Waters ACQUITY UPLC Quick Start Guide.pdf

ACQUITY UPLCACQUITY UPLCTMTM Quick Start Guide Quick Start Guide Waters Asia Preparing the System Preparing the System 1.To enter the ACQUITY console.1.To enter the ACQUITY console.At this point,a system has been generated along with the project.Before we create methods,we need to enter the ACQUITY console to prepare the system.To enter the ACQUITY console,we need to enter Inlet Editor.Confidential 2 1.Select the inlet method from the MassLynx shortcut bar.2.Choose the second tap“ACQUITY Additional Status”.You can monitor current status of the ACQUITY Binary Pump,ACQUITY Sample Manager and the ACQUITY TUV detector.3.To enter the ACQUITY Console,Click the ACQUITY Consol Icon form the Sample Manager control panel.The ACQUITY Console will appear.March 05 Waters Asia Notice the picture of the system.To the left of the ACQUITY system image is ACQUITY UPLC System menu tree that can be expanded to illustrate each module and their interactive displays.To the right of the picture the system,there are flow and pressure read backs along with the percentage indicator bar.Notice the Green System Status Button in the lower left side of the screen.To the right of the screen,notice the buttons for flow and stop flow.March 05 3Confidential Waters Asia 2.How to prepare for ACQUITY Binary Pump.2.How to prepare for ACQUITY Binary Pump.Wet Priming the binary solvent manager Wet Priming the binary solvent manager Priming is a timed operation that replaces solvent in the path from the reservoirs to the binary solvent manager.During priming,the vent valve moves to Vent position to ensure minimal backpressure.The flow rate during priming can be 8 mL/min total(4 mL/min each for pumps A and B).Prime the binary solvent manager when performing these tasks:Changing reservoirs or solvents Preparing a new system or binary solvent manager for use Running the system after it has been idle for more than four hours 1.Choose the CONTROLPrime A/Bthe CONTROLPrime A/B Solvents function.2.Select A1/B1 as a pair with a time of 3 minutes and press start.Each line will prime at a rate of 4 ml/min.Once completed,wet prime the A1/B1 lines.March 05 4Confidential Waters Asia 3.Preparing the sample manager 3.Preparing the sample manager Prepare the sample manager for operation after you prepare the binary solvent manager.Preparing the sample manager involves these steps:1.Priming 2.Characterizing the seal 3.Characterizing the needle and sample loop volumes 4.Loading sample plates A.Selecting weak wash and strong wash solvents A.Selecting weak wash and strong wash solvents Tip:Tip:For best performance,follow these guidelines when selecting wash solvents.Otherwise,performance may be reduced,specifically Area/Height RSD and Linearity.This does not mean that all other solvent combinations are prohibited.Other combinations can be run with lower performance expectations or by manipulating default injection parameters.Use a weak wash solvent based on the sample and mobile phase chemistries of your application,making sure all solutions/buffers are miscible and soluble.The following table lists some weak and strong wash solvents recommended for use with certain mobile phase conditions.High sample concentrations may require other weak wash solvents.For best results,weak wash solvent should be equivalent to the initial gradient condition mobile phase composition(isocratic)Suggested weak wash and strong wash solvents Suggested weak wash and strong wash solvents Chromatographic Condition Chromatographic Condition Weak Wash Solvent Weak Wash Solvent Strong Wash Solvent Strong Wash Solvent Buffered aqueous,100%H2O or 0 to 25%50 to 100%methanol or reversed-phase methanol or acetonitrile acetonitrile Tip:Tip:For best performance,the weak wash solvent should be similar or identical to your isocratic or initial gradient solvent considerations,excluding buffers.Buffers are not recommended for use in wash solvents.These guidelines should also be noted:March 05 5Confidential Waters Asia B.Priming the sample manager B.Priming the sample manager The priming process fills the sample needle with solvent,changes the solvent,and/or purges air from the lines.You prime the sample needle and/or sample syringe to accomplish these tasks:Prepare the sample manager for operation Rinse the internal needle,the external piercing needle,and the injection port Remove bubbles from the lines Guidelines:Guidelines:Ensure that the priming solvent is correctly composed.It must also be high in quality and miscible with the other solvents.Use filters in all solvent reservoirs,and ensure the volumes of solvents are sufficient for priming.For example,allow approximately 20 mL for each solvent(A and B)when performing five primes.Requirement:Requirement:The sample manager must be primed before you attempt to characterize the seal.To prime the sample manager:To prime the sample manager:1.In the console,select Sample Manager from the system tree.2.Click Control Prime syringesControl Prime syringes March 05 6Confidential Waters Asia 3.Prime Syringes dialogue box will be appeared Confidential 7 4.Select“Sample syringe and wash syringesSample syringe and wash syringes”5.Type the number of primes in the Number of cycles text box(the default is 1the default is 1).Recommendation:Recommendation:Waters recommends 5 to 7 primes when you are changing solvents.6.Click OKOK.When the system status is“Idle,”priming is finished.Tip:Tip:Each prime takes approximately 2 to 4 minutes.C.Washing the sample manager needle C.Washing the sample manager needle Washing the needle is an optional procedure that flushes strong and/or weak wash solvent through the needle and injection port.Washing the needle removes contaminants from the inside and outside of the needle,the external piercing needle,and the injection port.You can also perform a needle wash to ascertain proper flow through the waste tubing and to confirm that the needle wash system is primed and properly operating.Requirement:Requirement:Do not use buffered solvents as wash solvents.Tip:Tip:Priming the system washes the sample needle,so whenever you prime the system,you can omit this procedure.Observing wash solvent recommendations Observing wash solvent recommendations Waters recommends that you observe these guidelines for washing the needle:To ensure that the strong wash solvent is completely removed,the system washes the needle with 200 L of weak wash solvent after you use strong wash solvent.You can increase,but not decrease,the default value of 200 L.Rule:Rule:Strong wash solvent does not contact the sample.Match the weak wash solvent to the mobile phase or to the initial conditions.March 05 Waters Asia If you dilute the samples,match the weak wash solvent to the sample diluent.Match the types of organic species in the wash solvents to those in the chromatographic application.To wash the sample manager needle:To wash the sample manager needle:1.In the console,select Sample Manager from the system tree.The sample manager information window appears.2.Click Control Wash Needle.Click Control Wash Needle.3.Wash Needle dialog box will be appeared.4.In the Strong Wash box,specify the volume for the strong wash solvent.Default:Default:0.0 L Tip:Tip:Using both a weak and strong wash solvent increases the wash time and solvent consumption because the system must be fully cleansed of the strong solvent before starting the next injection.5.In the Weak Wash box,specify the volume for the weak wash solvent.Default:Default:200.0 L Recommendation:Recommendation:200 to 500 L 6.Click OKOK.The needle wash begins.7.When needle washing is complete,the status returns to“Idle.”March 05 8Confidential Waters Asia 4.Preparing the detector 4.Preparing the detector If your system includes a TUV,PDA,or mass detector,prepare it for operation by following the procedures in this section.To start the TUV Detector:To start the TUV Detector:1.Ensure the detector cell is filled with solvent and free of air bubbles.The detector may not initialize correctly if the cell contains air.2.Press the power switch on the door to power-on the detector.The detector beeps three times and runs a series of startup tests,while the power and lamp LEDs blink.Initialization usually requires approximately 2 minutes,and lamp warm-up requires approximately 3 minutes.3.When the lamp LED shows constant green,start MassLynx.You can monitor the console for messages and visual signals.For best results,allow 30 minutes to equilibrate the detector and stabilize the baseline.Confidential 9 4.You can choose your wavelength March 05 Waters Asia Create the LC method and the detector method.Create the LC method and the detector method.Click on the Inlet MethodInlet Method icon in the Instrument panelInstrument panel of the Confidential 10 A.A.Setup the Binary solvent Manager.Setup the Binary solvent Manager.Click on the InletInlet icon.Click on each tab and make the appreciated entries MassLynx top level screen 1using the following parameters:March 05 Waters Asia Confidential 113.Modify ACQUITY Binary Solvent Manger dialog box appears .SolventsSolvents tap;nt sources for pump A(A1 or A2)and pump B(B1 or B2).st of r remove solvent names from the list.Click to open the 4Select the solveEnter a solvent name or select the solvent you want to use from the liavailable solvents.TipTip:You can add oEdit Solvent Name List dialog box and edit the list.March 05 Waters Asia March 05 125.Set the low and high pressure limits for the pump(s).The range for each setting extends from 0 through 15000psi,or you can use customized pressure units 6.Seal WashSeal Wash the seal wash.Choose a value from 0.5 7.Gradient tablGradient tablu can set the following parameters for gradient events:Time(min)R0.10 through 999.99 minutes Flow(mL/min)R0.000 through 2.000 mL/min%A Range:0.0 through 100.0,%B Range:0.0 through(100.0-%A)Curve:Sets the rate at which the solvent changes to the new proportions and/or flow rates For isocratic operation,the first row describes the flow and composition 8.Input the Run TimeRun Time.;Specify the duration forthrough 999.9 minutes.e e:Yoange:ange:Confidential Waters Asia Confidential 13B.B.Setup the Autosampler Setup the Autosampler 1.Click on the Autosampler Autosampler icon.Click on each tab and make the appreciated entries using the following parameters:2.Select Partial Loop or Full Loop.March 05 Waters Asia Confidential 14To specify the interval between one sample injection and the next sample loading,enter a value(in minutes)for Loop Offline.Ensure the value you enter is not less than the time needed for the current sample to leave the loop.Range:0.01 through 999.99.To decrease delay volume during the separation,isolate the loop from the flow stream by selecting Disable.3.Click the Advanced buttonAdvanced button to override the automatic operation of certain features.In the Advanced Settings dialog box,select any of the following options.This is where you can include custom parameters like syringe speed and air gap:rfill Factor Full Loop OveSpecify a value from 1.0 through 4.0 times,or leave it as Automatic.The Automatic default value is 3xdefault value is 3x(this value changes with smaller loops).Use the following information for overfill calculations:Syringe Draw Rate Specify a value from 2 through 1000 uL/min,or leave it as Automatic.Default:200Default:200.TipTip:Viscous samples may require a lower draw rate.Needle Placement Specify a value from 0.0 through 999.9 mm,or leave it as Automatic.Default:2.0Default:2.0.Air gaps Specify a pre-aspirate and post-aspirate value from 0.0 through 999.9 uL,or leave it as Automatic.Defaults:4.0 and 4.0Defaults:4.0 and 4.0.March 05 Waters Asia Confidential 154.Enter a solvent name or select the solvent you want to use from the list of available weak or strong solvents.TipTip:You can add or remove solvent types from the list.Click o open the Edit Solvent Name List dialog box and edit the list.Wash Solvent Volume:Enter the volume to use for weak and strong wash.Range:0 through 5000 uL.5.Set the following options for your column or sample.Column Select Off or specify a target column temperature from 0.0 through 65.0 Sample Select Off or specify a target sample temperature from 4.0 through 40.0.March 05 Waters Asia Confidential 16C.C.SeetSeet1.Click on thick on each tab and make the appreciated entries using the following parameters:tup the D ector tup the D ector e Detector Detector icon.Cl 2.Use this tab to set parameters for monitoring wavelengths.Channel B Tab appears only when you use dual wavelength mode.March 05 Waters Asia March 05 173.Lamp On Lamp On:Clear this check box to turn the lamp off.This should be done only for shutdown methods.(This function is available for Channel A Tab only.)4.Wavelength(nm)Wavelength(nm):The detector operates in single or dual wavelength mode.In single mode,specify a wavelength for Channel A from 190 nm through 700 nm,using increments of 1 nm.For dual mode,set any two wavelengths from 190 through 700 nm on their respective channel tabs.5.Sampling Rate(points/sec)Sampling Rate(points/sec):1 or 2 for dual mode;1,2,5,10,20,40 for single mode.You can set this parameter only on the Channel A tab.6.Filter Time Constant(sec)Filter Time Constant(sec):Program a filter time in seconds.Options:ow,Normal,Fast,Other.disable all filtering,select No Filter.Slow,Normal,and Fast are preset values that depend on the wavelength mode and data rate.Select Other to enter any value in the range of 1.0 through 50.7.Data modeData mode:Absorbance A,Max plot(A,B),Ratio A/B,Difference A-B for dual mode;Absorbance only for single mode.upon inject.9.Auto Zero On Wavelength ChangesAuto Zero On Wavelength Changes:Disable,Offset to Zero,Maintain Baseline.If you select Offset to Zero,the absorbance value on this channel will adjust to 0.000 when wavelength changes as a timed event.If you select Maintain Baseline,the absorbance level will be preserved when the wavelength changes as a timed event.No filter,SlTipTip:To8.Auto Zero on inject start(checkbox)Auto Zero on inject start(checkbox):If you check this box,the absorbance value on this channel will adjust to 0.000AUConfidential Waters Asia Confidential 18Create Create 1.Go t2.AddsSama sample list a sample list o the main MassLynx page.the ConAConA and Smaple TypeSmaple Type columns to the sample list(right click on the ample list,choose Customize Display)or choose Customize DisplayCustomize Display form the ple Field menu.3.Customize Field Display dialog box appears:Click the menu you want.March 05 Waters Asia Confidential 195.bottle can automatically enter the plate type and vial position.The Autosampler bedlayout will be accessed at this point.We will be using the 48-vial holder,2mL 4.Type the File Name.Choose the Inlet File and Mass File through right Click and select Browser.Browser.Right clicking on the 6.Autosmapler bedlayout appears.Click your vial position and then color is changed to green.March 05 Waters Asia Confidential 207.Righ