During step 1, check that the solvent moves into the solvent line correctly and it goes out the Purge Valve. The 1260 Infinity II Binary Pump is ideally suited for a broad range of UHPLC applications using columns with conventional, superficially porous or sub-2-micron particles. . Replace piston seals. 05 MPa or 0. After purging the pump heads, pu rge the ProFlow flow meter for about 10 minutes. Wash out the blockage. The 1290 Infinity II Flexible Pump is a quaternary pump that provides UHPLC gradient formation and solvent blending from up to four solvents. Clog removal. If seal wash option is installed, replace wash seals and gaskets. ID stainless steel tubing kit • Galaxie Chromatography Software-0. Solvent Degasser – removes air gases from the solvents as they flow to the HPLC pump. Gradually increase system flow up to the required flow rate,. Then wash them out with water and re-assemble them on the pump head. Next, it purges the flow lines up to the column with the mobile phase at the starting gradient concentration and then pumps it through the column at a low flow rate to protect the column. Monitoring your HPLC system for potential issues can help to. . When I used the pump C last week the pressure suddenly fell down from 300 PSI to 0 (with 100% methanol, 0. 2:32. , 5-10 mL/min. HPLC is an abbreviation for High Performance Liquid Chromatography. HPLC pumps are designed to pump liquid, not air. Replace piston seals. Purge each mobile phase that will be used by selecting “Purge” on the PumpModule tab on Chromeleon. 5 01. To manually purge the reference side of the 2414 flow cell, press the Shift + 1 button (Purge) on the screen while the pump is flowing mobile phase. HPLC Starter Kit G4201-68707 195 HPLC Starter Kit G4202-68707 195 HPLC System Tool Kit 196 Bottle Head Assembly 197 Solvent Cabinet 198. The manual covers the overview, installation, operation, and optimization of the pump, as well as troubleshooting and maintenance tips. 이동상을 준비하고 pump를 ON하기 전에 purge를 실시하는데, 3분 정도 purge를 하도록 설정돼있습니다. Turn the handle counterclockwise, about 1/2-turn, to open the valve, and pull back on the plunger to remove air bubbles. Figure 2. Your purge valve is a great tool for flushing and priming your HPLC pump system. ) If system has check valve, loosen valve to allow air to escape. Figure 1: Reciprocating piston pump during filling stroke. The pump inlet port is closed to allow the pump to run with the gas ballast valve in the open position, which helps to purge the pump. off. Agilent University. This video shows the operation a HPLC purge valve. Purge pump at high flow rate (e. • Mast kit with mixer, purge valves and manual injector • Pre-formed 0. Rough pumps stay on so the system remains under vacuum. 7:30. Featured. 2. Maintaining your LC columns. After purging is done click [Pump on] button / to deliver the solvent for at least 20 to 30min to rinse the flow line . Purging air from pump system: Change solvent line to a bottle containing 100% isopropanol. Dear Huq, check out the whole tubing starting from Pump to the Column inlet for any air trapped, purge the pump for few minutes. g. 02 in. First statement: Don’t let it run dry. 2 Push the “conc” button on LC20AD. Use only HPLC-grade solvents and water filtered through 0. 3601-EN-5. Prepare the Sampler • Avoid using needle wash and seat backflush with organic solvents with buffer applications,Most solvents need to be degassed, so U/HPLC systems typically include a solvent degassing system. Open the purge valve and purge the system for 5 min with isopropanol at a flow rate of 2 mL/min. Our industry leading LC-40D X3 pump ensures precision and accuracy at all flow rates. If the VP button does not show mobile phase, press “CE” followed by “Function” until “System” is shown on the screen. 6 Calibration Procedure of HPLC: 6. 액상미립자측정기에서의 purge는 측정완료된 샘플용액을 제거하기 위해 사용하는 것 같다. pump에 대한 자세한 설명은 아래 링크를 참고해주세요! ANSWER. operating pressure (9,99 ml/min max. Cause a:Air bubble in pump head. Prepare the solvent: Begin by ensuring that the solvent is filtered and degassed. If problem persists, flush system with 100% methanol or isopropanol. This is needed to purge any air from the tubing, introduce fresh mobile phase to each line and then to VERIFY that each channel delivers the reported amount of fluid to the column (measure it). The good news is that there is a simple solution: open the purge valve, turn the flow rate to five mLs a minute, let it sit there for 10 to. Flush the pump head with an appropriate solvent using the purge function; Turn off the pump flow and wait until the system pressure is down to zero; Analysis. 5. Pump 100% isopropanol through the pump for 3 minutes. g. Parallel dual micro plunger (10 µL) construction for low pulsation and stable baseline. What is purging in HPLC? Answer: “When setting up an HPLC system, the aim of the purge is simply to flush through all the lines so that any remaining solvent in them from a previous analysis or wash is replaced with the new mobile phase. Then, turn it slightly past the initial resistance (check this). Here’s a trick: you can set a minimum pressure on your HPLC pump. The reduction in condensation risk can allow the pump to achieve its vapor duty at. For modular autosamplers purge from front panel. Read this guide thoroughly before using the product and operate the product in accordance with the instructions in this manual. Press the EDIT/ ENTER key to start purging. off. You may. We have been experiencing some pressure problems on our hplc lately and the problem is mostly fluctuations in the pump head. 9. 1. Pump a 20% nitric acid/water solution through the pump for 3 minutes. Pump 100% isopropanol through the pump for 3 minutes. Blocked channels can often be released by purging with a high flow with warm water (60 to 70 °C, HPLC grade). The historical evolution of HPLC pumps and their characteristics has been reviewed in many textbooks (1–3), as well as journal articles (4–6). (Prime each pump head separately. The source of column clogging comes from particulates in a number of places:We have been experiencing some pressure problems on our hplc lately and the problem is mostly fluctuations in the pump head. 2. Search. Purge valve Purge functionFig. Optional solvent selection valves can be added for solvent. This video shows the proper procedure to safely remove and replace the plunger seal on the Shimadzu LC-40 Series Nexera pump. 3. Check for flow. - filter non-HPLC grade solvents - filter buffer solutions - Install an in-line filter between auto-sampler and column (removes pump seal debris, ALS rotor debris, and sample particulates). Weigh the sample into a syringe. To purge the 2414 via Empower, use the Purge. A = 100%). 0046. A purge valve with The pump assembly includes a pump head with a passive inlet valve and an outlet valve. • Purge each channel for at least 10 min at 5 mL/min. Fig 2. This pump enables quaternary solvent delivery at pressures up to 1300 bar and flow rates up to 5 mL/min. You. Set the flow rate to 5 ml/min d. The target url is a user manual for the Agilent 1290 Infinity Quaternary Pump G4204A, a high-performance liquid chromatography (HPLC) device that can handle a wide range of flow rates and compositions. L/mni for 5mni . The pump offers a dynamic flow rate range from 1 to 50 mL/min at up to 420 bar, allowing workflows on. Remove sample vials from the auto-sampler tray g. Delivering Scientific Insights to Improve Human Health and Well-Being. Maintenance Kits for UltiMate™ 3000 Pumps. 1 uL/min to 10 mL/min with accuracy of ± 1 % or ± 2 µL/min (whichever is larger), and precision < 0. Contacting Us Vanquish Pumps (VH-P10) Operating Manual Page 3 Contacting Us There are several ways to contact us: Ordering Information For ordering information or sales support for HPLC products, contactSolving Problems That Matter - Batteries. Excellent gradient performance resulting from the highly accurate solvent delivery by the 5110/5160 pump with Hitachi’s unique high-speed. When they fill with air, they lose their prime and stop • Effects Column Life pumping [26]. Overall Routine Pump Maintenance 1. A damping unit is connected between the two piston chambers. If problem persists, flush system with 100% methanol or isopropanol. 5. PENDAHULUAN Penelitia n yang berkaitan dengan tumbuhan berguna Indonesia. For more information on purging the pump, see the reference. Open the Purge Valve of your pump. A pressure sensor monitors the pump pressure. 3601-EN Revision 5. Catalog number: 5040. Place the solvent bottle back into LC eluent. 0 • August 2022 *4820. Pump & Maintenance Kits for HPLC Part Number: 01018-22707. 1, 3, and 4. 8:42. The manual covers installation, operation,. 6 mm ID columns over the flow rate range up to 5 mL/min. Set the pump to 100% 'A' at 1. If problem still persists, contact system manufacturer. ) If system has check valve, loosen valve to allow air to escape. |The 1290 Infinity II Bio Flexible Pump. In stainless steel (SS) the passivation process uses nitric acid or citric acid to remove free iron from the surface. 00 ml/min, turn the pump back ON and close the prime purge valve to allow the liquid to flow through solvent lines, through the degasser channel, through the pump. Vanquish Binary Pump N: Up to 1500 bar (22,000 psi) and 100 µL/min with active flow control. ), prime system if necessary. • After few seconds, the display will show, Flow 1. HPLC is an abbreviation for High Performance Liquid Chromatography. Plunger indicator rods Show position of each pump head plunger. It is recommended to change your HPLC pump’s piston seals during planned preventative maintenance. g. When launching a long and continuous analysis sequence, for example 40 samples (analysis of each sample. Passivation is a metal finishing process to prevent corrosion and resist rust. Flushing and purging all solvent lines and fluidics is the first step that should be done if unstable system pressure. Leak check: the Vice President HPLC, Fraser McLeod. The name of the game in HPLC is repeatability. Press “Prime” button (or pump at max flow), continue to draw on syringe until no bubbles are seen. Equal functions for Right Pump found just below. g. Draw syringe back to prime. If there is no pressure drop, the blockage is. We routinely see HPLC systems with white fluffy crystals built up around the pump heads, pistons and various fittings from lack of maintenance on a daily basis. The sensor signals the control board when the vacuum is below a preset level. Check if application conditions are within the column specifications (e. We have been experiencing some pressure problems on our hplc lately and the problem is mostly fluctuations in the pump head. . Initial System inspection It is always worthwhile to spend a few minutes before starting analysis on assessing the system suitability for the analysis task in hand. 23 MPa. "Chromatography" is a technique for separation, "chromatogram" is the result of chromatography, and "chromatograph" is the instrument used to conduct chromatography. The target url is a user manual for the Agilent 1290 Infinity Quaternary Pump G4204A, a high-performance liquid chromatography (HPLC) device that can handle a wide range of flow rates and compositions. When you vent the system, the components are turned off and the rough pump turns off. Open the purge valve on the Agilent Infinity 1260 isocratic pump b. Learn essential HPLC maintenance tips. 결과 및 정리. The Vanquish HPLC and UHPLC systems offer uncompromised performance, robustness, and ease of use. . ), prime system if necessary. Figure 7: Pump tab. If tiny air bubbles are observed in the lines continuously, it’s. Honoring the Past, Focusing on the Future: A Conversation with Jess Lee. Press “Menu” till the purge appears on the screen. ) If system has check valve, loosen valve to allow air to escape. 4 Open the purge valve and purge the system for 5 minutes with isopropanol at a flow rate of 2 ml/min. The quantity is such that the compression ratio of the pump is reduced to 10:1 max. Confirm that the lever is in the INJECT position, insert the syringe all the way into the injection port, and quickly switch the valve to the LOAD position. Make sure that the AVR7-3 inject valve is in the Purge position. If using MassLynx or Empower software, refer to the PDFs entitled Priming the 1525 Binary HPLC Pump Using MassLynx or Priming the 1525 Binary HPLC Pump Using Empower. Open the Purge Valve by turning it counterclockwise b. g. Flushing and purging all solvent lines and fluidics is the first step that should be done if unstable system pressure. How do i manually prime/purge it? Because there are some air bubbles, and if they get inside the needle, its gonna be bad news bears ( i think). b Select Set up pump. The manual covers the overview, installation, operation, and optimization of the pump, as well as troubleshooting and maintenance tips. 2 shows the flow line during LC-2010 analysis. Meet Your New Lab Ally - the Alliance iS HPLC System. 6. Retention times can shift from one run to the next – you need to know when it is ok and when it signifies a problem. Purging the solvent pumps 1. Flush the pump with 100% filtered, distilled water for at least 3 minutes. 1 ml/min and connect the size exclusion column (4. Wide peaks are generated either by substantial influence on the part of the HPLC system (bad capillary connections, void volumes, too large detector cells or ill. 1290 Infinity II High Speed Pump User Manual. Open the purge valve by turning the knob on the front of the pump to the left until you feel some resistance. 3. 7. If the preceding step does not solve the issue, remove the purge valve and start the pump. some time this may be due to filters in the channels; please remove your filters (a inlet filter from your solvent reservoir bottle to the HPLC pump ) and sonicate in the Methanol. In fire and explosion protection engineering, an inert (ie, non-flammable) purge gas (like nitrogen, helium, argon, etc) is introduced into an enclosed system (eg, a container or process vessel) to prevent the formation of a fuming. Prepare the Sampler • Avoid using needle wash and seat backflush with organic solvents with buffer applications, Most solvents need to be degassed, so U/HPLC systems typically include a solvent degassing system. Second, the dissolved gasses must diffuse through the membrane (flat film or tubing wall). Consumables likely need to be replaced on aqueous pump. UltiMate 3000 Series: SD, RS, BM, and BX Pumps Operating Instructions Page I Declaration of Conformity (Original Declaration of Conformity) Product: Thermo Scientific Dionex UltiMate 3000 - Pump Types: ISO-3100SD, ISO-3100BMWe have been experiencing some pressure problems on our hplc lately and the problem is mostly fluctuations in the pump head. 0*Today, we're diving into the exciting world of lab consumables and equipment, specifically focusing on the Shimadzu HPLC pump. Put the Pump A in your buffer. degasser ports and/or pump head multichannel valve or pump head inlet, as needed, to draw liquid through until all lines are filled. † Automated instrument features like switching mixer in/out, automaticVarian Modular Analytical HPLC Systems Figure 1. Pump intelligence - Active Damping - with high pump resolution (~300 pL minimum step) 3. Pump 15 min at this pressure to wear the seals in. Troubleshooting Strategy and Processes 5. Use 2 um frit for 3. Pump heads with maximum flow rates of 10 and 50 ml/min are available. 88 Service Handbook for 1050 Series of HPLC Modules - 11/2001 Pumps: General Information Specifications Specifications Table 2 Specifications of 1050 PumpsHi,when I do wet prime/dry prime in hplc ? Thank you. 8 3. Column frits are a common point of clogging. 결과 및 정리. The 1260 Infinity II Flexible Pump is a quaternary UHPLC pump that provides gradient formation and solvent blending from up to four solvents. • Press “Menu” till purge appears on screen. 7. If a column shows a pressure increase in a short period of use (small number of injections), there is usually an underlying problem. Operable in constant flow or constant. Documents & Downloads. Fig. 05 0 0. 4. Pump Valves for HPLC. (Prime each pump head separately. If the column was used with buffered. If the pressure drops off suddenly (by more than a few kgf/cm 2), the backpressure tube is clogged. Purge the pump, check general functionality (for instance with a diagnostics test), read pre-compression value from pump panel and compare against values from manual (Thermo Scientific Dionex pumps only). 6 x 300 mm, 3 µm, 150 Å). 2. Perform sample clean-up (i. Operation of DAD: [][]Switch ‘ON’ the power of Quaternary Pump, Autosampler, Thermostatic Column Compartment, Detector, CPU, Monitor if already not switched on. Leak at pump 18 Selectivity Differences in selectivity 19 Baseline Disturbance at void time 20. 2. 6. 2 1. Proper storage of HPLC columns will increase longevity and performance. PROCEDURE: Flush the system using one of the cleaning mixtures as described in the cleaning. e. 8. After purging is done click [Pump on] button / to deliver the solvent for at least 20 to 30min to rinse the flow line . The pump (s) should be run (column disconnected and bypassed) run with a solution of pure HPLC grade water to try and rinse the material off. Mobile phase is passed through porous polymer tubing placed in a vacuum chamber. Agilent recommends a purge of at least 30mL in total for the 1200 systems. Delivering Scientific Insights to Improve Human Health and Well-Being. Peak shape C. Loaded 0%. The Agilent 1260 Infinity II Flexible Pump is a quaternary UHPLC system pump that provides gradient formation and solvent blending from up to four solvents. For more mobile phase flexibility in any ultrahigh speed and ultrahigh resolution application, the Thermo Scientific™ Dionex™ Ultimate™ HPG-3400RS Rapid Separation Binary Pump offers more solvent capability. The dual piston design has minimal pulsation for stable baselines. 물론 이건 내 생각이다! 여튼 purge. However, even if the degassing system works properly, some air bubbles might stick in solvent lines or U/HPLC system fluidics. Make sure that you know which solvent is linked to which pump. The green mark will change into the A2 and B2 side 9 Repeat step 8 with the next 3 valves. Unit conversion calculator for converting commonly used unitsSJ: There are two diffusion paths involved in inline degassing (ignoring vacuum side diffusion). Sonication: ultrasonic baths as a stand-alone. Make sure to confirm that the purge valve isHow to prevent your HPLC from running dry. 3. If it does not, check the state of the Solvent Filters and replace them. So, let's get started and purge away any confusion! Purging, in the context of lab equipment, is the process of flushing a lot of. 1 Calibration of the Pump is to be done with the help of distilled water at 25°C±2°C. If I run the system at 0. Steps to follow: Store the column in an appropriate solvent (check column information sheet) When you remove the column, add a column union so the detector gets flushed as well in the following steps. A low pressure gradient (LPG) for reliable blending of up to four eluents. Before using an HPLC system, you should prime all of the lines in your HPLC pump. HPLC pumps are designed to pump liquid, not – Improve Peak Shape for Ionizable Compounds air. thermoscientific Vanquish Pump H VH-P10-A-02 Operating Manual 4820. Cap seal for purge valve knob; Cleaning swabs; Figures. Open the Purge Valve of your pump and purge each solvent line, one by one, with HPLC-quality Isopropanol (IPA). Attach syringe to prime/purge valve. High Priced Liquid Chromatography. This video shows the operation a HPLC purge valve. Note: Follow the instructions in the Ready Check Results window and close both purge screws. Blockage of. Perform seal wear-in procedure. b. Set maximum flow gradient to 0. Between samples when your compound is very sticky (apolar) to help clean the column. Remove and disassemble the pump head. Read Less. 13), a precisely defined quantity of air (“the gas ballast”) is admitted into the pumping chamber of the pump. Prepare the 20 ppm of caffeine in HPLC water by diluting 2 ml of stock solution to 50 ml with HPLC water and inject the triplicates of 5 µl, 10 µl, 20 µl, 50 µl and 100 µl of sample preparation. First, disconnect connection (1) in Figure 1 and pump the solvent (to check the backpressure tube 4)). • Purge each channe lwtih 25–3m. . Using a solvent filtration kit or an online degasser will remove any dissolved. For reverse-phase columns, wet the column using a mobile phase like methanol and bring it to normal flow rate. Label bottles correctly with bottle content,. It accommodates flow rates from 0. If you cannot see a leak at the bottom of the pump head when you try to purge, then most likely a leak is not the cause of the issue. 05 min is normal however the behavior of the method should be used to determine what is a normal variation. Sonication: ultrasonic baths as a stand-alone. Up to 6 FRC-40’s can be configured. Select this procedure when you want to prime and purge any 1525 Binary Series. 5:26. Properly shut down all the HPLC modules from the front (LC-20 and LC-30) or backThe 1260 Infinity II Preparative Binary Pump is an affordable high-pressure gradient pump for LC purification. [][]Clean the solvent diffuser/filter once in a week. The auto purge function was achieved by devising the mechanism of flow lines including the high-pressure 6-way valve and by modifying the solvent delivery unit check valve. 1:55. (Prime each pump head separately. Turn on the pump flow rate to 3 mL/min through your CDS Software, Lab Advisor or through the Instant Pilot and open. Algae Growth in HPLC Systems 54 Prevent Blocking of Solvent Filters 55 Normal Phase Applications 57 4 Optimizing Performance 59. , 5-10 mL/min. 9-0. Purge pump at high flow rate (e. Hello Kancheri this is the purge valve for Agilent 1260 G1312-60071 | Agilent You can check in the manual at page 154 on HOW to replace purge valve with this. You. Feeding the degasser with an inferior mobile phase will create inferior results. Replace the part that causes the obstruction. , 5-10 mL/min. flow rate) One of the most important performance measures for HPLC is retention time reproducibility. The Vanquish HPLC and UHPLC systems offer uncompromised performance, robustness, and ease of use. 2 μm filters. Initiate solvent flow by pushing the “PUMP ON/OFF” button on the front of the pump, then press “ENTER”. Perform search. Prepare the Pump • Use fresh or different mobile phase (as required). the purge valve, and the actual pump mechanism. The impact of this type of clogging will be gradual, not sudden. Prime the HPLC pumps under the following conditions: - starting the pumps for the first time after it has been idle for more than 24 hours. HPLC pumps incorporate single or dual piston, syringe, or diaphragm pump designs. Please refer to the section “Exchanging the Purge Valve Frit or the Purge Valve” on page 126 in this manual. Replace PTFE frit in the purge valve. 19 Section 3: Pump Maintenance 20 Purge Valve 21 Outlet Ball Valve 21 Active Inlet Valve (AIV) 22 Seals and Pistons 25 Pump Head 26 Parts and Supplies 27 Solvent Inlet FiltersChromaster HPLC 5110 pump with auto-purge valve. 6. After the pump, the mobile phase encounters the autosampler. Consumables likely need to be replaced on aqueous pump. Over time, these frits will accu. Open the manual purge valve or use the purge command, depending on the pump type. closed. This pump operates at pressures up to 600 bar and flow rates up to 10 mL/min. From my little experience i learnt that it may be due to air trapped in. 6. Purging pump and autosampler for i-Series Step 4. We have been experiencing some pressure problems on our hplc lately and the problem is mostly fluctuations in the pump head. Press the purge buttons again to. After the valve sonication procedure, it is suggested to purge the pump with water at 60 degrees, keeping the purge valve open. Purge pump at high flow rate (e. 2 Wellplate Autosampler Purge Kit Features Features The new Wellplate Autosampler Purge Kit offers the following features: • Fits for Agilent 1100 Series Wellplate Autosampler (G1367A and G1368A (thermostatted)). if the column gets blocked or leaks occur † Automated electromagnetic purge valve for convenient system setup Agilent 1200 Series fraction collectors Designed for lowest delay volumes to minimize peak dispersion and carryover. A damping unit is connected between the two piston chambers. Purge the system to remove air bubbles and make sure the solvent is the same solvent when the GPC column is placed. The technical specifications of all pumps meet the highest. By the end of this video you will learn how to safely and quickly swap that suck. If you've ever wondered how to purge this piece of equipment, you're in the right place. Purge the mobile phase reservoir by opening the purging valve. Power off the pump and remove the waste tubing from the purge valve. Choose between 3 configurations: A 2 × 2 binary high pressure gradient (HPG) for high accuracy blending of up to two eluents from two selectable solvents each. 1260 Infinity Binary Pump User Manual 11 Introduction to the Binary Pump 1 Instrument and OperationThe pump assembly includes a pump head with a passive inlet valve and an outlet valve. If a purge valve is not functioning correctly, it can lead to the vehicle losing power or stalling unexpectedly. A purge valve including a PTFE frit is fitted at the pump outlet for convenient priming of the pump head. 2 Set the flow of the pump at 1ml/min. Prime/Purge the pumps until no more air bubbles leave the pump heads. 1 Procedure for Operation of HPLC pump: • Ensure that all the cables and power cords of the pumps are connected properly. At the extreme, air in the pump will cause the pump to stop delivering mobile phase to the column. 1-15 of 17. Open prime/purge valve (in the case of Chrom Tech pumps, you turn the knob counterclockwise one to two turns. Manual - isocratic HPLC System G1380AA standard configuration • G1310A isocratic pump with solvent cabinet • G1328A manual injector (MI) • G1314A variable wavelength detector (VWD) with standard flow cell (10 mm path length, 14 µl volume, 40 bar maximum pressure) • G2220AA 2D-Value Solution ChemStation other modules that can be addedFailure to use fresh clean pickup filters in your HPLC may lead to total contamination of the flow path (column, pump, A/S and detector(s)). Secondary Pump Head Parts 216 Purge Valve 220 Cover Parts 222 Leak Parts 224 Fuses 224 Accessory Kit 225 Tools 226 HPLC System Tool Kit 228. 1. If adequate purging cannot be achieved with auto-purging or a suction tube does not contain liquid, for example, use the following procedure to manually purge with a syringe. With regular pump usage, this is recommended every six months. Easy to use fraction collector and Versatile Liquid Handler. 5 um columns, use 0. A purge valve including a PTFE frit is fitted at the pump outlet for convenient priming of the pump head. 5 0. Treatment 1: Open a purge valve. Turn the black valve anticlockwise to purge the mobile phase. Afterwards. 17: Ready Check Results window appear s after the NC pump blocks purge is executed 2. Some bubbles may be generated from the suction filter head dipping in the mobile phase. 02 to 0. Initial investigation should consist of purging the pump with fresh eluent to confirm no air is present within the pump, as the presence of air within the pump may cause fluctuations in pressure. Close the purge valve, set the flow to a value that gives a pressure of 350 bar.