To lay down the procedure for operation of gas chromatograph (agilent) system.
1.0 PURPOSE:
1.1 To lay down the procedure for operation of gas chromatograph (agilent) system.
2.0 SCOPE:
2.1 This SOP is applicable for Quality control department.
3.0 RESPONSIBILITY:
3.1 Assistant – QC
4.0 ACCOUNTABILITY:
4.1 Executive – QC
5.0 PROCEDURE:
5.1 Check that the power supply to the instrument is switched ‘OFF’ before cleaning. Clean the instrument with a clean dry cloth every day. Occasionally wet cloth dipped in dilute soap solution may be used. Precautions to be taken to clean the instrument immediately with dry cloth to remove the moisture.
5.2 GC System Configuration is as follows:
5.2.1 Agilent GC System, agilent 6890 Series
5.2.2 Agilent Head Space Sampler, agilent 7694E
5.2.3 Waters Millennium32 Software.
5.2.4 Agilent 6890 series Injector
5.3 Procedure to operate agilent 6890 series gas chromatograph:
5.3.1 Turn on the gas supply; set the gas regulator such a way that a line pressure of about 4 kg/cm2 is maintained. Switch on the instrument; switch is located at the front, bottom, and right corner of the instrument. Open the oven door by lifting gently the oven door latch, situated just below the oven door to the left of the instrument.
5.3.2 Entering new values on the instrument.
5.3.3 Press required key to call up a control table. It appears in the display. The first line is a title that identifies the table.
5.3.4 Inspect the set points in the table, move the cursor keys [? and ?] to move the cursor [<] to the line you wish to change, type the new value, and press [Enter]. Repeat this process for other set points as required.
5.3.5 Installing column
5.3.6 Capillary column: you must prepare your new capillary column before installation
5.3.7 Preparing the capillary column:
5.3.8 Place a capillary column nut and ferrule on the column.
5.3.9 Score the column using a glass-scribing tool. The score must be square to ensure a clean break.
5.3.10 Break off the column end by supporting it against the column cutter opposite the scribe. Inspect the end with magnifying lens to make certain there are no burrs or jagged edges.
5.3.11 Wipe the column walls with a tissue dampened with isopropanol to remove fingerprints and dust.
5.3.12 Installing capillary columns.
5.3.13 Always install capillary columns at back inlet (split/splitless injector), and connect the detector end at Front detector. If the any other capillary column is installed on that position, then remove the installed column, by turning the column nut anticlockwise at both inlet and detector with a wrench, then further loosen the column nut with fingers at both inlet and detector fittings.
5.3.14 Position the column so it extends 4 to 6 mm above the end of the ferrule. Mark the column with glass marking pen or suitable correction fluid at the point even with the column nut.
5.3.15 Place the capillary column on the hanger so that the position that best centers the column in the oven. Column ends should make smooth curves to the inlet and detector fittings. Do not let any section of the column come in contact with the oven surfaces.
5.3.16 Insert the column in the inlet and slide the nut and ferrule up the column to the inlet base. Finger tighten the column nut until it starts to grab the column.
5.3.17 Adjust the column position so that the mark on the column is even with the bottom of the column nut.
5.3.18 Tighten the column nut an additional 1/4 to 1/2 turn so that the column cannot be pulled from the fitting with gentle pressure.
5.3.19 Assemble a brass nut and graphite vespel ferrule on to liner/adapter.
5.3.20 Insert the adapter straight into the detector base as far as possible.
5.3.21 Hold the adapter in this position and tighten the nut finger tight.
5.3.22 Use a wrench to tighten the nut an additional 1/4 turn.
5.3.23 Position the column so it extends above the end of the ferrule by 68 mm. Mark the column with a marker at a point even with column nut.
5.3.24 Insert the column in the detector. Slide the nut and ferrule up the column to the detector base. Finger tighten the column nut until it starts to grab the column.
5.3.25 Adjust the column position so that the mark on the column is even with the bottom of the column nut.
5.3.26 Tighten the column nut finger tight.
5.3.27 With draw the column about 1 mm. Use the wrench to tighten the nut an additional 1/4 turn.
5.3.28 After the column is installed at both inlet and detector, establish required flow of carrier gas through the inlet. Heat the oven, inlet, and detector to operating temperatures. Allow them to cool, and then retighten the fittings.
5.3.29 If the column is already installed once, then follow the steps from 5.4.
5.3.30 Installing Glass packed column at purged -packed inlet.
5.3.31 Always install the Packed columns at Front Inlet (Purged packed Injector) and connect the detector end at Front Detector. If the any other column is installed on that position, then remove that column, by turning the column nut anticlockwise at both inlet and detector with a wrench, then further loosen the column nut with fingers at both inlet and detector fittings.
5.3.32 Glass packed columns must be installed simultaneously at the inlet and the detector and must be installed parallel to the oven door.
5.3.33 To the end of the column that to be connected to the inlet, allow at least 50 mm of empty column to prevent an inserted syringe needle from contacting either the glass wool plug or column packing.
5.3.34 At the detector fitting, remove the adapter, if installed. There must be at least 40 mm of empty column to prevent the bottom end of the jet from touching either column packing or the glass wool plug.
5.3.35 Assemble a brass nut and graphitised vespel ferrule onto each end of the column.
5.3.36 Insert the column into the inlet until it bottoms. Insert the column into the detector fitting, but do not force it. It may be necessary to start the long end of the column in the inlet at a slight angle to clear the oven floor.
5.3.37 With draw the column, 1 to 2 mm from both the inlet and detector.
5.3.38 Tighten both column nuts finger tight.
5.3.39 Tighten both column nuts 1/4 turn with a wrench.
5.3.40 Set the flow through the column and raise the inlet, detector, and oven to operating temperature. Then set the oven to ambient and allow it to cool.
5.3.41 Use the wrench to tighten the nut an additional 1/2 turn. Tighten further as necessary to prevent leakage, but do not over tighten the column.
5.3.42 Conditioning the columns
5.3.43 If the installed column is new, it should be conditioned before use.
5.3.44 Conditioning the capillary columns
5.3.45 Turn off the detectors. Shut off the detector support gases. It is especially important to shut off hydrogen. This can be done by pressing the [Back Det], detector control table parameters are displayed. Then turn required parameters to off mode by pressing [Off] at the required cursor position.
5.3.46 Connect one end of the column to inlet. But do not connect the detector end.
5.3.47 Cap the detector(s) fittings with the no-hole ferrule and column nut.
5.3.48 Establish a stable flow of carrier gas by setting the pressure to value in psi given in the Table-1.
5.3.49 Let gas flow through the column at room temperature for 15 to 30 minutes to remove air. This can be done by pressing [Col 2] and moving the cursor to pressure parameter and set the required value by using numeric keys and press [Enter].
5.3.50 Program the oven temperature from room temperature to the maximum temperature for the column. Increase the temperature at a rate of 10-15°C/minute and hold at the maximum temperature for 30 minutes. This can be done by pressing the [Oven] and enter the required parameters by using cursor, numeric and [Enter] keys.
5.3.51 If you will not be using the conditioned column immediately, remove it from the oven. Cap both ends to prevent air, moisture, and other contaminants from entering the column.
5.3.52 Conditioning the Packed columns:
5.3.53 Turn off the detectors. Shut off the detector support gases. It is especially important to shut off hydrogen. This can be done by pressing the [Front Det], detector control table parameters are displayed. Then turn required parameters to off mode by pressing [Off] at the required cursor position.
5.3.54 Connect one end of the column to inlet. But do not connect the detector end.
5.3.55 Cap the detector(s) fittings with the no-hole ferrule and column nut.
5.3.56 Establish a stable flow of carrier gas by setting the a flow of 20 to 30 ml/minute for 2 mm columns and 50 to 60 ml/minute for 4 mm columns.
5.3.57 Let gas flow through the column at room temperature for 15 to 30 minutes to remove air. This can be done by pressing [Col 1] and moving the cursor to flow parameter and set the required value by using numeric keys and press [Enter].
5.3.58 Program the oven temperature from room temperature to 30°C less than the maximum temperature for the column. Increase the temperature at a rate of 10-15°C/minute and hold at the conditioning temperature for overnight. This can be done by pressing the [Oven] and enter the required parameters by using cursor, numeric and [Enter] keys.
5.3.59 If you will not be using the conditioned column immediately, remove it from the oven. Cap both ends to prevent air, moisture, and other contaminants from entering the column.
5.3.60 Close the oven door.
5.4 Procedure to operate agilent 7694 head space sampler:
5.4.1 Switch on the instrument, which is located on the rear, bottom side of the instrument.
5.4.2 Entering new values by key board operation
5.4.3 Press a key to go to the first item in the menu of the parameters connected with each key. The cursor keys (?,?) scroll through the menu items for each control function.
5.4.4 The value displayed with each parameter is the current value of the set point for that parameter. To view a set point, you must first press the appropriate key on the keypad and then scroll through its list of set points.
5.4.5 To set a set point, key in the value on the numeric keypad. A flashing * will appear to the left of the new value as you enter it. Pressing Enter makes the new value part of the active method.
5.4.6 Press start/stop key to start the method.
5.4.7 Press Start/Stop key again in you want to stop the analysis, instrument ask for confirmation, press Start/Stop key again to stop the analysis.
5.4.8 Default parameters:
5.4.9 Unless other wise directed in the analytical method feed the following default parameters:
5.4.10 Set the Zone temperatures as follows,
5.4.11 Press [Zone Temps] key, Oven temperature is displayed, set the oven equilibration temperature as “80”.
5.4.12 Press cursor key [?], Loop temperature is displayed, set the Loop temperature as “85”.
5.4.13 Press cursor key [?], Transfer line temperature is displayed, set the Transfer line temperature as “100”.
5.4.14 Set Event times as follows,
5.4.15 Press [Event times] key, GC cycle time is displayed, set the time in minutes required to complete the GC Cycle, which include GC Run time and cool down time.
5.4.16 Press cursor key [?], Vial equilibration time is displayed, set the time in minutes the vial spends heating in oven as “60”.
5.4.17 Press cursor key [?], Pressurize time is displayed, set the time in minutes gas enters the vial through needle as “0.5”
5.4.18 Press cursor key [?], Loop fill time is displayed, set the time in minutes the gaseous contents of vial pass through loop to vent as “0.01”
5.4.19 Press cursor key [?], Loop equilibration time is displayed, set the time in minutes gas spend in loop, before injection as “0.05”
5.4.20 Press cursor key [?], Inject time is displayed, set the time in minutes, gases in sample loop are swept into the GC as “1.0”
5.4.21 Set the Vial Parameters as follows:
5.4.22 Press [Vial Parameters] key, first vial No is displayed, set the position No of the vial in the tray that to be analyzed first.
5.4.23 Press Cursor key [?], Last vial No is displayed, set the position No of the vial in the tray that to be analysed last.
5.4.24 Press cursor key [?], Shake level is displayed, set the agitation level of oven carousel as “0(off)”. Shake level 1 is low shaking and level 2 is high shaking.
5.4.25 Store method: Press [Store Method] key, to save the current changes, press a number (1-4) to store the method under that method number.
5.4.26 Load method: Press [Load Method] key to load the method, press the method number that contains required method.
5.4.27 Tray Advance: Press [Tray Advance] key to move the tray to required position.
5.4.28 Preparation of sample vial:
5.4.29 Place the cap upside down on the table.
5.4.30 Place the required quantity of sample / sample preparation in the vial. The vial should not be more than 50% full.
5.4.31 Place the cap over the vial opening.
5.4.32 Place the crimper over the cap and vial.
5.4.33 With slow and steady pressure, squeeze the crimper handles to seal the vial.
5.4.34 Remove the crimper.
5.4.35 Check that the seal is adequate by trying to twist the cap on the vial. It should be difficult or impossible to turn the cap. If you can easily turn the cap on the vial, re-crimp the cap.
5.4.36 Discard the vial if it fails the test.
5.4.37 Procedure:
5.4.37.1 Use the following procedure for all methods that require head space sampler.
5.4.37.2 Set the switch on the back of the horizontal loader to the proper vial size setting.
5.4.37.3 If 10-ml vials are used, insert the adapters in both sample tray and in oven. Ensure that these adapters are removed when 20-ml vials are used.
5.4.37.4 Place the Vials containing standard preparation are placed in the vial tray and programmed to inject the headspace of the vial, and record the response. Check the reproducibility.
5.4.37.5 Place the Vials containing sample preparation are placed in sequence and programmed to inject the head space of the vial, record the peak responses.
5.4.37.6 Unless otherwise directed in the analytical method, use 10-ml vials.
5.5 Procedure to operate agilent 6890 series injector:
5.5.1 Use this procedure if liquid Auto injector is used for the analysis. Place the agilent GC auto sampler in the front inlet port if the front inlet is used for the analysis and connect the plug to Injector 1 (Default Front) which is located on the back side of the Agilent GC auto sampler controller. Place the Agilent GC Auto sampler in back inlet port if back inlet is used for the analysis and connect the plug to Injector 2 (Default Back), which is located on the back side of Agilent GC Auto sampler controller.
5.5.2 Switch on the instrument, which is located on the rear, bottom side of the agilent GC Auto sampler controller.
5.5.3 Preparation of sample vial:
5.5.4 Place the cap along with septum upside down on the table.
5.5.5 Place the required quantity of sample / sample preparation in the vial.
5.5.6 Place the cap over the vial opening and screw it tightly or slug it tightly.
5.5.7 Place the vials containing sample solutions to be injected only in the vial tray labeled from 1 to 8.
5.5.8 Place the required quantity of the rinsing solvent (Used for the dilution of samples) in a vial, close it with a screw cap fitted with septum.
5.5.9 Position the rinsing solvent vial in Position A in the vial tray. If the second rinsing is required, then position the vial containing second rinsing solvent in the Position B.
5.5.10 Place the empty vial in the position ‘WA’ on the vial tray.
5.6 Procedure to create methods in agilent 6890 GC:
5.6.1 Create the methods if it is not already stored in GC
5.6.2 Press the Oven button to access oven control table and enter the required program as per analytical method. Where the method is isothermal run, enter the oven temperature for the isothermal run. Enter the number of minutes (init time) that the oven to stay at this temperature. If rate1 is not 0, enter 0 for the isothermal run. If the method is in programmed mode then set the Oven program accordingly. Enter the starting temperature (Temp). Enter the time (Init time) the oven to stay at Temp. Enter the rate (Rate 1) at which the oven temperature is to increase. Enter the final temperature (Final temp1). Enter the time (Final time 1) the oven is to hold Final temp 1. To end the oven program after Ramp program after ramp1, set rate2 to 0 or if necessary set the next ramp similarly.
5.6.3 Press on Front inlet or Back Inlet. Scroll to the Mode line. Press Mode/Type to see the inlet mode menu. Move the cursor to inlet mode required. Press Enter button. The inlet table may change depending on the choice. Scroll to Temp, Type the temperature required. Press Enter button. Set the remaining parameters as required.
5.6.4 Press Front or Back injectors, current injector options will be displayed, enter required injection volume 1 microlitre to 5 micro liters. Set the syringe size to the capacity of the micro syringe used. Feed the required number of rinse cycles for the syringe in corresponding fields for Pre-injection and Post-injection. Press [Method} button and scroll to the method number you wish to use.
5.6.5 Press Col1 or Col2 for column1 and column2 respectively, current column parameters are displayed,. Set the carrier flow rate or linear velocity to required value. Press Config, and then Col1 or col2 and set the column dimensions, mode.
5.6.6 Press Front detector, feed the detector temperature. Set the hydrogen flow at 35 ml/minute, airflow at 300 ml/ minute and the make up at 25 ml / minute constant column make up flow. Keep all the parameters along with flame and electrometer are set to on mode.
5.6.7 Press Signal1 and set the detector parameters as required.
5.6.8 To store a method, Press Method and scroll to the method number you wish to use. Press the store key. Confirmation will be asked and press Enter button. If the method already exists, then another confirmation will be asked, Press Enter button to overwrite.
5.6.9 To load a stored method, Press Method button to access the stored methods. Scroll to the method you wish to load, Press load button.
5.7 Procedure to create and run a sequence in agilent 6890 GC:
5.7.1 Press Seq button to open sequence control table.
5.7.2 Scroll to the sub sequence method # line.
5.7.3 Enter a method number. Use 0 for currently active method, 1 to 5 for the stored methods, or off to end the sequence.
5.7.4 Press Mode/Type button and select one of the three-injector types.
5.7.5 Supply the rest of the subsequence parameters like # of injections per vial (the number of repeat injections from each vial and Samples (the range (first-last) if sample vials to be analysed.
5.7.6 Press Store button and then Seq button to store the sequence. Enter the identification number of the sequence; press Enter to store the sequence.
5.7.7 To load previously stored sequence, Press Load button and then Seq button. Press the identification number of the sequence; press Enter to load the sequence.
5.7.8 To Run a sequence press Seq Control button. Scroll to the start sequence line and press Enter.
5.7.9 The sequence continues to run until all steps are completed. Make sure that the Millennium data station is in “Waiting for Injection” status before running the sequence.
5.7.10 To stop a sequence, scroll to the Stop sequence and press Enter button.
5.8 To reprocess the chromatogram:
5.8.1 Open the project window and select the channels, click update.
5.8.2 Select and double click the required file name.
5.8.3 Review window displays on the screen.
5.8.4 In review window open file menu, click open and select method set.
5.8.5 Select the appropriate method set and click “ok”.
5.8.6 Select the process and click integrate.
5.8.7 After integration completed select the file, choose save and click result.
5.8.8 Close the review window.
5.8.9 Press update icon at particular project window.
5.8.10 Select the required file name.
5.8.11 In main tool bar open tools and click print.
5.8.12 Print window displays on the screen.
5.8.13 Select appropriate report method and click ok icon.
5.8.14 After collect the report enter the GC logbook.
5.9 After analysis completed close the project window and log off the millennium and switch of the detector and computer
5.10 Flush the column with water and methanol for 60minutes each with a flow rate of 1.0ml/min.
5.11 Disconnect the column from the unit after washing and store in an appropriate place.
5.12 Switch off the instrument.
END OF DOCUMENT
Gas chromatograph,quality control, oven surfaces
