data sheet NanoLC-Ultra™ system - Eksigent

data sheetNanoLC-Ultra systemThe new NanoLC-Ultra is Eksigent’s thirdgeneration system, delivering superior gradientprecision at pressures up to 10,000 psi.Introducing the NanoLC-Ultra family of highpressure HPLCs for proteomics researchThe higher pressure capability of the NanoLC-Ultra platformallows the use of longer columns and/or columns packedwith smaller particles for higher resolution separations ofcomplex (digested) protein samples. Moreover, the NanoLC-Ultra family features self-priming and self-purging pumps foreasier changing of solvents, and a temperature-controlledcolumn compartment for maintaining exceptional retention timereproducibility.Eksigent’s proven Microfluidic Flow Control (MFC)technology guarantees reliable and reproducible results, whileintegration with major mass spectrometer software packagesmakes the system a seamless addition to almost any nanoLC-MS workflow.A range of different NanoLC-Ultra system configurations areavailable for various proteomics applications—from singlegradient systems for direct sample injection, to dual gradientsystems for multidimensional separations, and dual columnset-up for increased throughput.The next generation NanoLC system fromEksigent promises to deliver high resolutionseparations, combined with excellent reproducibility,resulting in improved identification of lowabundance peptides/proteins with nanoLC-MS.

Key advantages of the new NanoLC-UltraThe NanoLC-Ultra system is a proteomics lab workhorse,delivering stable, reproducible gradient separations forpeptide/protein identification, phosphopeptide analysis,biomarker discovery, metabolomics and more.Splitless, nano-scale flow controlEksigent’s Microfluidic Flow Control (MFC) technology (seeFigure 1) uses fast flow measurement in combination withautomatic feedback to a regulated pressure source to generateaccurate and reproducible flow rates with a resolution betterthan 1 nl/min. By combining true microfluidic pump controlwith direct feedback of flow rate, the system eliminates theflow inaccuracies and plumbing problems caused by splitterbasedsystems. The result is extremely reproducible gradientsdown to 50 nl/min, without the use of flow splitting, andobserved gradient reproducibility typically better than 0.5%RSD (see Figure 2). And the self-priming/self-purging designmakes the NanoLC-Ultra pump even easier to set-up and run.10,000 psi maximum pressureUltra high pressure LC has been successfully applied in smallmolecule analyses using 2.1 mm ID columns for increasedseparation efficiencies and/or analysis speed. For theidentification of proteins using LC-MS in proteomics, NanoLCis required because of the increased sensitivity obtained bynanospray mass spectrometry.Figure 1. Microfluidic Flow ControlWith MFC technology, direct pumping is used to deliveraccurate, repeatable gradients. Each pressure sourcedelivers fluid with an actual flow rate monitored using theflow modules in each channel. A CPU closes the loop bymaking any necessary adjustments to the flow rate usingelectro-pneumatic controllers.The operating pressure of 10,000 psi obtained with theNanoLC-Ultra system enables the use of longer columns,or columns packed with smaller particles, for the analysis ofincreasingly complex proteomics samples (see Figure 3).Retention time reproducibilityMaintaining a constant column temperature is requisite toobtaining the best possible retention time reproducibility.To address this requirement, NanoLC-Ultra systems areequipped with a temperature-controlled column compartmentwith integrated 10-port valves, that can be regulated to atemperature up to 40°C.Elevated temperatures also can be utilized to decrease columnpressure and/or improve separation efficiency. For advancedapplications like sample pre-concentration or dual-column setup,the NanoLC-Ultra system offers up to two 10-port nanovalves rated at 10,000 psi that are integrated into the columncompartment. These 10-port valves have an extremely low portto-portvolume of

Peak parkingBecause of the excellent control over the flow rate offered byour MFC technology, the NanoLC-Ultra systems can operatein a “peak parking mode” where the flow rate is temporarilydecreased to facilitate longer MS interrogation of an analyteof interest. The result is extended MS/MS analysis acquisitiontime that improves the MS identification of co-eluting and lowabundance peptides.Mass spec software integrationWith Eksigent’s Control Software, users can rapidly setupexperiments and create or modify methods. Specializeddiagnostic tools make the system easy to learn, maintain andoperate. The software continuously monitors pressure andflow rate, and provides real-time views of gradient compositionand column backpressure. Plus, Eksigent’s software engineersplaced special emphasis on integrating control of the NanoLC-Ultra systems into the major mass spectrometry softwarepackages. Drivers are currently available for Thermo Electron’sXcalibur® (see Figure 4), Bruker Daltonics’ HyStar® andABI/Sciex Analyst® software.Figure 4. Software integration under Thermo Electron’sXcalibur.Figure 3. High resolution separation of a BSA tryptic digest.Separation of a BSA tryptic digest on a 75 µm ID x 80 cm column packed with 3 µm C18 stationary phase.Back pressure was 9,000 psi. The peak capacity for this separation is approximately 250.

traptrapFlexible system configurations for proteomicsCurrently, four configurations of the NanoLC-Ultra system areavailable for proteomics researchers:• NanoLC-Ultra 1D,• NanoLC-Ultra 1Dplus,• NanoLC-Ultra 2D,• and NanoLC-Ultra 2Dplus.The NanoLC-Ultra 1D system contains one binary gradientpump, allowing for direct on-column injections. The NanoLC-Ultra 1Dplus system adds an isocratic loading pump for sampleenrichment experiments in which sample is loaded onto a trapcolumn prior to analysis on a NanoLC column.NanoLC-Ultra 2D systems include all of the features of the1D, plus the added capability of automated 2-dimensionalchromatography. The NanoLC-Ultra 2D (two gradients) systemand NanoLC-Ultra 2Dplus system (two gradients and oneisocratic loading pump) are designed for advanced applicationssuch as on-line 2D LC (see Figure 5) or high throughput setupsusing two nanoLC columns.All NanoLC-Ultra 2D and 2Dplus systems are available withone or two built-in 10-port valves. NanoLC-Ultra 2D and2Dplus system can be configured to run in either 2-dimensionalseparation mode or as two parallel, completely independentbinary gradient HPLC systems.AS-2 autosamplerThe new AS-2 Autosampler stacks together with the NanoLC-Ultra and both are operated by our Eksigent Control Softwareor through integration with the MS software. The flexible andeasy-to-use autosampler employs a 2.4 ul volume needle forinjection of small volumes without sample losses. The needleis coated with a proprietary inert coating on both the insideand outside to prevent sample losses due to adsorption.The sample compartment is completely enclosed to ensuretemperature control down to 4°C. Capacity is two microtiterplates or 96 vials. Using two different wash positions, theneedle is washed inside and outside to reduce carry-over.The 6-port injection valve has port-to-port volumes of

Ordering informationp/n system 10-port valves loading pump gradient 1 gradient 2950-00055 NanoLC-Ultra 1D N/A N/A 50-500 nl/min N/A950-00056 NanoLC-Ultra 1Dplus 1 1-30 µl/min 50-500 nl/min N/A950-00057 NanoLC-Ultra 2D 1 N/A 1-10 µl/min 50-500 nl/min950-00058 NanoLC-Ultra 2D 1 N/A 50-500 nl/min 50-500 nl/min950-00059 NanoLC-Ultra 2D 2 N/A 1-10 µl/min 50-500 nl/min950-00060 NanoLC-Ultra 2D 2 N/A 50-500 nl/min 50-500 nl/min950-00061 NanoLC-Ultra 2Dplus 2 1-30 µl/min 50-500 nl/min 50-500 nl/min620-00142 AS-2 autosampler N/A N/A N/A N/ASystem specificationsNanoLC-UltraDimensions 14” x 24” x 18” (W x D x H)WeightWorking temperature 15º to 30ºCPowerMaximum pressureStandard flow rate rangesLoading pumpNano gradientMicro gradientFlow rate accuracy

eksigent technologiesheadquarters5875 arnold roaddublin, california 94568tel: 925 560 2600fax: 925 560 2700new jersey office & laboratory11 deer park, suite 204monmouth junction, new jersey 08852tel: 732 274 9190fax: 732 274 9161email: info@eksigent.comwebsite: www.eksigent.comEksigent is creating new possibilities for life scienceresearch, drug discovery & development, and medicaldevices with its innovative MicroFlow and NanoFlowfluid delivery systems.Eksigent’s LC systems deliver dramatic increases inanalysis speed, throughput, and sensitivity. Eksigent’sdrug delivery systems bring new levels of precision toportable drug delivery.Today, leading research, pharmaceutical, and biotechnologyfirms around the world use Eksigent’s innovativesolutions.© 2008 Eksigent Technologies. All rights reserved. Information subject to change without notice.Eksigent is a registered trademark of Eksigent Technologies. NanoLC, NanoFlow, and MicroFlow aretrademarks of Eksigent Technologies. All other trademarks are property of their respective owners.Patents pending worldwide.Printed in the USA. NU-DS-0208-1000