Teledyne ACCQPrep HP125 Installation And Operation Manual Download Page 131

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ACCQPrep HP125

Appendix A

A-3

ACCQPrep HP125

Appendix A

Figure A-1 Tubing diagram of the AutoSampler connected to the ACCQPrep

Auto

Sampler

Power

External

Detector

USB

Auto

Sampler

Waste

Level

Sense

B
IN

A
IN

Waste

Drain

Solvent Level Sensing

Solvent Inlet

Circuit Breaker

Auto

Sampler

Power

USB

Ethernet

Auto

Sampler

Waste

Level

Sense

B
IN

A
IN

Waste

Drain

Solvent Level Sensing

Solvent Inlet

Main

Power

Main

Power

Auto Injector

Optional

Column Valve
Optional

To Injection
Valve

To bottom
column port

Needle Wash

Not Normally Used

Not Used

Optional

External Gas

Not Normally Used

Solvent inlet

valves

optional

If not used,

connect directly

to A and B

Repeat tubing for A1 Solvent as needed

Needle Wash
Inlet Line

Check Valve

Auto Sampler Probe
Wash Pump

1 – Wash Inlet
2 – From Wash Station
3 – To Wash Station
4 – Waste

Waste

WASTE

Fraction Collection

A1 Solvent

Wash Station

Solvent Level
Sensing

Solvent Lines

Sample

Waste line

Wash station
supply

Do not extend length.

Summary of Contents for ACCQPrep HP125

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Page 3: ...ovided by the instrument may be impaired WARNING WARNING If you are using flammable solvents or chemicals with this system vapor concentration levels may exceed the maximum exposure levels as recommended by OSHA Guide 1910 1000 To reduce those levels to a safe exposure Teledyne Isco recommends that you place the system in a laboratory hood designed for the purpose of ventilation This hood should b...

Page 4: ...in the trian gle is a warning sign alerting you of dangerous voltage inside the product Symboles de sécurité Ce symbole signale l existence d instructions importantes relatives au produit dans ce manuel Ce symbole signale la présence d un danger d électocution Warnungen und Vorsichtshinweise Das Ausrufezeichen in Dreieck ist ein Warn zeichen das Sie darauf aufmerksam macht daß wichtige Anleitungen...

Page 5: ...ety Warnings 5 For Additional Information Technical assistance for the Teledyne Isco ACCQPrep HP125 can be obtained from Teledyne Isco 4700 Superior St Lincoln NE 68504 Phone 800 775 2965 or 402 464 0231 Fax 402 465 3001 E mail IscoService teledyne com ...

Page 6: ...ACCQPrep HP125 Installation and Operation Guide Safety Warnings 6 ...

Page 7: ...ng the AutoSampler 2 10 2 4 1 Establishing the Fluid Connections 2 16 2 5 Column Installation 2 19 2 5 1 Column mounting 2 19 2 5 2 Column Configuration 2 21 2 5 3 Column Select Valve Optional 2 21 2 6 Installation Troubleshooting 2 22 Section 3 Configuration 3 1 Configuration of the ACCQPrep 3 1 3 2 Network Configuration 3 2 3 3 User Preferences 3 2 3 4 Prep HPLC 3 3 3 4 1 Define a Column 3 4 3 4...

Page 8: ... 26 5 3 1 Editing a method 5 26 5 3 2 Alternative Ways to Create Method Files 5 30 5 3 3 Defining a Gradient 5 30 5 3 4 Real time Gradient Editing 5 31 5 3 5 Bypassing the Injection Loop 5 31 5 3 6 Monitoring the Purity Measure 5 32 5 3 7 Viewing runs 5 32 5 3 8 Manual Control of the ACCQPrep 5 35 Section 6 Maintenance 6 1 Introduction 6 1 6 1 1 Cleaning 6 1 6 1 2 Collection Rack and Tray Cleaning...

Page 9: ...6 22 6 8 4 Replacement of Ion Source Housing 6 23 6 8 5 Overpressure Error 6 23 6 8 6 Check Valve Cleaning 6 23 6 8 7 Replacing Check Valves 6 24 6 8 8 PurIon Cone Cleaning 6 24 6 8 9 Capillary Inlet Cone Removal 6 25 6 8 10 Capillary Inlet Cone Cleaning 6 25 6 8 11 Capillary Inlet Cone Installation 6 26 6 8 12 PurIon Troubleshooting 6 26 Appendix A A 1 Diagrams for the AutoSampler and ACCQPrep A ...

Page 10: ...ACCQPrep HP125 Table of Contents 4 ...

Page 11: ...high flow rates for maximum throughput or added tolerance for high back pressures as columns age WARNING Avoid hazardous practices If you use this instrument in any way not specified in this manual the protection provided by the instrument may be impaired this may increase your risk of injury The ACCQPrep HP125 is available in different configurations to meet your needs An optional Solvent Selecto...

Page 12: ... ELSD This system has the same high performance features as the ACCQPrep HP125 system but includes an internal evaporative light scattering detector ELSD During operation this detector can be com bined with the UV 200 400 nm or UV vis 200 800 nm detection to isolate visible and UV absorbing compounds as well as compounds with little or no chromophores ACCQPrep HP125 with optional PurIon Mass Spect...

Page 13: ...ch panel and up to ten network devices The touch panel shares control with all connected devices The system performs the most recent command from any control input 1 3 2 FileStorage To support operation from a variety of direct and network con nections the software and all files are stored in the ACCQPrep HP125 This ensures that your compound purification methods and run history files can be viewe...

Page 14: ...0 relative humidity maximum at 20 to 40 C non condensing Flow Rate Range 1 125 mL min ELSD peak collection limited to 4 125 mL min Flow Rate Accuracy tested with water at 6 9 bar or 100 psi 2 Pressure Limit 414 bar 6000 psi derated linearly from 6000 psi at 75 mL min to 4000 psi at 125 mL min Pressure Accuracy 5 of full scale Gradient Formation Binary gradient 2 solvent inlets Optional Solvent Sel...

Page 15: ...tection 10 1200 Dalton for S model 10 2000 for L model 1 Dalton Resolution Electrospray Ionization ESI or optional Atmospheric Pressure Chemical Ionization APCI Simultaneous positive and negative ionization for both S and L models Note 1 All specifications are subject to change Table 1 3 Component Materials List Chromatographic Tubing 316 stainless steel tubing PEEK and Fluoropolymer Drain Tubing ...

Page 16: ...ACCQPrep HP125 Section 1 Introduction 1 6 ...

Page 17: ...em The ACCQPrep chassis including AutoInjector Solvent Selector Valve Module and Column Selector Valve Module but without the pump assembly weighs up to approximately 64 pounds 29 kg Handles are provided on each side of the instrument to facil itate lifting CAUTION A two person lift is recommended to prevent injury The pump assembly weighs 25 pounds 11 5 kg resulting in a final weight of 93 pounds...

Page 18: ...ump 1 Remove the two pump retaining screws in the pump com partment 2 Align the pump assembly horizontally with the two notches facing the opening of the pump compartment Figure 2 2 Figure 2 2 Location of notches on the pump sled 3 Slide the pump assembly into the pump compartment It may require a little force since it is metal on metal sliding together If the force seems excessive ensure there is...

Page 19: ...f the pump First connect the black connection on the left side Ensure the tab on top of the connector is facing up before pushing it into the mating connector Figure 2 4 Figure 2 4 Black connection with tab facing up Then connect the white connection on the right side Ensure the tab on top of the connector is facing up before pushing it into the mating connector Figure 2 5 Figure 2 5 White connect...

Page 20: ...6 Location of threaded holes in the base of the unit and the hex wrench 2 2 2 Solvent Line Installation 1 The left two pump heads form the A solvent pump and the outlets are connected to a tee The outlet of this tee con nects to the left bulkhead union Tighten with the 1 4 open end wrench 2 The right two pump heads form the B solvent pump and the outlets are connected to a tee The outlet tee on th...

Page 21: ... and FEP fitting and make small turns as to not bend the tubing Figure 2 8 Figure 2 8 Wrench used to tighten the FEP inlet line 5 Once complete install the faceplate onto the unit by snap ping the side mounted connectors into place 2 2 3 Fluid Line Installation Note The solvent lines are installed at the factory and include labels identifying the lines A1 A2 and A3 for the A solvents and line B1 B...

Page 22: ...ELSD gas port Nitrogen input line circled The ACCQPrep ELSD system has several additional connections 1 Install the ELSD P trap vent assembly found in the acces sory package PN 3 250004 by placing the short tube into the port on the rear panel orienting the vent as shown and tightening the fitting nut finger tight plus a small amount more with a wrench Figure 2 10 Figure 2 10Back of ACCQprep with ...

Page 23: ... to the P trap drain vent assembly above the instrument and place 10 mL of liquid such as isopropyl alcohol into the tube Make sure the fluid level in the tubing doesn t exceed the level of the instrument case top If the tubing is raised too fast fluid may flow out the top of the vent tube causing a spill Lift it high enough so the fluid enters the P trap drain Figure 2 11 Figure 2 11Keep liquid l...

Page 24: ...t tube If the exhaust is restricted some of the gas flow could be diverted out the P trap drain resulting in reduced sensitivity and the need to route the P trap drain to an exhaust In normal operation the exhaust gas is dry and does not require a waste collection container CAUTION Discontinue use of the ACCQPrep if liquid is present at the Pump Drain Contact Teledyne ISCO technical service for as...

Page 25: ...t can be added in the CONFIGURATION screen This action can be completed at a later time 3 Once priming is complete a new screen will appear direct ing you to turn the valve from thePRIME position to the RUN position Note The system will prime both pumps simultaneously There is no method to prime A and B individually 2 3 Installing the Manual Injection Port Manual sample injection can be performed ...

Page 26: ...pump Ensure that the arrow on the check valve points toward the waste container This check valve ensures sample doesn t run from the port when the injection syringe is removed 2 4 Installing the AutoSampler These instructions assume that the AutoSampler was purchased with a new ACCQPrep In that case the AutoInjector portion of the AutoSampler is already installed in the ACCQPrep If not please refe...

Page 27: ... holes in the Y axis carriage 4 Using the two plastic thumbscrews secure the Z Drive to the carriage and finger tighten Figure 2 17 Figure 2 17Z Drive secured to the Y axis carriage 5 Rotate the Z Drive rotor back and forth to ensure that the Z Drive moved up and down freely Figure 2 18 Note If the rotor does not move freely check that the other cables are not interfering with the movement of the ...

Page 28: ... ports If disconnected in the future it is important to con nect the wash well waste tube upper tube on wash well labeled 2 to port 2 of the peristaltic pump and connect the wash well supply tube lower tube on wash well labeled 3 to port 3 of the peristaltic pump 8 Connect the tube labeled Wash Supply to the tube labeled 1 connected to port 1 of the peristaltic pump on the back of the Autosampler ...

Page 29: ...ure 2 20AutoSampler Module peristaltic pump with tubing correctly attached 9 Connect the tube labeled Wash Waste to the tube labeled 4 connected to port 4 of the peristaltic pump on the back of the Autosampler Module The free end of the tub ing can be placed in the waste container or tee d into an existing waste line Figure 2 21 Figure 2 21Wash and waste tubing connections and labels ...

Page 30: ...with the supplied tee s Auto Sampler Power External Detector USB To Auto Sampler Waste Level Sense B IN A IN A1 A1 A2 A3 B1 B2 B3 A2 B1 B2 B3 B4 J A3 Waste Drain Solvent Level Sensing Solvent Inlet Circuit Breaker Auto Sampler Power USB Ethernet To Auto Sampler Waste Level Sense B IN A IN A1 A1 A2 A3 B1 B2 B3 A2 B1 B2 B3 B4 Waste Drain Solvent Level Sensing Solvent Inlet Main Power Main Power Auto...

Page 31: ...gure 2 23 See Appendix A Figure A 2 for a larger diagram 14 The short USB cable connecting the USB B port on the top portion of the back panel of the Autosampler Module to the USB A port on the lower portion of the back panel of the Autosampler Module should already be pre installed If not make connection as seen in Figure 2 23 Auto Sampler Power External Detector USB To Auto Sampler Waste Level S...

Page 32: ...Prep 2 4 1 Establishing the Fluid Connections To establish the necessary fluid connections 1 Connect the sample probe fitting to the inlet port 2 of the injection valve on the ACCQPrep Figure 2 25 Figure 2 25Probe fitting connected to inlet port 2 2 Secure the probe tubing to the side of the ACCQPrep by removing the housing screw fitting the tubing bracket to the screw and replacing the screw Figu...

Page 33: ... bracketed solvent injections In this tech nique the injected sample is separated from the chromato graphic solvents by a small amount of air and a bracket solvent during the sample loading and injection process Figure 2 27Location of the test tube and the wash well on the AutoSampler To Power on the System The AutoSampler gets its power from the ACCQPrep 1 Turn ON the power switch on the AutoSamp...

Page 34: ...Sampler Wash Station 1 Ensure the wash station supply line is placed into a suit able solvent typically the strong chromatographic solvent and the waste line is routed to a suitable container 2 Go to TOOLS AUTOMATION MANUAL CONTROL and select the START WASH button Figure 2 30 Figure 2 30Screen of Start Wash button 3 Ensure the wash solvent is pumped into the center portion of the wash station and ...

Page 35: ...ive HPLC systems but these may be difficult to mount with the included hardware Larger diameter columns can also be used on the system at below optimal flow rates and will require the column to be supported outside of the instrument Column packing materials of 5 or 10 µm work well on the ACCQPrep due to its high pressure capa bility but any packing can be used if compatible with the flows and pres...

Page 36: ...y slip the mount into one of the two column mounting tracks on the side of the ACCQPrep Figure 2 33 Figure 2 33Column track on the ACCQPrep 2 Remove the union connecting the column fluid supply line to the column effluent line 3 Connect the line from the injection valve to the inlet of the column and connect the remaining tubing to the column outlet Figure 2 34 ...

Page 37: ...plete set of column mounts can be obtained P N 60 5234 790 In addition an assortment of tubing lengths and nuts can be obtained P N 60 5239 025 2 5 2 Column Configuration The ACCQPrep software supports default methods for individual columns This allows easy selection of default methods with a single key touch This process is described in section 3 4 1 2 5 3 Column Select Valve Optional The ACCQPre...

Page 38: ...ld like to flush the internal tubing of the ACCQPrep between solvents such as when changing from a low pH to a high pH one of the column positions such as position 4 can be connected with a length of tubing For ease of use the column defined for the position could be called Flush When changing from a low pH to a high pH solvent a line can be placed into the AutoSampler queue to use a method with n...

Page 39: ... Check the USB cable connection from the lower rear panel of the AutoSampler to the USB connector on the upper left corner of the rear panel while viewing the AutoSampler from the rear panel Recycle the power on the ACCQPrep The PeakTrak software only checks for the presence of the AutoS ampler on power up If the AutoSampler is powered up after the ACCQPrep the software will not detect the presenc...

Page 40: ...up waste lines 7 ELSD PurIon connections and gas supply if applicable 8 Connect and route drain lines 9 Position the system 10 Installation of the collection tube racks 11 External detector optional 12 Turn on the power 13 Configure the system 14 Prime the solvent lines 15 Prime AutoSampler wash station if installed 16 System verification Certification of Section 2 Completion Installer Name print ...

Page 41: ...the solvents are selected on the MAIN RUN screen and the drop downs are not shown in the CONFIGURATION screen The CONFIGURATION screen will still show the ADD SOLVENT and REMOVE SOLVENT buttons to edit the list If you do not see your solvent name on the list click on ADD SOLVENT and type in the solvent name There is also an option to remove solvent names you no longer use Solvent Level Sensing ENA...

Page 42: ...y save data files right to the network To connect to a network you will need access to IP addresses and network information from your IT department For more information on network guidelines see TN28 Networking Guidelines for CombiFlash Products available at www isco com 3 3 UserPreferences The following options are found on the USER PREFERENCES tab and can be found under TOOLS CONFIGURATION USER ...

Page 43: ...IGURATION PREP HPLC Loop Volume Ensures that the samples entered do not exceed the loop volume amount per injection A warning will display if half of the loop volume is exceeded but it will not limit the injection amount that can be programmed This warning can be ignored Column Position This allows you to define a column s used on the instrument If you do not have the Column Selector Valve Module ...

Page 44: ...lumn to a suggested stor age fluid concentration 6 The pressure limit is set by default to 3000 psi 207 bar which is a typical limit for preparative HPLC columns but can be changed for each column to a suitable pressure not to exceed the system pressure limit of 6000 psi 7 Press SAVE to save the default method associated with the column To customize the just created method press DEFINE METHODS fol...

Page 45: ... created If multiple methods are associated with a single column these methods can be easily accessed whenever a particular column is selected These methods will be shown in the column selection drop down menus in the order the methods were created To create the new method 1 Select the column the new method will be associated with 2 Press the DEFINE METHODS button Figure 3 3 ...

Page 46: ... convenient way of minimizing method development Typically a universal gradient separation can be performed on an analytical HPLC system Based on the retention time of the analytical separation a method with an ideal narrow range gradient such as 20 30 B can be selected for optimal separation of a target peak When creating a set of focused methods keep in mind that the methods will be displayed in...

Page 47: ...separation are stored with the chromatographic data so no information is lost In addition the method and injection parameters used during a separation can be extracted from the data file and saved as a named method for future reuse 4 2 Method Selection Several options can be used to select a method Use a pre programmed column that hasseveral methods associated with it Once the column is selected y...

Page 48: ...e PurIon operating conditions This allows the use of default or commonly used methods while allowing customization of the separation for both masses used for fractionation or mass loading of sample sent to the PurIon for detection Low mass load is the default and sends the minimum amount of sample to the PurIon for detection Due to the high concentrations of sample in a preparative separation this...

Page 49: ...n AutoInjector Module is present If an Auto Sampler Module is present you will select the AUTO SAMPLER dropdown Note If you want to manually inject your sample without using the AutoInjector Module you will have to replace the sample probe fitting with a Luer injection port supplied or 22 gauge needle port optional Figure 4 1 Selecting for multiple injections 4 In the dialogue box enter the total ...

Page 50: ...y changes made to the method will be included on any of the remaining injections This includes automatic changes such as using the peak hold feature or automatic run length extension For the most reliable operation it may be best to disable these automatic features Mid separation flow rate changes will occur at the beginning of the next separation Please be aware that changes of flow rate will not...

Page 51: ...an be named A03 I1 the second can be A03 I2 and the third can be named A03 I3 8 Select the file you wish to display 9 The remaining injections can be immediately viewed by pressing the left or right arrows at the lower left and right of the file viewer screen This function can also be used to immediately view files before or after the currently viewed file even if they are not part of the same inj...

Page 52: ...nal injection will aspirate a small amount of air Small amounts of air will not damage the HPLC column If the sample container held more sample than programmed there may be some sample remaining in the probe after the final injection For this reason it is recommended that users program the injection sequence with about 0 5 mL more sample volume than the actual sample amount provided in the sample ...

Page 53: ...screen On this screen you can perform a manual injection by removing the Luer fitting from the inject loop or you can perform multiple injections on a single sample from the AutoSampler Module A queue will be created automatically Selecting the QUEUE tab will allow you to set up a separation sequence for multiple samples Note If you are performing a manual injection the Luer fitting or a needle po...

Page 54: ...ethod parameters are saved with each separation If you would like to save the modified method for later use select SAVE AS before you exit the method editor and create a unique name for the new method Sample Position This field indicates where the sample for this separation is located If the sample size is too large for a single vial create a second line in the queue and access the second vial Sam...

Page 55: ...splayed at the top and bot tom of the screen which allow access to hidden rows 6 Once the entire queue has completed the RESULT screen is displayed with the last separation To easily view previous separations select the separation on the QUEUE screen or select FILE OPEN to view previous separations If a sample had multiple injections the run sequence will be dis played within the FILE OPEN dialogu...

Page 56: ...o the sample probe to minimize the mixing of the wash solvent and the soon to be loaded sample This also minimizes the potential of the sample coming out of solution 6 The sample probe is moved to the sample container 7 A portion of the programmed injection amount is aspirated into the sample probe displacing most of the wash solvent in the sample probe This compensates for the volume of the sampl...

Page 57: ...ong solvent remaining in the loop from affecting the current separation 2 The sample probe is placed into the wash station 3 About 1 5 mL of the solvent is pulled from the wash station through the probe to remove air from the probe line This minimizes injection volume errors due to air in the probe Since the loop is still in the column flow path this aspi rated fluid bypasses the loop and is sent ...

Page 58: ...the probe with the following sequence 1 The inject valve will be moved to bypass to prevent contamination of the loop during the cleaningprocess 2 Air is drawn into the probe to eliminate any remaining solvent in the probe 3 The probe is placed into the wash station The wash station pump will flush wash fluid over the exterior of the probe while 10 mL of wash fluid is drawn into the probe to wash ...

Page 59: ...s to scratch the valve sealing surface Filter the sample with a 20 µm or finer filter Dissolve sample in sufficient solvent so that evaporation of solvent doesn t cause formation of crystals Manual injection using a hypodermic needle with pointed tip can scratch the internal sealing surface resulting in leaks When using a needle injection port only use a 22 gauge blunt tip needle When using a luer...

Page 60: ...t points and place into fluid to determine location of problem If loose fitting tighten or replace if needed If plugged replace part or connect to a high pressure pump to dislodge plug Incorrect wavelength used assuming no other detectors than UV UV vis Compound doesn t absorb light B Solvent not being delivered Check solvent level connections check valves confirm solvent lines in correct bottles ...

Page 61: ...aks elute too late Solvent B not being delivered properly Check solvent level inspect or clean pump B check valves confirm solvent lines in correct solvents look for leaks of air into the lines Check for proper flow rate of both solvents If flow of one of the solvents is only half of the expected flow prime the system again since one of the pump heads isn t primed If still unsuccessful open the fr...

Page 62: ...iscussion on ELSD operation in section 5 2 7 before adjusting the ELSD operating temperatures If the system detects peak shortly after the first separation of the day and then fails to detect the drift tube temperature may be too low to vaporize the solvent aerosol or the spray chamber too warm to condense out excess higher boiling point solvents Refer to the discussion on ELSD operation in sectio...

Page 63: ... small increase in dispersion in the alignment compensation tubing PurIon detection problems Little or no mass spectrum detected on method devel opment screen but compound should be visible Sample is dissolved in a solvent not suitable for MS injection such as DMSO or DMF Dissolve in methanol or acetonitrile and try again Try other ionization parameters such as switching to Robust Verify that the ...

Page 64: ...into the system f Filter samples in the future to prevent problems Fractions not correct volume Air trapped in pump head Reprime and test again Collect all with 100 A and 100 B If only 1 solvent shows filled fractions one of the heads is not priming Remove the front panel and loosen the fitting of each individual head while running the pumps in manual control Air leak in inlet lines Remove front p...

Page 65: ...tings are dis played on the METHOD EDITOR window Many of the PEAKTRAK WINDOWS contain these main elements or a subset of them Note The available commands and options will change according to the active window the state of the system and the current file 5 1 2 Method File PeakTrak controls the separations performed by the system through a METHOD FILE You do not have to create a complete method to p...

Page 66: ...g screen There can be a DEFAULT METHOD for each size and type of column defined in the config window DEFAULT METHODS provide a starting point for your separation or purifi cation From these initial settings you can perform a purification run or you can modify the settings for your next run Subsequent runs will use the settings in the active temporary window If you have modified the settings you ca...

Page 67: ...is opens the SET DATA PATH window Use this window to specify the default storage folders for users These folders contain multiple METHOD FILE and RUN FILE resources created by system users Save Method Choose this menu command to save the settings of the current method file to the system s internal storage if the file is already named If not already named you will be prompted to give the method a n...

Page 68: ... window so you can select the method file After selecting the file click the OPEN button to save the METHOD FILE on the system s internal hard drive Export Method Choose this command to save a METHOD FILE in a location other than the system s internal hard drive This can be done to archive METHOD FILES or to transfer the method to another system using the menu IMPORT METHOD command From the touch ...

Page 69: ... settings This command opens the IONIZATION SETTINGS window to allow creation of custom ionization parameters that can be saved for various compound classes and projects This command is not available from a remotely connected computer via a web browser Operate This command sets the status of the PurIon mass spectrometer from standby to an operational mode by applying power to the heaters and dynod...

Page 70: ... safe place Passwords protect unauthorized modifications to User management System date and time System password Before any of the above can be accessed you must enter a password Password protection can be disabled so all users can modify the entry User Management Choose this command to open the USER MANAGEMENT window You can use this window to add or remove users from the system To add a user cli...

Page 71: ...uses the administrator password Software Update Choose this command to open a window used to install patch files to update your ACCQPrep system software Export Log Files During operation the system writes operating data to a log file Teledyne Isco service personnel can interpret this data to optimize system performance or to troubleshoot difficulties Teledyne Isco recommends that you use the menu ...

Page 72: ...window displays the RUN SETTINGS RUN CONTROL buttons and PEAK COLLECTION buttons After a run the left side of the window changes to show the collection rack map or the method parameters in a PEAK COLLECTION DATA pane Gradient Plot The gradient plot area depicts the current gradient that will be used for the run The X axis of the plot displays the run time in minutes The X axis scale can be adjuste...

Page 73: ...PurIon S and PurIon L systems only A dot in the button indicates whether the positive or negative ionization spectrum is displayed UV The UV UV Vis on ACCQPrep systems with a UV Vis detector button displays the UV spectrum on systems This spectrum hides the flow rate and solvent selection controls Pressing the UV control again causes the spectrum to disappear When the spectrum is displayed touchin...

Page 74: ...lled an isocratic hold Note Pausing the run extends the run length While in the paused state you can resume the gradient by clicking the PLAY button or stop the run by clicking the STOP button If you resume the run the system continues the gradient curve from the B when the system was paused STOP This button suspends the entire run Unlike the Paused state the pump peak detection and fraction colle...

Page 75: ...tton to collect only eluted peaks in the fraction collection tubes NONE Click this button to divert all eluent to the waste port This is sometimes used to divert all peaks except the peak of interest to waste This action can also be accomplished with the Initial Waste or Time Windows functions Peak Collection Data Peak Collection data is displayed in the MAIN window after a run You may also open a...

Page 76: ... system s internal storage EXIT Click this button to close the METHOD EDITOR and return to the MAIN WINDOW COLUMN DATA Click this button to open the COLUMN DATA window This window reports information about the column selected on the main screen Run Settings The Run Settings section contains the following Column The name of the currently selected column is displayed If a Column Selector Valve Modul...

Page 77: ...ents or notes for the run These comments will be saved with the run and will appear in TXT and PDF reports Column identification information is automatically placed in this field Gradient Plot Area The gradient plot area depicts the current gradient profile that will be used for the run The X axis of the plot displays the run time in minutes The X axis scale can be adjusted by typing or selecting ...

Page 78: ...omplete instructions on defining gradient curves see Section 5 3 3 Defining a Gradient The gradient table depicts the points that define the gradient curve in a tabular format From this table you can change Solvent the duration and B concentration of any point on the curve To do so select the new value for a gradient point Changes that you make to the table will be reflected in the gradient plot a...

Page 79: ...indow contains option buttons to enable and disable various peak detection options When an option is enabled the window also allows you to modify the setting details for that peak detection option All enabled options will be displayed on the chromatogram The options and settings for the ACCQPrep systems are described below Note You can select up to four peak detection options on the ACCQPrep syste...

Page 80: ... 45 seconds you should enter a peak width of 1 minute Sometimes very small peaks need a larger peak width setting since their small height results in a smaller slope than larger peaks of the same width All Wavelength Detection Enable this option to detect peaks within a user selected range of wavelengths When enabled click the DETAILS button to configure additional settings These settings include ...

Page 81: ...or option is not available on ACCQPrep systems with ELSD or PurIon detectors ELS Detector Enable this option to use the evapo rative light scattering ELS detector on an ACCQPrep system When enabled click the DETAILS button to configure additional settings Sensitivity Can be selected as NORMAL to keep larges peaks on scale or HIGH to detect small peaks The default setting is HIGH to ensure most com...

Page 82: ...m This information includes The Number of times used First used on and Last used on information can help you determine when the column should be replaced The Last fluid used will help you determine if any solvent remaining in the column will be miscible with the solvent currently used in the system To view the Column Data window open the METHOD EDITOR window then click the COLUMN DATA button 5 2 9...

Page 83: ...a Save As command use this box to name the file OPEN SAVE DELETE This button performs the listed action Load Previous Run from Detected Rack This option appears when using the FILE OPEN command If you click this option the system reads the RFID tag on a single rack and displays the last RUN FILE collected on the detected rack on this instrument This feature is useful you are unsure of the rack s c...

Page 84: ...tive ionization Ion Settings Allows selection of different ion settings to maximize the intensity of the desired molecular ion The factory selections include Robust for compounds that do not easily ionize Typical works well for most compounds The Fragile setting is used for those compounds that are delicate or easily fragment User defined ion settings may also be loaded Ion Finder Identifies poten...

Page 85: ... to a narrower range causes the system to relabel the ions to identify ions that may not have been intense enough to be labeled on the broader range graph Spectrum Name the file name for the mass spectrum is only needed if you want to save the mass spectrum to a file for later viewing Save Data saves the mass spectral data to the internal hard drive Save as PDF save the displayed spectrum on eithe...

Page 86: ...just various settings to improve the ionization of a particular compound by reducing fragmentation or adduct formation Mass Spectrum Graph this displays the mass spectrum There are two mass spectra displayed Blue displays the maximum of the current injection Red displays the maximum of the previous injection for comparison When a new injection is performed the blue spectrum replaces the red spectr...

Page 87: ...hanges and create a new ion settings file This new file can be selected in ionization settings or method development Additionally this file can also be selected when setting up a separation run Factory Settings restore the typical Robust and Fragile ion settings to the factory default values Probe The probe nebulizes and ionizes the sample There is a choice of ESI electrospray interface or APCI at...

Page 88: ...ent in a tube select the OPTION button and enter the tube number This feature can be useful when trying to recover a compound that has precipitated or crashed somewhere in the fluid path Prime A Click the PRIME A button to pump 100 of solvent A at the selected flow rate Prime B Click the PRIME B button to pump 100 of solvent B at the selected flow rate PRIME XX B Click this button to pump a mix of...

Page 89: ...llow the on screen instructions for entering the address queue and printer type information After successfully printing a test page the network printer will be available for printing using the FILE PRINT menu command and the AUTOMATICALLY PRINT AT END OF RUN feature Network File Save Configuration When configured this feature allows the system to access a network directory from which it can save r...

Page 90: ...ed Basic Method Settings The basic settings allow you to control the B gradient mix flow rate solvents and run length These settings are part of the MAIN window These settings are the most frequently changed settings when developing or improving purification methods The B gradient mix and run length are shown on the plot area Refer to Section 5 3 3 Defining a Gradient for more details The other se...

Page 91: ...l other fluids to the waste port The None option will divert all fluids peak and non peak to the waste port The None option is useful for developing custom methods that perform a column wash system cleaning or similar function where solvents and elute do not need to be collected When using the All or Peaks options you can specify the maximum Peak and Non peak volumes to be collected in the tubes B...

Page 92: ...baseline The slope detector will detect peak widths ranging rom about 0 2 to 2 times the peak width setting For example if you entered a peak width of 1 minute the range would be 12 seconds to 2 min utes For best operation the peak width should be set to just over the average peak width being separated For instance if the average peak width is 45 seconds you should enter a peak width of 1 minute F...

Page 93: ...settings Signal Gain You may modify the gain to scale the detector signal Slope Based and Threshold Refer to these features described in λ1 and λ2 above Spray Chamber Temperature The default settings are 30 C normal phase default methods and 15 C reverse phase This setting may be adjusted from 10 to 60 C so that detection can be optimized for the solvent system in use This setting is limited to a ...

Page 94: ... than editing a method file on the ACCQPrep system there are more ways to create method files Importing Method Files from another System If a method file is located on another ACCQPrep system you can use the FILE EXPORT METHOD menu command to save the method file on an external storage device From the touch screen panel this storage device is a USB Flash Drive connected to the USB port below the d...

Page 95: ...n of the Prep injection loop During normal operation the loop is placed into the flow path ahead of the column after the equilibration is completed It remains in this position for the remainder of the separation This feature allows you to remove the loop from the flow path and is useful for manually stacking injections when doing an isocratic separation It will require you to perform suffi cient m...

Page 96: ...ng a second wavelength it may be possible to reveal the second compound Because of the slight shift in time and the different absorbance properties of the two compounds the changing ratio during the detected peak would reveal the impurity Therefore one can therefore assume that if the ratio is not constant for the entire duration of the peak the compound eluting may not be pure The SHOW SPECTRAL P...

Page 97: ...ave a PDF file from the touch panel display insert a USB Flash drive in the USB port below the display panel SAVE AS TXT Click this button to save the run summary in an ASCII text file format When viewing the run from a remote personal computer the summary can be saved to any connected storage device If you are attempting to save a TXT file from the touch panel display insert a USB Flash drive in ...

Page 98: ...PREVIOUS and NEXT RACK buttons are active Click these buttons to scroll through the available racks The currently displayed rack is identified by letter which is shown below the rack Reading the Chromatogram The plot area displays the following The red absorbance trace produced by the system s peak detector Absorbance units also shown in red that correspond to this trace are shown on the left Y ax...

Page 99: ...cific tube number Raw Lamp Energy The left side of the Manual Control window shows a Raw Lamp Energy gauge This provides an indication of the UV light mea sured by the optical detection system at 254 nm High lamp energy green means that the flow cell easily passes through a sufficient UV light source Lower lamp energy yellow or red means that either the light source is weak or that the flow cell i...

Page 100: ...ACCQPrep HP125 Section 5 PeakTrak 5 36 ...

Page 101: ...ive properties The racks and tray must be kept clean to dissipate static electricity The collection tube racks and tray are made of conductive plastic Dirt film or coatings might prevent their ability to dissipate static electricity To avoid problems that possibly result from an electrostatic discharge clean the racks and tray monthly Use distilled water with a mild detergent For tougher stains us...

Page 102: ...ys wait at least one minute after placing the system in STANDBY before removing the AC mains power cord 6 2 1 Tubing Inspection WARNING Risk of fire or equipment damage Faulty tubing fittings and drains may allow organic solvents to pool in unsafe areas creating a potential for dangerous levels of flammable vapors Improper draining may damage the instrument s internal components Perform a tubing i...

Page 103: ... high percentage of solvent B to flush residual compounds from the column internal tubing and flow cell Refer to section 6 4 Allow Cone wash to run to completion PurIon systems only to wash residual compounds from the fluid interface probe and to clean the cone area Allow Valve Wash sequence to run to completion to wash residual compounds from the injection valve and ELSD if installed flow path Mo...

Page 104: ...ance trace Frequent messages recommending flow cell cleaning Figure 6 3 Typically chemists STOP and then TERMINATE the run after the last compound elutes This action skips the post separation flush If any of the above conditions appear consider allowing some of the runs to continue through the flush or run a high percentage of B solvent through the system for a few minutes at the end of each day I...

Page 105: ...rest to waste Help Click this button to display the flow cell cleaning on line help topic To perform a quick cleaning 1 After clicking the CANCEL RUN button select the TOOLS MANUAL CONTROL menu option 2 From the MANUAL CONTROL window note the Raw Lamp Energy level at 254 nm The Raw Lamp Energy gauge has two ranges red and green Figure 6 3 Figure 6 3 Raw lamp energy gauge Red Lamp energy is obstruc...

Page 106: ...system pumps solvent the raw lamp energy should gradually improve Pump solvent for two to five minutes or until the indicator reaches the far right of the green range If after five minutes the indicator is still in the red range repeat the cleaning steps using an alternative solvent Or complete the steps in the Monthly Flow Cell Cleaning procedure section 6 4 3 Note If the numerical values of the ...

Page 107: ...g instructions are for the removal and replacement of the injection valve rotor PN 209009917 ACCQPrep systems with a serial number starting with 218H or higher may have spare rotors included with the system For these units access the pump compartment by gripping the edges of the front panel and pulling forward on both sides Figure 6 8 Inside the front cover an envelope contains spare rotors a hex ...

Page 108: ...ide to side motion until removed 3 When the stator is removed use your finger or a small tool to pry the black plastic rotor out of its recess Figure 6 6 Figure 6 6 Removing the rotor 4 Usually the failure mode is a visible scratch between the normal passages on the rotor Figure 6 7 Sealing surface of the stator Gap will appear here Screws Rotor Sealing surface ...

Page 109: ...alve body evenly The rotor seal is accomplished by a spring so excessive screw torque is not necessary 6 5 2 Pump Seal Replacement 1 Turn off the power to the system 2 Access the pump compartment by gripping the edges of the front panel and pulling forward on both sides Figure 6 8 Figure 6 8 Front panel of the ACCQPrep 3 Remove the two screws securing the pump assembly in place using the included ...

Page 110: ...igure 6 10 Note The seal back up washer may remain on the piston Remove the washer from the piston if it did not stay in the pump head Figure 6 10Removing the pump head 10 Carefully pull the pump spacer block forward and off the guide pins Pull straight and slowly to prevent damage to the piston Also remove the guide bushing from the piston if it did not stay in the pump spacer block Figure 6 11 ...

Page 111: ...he sealing sur face and create a leak Figure 6 12Removing the seals 12 Use the scouring pad included in the seal replacement kit to clean the piston Gently squeeze the piston within a folded section of the pad and rub the pad along the length of the piston Rotate the piston frequently to assure the entire surface is scrubbed After scouring use a lint free cloth dampened with alcohol to wipe the pi...

Page 112: ...on removal tool so that the spring energizer is visi ble when the seal is fully seated on the tool Insert the tool into the pump head Figure 6 14 Note Be careful to line up the seal with the cavity while inserting Figure 6 14Replacing the seal 14 Carefully replace the pump spacer block making sure the O ring is properly installed Figure 6 15 ...

Page 113: ...s are on top Figure 6 16 Note Push onto guide pins straight and slowly to prevent damage to the piston Do not force the self pump spacer block or pump head into place Figure 6 16Reinstalling the pump head 16 Reinstall fasteners As you tighten alternate side to side until snug Turn 1 flat past snug using a 3 16 hex key Fig ure 6 17 Note Be sure the Inlet Check Valve Housing is on the bottom and the...

Page 114: ...de methanol or acetonitrile and water Suggested Conditioning Parameters Using a suitable column run the pump with a 50 50 solution of Methanol Water for 30 minutes at a flow rate that results in a pressure of 1000 psi Then run the pump for another 15 minutes at a flow rate that results in 3000 psi 6 5 4 Check Valve Replacement The following instructions are for the removal and replacement of the p...

Page 115: ...ot tles Place a tray or absorbent pad beneath the Y fit ting below each pair of pump heads Loosen the fitting entering the Y fitting to allow the solvent to drain from the supply lines 4 Remove the two screws securing the pump assembly in place shown in Figure 6 19 using the included 3 16 hex key Figure 6 19Location of the pump assembly mounting screws 5 Remove the two stainless steel outlet lines...

Page 116: ...re 6 21Pump assembly 8 With the pump assembly pulled out remove the inlet fit tings connected to the bottom of the pump head Using a 1 4 wrench remove the stainless steel outlet lines on the top of the pump head Figure 6 22 Figure 6 22Location of fittings Outlet lines Inlet lines Pump drive connector Outlet lines removed Inlet lines removed ...

Page 117: ...eck valve cartridge into the housing Make sure the arrow on the body of the check valve cartridge is pointing upwards in the direction of the flow Figure 6 24 Figure 6 24Check valve housings top and inlet check valve and housing bottom Note On an outlet check valve the arrow is reversed 11 Reinstall the check valve housings into the pump heads and tighten Reconnect the Y fittings removed in step 5...

Page 118: ... to be replaced Peristaltic Tubing Replacement The peristaltic tubing P N SP7477 Figure 6 25Back of the AutoSampler 1 Removed the outer band of the peristaltic pump Figure 6 26 Figure 6 26Removing the outer band 2 Lift up to remove the peristaltic tubing from the pump repeat on the bottom Figure 6 27 Figure 6 27Removing the tubing ...

Page 119: ...c cleaning of the spray chamber will keep the ACCQPrep operating at maximum performance Use the following steps if spray chamber cleaning is recommended 6 7 1 Cleaning the ELSD Detector 1 From the run screen select the METHOD EDITOR 2 In the Peak detection window select the ELS DETECTOR radio button then select DETAILS 3 Set the spray chamber temperature to 40 C 4 Exit the Edit Method screen and a...

Page 120: ... to 40 mL of ace tone Lift the end of the tubing as needed to transfer most of the liquid into the spray chamber Make sure the fluid level in the tubing doesn t exceed the level of the instru ment case top If the tubing is raised too fast fluid may flow out the top of the vent tube causing a spill Hold the tubing up for at least 1 minute after the fluid has been transferred to the spray chamber Fi...

Page 121: ...e while it is in the 100 mL beaker below the p trap drain line level 11 Use the DETAILs menu to set the spray chamber to 60 C 12 Fill the p trap with up to 40 mL of Isopropyl Alcohol as before and drape the p trap drain line of the top of the unit and let it set for at least 20 minutes 13 Allow the Isopropyl alcohol to drain out of the unit by drop ping the end of the p trap drain line while it is...

Page 122: ...s 3 Finger tighten the 28 PEEK fitting at the top of the source housing 6 8 3 Cleaning the Ionization Source Capillary Plugging of the capillary either in the ESI or APCI probe causes the pressure of the carrier fluid from the fluid interface to exceed the maximum operating pressure indicated by Error 310 or Error 316 This error can be avoided by filtering directly injected samples through a 0 45 ...

Page 123: ...g If the error occurs during a run select CONTINUE WITHOUT PurIon and the purification can continue without the PurIon signal or peak detection but will rely on any other detector selected such as UV or ELSD This allows the run to be completed before trouble shooting the plug Use the menu item MS Manual control Press the START CARRIER PUMP button Watch the pressure on the ribbon gauge Loosen the f...

Page 124: ...ure then reduces the flow rate to 0 2 mL minute under typical operating conditions for the fluid interface The operating mode can be determined by listen ing to the pump operate While running at 50 mL min the pump motor speed is continuous Once operating pressure is reached the pump has a rapid refill stroke approxi mately that occurs 8 seconds 6 8 7 Replacing Check Valves 1 Install new check valv...

Page 125: ... place your fingers on the top surface of the cone and turn counterclockwise to unscrew the capillary inlet cone Many times this is sufficient to remove the capillary f you are unable to unscrew the part manually use an adjustable wrench on the flats of the cone to unscrew The flats are not large and may be difficult to see and keep a wrench seated on the flats 5 Remove the O rings from the capill...

Page 126: ...ni pressure must be below 5 5E 3 mbar before the PurIon turbo pump will operate If trying to place the PurIon in operate without turning on the roughing pump this message will appear This error generally occurs if the user forgets to turn the roughing pump back on after cleaning the capillary or changing the pump oil The PurIon has been shut down It will be unavailable for use until the NextGen ha...

Page 127: ...ry cone entrance to the vacuum area See cleaning the cone capillary 6 8 3 A plug has occurred in the PurIon fluid lines The plug must be corrected to allow continued operation Error 316 This error occurs if the tubing is plugged while the method development screen is in use See cleaning the probe capillary 6 8 3 The ionization probe ESI or APCI isn t fully seated into the ion source housing Please...

Page 128: ...ACCQPrep HP125 Section 6 Maintenance 6 28 ...

Page 129: ... The following diagrams are for connecting the AutoSampler to the ACCQPrep system These diagrams include A tubing diagram of the connections from the AutoS ampler to the ACCQPrep Figure A 1 A wiring diagram of the USB connections from the AutoSampler to the ACCQPrep Figure A 2 ...

Page 130: ...ACCQPrep HP125 Appendix A A 2 This page is intentionally left blank ...

Page 131: ... B3 B4 Waste Drain Solvent Level Sensing Solvent Inlet Main Power Main Power Auto Injector Optional Column Valve Optional To Injection Valve To bottom column port Needle Wash Not Normally Used Not Used Optional External Gas Not Normally Used Solvent inlet valves optional If not used connect directly to A and B Repeat tubing for A1 Solvent as needed Needle Wash Inlet Line Check Valve Auto Sampler P...

Page 132: ...ACCQPrep HP125 Appendix A A 4 This page is intentionally left blank ...

Page 133: ...diagram of the Autosampler connected to the ACCQPrep Auto Sampler Power External Detector USB To Auto Sampler Waste Level Sense B IN A IN A1 A1 A2 A3 B1 B2 B3 A2 B1 B2 B3 B4 A3 A4 Waste Drain Solvent Level Sensing Solvent Inlet Circuit Breaker Main Power ...

Page 134: ...ACCQPrep HP125 Appendix A A 6 This page is intentionally left blank ...

Page 135: ...A 7 Appendix A ACCQPrep HP125 This page is intentionally left blank ...

Page 136: ...A 8 This page is intentionally left blank ...

Page 137: ...urement Control and Laboratory Use EN 61010 1 2010 Safety Requirements for Electrical Equipment for Measurement Control and Laboratory Use EN 61010 2 2015 Safety requirements for electrical equipment for measurement control and laboratory use Part 2 081 Particular requirements for automatic and semi automatic laboratory equipment for analysis and other purposes EN 61000 3 2 2014 Limits for harmoni...

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Page 139: ...esent the concentration of the hazardous substance in this component s any homogeneous pieces is lower than the ST standard limitation X烉堐䣢宍㚱㭺㚱䈑峐军宍悐ẞ䘬㝸ᶨ 峐㛸㕁䘬慷崭 ST 㞯Ⅾ奬䘬旸慷天㯪ˤ ẩ 㬌炻㟡旭ね ᶲ堐ㇻ X 䘬㛗徃埴徃ᶨ㬍宜㖶ˤ X Represent the concentration of the hazardous substance in this component s at least one homogeneous piece is higher than the ST standard limitation Manufacturer may give technical reasons to...

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Page 141: ...ue to corrosion misuse accident or lack of proper LQVWDOODWLRQ RU maintenance This warranty does not cover products not sold under the Teledyne 6 2 trademark or for which any other warranty is specifically stated No item may be returned for warranty service without a return authorization number RMA issued by Teledyne 6 2 This warranty is expressly in lieu of all otherwarranties and obligations and...

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