Ultrafast separations beyond high pressure driven UHPLC separations beyond high pressure driven...

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    Ultrafast separations beyond high pressure driven UHPLCNUCLEOSHELL particles

    0.6m

    5 m

    0.5m

    2.7 m

    0.6m0.5m

    shell of porous silica

    shell of porous silica

    3.8m1.7m solid core of silicon dioxide

    Core-shell technology

    Solidcoreofsilicondioxide,homogeneousshellof porous silica Highestefficiencycomparedtototallyporoussilicaparticles Particlesize2.7m(core1.7m)and5m(core3.8m),poresize90,specificsurface

    130(2.7m)and90(5m)m2/g;lowbackpres-sureenablesuseonconventionalLCsystems

    Pressurestabilityupto600barNUCLEOSHELL modificationsTheprogramofNUCLEOSHELLsurfacemodifica-tionsnowcomprisesthefollowingphases:

    NUCLEOSHELL RP 18 NUCLEOSHELL RP 18plus NEW! NUCLEOSHELLPhenyl-Hexyl NUCLEOSHELL PFP NUCLEOSHELLHILICSeveral approaches have beenmade to achieve fastseparations without losing chromatographic perfor-mance. HPLC columns packed with particles

  • 3www.mn-net.com

    NUCLEOSHELLCore-shell silicaDemands on HPLC separations are constantly in-creasing with respect to separation efficiency, de-tection limits, and the time requirements for each analysis. Core-shell technology sets new standards for analyses in research and quality control.

    Rs =N4

    ki+1ki

    1

    Rs =Resolutiona =Selectivityki =RetentionN =TheoreticalplatesN 1/dPdP =Particlesize

    Resolution RS as function of particle sizeColumns: 50x4mmeach

    NUCLEOSHELL RP 18, 2.7 m NUCLEODURC18Gravity,3m NUCLEODURC18Gravity,1.8m

    Eluent: acetonitrilewater(60:40,v/v)Flowrate: 1mL/minTemperature: 25CDetection: UV,254nmPeaks: 1.Naphthalene2.Ethylbenzene

    Better resolution at lower back pressureand shorterretention time

    1

    2

    22

    11

    0.0 0.5 1.0 1.5 2.0 2.5 min

    RS = 2.47104 bar

    RS = 2.17190 bar

    RS = 1.7188 bar

    MN Appl. No. 125270

    Electron microscopic image of NUCLEOSHELL particlesUtilizing a proprietary process of synthe-sis, NUCLEOSHELL particles exhibita distinct narrow particle size distribu-tion (d90/d10 ~1.1). Columns packed withNUCLEOSHELL core shell particlesfeature exceptional separation efficien-cieswith theoretical plate numbers easilycomparabletototallyporoussub2micronparticles.

    Van Deemter plots

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    Term A:Term B:Term C:

    Eddy diffusion Longitudinal diffusion Mass transfer

    Columns:Eluent:Temperature:Sample:

    50 x 4.6 mmCH3CN H2O (70:30, v/v)

    Acenaphthene25 C

    Linear velocity u [mm/s]

    Plat

    e he

    ight

    h [

    m]

    h = A + Bu--- + C u

    NUCLEOSHELL RP 18

    NUCLEODUR C18 Gravity, 1.8 m

    Kinetex C18

    Ascentis Express C18

    Poroshell 120 EC-C18NUCLEODUR C18 Gravity, 3 m

    MN Appl. No. 125500

    Pressure drop

    0

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    600

    0 2 4 6 8 10 12

    Kinetex C18

    Ascentis Express C18

    Poroshell 120 EC-C18

    NUCLEODUR C18 Gravity, 3 m

    dp2 LC up =

    NUCLEOSHELL RP 18

    Columns:Eluent:Temperature:

    50 x 4.6 mmCH3CN H2O (70:30, v/v)25 C

    Pres

    sure

    [bar

    ]

    Linear velocity [mm/s]

    = pressure drop= flow resistance (nondimensional)= column length= viscosity= linear velocity = particle diameter

    pLCu dp

    NUCLEODUR C18 Gravity, 1.8 m

    MN Appl. No. 125510

    In direct comparison with the con-ventional sub 2 micron phases,NUCLEOSHELL columns only gen-erateabout60%ofthebackpressureandcanbeoperatedwith themajor-ity of conventional HPLC systems.In order to develop the maximumperformance of NUCLEOSHELL columns, we recommend reducingextracolumnvoidsbyusingsuitablecapillaries(

  • info@mn-net.com4

    Features of core-shell silica particlesStability under acidic and basic conditionsColumn: 50 x 4.6 mm NUCLEOSHELL RP 18, 2.7 m

    50x4.6mmKinetex2.6mC18 Eluent: acetonitrile1%TFAinH2O,pH 1(50:50,v/v)Flowrate: 1.3mL/min;temperature80CDetection: UV,254nmSample: anthracene

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    0 5000 10000 15000 20000 25000 30000

    pH 1

    NUCLEOSHELLKinetex

    Column volumes

    % in

    ital r

    eten

    tion

    (ant

    hrac

    ene)

    Columns: 50 x 4.6 mm NUCLEOSHELL RP 18, 2.7 m 50x4.6mmAscentisExpressC18,2.7m 50x4.6mmPoroshell120EC-C18 50x4.6mmKinetex2.6mC18

    Eluent: 20mmol/LNaborate10mmol/LNaOHmethanol,pH10(21:49:30,v/v))

    Flowrate: 1.5mL/min;temperature40CDetection: UV,220nmSample: toluidine

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    0 5000 10000 15000 20000

    pH 10

    Ascentis ExpressNUCLEOSHELL

    Poroshell 120Kinetex

    Plat

    es (t

    olui

    dine

    )

    Column volumes

    MN Appl. No. 125520/125530

    The above figure shows a column stability test ofNUCLEOSHELLRP18 atmobile phase levels pH1andpH10comparedwiththreecompetingphases.

    Acriterionforthelong-termstabilityofthecolumnatpHextremesisthepercentagedecreaseofinitialretentionandinitialplates,respectively.Thecolumncanalsobeoperatedatelevated temper-atureswithout loss in retention behavior, efficiency orpeaksymmetry.

    Temperature stabilityStability test:Column: 50x2mmNUCLEOSHELLRP18,2.7mEluent: A)10mmol/Lammoniumformatemethanol

    (9:1,v/v)+120Lformicacid,~pH4 B)10mmol/Lammoniumformatemethanol(1:9,v/v)+120Lformicacid,~pH4 0100%Bin7min

    Flowrate: 0.5mL/minTemperature:100 CDetection: UV,220nmPeaks:1. Phenol2.Naphthalene

    0

    1

    2

    2 4 6 8 min

    38 h34 h30 h26 h22 h

    MN Appl. No. 125400Efficiency test:Eluent: acetonitrilewater(60:40,v/v)Flowrate: 0.33mL/minTemperature:25CDetection: UV,254nmSample: anthracene

    HETP[m] AsymmetryStart(t=0) 5.2 0.98End(t=40h) 5.2 1.01

    Batch-to-batch reproducibilityColumn: 50x4mmNUCLEOSHELLRP18,2.7mEluent: methanol25mmol/LKH2PO4 pH 7

    (70:30,v/v)Flowrate: 1mL/minTemperature: 40CDetection: UV,254nmPeaks:1.Uracil2.Toluene3.Ethylbenzene4.Acenapthene5.Amitriptyline6. o-Terphenyl7.Triphenylene

    0 2 4 6 8 10

    3

    4

    56

    721

    min

    Batch 1

    Batch 2

    Batch 3

    MN Appl. No. 125410

    UniformlyshapedNUCLEOSHELLparticlescombinedwith optimized bonding technology safeguard tightlypackedcolumnsfor100%reproducibleresults.

  • 5www.mn-net.com

    NUCLEOSHELLLoading capacityNUCLEOSHELL columns allow reliable quantifica-tioninawideanalyticaldetectionrange.Retentiontimeandpeakwidthat50%heightremainconstantwithin-creasingcolumn loadalthoughcore-shellparticlesaresuspectedofshowingaslightly lower loadingcapacitycomparedtofullyporoussilicamaterials.

    Loading capacityColumn: 50x3mmNUCLEOSHELLRP18,2.7mEluent: acetonitrile25mmol/LKH2PO4, pH 3

    (70:30,v/v)Flowrate: 0.66mL/min,temperature30CDetection: UV,285nmPeaks:1.Valerophenone

    Normalized column parameters

    0 2 4 6 8 10

    Peak width(at 50 % peak height)

    Retention time

    Efficiency(at 10 % peak height)

    Load on column [g]

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    0.2

    0.4

    0.6

    0.8

    1

    1.2

    Peak capacityThepeakcapacityisameasureofthenumberofsampleanalytes thatcanbeseparatedonHPLCcolumnspertimeunit.Narrowpeaksincreasethepeakcapacityandefficiency of analytical columns. The example shows,that in comparison with totally porous NUCLEODUR silica (1.8m)NUCLEOSHELL provides33%higherpeakcapacity.

    Peak capacityColumns: 100x4.6mmeach

    NUCLEOSHELL RP 18, 2.7 m NUCLEODURC18Gravity,1.8m NUCLEODURC18Gravity,3m NUCLEODURC18Gravity,5m

    Eluent: A)acetonitrile,B)water,40100%Ain4minFlowrate: 1.5mL/min;temperature25CDetection: UV,230nmPeaks:1.Acetophenone2.Benzoin

    3. Propiophenone 4.Butyrophenone

    5.Benzophenone6.Valerophenone

    0 1 2 3 4 min

    3 4

    56

    2

    1

    MN Appl. No. 125540

    Max. pressure [bar]

    Resolution(4,5)

    NUCLEOSHELL, 2.7 m 255 5.45NUCLEODUR,1.8m 450 4.14NUCLEODUR,3m 214 2.97NUCLEODUR,5m 142 2.30

    100125

    163

    216

    0

    50

    100

    150

    200

    250

    nc:tg:W:

    NUCLEODUR5 m 3 m

    NUCLEOSHELL

    Wtgnc = 1 + ( )

    peak capacity

    Comparison of peak capacities

    gradient timepeak width (at baseline)1.8 m 2.7 m

    n c (n

    orm

    aliz

    ed)

    Method transfer of 5 m particle columnsNUCLEOSHELLisalsoavailablein5mparticlesizetoofferallbenefitsofcore-shelltechnologytoallappli-cationswhichareboundtoparticlesize.

    Cephalosporin antibiotics Comparison of 5 m core-shell and totally porous phasesColumns: 100x4.6mmeach

    A) NUCLEOSHELL RP 18plus, 5 m B)NUCLEODURGravityC18,5m

    1

    2

    3

    4

    0.0 2.5 5.0 7.5 10.0 12.5 min

    Eluent: methanolwater+0.1%formicacid(35:65,v/v)

    Flowrate: 1.3mL/minPressure: 182 bar, 219barTemperature: 25CDetection: UV,254nmInjection: 4.0L

    Peaks: Ret.time[min]

    Asymmetry(EP)

    Plates (EP)

    A B A B A B1 Cefotaxime 1.30 1.96 1.19 1.12 6800 22182 Cefoxitin 2.14 4.72 1.22 1.20 6599 34713 Cefamandole 2.97 6.57 1.24 1.25 6259 33674 Cefalotine 5.33 13.73 1.32 1.61 6948 3672

    MN Appl. No. 126630

    9 g

    0 1 2 3 min

    3 g

    0.9 g0.3 g

    0

    200

    400

    600

    800

    0 2 4 6 8 10

    R2 = 0.9991

    Area

    Load on column [g]

  • info@mn-net.com6

    NUCLEOSHELL RP 18 Key features:

    Basedoncore-shellparticletechnologyforfastandefficientHPLC

    SuitableforLC/MSandHPLCatpHex-tremes(pH111)

    Superiorbasedeactivation,idealformethoddevelopment

    Technical characteristics: Octadecylmodification,multi-endcapped;poresize90,availableparticlesizes2.7mand5m,carboncontent7.8%for2.7m,6.1%for5m

    Recommended application: Overallsophisticatedanalyticalseparations,e.g.,analgesics,anti-inflammatorydrugs,antid