Automatic Resampling Using Chromeleon AutoDilution
Contents
1. 2011 ies Automated Dilutions Using Chromeleon AutoDilution and Two Injection Loops 2011 Thermo Fisher Scientific Inc formerly Dionex Corp Technical Note 84 LPN 2255 www dionex com en us webdocs 73254 TN84 ChromeleonAutodilution 05May2010 LPN2255 01 pdf accessed Nov 28 2011 4 AS DV Operator s Manual 2009 Thermo Fisher Scientific Inc formerly Dionex Corp Document No 065259 www dionex com en us webdocs 73226 Man IC AS DVoperators May2010 DOC065259 03 pdf accessed Nov 28 2011 Legal Notices 2011 Thermo Fisher Scientific Inc All rights reserved PEEK is a trademark of Victrex PLC All other trademarks are the property of Thermo Fisher Scientific Inc and its subsidiaries This information is presented as an example of the capabilities of Thermo Fisher Scientific Inc products It is not intended to encourage use of these products in any manners that might infringe the intellectual property rights of others Specifications terms and pricing are subject to change Not all products are available in all countries Please consult your local sales representative for details Part of Thermo Fisher Scientific In addition to these offices Thermo Fisher Scientific maintains a network of represen tative organizations throughout the world Thermo Fisher Scientific San Jose CA USA is ISO Certified TN111_E 12 11SV LPN2961 SCIENTIFIC2. 2mm Suppressor Current 23 mA ECD Total Step om2 Oms ECD Total Average Off Channel Pressure Step 0 20 s ChannelePressuresAverage Or Concentration 37 00 mM Curve 5 Flow 0525 mar 0 000 Wait CycleTimeState Autozero Valve _2 state B ECD_1 AcqOn ECD_Total AcqOn Channel Pressure AcqOn 30 000 ECD _1 AcqOff DeliverRinse Valve _2 state A ECD_Total AcqOff Channel Pressure AcqOf Volume 1000 End Figure 5 Example of Small Loop PGM commands e When using a Dionex AS DV Autosampler use a rinse step to remove the residual sample from both sample loops Without the rinse sample carryover will occur in the sample run Add the command DeliverRinse Volume 1000 to the end of the run which does not increase the overall run time e The AutoDilution commands and conditionals are created in the Large Loop PGM using postacquisition steps and the Create Purification Samples wizard which is described in detail in the following section e Create the sequence as the original run using the Large Loop PGM for all the samples Reminder Any reruns that are needed will be run from the same vial as the original run using the Small Loop PGM Using the Create Purification Samples Wizard The AutoDilution process is added to the Large Loop PGM file by adding a postacquisition step The Create Purification Samples postacquisition step is available only if the timebase is connected to the related server the s
3. dilution runs can be stored in either the original sequence or a separate sequence for easy identification of reanalysis results The Chromeleon software s AutoDilution using the Dionex AS DV Autosampler with two sample loops of different size offers a quick easy reliable and economical method to perform sample analysis thereby eliminating labor intensive sample dilution The Dionex AS DV Autosampler in combination with a Thermo Scientific Dionex ICS 900 ICS 1100 ICS 1600 or ICS 2100 system is the cost effective solution for environmental water and wastewater analysis The use of AutoDilution requires the Chromeleon AutoDilution license P N 069725 Column Thermo Scientific Dionex lonPac AS25 2 x 250 mm Eluent Source Dionex EGC KOH Cartridge Eluent 37 mM KOH Flow Rate 0 25 mL min Inj Volume A 25 pL B 2 5 pL Column Temp 30 C Detection Suppressed conductivity Thermo Scientific Dionex ASRS 300 Suppressor 2mm recycle 23 mA Chromatograms A Original sample B AutoDilution of Sample A Peaks 1 Chloride 97 mg L 2 Sulfate 50 900 1 US 2 A 0 50 nN Minutes aN N 29104 Figure 1 AutoDilution of a San Jose municipal water sample using two loops with the Dionex AS DV Autosampler This study presents an example of automatic reinjection utilizing a smaller secondary loop fill and injection using an optional 10 port valve available on the Dionex AS DV Autosampler and Dionex ICS 2100 system F
4. runs can be stored in a separate sequence for easy identification e Increased productivity saved time and reduced cost www thermoscientific com For more information on methods to automate dilution using the Dionex AS Autosampler refer to Dionex TN 81 TN 83 and TN 84 1 In Dionex TN 84 AutoDilution by the Dionex AS Autosampler is discussed using two loops with the Dionex ICS 1100 1600 2100 systems and in the Automation Manager for the Dionex ICS 5000 and ICS 3000 DC Detector Chromatography Compartment In Dionex TN 81 AutoDilution by the Dionex AS Autosampler is achieved by injecting smaller amounts in the existing loop using partial loop injections In Dionex TN 83 the Dionex AS Autosampler performs vial to vial dilutions using the Sample Prep module 1 Automated Dilutions Using Chromeleon AutoDilution and the Partial Loop Injection Capability of the ICS 3000 AS Autosampler 2009 Thermo Fisher Scientific Inc formerly Dionex Corp Technical Note 81 LPN 2205 www dionex com en us webdocs 70792 TN81 Chromeleon AutoDilution 06Mar09 LPN2205 pdf accessed Nov 28 2011 2 Automatic Vial to Vial Dilutions Using Chromeleon AutoDilution and the Partial Loop Injection Capability of the ICS 3000 AS Autosampler with the Sample Prep Option 2009 Thermo Fisher Scientific Inc formerly Dionex Corp Technical Note 83 LPN 2245 www dionex com en us webdocs 70900 TN83 Automatic Dilutions 17Mar09 LPN2245 pdf accessed Nov 28
5. will reinject any samples that are above the threshold limit using a small sample loop 2 5 pL injection volume On the first line in the Large Loop PGM Figure 4 add the command Valve_2 StateB This command will rotate the 10 port injection valve to the LOAD position for the large loop At time 0 000 after the AcqOn statement enter the command Valve_2 StateA This command will flip the 10 port valve to the INJECT position and transfer the contents of the large loop onto the column To the first line in the Small Loop PGM Figure 5 add the command Valve_2 StateA This is the opposite of the Large Loop PGM command and will rotate the 10 port injection valve to the LOAD position for the small loop At time 0 000 after the AcqOn statement enter the command Valve_2 StateB This again is the opposite of the Large Loop PGM and will flip the 10 port valve to the INJECT position for the small loop and transfer the contents of the small loop onto the column Valve 2 State B Valve_2 State A DeliverRinse Volume 1000 Valve 2 State B Figure 4 Example of Large Loop PGM commands Pressure LowerLimit 200 psi Pressure UpperLimit 3000 psi SA Equate SA CR TC On DeliverSpeed 4 0 ml min DelayVolume P25 pe FlushFactor 10 Valve 2 state A DeliverSample EndSamplePrep Data_Collection Rate SPU sval CellTemperature Nominal 3 5 0 icc ColumnTemperature Nominal 30 0 C Suppressor Type ASRS_
6. Loads Small Loop E sien Table 1 Functions of the 10 Port Valve Positions Detector Auto Sampler Suppressor Analytical Column Guard Column Small Loop Figure 3A Valve position B load Large Loop or inject Small Loop Detector Auto Sampler Suppressor Large Loop Analytical Column Guard Column Small Loop Figure 3B Valve position A load Small Loop or inject Large Loop Plumbing the 10 Port High Pressure Valve Figures 3A and 3B show the detailed plumbing connections for the 10 port valve e Connect the line from the pump to Port 1 e Connect the Small Loop to Ports 4 and 8 e Connect Ports 2 and 3 together with a short piece of tubing e Connect the Large Loop to Ports 6 and 10 e Connect the sample line from the autosampler to Port 5 e Connect the column line to Port 7 e Connect the green PEEK tubing waste line to Port 9 Creating Program PGM Files Examples of Large Loop and Small Loop PGM commands Figures 4 and 5 show the program commands used in this application The following examples use a 10 port valve on the Dionex ICS 2100 system To create the program file e Rename the current PGM to Large Loop This PGM will contain the postacquisition step and will be used on all initial runs using the large loop 25 pL injection volume Create a copy of this file named Small Loop see the Chromeleon User s Manual for instructions on how to create a copy of a file This PGM
7. Loop and Small Loop Two QNT files will need to be generated one for the Large Loop used in the original run and one for the Small Loop used with the rerun samples Optionally a calibration can be run for the Large Loop and a dilution factor of 10 operators need to confirm the loop volumes can be entered for the dilution samples in the sequence If a dilution factor is used the loop volumes should be confirmed in the following procedure e Obtain a 1000 mg L ppm bromide standard for anion determinations or a 1000 mg L ppm lithium standard for cation determinations e Prepare a 10 mg L ppm stock standard e Create a sequence using the Large Loop PGM file and run four injections of the 10 mg L ppm standard e Verify that the RSD is 5 or better e Create a sequence using the Small Loop PGM file and run four injections of the 10 mg L ppm standard e Use this equation for the dilution factor Average Area Large Loop Dilution Factor Average Area Small Loop e Use this equation to calculate the injection volume and enter it in the sequence in the Sample Properties dialog Volume of the Large Loop Volume of the Small Loop Dilution Factor The Chromeleon software s AutoDilution offers a quick easy and reliable method to deliver samples in the cali brated range of the instrument e Automated check for samples that exceed threshold e Automated rerun of samples with a small injection loop e All dilution
8. Technical Note 111 e Environmental e Water e Wastewater e Automation e Dilution e Sample Preparation SCIENTIFIC Automatic Resampling Using Chromeleon AutoDilution Two Injection Loops and the Dionex AS DV Autosampler Barbara Shao Kris Bahten John Madden Terri Christison Fei Pang and Frank Hoefler Introduction In environmental water analysis it is often necessary to establish multiple calibration curves to accommodate samples varying in concentration or to dilute concentrated samples to fit the standard curve AutoDilution an important feature of Thermo Scientific Dionex Chromeleon software provides a less time consuming approach to analyzing out of calibration range samples by utilizing a smaller sample loop compared to the one used for standards Operating AutoDilution using the Thermo Scientific Dionex AS Autosampler was previously demonstrated in Thermo Fisher Scientific Inc formerly Dionex Corp Technical Notes TN 81 83 and 84 1 However some customers prefer the Dionex AS DV Autosampler which requires additional commands in the instrument program Here AutoDilution is demonstrated using the Dionex AS DV In AutoDilution samples are first analyzed using a full loop injection from a large loop while the Chromeleon software monitors the sample concentration If the sample concentration is outside the specified range AutoDilution will automatically reinject the sample using a smaller loop All
9. ariable without an extension e g seq name the Chromeleon software will not create a new sequence but append the newly created samples to the existing samples in the current sequence instead The Sequence Properties dialog is also used to specify running a previously created sequence containing standards that need to run before the dilutions The sequence an operator selects will automatically run before the dilution sequence If desired standards can be selected to run before or after the diluted samples Sees Oe isa ie I TE bamia Fer Lib peu eqs ee ee es ree E E fe gee ee eee eee ees Peo ee ee ee gee ee el ne Const fe Figure 10 Sequence Properties dialog box Sample Properties In the Program box on the Sample Properties dialog Figure 11 enter the PGM file to be used for the rerun samples Open the dialog box and select the Small Loop PGM file that was created earlier Also enter the QNT method which is generally the same QNT method used for the original samples Enter the dilution factor in the sequence if the same QNT method is used to quantitate the dilution samples Dilution factors are discussed in the Confirming the Loop Volumes section If the operator has run a calibration for the Small Loop select this QNT file here The Type field indicates the Sample Type of the samples to be created The sample type is always Unknown The entry cannot be edited The Status field indicates the status of t
10. erver is running and an AutoDilution license is installed The program for AutoDilution is created using a built in wizard that guides users through the main steps required to create an AutoDilution program The wizard automatically shows dialogs needed to create the program To create the conditionals for the AutoDilution program open the Large Loop PGM and click on the postacquisition steps icon to add a postacquisition step Click on Click on this line to add a new step and select Create Purification Samples to start the wizard program The dialogs used in the wizard are shown below e Number of Target Compounds e Target Compound X e Target Compound X Tests e Special Options e Conditional Program Selection e Sequence Properties e Sample Properties e Extended Sample Properties Number of Target Compounds Create Purification Sena Harmer ot Tarett Canisaenh Erter ha number of tag comport f pag Figure 6 Number of target compounds dialog box In the first dialog enter the number of target compounds that are searched in the AutoDilution process up to a maximum of 10 target compounds Figure 6 Target Compound X On the second wizard page Target Compound X shown in Figure 7 defines how the target compounds are identified Any connected detector can be selected from the drop down list Peaks may be selected by name if operators have already defined and entered the QNT method in the sample list the names ente
11. he samples to be created The sample type is always Single The entry cannot be edited If a dilution factor is used it can be entered in the Sample field list by double clicking the respective line Chat Purittieainn Sareea Sasmal Pre peri aes Camii Prii z ETI hopem FGM VONESA ERACI beat AIE ME D Loop Gai meod HT EAE BE ea PERT a Pct Frai 82 dell E Figure 11 Sample Properties dialog box Extended Sample Properties The last wizard dialog box is the Extended Sample Properties Figure 12 In the Extended Sample Properties dialog select the tray and vial locations for the diluted samples For this method the small loop injections are made from the same vial as the Large Loop undiluted run Select Use analytic sample tray and Same as analytic sample vial The number of preparative injections for both methods is one The postacquisition step creation is now complete and ready to run Tea Fito naha soem ae I Lem towed tomy z C Dee bam ii deir oian Wal FS eee ime are sachet veal eevee if ra a betel te Gerd bom utes chet ocd Rhames of pirga kran rari fedan ft 8 Tp number iregi voller mj y tea m Figure 12 Extended Sample Properties dialog box It is important to calibrate both loops or to run calibration standards on both loops to produce the most accurate dilution results A calibration curve can be created for both loop sizes by running a calibration for each PGM Large
12. igure 1 Sample Results from Large Loop Reanalyze Sample Using Small Loop PGM Figure 2 Flow chart of the AutoDilution process Process Overview Figure 2 shows the flow chart of the AutoDilution process First the sample is injected using the large loop The Chromeleon software automatically determines if any of the target compounds are above the specified threshold level then automatically reinjects the sample using the small loop at the end of the sequence The diluted injections can be stored in either the original sequence or a separate sequence for easy identification of reanalysis results The Chromeleon software automatically specifies the sequence parameters programmed by the conditionals in the Create Purification Samples wizard Equipment Dionex I CS 2100 system or any Dionex ICS series system Dionex AS DV Autosampler equipped with a 10 port valve P N 068921 Chromeleon 6 8 AutoDilution License P N 069725 Dionex AS DV Autosampler Operation Specific procedures to operate the Dionex AS DV Autosampler with AutoDilution are described here For detailed operations refer to the Thermo Fisher Scientific Inc formerly Dionex Corp AS DV Autosampler Operator s Manual Sampling The Dionex AS DV carousel holds 50 vials from which the Dionex AS DV Autosampler samples in any order with sin gle or multiple injections from each vial Before loading the carousel always press Carousel Release butt
13. nually add the InjectPosition and DeliverRinse commands to the Command Parameters table Use this mode to define the volume and position where the rinse comes from e g DeliverRinse Volume 100 Position 10 Enter these rinse commands at the end of the program as shown Estimating the Delivery Volume The Dionex AS DV Autosampler calculates the delivery volume using the formula shown in the Total Volume Delivered Estimator Total Volume Loop volume 25 pL Delay Volume 125 pL x Flush Factor 8 1200 pL The large loop 25 pL volume is used to estimate the delivery volume in both cases Sample Loops Dilutions Using Both Large and Small Injection Loops and a 10 Port Valve In this method two sample loops on a 10 port valve are used for AutoDilution Table 1 The original sample is loaded through Valve 2 in State B and injected through the large loop 25 uL at State A using the Large Loop Program PGM The smaller injection volumes are achieved using a small loop 2 5 pL injection in reverse loading in State A and injecting in State B The Chromeleon software monitors the sample concentration while injecting the sample using Large Loop PGM and reinjects the samples using the Small Loop PGM 2 5 pL loop if the concentration is over the specified threshold limit Position B Function Valve_2 StateB Position A Function Valve_2 StateA Injects Large Loop contents Large Loop Ta i i Loads Large Loop Small Loop
14. on to reset the vial memory After loading the vials in the carousel press the Carousel Release button again to engage the carousel Specifying a Rinse The Rinse command delivers rinse liquid from a vial to the 10 port valve through the open valve position and then to waste In AutoDilution the Rinse function is used to rinse any residual sample from the small loop when injecting through the large loop or vice versa To perform a rinse cycle the Dionex AS DV Autosampler needs the position of the vial containing the rinse liquid the rinse volume and the time specified to start the rinse In the Chromeleon Server Configuration program first specify the position of the vials containing water for rinsing such as Vials 45 to 50 In the Program Wizard for the Dionex AS DV Autosampler rinsing is programmed through the Basic Edit or Advanced Edit modes To use the Basic Edit mode first select the mode then select the Rinse check box and enter the Volume Using Basic Edit mode the rinse is always delivered from the vials in the range specified in the Chromeleon Server Configuration program Always enter the rinse commands at the end of the program so that both loops will be rinsed prior to the next injection as shown in the Creating Program PGM Files section If the first vial is empty the rinse solution will be withdrawn from the next vial specified in the range until all rinse vials are empty To use the Advanced Edit mode ma
15. red in the QNT Method are listed for selection retention time greatest height or greatest area The Mass Spectrometry Settings are not used for this application Lrede Poritication Senpi largel Cempeand 1 ta Teg Darel Darr Channel hin Falei Pagh ye hiara Piar Dini blirit Timihirety enra Targi fikiri a EA mii Bcd Ti Aig Heef ja mra hii ij Adii f mme mi mig binigti im rs fel arh miee are a 5 i Fiar ttre atmi bae Figure 7 Target Compound X dialog box Target Compound X Tests The Target Compound X Tests dialog in Figure 8 defines the test used to determine if the sample is rerun at a dilution Custom tests can also be created using many of the standard Chromeleon software variables but are not covered in this document For AutoDilution the Amount Check is selected This test checks whether the amount of the target compound is above the threshold amount Double clicking on the test name will open the Amount Check Test dialog box This dialog box allows operators to select the channel from which they can then check the amount of the target compound from a drop down list The threshold is the concentration value limit for the target compound Any samples that have a target compound reported result report units are ignored above this concentration will be added to the dilution sequence If for example chloride is selected as a target compound and 10 is selected as the th
16. reshold any samples that have reported results above 10 ppm or any concentration units from the QNT file will fulfill this test and be added to the dilution sequence Paii FE a dahaki o ce beets hey lo ceed Tan Type T Fateh C tows perar g blpip e E aip Ef dered check i oO Peron uT ECO I Pestak Amain Chek Teri Bach hias Circ Hee Figure 8 Target Compound X Test dialog box Special Options Use of a conditional program Figure 9 is optional If mul tiple dilution levels will be used each in a separate PGM they can be specified in the conditional program dialog Checking this box will open the Conditional Program wizard page on which operators can define which PGM files will be used with the sample in the dilution sequence Because all reruns will use only a single PGM file Small Loop pgm a conditional program is not needed Uncheck both boxes Create Purification Sempies Special iptions Lien Donien Pte acc Dirim AST heeded ben Tg Compared 1 ee na Figure 9 Special Options dialog box Sequence Properties The Sequence Properties dialog box Figure 10 is where the name of the dilution sequence is entered Enter a name that will be readily recognized as containing the diluted samples for the original run A new sequence can be created for the rerun samples or they can be appended to the end of the current sequence If operators use the v
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