. Suspected error in calibration.

A condensed summary follows:

The computer, printer and balance should be on. If not, allow the balance to warm up forthirty minutes.

With the 20 gram weight, check the balance calibration and record weight on the BalanceCalibration Log. See Quality Manager if balance is not in calibration.

Log on to the computer by pulling down the Password menu and selecting Logon . If you donot have a password, see Quality Manager.

From the WorkList, select the pipetter that is to be calibrated. Pull down the Device menu and select Run Calibration . From the meters on the wall, enter the current temperature, barometric pressure and relative

humidity. Begin the calibration by clicking on Start Run button. Place container with enough liquid to cover the bottom on to the balance. Tare the balance and click on the Tare Wgt button, which sends weight over to the

computer Calibration Run Table. Important: Tab over to Sample Wgt button. Pipette a volume into the container. Once a stable weight has been obtained, press Enter, which sends the weight over to the

computer Calibration Run Table. Continue until all volumes have been recorded. Using AbortRun, VoidSample and

Redo as necessary. At the completion of the calibration, either click on Accept Run or Abort Run. Print a hard copy of the report, sign and file next to the balance or with the Quality Manager. Another pipetter may be chosen by pulling down the device list. Once the calibration(s) are completed, Click on the Done button. Log off the computer by pulling do down the Password menu and selecting Logoff .

If pipetter performance is acceptable, the printed report is filed next to the balance or givento Quality Manager. If pipetter performance is not acceptable, immediately remove from service and notify

Quality Manager.


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DISPENSETTE CHECK

1) At the beginning of each week, solution dispensettes will be checked for volume accuracy.Materials needed for this check include volumetric flasks and the dispensette containing the

solution to be dispensed. Dispense the solution into the flask and note the volume in theflask. The bottom of the meniscus should be at the line of the volumetric flask.

Solution Dispensette Vol. Volumetric Flask MeCl 2 w/IS 10 ml 10 mlBuCl w/IS 5 ml 5 mlEtOH w/IS 2 ml 2 ml

2) Using the Weekly Dispensette Volume Check log sheet, record the date, the initials of thechemist performing the check, the solution and volume dispensed, and the size volumetricflask used. Document whether or not the volume dispensed equales the volume of the flask.

a. If the volume dispensed meets the volume of the flask, the dispensette may be used.b. If the volume dispensed does not meet the volume of the flask, advise a supervisorand remove the dispensette from service or inspect, clean, or adjust it as needed.

i. The procedure for readjusting the dispensette can be found in the Calibrationsection of the Dispensette Operating Manual.

ii. If adjustments are required, document these in the Comments section of theWeekly Dispensette Volume Check log.

3) If dispensing the solution from the dispensette becomes difficult, the dispensette may requirecleaning.

a. Follow the cleaning procedure found in the Dispensette Operating Manual.4) If an air bubble forms in the dispensette filling valve, unscrew the dispensette from the bottle,

remove the filling tube and insert a glass pipette to remove any liquid around the valve. If this does not remove the air bubble, the dispensette may need a more thorough cleaning.5) Do not interchange dispensettes between different solutions.

a. If it is necessary to change the type of solution in the dispensette, clean thedispensette thoroughly and document any changes on the logsheet.

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THERMOMETER CALIBRATION CHECK

1. Annual Calibration Check1.1. Reference Thermometers

1.1.1. NIST traceable reference thermometers are maintained under the direction of the

Quality Manager.1.1.2. Calibration of appropriate reference thermometers is verified annually by anoutside vendor.

1.1.3. Records are maintained by the Quality Manager.1.2. General Use Thermometers

1.2.1. Calibration of laboratory mercury and alcohol thermometers will be verifiedannually against a NIST traceable thermometer.

1.2.2. General use thermometers should read within +/- 2 divisions of the NISTthermometer.

1.2.2.1. If the general use thermometer does not meet the designated range, it will bereplaced or repaired.

1.2.2.2. In the event that replacement or repair is not an option, the amount of deviation from the NIST thermometer is recorded on applicable log sheets and

laboratory thermometer readings are adjusted accordingly.1.2.3. Records of annual thermometer calibration checks will be maintained in the

laboratory.

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DRUG CASE REVIEW GUIDANCE

1. Administrative and technical review will be performed by the analyst and the reviewer priorto release of a case report.1.1. The primary responsibility for ensuring the technical and administrative correctness and

completeness of the case file lies with the analyst of record.1.2. Case review by someone other than the analyst of record does not shift the responsibilityfor the scientific findings from the analyst to the reviewer.

2. The purpose of case review is2.1. To ensure that the conclusions of the analyst are reasonable and within the constraints of

scientific knowledge2.2. To ensure that the conclusions detailed in the final report accurately and fairly represent

the content in the case file2.3. To ensure that a the case file contains adequate information so that another competent

analyst can independently evaluate and interpret the data2.4. To ensure that proper laboratory procedures were followed

2.5. To ensure that the report is editorially correct and free from typographical errors3. Types of review

3.1. Administrative review3.1.1. A procedure to check for consistency with laboratory policy and for editorial

correctness.3.2. Technical review

3.2.1. Review of notes, data, and other documents which form the basis for a scientificconclusion.

4. Technical and administrative review require an extensive and careful review of the case fileand final report including but not limited to the following4.1. Does each page of the case file have a case number and analyst signature or initials?4.2. Are all entries in the case file clear and legible?4.3. Are corrections handled properly (single strike through with initial)? There should be no

write-overs.4.4. Transmittal sheet and chemist report

4.4.1. Are the following items consistent on the final report4.4.1.1.Case name4.4.1.2.IFS case number4.4.1.3.Agency number: service number, tag number, etc.4.4.1.4.Agency name4.4.1.5.Date received4.4.1.6.Received by4.4.1.7.Delivered by4.4.1.8.Item received (transmittal sheet)/evidence submitted (final report)

4.4.2. Is all suspected drug evidence which is listed as an exhibit on the transmittal sheetaccounted for on the final report?

4.4.3. Are heavy non-drug items which are listed on the transmittal sheet accounted foron the final report?

4.4.4. Is the internal evidence transfer complete from Evidence Registration to theanalyst?

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METHOD VALIDATION GUIDANCE

Validation and verification of new testing methods is summarized in the Quality Manual. TheSupervisor and/or Section Chief will design a specific testing protocol to validate the accuracy of a new method or to verify the accuracy of a method transferred from one instrument to another.

I. Transfer of a Method to New Instrumentation

First, proper calibration and operation of new instrumentation is verified and documentedusing the applicable instrument calibration process. Secondly, analytical results arecompared between the new and old instrumentation.

Where possible, side by side operation of old and new instrumentation is compared usingthe same samples or extracts. Standards, internal standards, and quality control samplesshould meet their respective criteria described in the analytical procedure. Testspecimens should also be run and should meet acceptability criteria established for

quality control samples or similar proficiency test results. Replicate analytical runsshould be made on a single day and also compared between multiple days; criteria shouldmeet those established for standards, internal standards, and quality controls; testspecimens should meet acceptability criteria established for quality control samples orsimilar proficiency test results. As applicable, analysts are trained in operation of newinstrumentation.

II. Development of a New Analytical Method

A supervisor outlines proposed steps for a new analytical method based upon literature,other established procedures, or knowledge and experience of the materials to beanalyzed. The target sensitivity and linear range is established based upon expectedsample conditions and instrument performance. Standard curves are run to assess theapplicability of the proposed analytical technique in comparison to the targeted analyticalneeds.

Once it is determined that sensitivity and range are appropriate, the method is evaluatedfor interference from expected matrices or co-occurring substances. The assay isevaluated for stability by repeating analysis of single prepared samples and duplicatepreparation of samples both within day and between day. Variability of standards,internal standards, quality control samples, and specimens are evaluated with respect toliterature values and/or similar established procedures. Criteria for acceptable standardcurves, internal standards, and/or quality control samples are established. A formalprocedure is written, applicable analysts are trained, and results are reviewed for stabilitywithin analyst. Method results may also be evaluated against external standards orspecimen results from reference laboratories; criteria for acceptability will be similar tothat established for quality control samples.

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WORKSHEET OF POSSIBLE VALIDATION STEPS

Submitting Unit:

Validation Title:

Prepared by/Date:

1. Type of validation (check applicable box):EquipmentModification of an existing protocolNew protocolOther:

2. Is this an original or revised validation plan?Original

Revised3. Describe the purpose or goal of the validation including the model and serial number of

any new equipment:

4. Attach a copy of the written procedure: current (if applicable) and new.5. The method is

QuantitativeSemi-quantitativeQualitative

6. Attach a validation plan with proposed timetable and list staff participating in thevalidation. The plan should lay out a course of action to assess the following, as

applicable, which will be included in the final validation report:a. Method validation is the process of testing an analytical method, technique, or

instrument to determine its suitability for meeting its intended purpose and todocument its reliability under expected conditions of use. Generally the validationprocess is expected to

i. Evaluate whether a new testing method meets identified analytical needsand current scientific practices;

ii. Compare the new test methods performance with existing laboratorymethodology;

iii. Describe the conditions under which a testing method will produce validresults;

iv. Predict possible sources of error;v. Determine limitations of a testing method; andvi. Establish baseline characteristics of the testing method (linearity,

accuracy, etc.) which serve as benchmarks to evaluate future method

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Method Validation Variables:

1. Evaluate whether a new testing method meets identified analytical needs and current

scientific practices.

2. Compare the new test methods performance with existing laboratory methodology.

3. Describe the conditions under which a testing method will produce valid results.

4. Predict possible sources of error.

5. Determine limitations of a testing method.

6. Establish baseline characteristics of the testing method which will serve as benchmarks to

evaluate future method performance. These characteristics may include:

a. linearity/linear range:

b. accuracy/precision: Accuracy is the ability to quantitate

SWIFS Controlled Substances Resources Documents v2.0 - reformatted 61 pages.pdf

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