Agilent 6430 Triple Quadrupole LC/MS System - Perlan Technologies

Agilent 6430 Triple QuadrupoleLC/MS SystemIdeal Quantitative LC/MS for UHPLC withDynamic MRM and Fast Polarity Switching,plus Unsurpassed Sensitivity with Chip LCSummaryThe Agilent 6430 Triple Quadrupole LC/MS System hasbeen specifically designed and engineered to be fullycompatible with ultra high pressure liquid chromatography(UHPLC) separations and as an ideal interface forAgilent’s proprietary HPLC Chip technology. The 6430Triple Quad is a feature-packed new addition to theAgilent triple quadrupole product family, with:• Reliable MassHunter software• MassHunter Optimizer for automatedcompound optimization• 1 ms dwell times with no collision cell cross talk• Dynamic MRM methods for up to 4,000 transitions• 30 ms polarity switching for fast analyses• Low attomole sensitivity in peptide quantitation withAgilent’s advanced HPLC-Chip Nano LC systemControlled by proven MassHunter software, the new 6430Triple Quad joins the Agilent 6410 Triple Quadrupole LC/MSSystem, an entry-level system for routine quantitative analysis,and the flagship Agilent 6460 Triple QuadrupoleLC/MS System with unrivalled sensitivity powered byAgilent Jet Stream technology, to create a powerful suiteof products to meet the quantitation demands of anyapplication or budget. The 6430 Triple Quad offers thesame combination of superior performance, exceptionalreliability, and excellent overall cost-of-ownership forwhich Agilent is famous.

Fast MRM Analysis and PolaritySwitching (Plus Sensitivity) —Perfect for UHPLCThe 1290 Infinity LC System for UHPLCoffers tremendous productivity gainsversus conventional HPLC. UHPLC withsub-2-micron columns can dramaticallyreduce separation times and maximizechromatographic resolution. This frequentlyresults in very narrow peakswhich may be less than a second wide.The new 6430 Triple Quad has beenengineered to achieve optimal quantitationof narrow UHPLC peaks.Reliable peak integration requires atleast ten data points across a peak,mandating MS cycle times as low as100 ms for a second-wide peak. Complex,multi-analyte assays require very shortdwell times. The 6430 Triple Quad cankeep up! It features extremely fast ionmonitoring and polarity switching. Theenhanced electronics in the 6430 TripleQuad enable 1 ms dwell times with nocollision cell cross talk. Further, theintroduction of dynamic MRM methodsallows dynamic selection of MRM transitionsduring a method, based onretention time windows, to optimizeinstrument cycle time and maximize collectionof high quality quantitative data. 1This allows hundreds of compounds tobe analyzed in a single LC/MS run.Samples containing compounds withboth positive and negative ion transitionsrequire fast polarity switching – the6430 Triple Quad can change polarityand begin acquiring data in just 30 ms.The improvement in polarity switchingtimes is derived from a new, resistivelycoated capillary that reacts very quicklyto voltage changes.In addition to fast analysis capabilities,the 6430 Triple Quad is more sensitivethan its predecessor – it has a standardsecond turbo pump in the Q3 region toenhance ion transmission by reducingchemical noise.Designed as an IDEAL SYSTEM for UHPLC/MS Analyses• 1200 bar pressure (up to 2 mL/min) for the fastest separations• 30 ms polarity switching for fast peaks• 1 ms dwell times for analyzing hundreds of compounds• Dynamic MRM methods for analyzing thousands of ion transitions• Improved sensitivityFigure 1: The Agilent 1290 Infinity LC System and the Agilent 6430 TripleQuadrupole LC/MS System. The 6430 Triple Quad is an ideal MS system forUHPLC separations.2

As part of a metabolic stability assay 2(Figure 2), several pharmaceuticalcompounds and their metabolites wereseparated using a 1.5 min method withthe Agilent 1290 Infinity LC System,using a sub-2-micron column and pressureof 1060 bar. The diclofenac iontransition was monitored along withfour other positive ion transitions andone negative ion transition. The cycletime for six ion transitions and positive/negativepolarity switching was lessthan 115 ms. This narrow peak wassampled 9 times, creating an excellentbasis for peak area determination - thearea RSD value for diclofenac was lessthan 5%.x10 33.83.432.62.21.81.410.6-MRM (294.0 -> 250.0)Diclofenac, narrowest peakAverage W 1/2= 0.37 sec9 points across WRel. Area RSD [%] = 7.9Area RSD [%] = 4.8Dynamic MRM Methods EnableComplex Multi-Residue AnalysesDynamic MRM methods dramaticallyreduce MS cycle times for complexmulti-analyte assays. MRM transitionlists are built dynamically during theLC separation based on a retention timewindow for each analyte. Using thedynamic MRM method capabilities ofthe 6430 Triple Quad, 224 pesticideswere analyzed in only 7 min as shownin Figure 3. The average peak widthswere about 1 sec wide, requiring anMS cycle time of about 100 ms forprecise and accurate quantitation of allcompounds. The minimum dwell timewas a very fast 2.5 ms. The excellentretention time precision of the 1290Infinity LC allowed very narrow retentiontime windows and reduced thenumber of required ion transitions atany point during the chromatogram tofewer than 20. The precision of thisvery fast pesticide assay was excellent,with an average peak area RSD of lessthan 5%.0.20.7 0.71 0.72 0.73 0.74 0.75 0.76 0.77Counts vs. Acquisition Time (min)W = 1.5 secCycle time = 114 msFig. 2: MRM chromatogram of diclofenac obtained using a flow rate of 1.5 mL/min. The graphicshows the peak width at half height (0.37 sec) and the number of data points collected acrossthe peak (9 data points).Figure 3. Fast analysis of 224-pesticides at the 500 ppt level in less than 7 min using the newAgilent 1290 Infinity LC and 6430 Triple Quad with dynamic MRM. Average peak area RSD was lessthan 5%.3

Robust, Reliable, SensitiveNanospray with HPLC-ChipThe highest possible sensitivity inLC/MS is achieved with the smallestcolumn diameters. Nano LC columnswith an i.d. of 75 microns offer anobvious route to high sensitivity buthave historically suffered from a lack ofrobustness and precision. Agilent’s revolutionaryHPLC-Chip/MS technologyprovides nanoscale LC on a plug-and-playmicrofluidic chip, minimizing leaks anddead volumes. The chip contains a fullyintegrated electrospray emitter forunprecedented spray stability.Figure 4. ChipCube with integrated valves, columns and spray tip.The 6430 Triple Quad with an HPLC-Chip/MS interface is ideal for targetedprotein quantitation using dynamicMRM methods. This system canachieve low attomolar sensitivity (seeFigure 5) in plasma protein digests.Figure 5. Attomolar detections limits for exogenous peroxidase protein spiked into immunodepletedhuman plasma, analyzed with the HPLC-Chip coupled with a 6430 Triple Quad.4

Dynamic MRM methods on the HPLC-Chip/6430 Triple Quad system can beused to quantitate hundreds or thousandsof peptides, as shown in theanalysis of the complex peptide digestshown in Figure 6. More than 2,000 iontransitions were monitored during thisLC/MS analysis.ConclusionsThe new Agilent 6430 Triple QuadLC/MS features seamless integrationwith the Agilent 1290 Infinity LCSystem and the proprietary AgilentHPLC-Chip/MS nanospray interface.This new system features:• Extensive sampling capabilities fornarrow UHPLC peaks• Fast 1 ms MRM dwell times• Dynamic MRM methods for complex,multi-analyte assays• Fast 30 ms polarity switching times• Attomole sensitivity for biomarkervalidation studies using theHPLC-Chip and 6430 Triple Quad• Rugged, reliable MassHuntersoftware control• Automated method optimization usingMassHunter Optimizer and PeptideOptimizer software• 21 CFR Part 11 support forvalidated methodsWith legendary Agilent robustness andreliability, and a commitment to highestquality service and support, the 6430Triple Quad is a productive new additionto the Agilent 6400 Series TripleQuadrupole LC/MS product family.Figure 6. 2,000 Dynamic MRM transitions from the tryptic digest of immunodepleted human plasmausing the HPLC-Chip/6430 Triple Quad system.References1. Peter Stone, Thomas Glauner, FrankKuhlmann, Tim Schlabach and KenMiller, “New Dynamic MRM ModeImproves Data Quality and TripleQuad Quantification in ComplexAnalyses,” Agilent publicationnumber 5990-3595EN, 2009.2. Anabel S. Fandiño, Edgar Naegele,“Metabolic Stability Study UsingCassette Analysis and PolaritySwitching in a UHPLC Triple QuadLC/MS System,” Agilent publicationnumber 5990-4469EN, 2009.5

www.agilent.com/chem/qqqThis item is intended for Research Use Only. Not for usein diagnostic procedures. Information, descriptions, andspecifications in this publication are subject to changewithout notice.Agilent Technologies shall not be liable for errors containedherein or for incidental or consequential damagesin connection with the furnishing, performance or use ofthis material.© Agilent Technologies, Inc. 2009Printed in the U.S.A. December 22, 20095990-4788EN