Agilent capillary electrophoresis system

Agilent capillaryelectrophoresis system

The Agilent CE system - new dimensions in capillary electrophoresisCapillary electrophoresis (CE), with its high efficiency and resolution, rapid analysistime, plus minimal sample and solvent requirements, is an established technique inmany laboratories. Its flexibility covers a broad range of applications in a wide varietyof industries, from drug discovery and development through quality control and ionanalysis.Agilent Technologies is the leading global partner for chromatography and capillaryseparations. When you select Agilent as your partner of choice you can be assuredof quality instrumentation, solutions and service.Agilent's CE system is the first choice forcapillary electrophoresis offering benefitssuch as:• HPLC-like reproducibility, sensitivity andlinearity• integrated software with graphical userinterface• pioneering new separation techniques,including capillary electrochromatography(CEC) and capillary electrophoresismass spectrometry (CE-MS )• extensive regulatory compliance tools• world wide service and supportChecklistþ HPLC-like sensitivity and linearityþ HPLC-like reproducibilityþ Self-correcting injection systemþ Enhanced integratorþ Replenishment systemþ CECþ Single vendor CE-MS solutionþ Multitechnique softwareþ Regulatory compliance toolsMultitechnique software withgraphical user interfaceMany users will already be familiar withthe Agilent ChemStation software, andnew users will learn quickly with thesimple graphical user interface. Thisresults in increased efficiency and areduction in training costs.

The Agilent high sensitivity detectioncell provides a 10-fold increase insensitivity, unsurpassed spectralfidelity and an extended linear range.All with a unique decoupled celldesign allowing simple capillaryreplacement.High sensitivitydetection cellHPLC-like sensitivity, linearity andreproducibilityThe diode-array detector incorporated inthe Agilent CE system offers quantitativeand qualitative analysis with excellentsensitivity, wide linear detection rangeand full spectral capabilities. In addition,the sensitivity and linear range can befurther enhanced with Agilents's extendedlight path capillaries or the high sensitivitydetection cell.75 µm i.d. capillaryThe Agilent CE system uses advancedtechnologies for instrument control suchas automated buffer replenishment.Combined with advanced data processingthis ensures HPLC-like reproducibility.CE-MSAgilent Technologies offers a fullyintegrated CE-MS solution with all systemcomponents and support coming fromAgilent. A unique feature of the system isits ease of use—the mass selectivedetector (MSD) control is smoothlyintegrated into the Agilent ChemStationsoftware, the Agilent spraying systemneeds no positional adjustments, and theunique design of the MS inlet makes theCE separation conditions independent ofthe MSD operation conditions.

CE solutions – for discovery,development and quality controlof drugsIn today’s demanding pharmaceutical industry, fast and robust separation methods areneeded in order to reduce the time to market for new drugs. Capillary electrophoresis,the powerful instrumental approach to electrophoresis, responds to these needs in allstages from drug discovery through quality control. CE offers fast separations withorthogonal selectivities to chromatographic techniques. In addition, the integration ofVitaminsCE into the US Pharmacopeia as a standard physical test—initiated in 1996, ensures itsacceptance worldwide.Agilent Technologies has made a long-term commitment to the pharmaceutical industry.Changing regulatory requirements are monitored all over the world to ensure you get theright product features and services—a large number of applications and solution kitshave already been developed as part of this process.OligonucleotidesAbsorbance [mAU]60ChecklistMethod:þ Fast separationþ Robustþ Low cost per analysisþ Regulatory compliance toolsþ Simultaneous quantification of maincomponent and trace contaminantsHardware:þ Agilent high sensitivity detection cellþ Replenishment for unattended operationþ CEC capabilityþ Single vendor CE-MS solutionSoftware:þ Multitechnique softwareþ Self-explanatory user interfaceþ Direct access to instrument controlþ Enhanced integrator for CE peaksþ Mobility calculation for improvedreproducibilityþ Integrated CE-MS controlþ Data accessibility for the next millennium50403020100without organicadditivewith organicadditive5 10 15 20 25 30 35OligonucleotidesThe analysis of oligonucleotides orantisense oligonucleotide therapeuticsby capillary electrophoresis is fast,reproducible and highly automated.This makes it ideally suitable for qualitycontrol applications in oligonucleotidecore facilities, as well as in many phasesof bio pharmaceutical drug development.ProteinsTime [min]

Absorbance[mAU]PeptidesDouble stranded DNA and PCR productsCapillary electrophoresis provides aconvenient method to determine thesize of double-stranded DNA and PCRproducts. The Agilent ChemStationautomatically generates a Ferguson plotfor size determination.864202.72.62.52.42.32.22.1-36 bp-51 bp-65 bp-75 bpLog BP-126 bp0.08 0.09 1/MT-179 bp-222 bp-350 bp-396 bp-480 bp-517 bp-676 bp-1,198 bp-1,605 bp8 9 10 11 12 13 14 15-2,654 bpTime [min]DNA“Regulatory authorities are increasingly requesting that the pharmaceutical industryassess the enantiomeric purity of their chiral building blocks, intermediates and finaldrug substances. This, coupled with the ever increasingdemands imposed on the analyst to reduce methodAbsorbance [mAU]development times, makes chiral CE a very attractivetechnique. It offers unprecedented chiral discrimination through its high resolving power, chiral selector500versatility and rapid automated method development.”400S-formDr. Melvin Euerby(Astra Charnwood, UK)300R-form2000.05 % area/area100Chirals00 10 20 30Time [min]Chiral drug compoundWhen used with the high sensitivitydetection cell the simultaneous quantificationof the main component and traceimpurities below 0.1 % is possible withoutsample overloading.Glycoproteins

CE solutions – the alternative for ionsCapillary electrophoresis is a true alternative to ion chromatography.Its main features include:• unmatched analysis time—enhancing your sample throughput• high resolving power—allowing analysis of complex samples in one run• typically < 1 ml of solvent required with only a small amount of waste— savingmoney and helping to protect the environment• no need for specialized columns, precolumns and membranes—easy instrumentsetup and maintenance• often no laborious sample preparation needed—just sample dilution• only nanoliter amounts of samples injected—allowing different methodsor analytical techniques to be applied to one sampleAbsorbance [mAU]Ionic contamination in primary andsecondary circuit water leading to metalcorrosion, is a major problem for thenuclear power generating industry.For this reason low levels of smallanions and cations must be monitored.CE provides a rapid and facile methodfor their determination in the lowppb range.Bromide ChlorideSulfateOxalate Nitrate NitriteChlorateFluorideFormate3 4 5 6 7 8 9Time [min]ChecklistPhosphateþ Ease of sample preparationþ Low sample consumptionþ Fast separationþ Minimal maintenanceþ Low cost of ownershipþ Low ppb sensitivityAcetatePower plantsAbsorbance [mAU]2.521.510.50-0.5ChlorideSulfateNitriteNitrate2 2.5 3 3.5 4 4.5Time [min](Data kindly provided bt Thomas Ehmann,Wacker Siltronic AG, Burghausen, Germany)Semiconductor industry“Capillary electrophoresis, as a microanalytical technique,features merits in the determination of contaminants on wafersurfaces. As the diameter increases, the financial value of thewafers increases exponentially, therefore it is mandatory toapply a great variety of different analysis techniques whenworking on one single wafer. This is possible with CE dueto the low sample volume. Moreover, by changing only thecomposition of the electrolyte, a broad variety of analytesin different matrices can be determined.CE covers a range from ionic to non-ionic analytes that areimportant in the semiconductor industry.”OxalateCitrateDr. Laszlo Fabry(Wacker Siltronic AG, Germany)FluorideFormate

Recycling for cost reduction and environmentalconcerns necessitates themonitoring of Kraft liquors for anioniccontent. The caustic nature of thesamples results in labor intensive maintenanceprocedures. CE analyses are threetimes faster and require minimal samplepreparation compared to ion chromatography.S 2O 32-Cl - SO 42-Green liquor,1:200 dilution in water2 2.5 3(Data kindly provided by Dwayne Van,Longview Fibre, Longview WA, USA)Pulp and paper industryS 2- SO 32-Time [min]Plating bath industry"In the plating bath industry, the monitoring of additives in bathsolutions or waste is essential for quality control and costsaving. The analysis by capillary electrophoresis is advantageousover ion chromatography with respect to samplepreparation, resolution and simplicity."Nobuo Ebina(Ebina Denka Kogyo Co., Japan)Absorbance [mAU]-15-20-25SulfateMalateHypophosphitePhosphateAbsorbance [mAU]-20-25-30-35-40-45ChlorideSulfateTartrateMalateCitratePyruvate SuccinateAcetateLactatePhosphate2 3 4 5 6 7 8Time [min]Food and beverages industry-30-35-40"Validation of CE for organic acid analysis in grape juice andwine has enabled us to replace other methods like ionchromatography (IC), or HPLC. Major advantages are, speed ofanalysis—six major organic acids in less than 13 minutes;instrumental simplicity; its adaptability in a routine QC lab—currently running 24 hours; no sample cleanup; low cost ofsupplies and maintenance, and minimal waste disposal."AcetateLactateNickel2 3 4 5 6Time [min]Steve Kupina(Canandaigua Wine Co., USA)

CEC – for chromatographic selectivityand highest efficienciesCapillary electrochromatography (CEC) is a fusion of the techniques of capillaryelectrophoresis and liquid chromatography, resulting in an increase in efficiencyby as much as an order of magnitude over conventional HPLC.This has three specific benefits:• separation of closely related compounds• shorter run times and increased sample throughput, and• substitution of gradient HPLC methods by isocratic CEC.CEC uses capillary columns packed withLC stationary phases. As in LC, CECseparations are achieved by the partitionof a solute between the mobile andstationary phases. The additional separationby mobilities resolves charged andneutral components.RadiusPressure drive“CEC’s vastly increased resolving powercoupled to the high speed analysis which thistechnique also offers, is essential in a modernpharmaceutical industry which demands theseparation of increasingly complex mixtures.“Absorbance[mAU]Dr. Melvin Euerby(Astra Charnwood, UK)In CEC, the pressure driven flow of LC isremoved and replaced by an electricallydriven flow—the electro osmotic flow(EOF). This means:• no pump with moving mechanical partsor seals,• elimination of eddy diffusion, resultingin increased efficiency, and• column length and particle size that arenot restricted by a pressure limit.ChannelParticleVelocity profileLCCECRobust, longterm operation of CECcolumns requires pressurization of bothcapillary ends. This is available as a corefunction of the Agilent CE system.Electro osmotic driveFlow velocityTime [min]Separation of an EPA PAHstandard by CECChecklistþ Microanalytical techniqueþ Separation of closely related compoundswithout gradientþ High sample throughputþ Chromatographic selectivity plusseparation mechanism by chargeAbsorbance [mAU]864263, 4 5 9 10178211121314151601 2 3 4 5 6 7 8 9Time [min]

CE-MS – one vendor, one software, one solutionCE-MS combines the short analysis time and high separation efficiency of CE with themolecular weight and structural information from the MS. The technique has beensuccessfully used for the analysis of biopolymers, drugs and drug metabolites, andagrochemicals, among others.Unlike other systems, Agilent Technologies offers a fully integrated solution—all thesystem components come from one vendor and are controlled by one softwarepackage, combined to give you one solution. In addition, Agilent's unique designfeatures surpass previous limitations of CE-MS.• The spraying system from Agilent is nowarranged at right angles and needs nopositional adjustments, enabling verysimple and stable operation and allowingthe use of moderate amounts of conventionalelectrophoretic buffers oradditives, for example phosphate orcyclodextrins,• the nebulizer is operated at groundpotential increasing the effective fieldstrength over the separation capillaryand making the CE separation conditionsindependent from the MS operationconditions,• the high-efficiency solvent dryingsystem ensures excellent sensitivity.Agilent's spraying systemPeptide separation with simultaneousUV and MS detectionMS: Abundance(M+2H) 2+ 530.9(M+3H) 3+ 433.1MW 1060.2 BradykininMW 1296.5 Angiotensin IMW 1672.9 NeurotensinMW 1046.2 Angiotensin IIMW 379.4 VYMMW 555.6 Leu-EnkephalinMW 573.7 Met-EnkephalinFully integrated and automated,Agilent's CE-MS softwareguarantees ease of use frommethod development and dataprocessingUV: Absorbance [mAU](M+3H) 3+ 558.5(M+2H) 2+ 523.8(M+H) + 380.3(M+H) + 556.2(M+H) + 574.2Time [min]

SpecificationsStep 4: Qualify system performanceduring routine operationAgilent’s software features automateddaily system suitability testing, with anoptional database with online qualitycontrol charts for documenting performanceparameters.Dimensions Width 42.5 cm, (16.8 in)Height 57.5 cm, (22.7 in)Depth52 cm, (20.5 in)Weight 52 kg, (115 lb)Environment Temperature: 5–40 °CHumidity: up to 80 %, at 30 °C (non-condensing)Power requirements Line voltage: 100/120/220/240 VAC ; +5 %, -10 %, 650 VALine frequency: 50 (48–55) Hz; 60 (57–66 ) HzPressure systemProgrammable with 0–50 mbar bidirectionalFlushing with 1 bar or with high pressure 2–12 bar bidirectionalVial pressurization with high pressure 2–12 barStep 5: Ensure data security, integrity andtraceabilityThe software has a password-protecteduser access. It saves instrument conditionsand logbooks together with raw datain checksum-protected binary files.Safety featuresCurrent leak detection; Low current limitSafety sensors at door and cover disabling high voltageDiagnostic functionsElectrophoresis power Voltage range: setable 0 to ± 30 kV supplyCurrent: setable 0–300 µAPower: setable 0–6 WOperation under constant voltage, current or powerProgrammable polarity switchInjection modesSelf correcting injection system with injection from both endsProgrammable range:up to 10,000 secondsPressure:0–50 mbarElectrokinetic:0–30 kVVoltage stabilityReplenishment functionalityAutosampler/fraction collectorReplenishmentVialsCapillary cassetteDetectorRaw data channelsSystem controlCE specific software48-position carousel. All vials are randomly accessible fromcathode and anode end of capillary. Temperature control withexternal waterbath with vial temperature from 10–40 o C(non-condensing conditions)Satellite station for refilling anode and cathode buffer vials with freshbuffer for automatic continuous operation and buffer selectablebuffer levelling100 µl sample vials, 1 ml or 2 ml buffer vials (polypropylene orglass) with resealing snap capsForced-air temperature-controlled with Peltier elementTemperature range:10 o C below ambient to 60 o C(± 0.1 °C) with a min. of 4 °CMinimum total capillary length: 33 cmCapillary compatibility365 µm o.d.Real time UV-Visible diode-array detector (190–600 nm)Wavelength accuracy:1 nmResponse time:0.1 to 10 s (8 choices)Light source:prealigned deuterium lampSignals:up to five signals simultaneously,full spectral acquisition withAgilent ChemStationDetector signals, voltage, current, power, capillary temperatureand pressure.Operating with graphical user interface under Microsoft âWindows 95 and Windows NT âTime programmable parameters: voltage, current, power, polaritypressure, inlet and outlet vialcapillary temperature,pre and post-run conditioningwith pressure and/or voltage,replenishment, fraction collectionMobility report, time corrected areas, pI calibration and bio polmersize calibration

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