G2229A - Agilent Technologies
Contents
1. e e e Agilent Nanoflow LC System for Mass Spectrometry MS i G2229A Quick Start Guide In this guide 1 Stacking the System 2 2 Preparing the System 4 3 Operation Tips and Hints 6 Part Information 8 Use this guide to help you install your Agilent Nanoflow LC System for MS This guide also provides valuable tips and hints for operation of the system Following these hints will ensure a successful run If you need to reorder parts please refer to the tables on the rear page ee Agilent Technologies Stacking the System 1 Stacking the System Micro Well Plate Sampler WPS G1377A Nano Pump G2226A Micro Degasser G1379A Solvent Cabinet 5062 8581 Preparing the System see page 4 5 Installation Step Unpack and install the WPS Unpack and install the nanoflow pump Unpack and install the micro degasser Unpack and install the solvent cabinet How to Proceed e Place the micro well plate sampler on the bench e Remove the ST safety foam e Connect the power cable e Connect the corrugated waste tube to the seat adapter and the solvent waste port from the leak plane e Place the nanoflow pump on top of the WPS e Connect the CAN cable between the pump and the WPS e Connect the power cable e Connect the waste tube to the EMPV of the pump e Place the micro degasser on top of the pump e Connect the power cable e Connect the solvent tube G1322 6 7300 between t2. MEM Control gt Nano Pump gt Purge task at the handheld controller e Set the flow to 2 5 ml min and purge each channel separately for 4 minutes e Switch to 50 50 A B and purge for additional 4 minutes Connect the e Turn ON the micro well plate sampler i capillaries i Plumbing the p Connect capillary 1 G1375 87322 see Figure 3 between flow sensor outlet and switching valve port 1 System Connect capillary 6 G1375 87323 to switching valve port 6 Replace the seat capillary 4 with capillary G1375 87316 provided in the accessory kit Activate the Micro mode by selecting Setting gt Nano Pump gt More gt Micro Flow at the handheld controller Set the flow to 4 ul min solvent composition 50 50 A B To monitor the flow and pressure at the handheld controller select Plot gt Select gt Nano Pump Pressure gt More gt Nano Pump Flow gt More Make sure these stabilize at least for 10 minutes before making the next connection Flow ripple Operating tips should be less than 15 Flow and pressure plots should look similar to Figure 4 see page This procedure can take up to 60 minutes If after this time the 6 pressure and the flow does not stabilize there are probably particles at the front end of the capillary Backflush the tubing to remove them 4 Nanoflow LC System for MS Quick Start Preparing the System Additional Installation Notes The choice of mobile phase affects chromatography ionization efficiency and sample recover
3. tend to get an air bubble at the bottom of the vial Air bubbles can cause injection problems e Conical polypropylene inserts 100 ul 5182 05449 are less opaque and less prone to persistent air bubbles at the bottom Nanoflow LC System for MS Quick Start 3 Operation Tips and Hints Pump Degasser Use primary flow rate for low solvent e Regular fluctuations indicate air within the high consumption pressure path After changing solvents purge each channel for Rotate EMPV valve once while under flow to 4 min remove dirt from the valve seat e Check pressure drop of solvent filter infront of Use clean solvent bottles and solvent the EMPV once a month e Never run without solvent inlet filters After sitting idle for a day or longer flush each e Use glass bottled solvents channel for a few minutes e System backpressure should be higher than 20 bar Filter solvents through 0 4 um filters The default settings compressibility flow sensor calibration are set for water in channel Irregular flow pressure fluctuations indicate A and acetonitrile in channel B partially blocked capillaries Well plate sampler WPS e The recommended solvent for automatic Use bottom sensing when working with low washing of the autosampler needle is 15 sample volume methanol 84 9 water 0 1 formic acid e For direct injection use bypass mode This e Use needle wash leads to a sample transfer time betw
4. 8 639 150 1 178 150 0 663 2000 550 0 270 350 0 172 8000 3 927 6 G1375 87322 G1375 87312 G1375 87315 G1375 87317 G1375 87316 G1375 87326 9 G1375 87323 6 G1375 87322 140 G2226 67300 G2226 90001 Edition 02 2003 Printed in Germany Part Number G2226 90001 Agilent Technologies Inc 2003 Agilent Technologies Deutschland GmbH Hewlett Packard Strasse 8 76337 Waldbronn Germany
5. een WPS e Check alignment once a month and column of 3 6 min 300 nl min e Ensure comparable pressure drop ina mainpass Prime flush pump at least once a week for one and bypass once a week minute Check that liquid is draining from the wash port while priming For more information on your Agilent Nanoflow System please check the Nano Pump User Manual G2226 90000 the Nano Pump Service Manual G2226 90100 or the WPS Reference Manual G1367 90002 Nanoflow LC System for MS Quick Start 7 Part Information Table 1 Fittings and Ferrules Fitting Type Name Description DoR Swagelok 1 16 SST fitting front and back p p ferrule gt Lite Touch 4 16 SST fitting Lite Touch 1 32 SST ferrule and lock ring Rheodyne M4 PEEK fitting Finger Tight Double winged nuts and 1 32 ferrules E Lite touch detector M4 1 16 SST fitting male Lite touch detector Lite touch detector SST ferrule PEEK sleeve Conditioning Part Number 10 pk 0062 2418 10 pk 0063 6593 10 pk 0065 4423 6 fitt 2 plug 5065 4410 10 pk 0065 4422 10 pk 0063 6593 10 pk 0063 6592 1 pk 0042 1396 Diameter Capillaries and Fittings for item numbers see Figure 3 Pressure drop for Part number 1pl min H30 bar 1 D C PFS 25 l Table 2 Item Fitting Type Material pm 2 PFS 100 3 B D PFS 100 4 PFS 100 4 PFS 15 5 C 6 D C PFS 25 6 D C PFS 25 Restriction Capillary FS 25 Length Volume mm nl 350 0 172 200 1 570 1100
6. he outlet port of the degasser and the solvent selection valve of the pump e Place the solvent cabinet on top of the degasser e Place the bottles in the solvent cabinet e At the bottle head assembly G1311 60003 replace the glass solvent inlet filter with a SST solvent inlet filter 1018 60025 e Connect the bottle head assembly to the inlet port of the degasser e Connect the tube from the peristaltic flush pump to the solvent bottle in the solvent cabinet Nanoflow LC System for MS Quick Start bottles bottle head assembly outlet ports inlet ports connecting tube EMPV solvent selection valve To Se peristaltic pump J waste tube to waste Figure 1 Stacking Overview solvent tubing 5m tube screw ferrules with lock ring a us solvent inlet filter SST Figure 2 Bottle Head Assembly Overview Nanoflow LC System for MS Quick Start Stacking the System Micro Well plate Sampler Preparing the System 2 Preparing the System Installation How to Proceed Step Prepare e Prepare 0 1 formic acid in water for channel A solvent 0 anid i Preparing Prepare 0 1 formic acid in ACN for channel B Sol t e Prepare 15 methanol 84 water 01 formic acid for the pan washing solvent of the WPS needle Purge e Turn ON the micro degasser and the pump Purging the e Connect the handheld controller to the pump Syst e Activate the purge mode by selecting View gt System gt
7. t the pump outlet e Check for plugged column capillaries if pressure increases more than 30 Capillaries Flush new capillaries before connecting to other components Wash both ends with organic solvent and be sure the connection is dry before connecting e Always install or retighten without flow Do not overtighten trap in module doors or bend with radius smaller than 4 cm e Avoid gaps within fittings e Use pH lower than 9 Vials The choice of glass versus plastic vials is Sample dependent If you experience sample recovery problems you may want to try a different type of vial Use the following hints as a guidance e Plastic vials are most commonly used e Polypropylene inserts and wide mouth vials are recommended e For best results use nanoflow rates from 0 1 ul min to 1 pl min e In micro mode abnormally high column flow variations are an indication of small particles within the system e When using buffer solutions flush the system with water before switching it off e Replace capillaries if they are bend just after the fitting or anywhere else with a diameter below 4cm e Compare capillary pressure drop to that listed in Table 2 Replace capillary if you have more than 30 deviation e Inspect suspicious capillaries under microscope Replace those with milky surface e Plastic capillary electrophoresis sample vials 300 ul 9301 0978 can work but they are Opaque and
8. y Typically mobile phase solvents are water and acetonitrile both with added organic acid e Formic acid causes less ion suppression than TFA e TFA gives better ion paring chromatography e 0 1 formic acid in both water and acetonitrile are good general purpose mobile phases for peptide analysis In the purge mode the flow goes to waste rather than through the analytical system You will not damage the system by using the purge mode at 2 5 ml min Purging the system Is necessary If e It is being used for the first time e It was switched OFF overnight or longer e The vacuum degasser lines are empty e You have changed to a solvent that is immiscible with the previous solvent e Before connecting wash both ends with organic solvent and flush before connecting new capillaries to other components e Avoid air gaps between fittings e Do not overtighten trap in module doors or bend capillaries with radius smaller than 4 cm e Always install and retighten without flow e Use pH lower that 9 Figure 3 Plumbing diagram main pass Figure 4 Stability of flow top and pressure bottom Nanoflow LC System for MS Quick Start 5 3 Operation Tips and Hints 3 Operation Tips and Hints System e The system pressure of your newly installed system should be 40 50 bar under typical conditions 300 nl min of water with a 50 x 0 075 mm 3 5 um column e For stable flow the system pressure must be higher than 20 bar a
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