www.mn-net.com

www.mn-net.comMACHEREY-NAGEL GmbH & Co. KG · Neumann-Neander-Str. 6–8 · 52355 Düren · GermanyFrance:MACHEREY-NAGEL EURLTel.: +33 388 68 22 68Fax: +33 388 51 76 88E-mail: sales-fr@mn-net.com

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MACHEREY-NAGEL

EN ISO 9001: 2008CERTIFIED

USA:MACHEREY-NAGEL Inc.Tel.: +1 484 821 0984Fax: +1 484 821 1272E-mail: sales-us@mn-net.com

Highest efficiency in HPLC by core-shell technology

NUCLE SHELL

HPLC methods for the future . . . we Meet your Needs

www.mn-net.com2

Ultrafast separations beyond high pressure driven UHPLCNUCLEOSHELL 2.7 µm

0.5 μm

0.5 μm

shell of porous silica

shell of porous silica

1.7 μm solid core of silicon dioxide

Core-shell technology Solid core of silicon dioxide, homogeneous

shell of porous silica

Highestefficiencycomparedtototallyporous silica particles

Particlesize2.7µm(core1.7µm); lower back pressure enables use on conventionalLCsystems

In the past several approaches have been made to achieve fast separations without losing chromatographic performance. The prevalent strategy is to use very small silica particles, commonly sized smaller than 2 µm. HPLC col-umns packed with those microparticles show very high efficiencies (plates/meter) and allow the use of smaller column sizes with the posi-tive side effect of significant solvent saving. However the downside of sub 2 micron particle HPLC is the generated high back pressure of the mobile phase during column runs which re-quire specifically designed and often expensive instrumentation (UHPLC suitable solvent deliv-ery and detection equipment).

Core-shell particle technology from MACHEREY-NAGEL is an alternate route to gain highest column efficiency and resolution at almost the same short run time but with much lower back pressure.

NUCLEOSHELL silica particles consist of a non-po-roussolidcoreof1.7µmdiameterandaporousouter shell of0.5µm thickness.Accordingly thetotaldiameteroftheparticleis2.7µm.Withconventionalfullyporousparticlesthemasstransferbetweenstationaryandmobilephaseusu-

allyresultsinpeakbroadeningathigherflowrates(C-terminvan-Deemterequation).Theshortdif-fusion paths in the core-shell particles reduce the dwelltimeoftheanalytemoleculesinthestation-aryphase,sothatevenathighflowvelocitiesofthe mobile phase, optimal separation results can be obtained.The Van Deemter plots on page 3 demonstratehowefficiencyisaffectedbyflowrate.Incompari-sonwith fullyporoussilicas, core-shellparticlesfromvariousmanufacturersmaintaintheefficien-cyoptimum(max.plates/m)overalongrangeofincreasinglinearmobilephasevelocity.

Theoretical column efficiency (optimal conditions)Silica phase dp

[µm]L [m]

HETP [µm]

Efficiency [plates/m]

L [mm]

N Rs Analysistime

NUCLEOSHELL 2.7 1 4 250 000 100 25 000 112 % 40 %NUCLEODUR® 1.8 1 4.5 222 222 100 22 000 105% 40%

3 1 7.5 133333 150 20 000 100% 60%5 1 12.5 80000 250 20 000 100% 100%

Benefits of core-shell technology Core-shell particles vs. totally porous silica gel

Short diffusion paths Fastmasstransfer(C-TermofVanDeemterequation)

Highflowvelocitywithoutpeakbroadening for fast LC

Narrow particle size distribution (d90/d10 ~ 1.1) Stable packing ColumnefficiencyNUCLEOSHELL~250 000 m–1 (HETP~4µm)

Minimized frictional heat

3www.mn-net.com

NUCLEOSHELLCore-shell silicaDemands on HPLC separations are constantly increasing with respect to separation efficiency, detection 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’

1αα–

Rs = Resolution a = Selectivityki’ = Retention N = Theoretical plates N ∝ 1/dPdP = Particle size

Resolution RS as function of particle sizeColumns: 50 x 4 mm

NUCLEOSHELL RP 18, 2.7 μm NUCLEODUR® C18 Gravity, 3 μm NUCLEODUR® C18 Gravity, 1.8 μm

Eluent: acetonitrile – water (60:40, v/v) Flow rate: 1 mL/minTemperature: 25 °CDetection: UV, 254 nmPeaks: 1. Naphthalene2. Ethylbenzene

Better resolution at lower back pressureand shorterretention time

MN Appl. No. 125270

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

Van Deemter plots

0

2

4

6

8

10

12

14

16

18

20

0 2 4 6 8 10

Kinetex™ C18Ascentis® Express C18NUCLEOSHELL RP18Poroshell 120 EC-C18

NUCLEODUR® C18 Gravity, 3 μm

Term A: Eddy diffusion Term B: Longitudinal diffusion Term C: Mass transfer

Columns: 50 x 4.6 mmEluent: CH3CN – H2O (70:30, v/v)Temperature:Sample: Acenaphthene

25 °C

Linear velocity u [mm/s]

Plat

e he

ight

h [μ

m]

h = A + Bu--- + C · u

NUCLEODUR® C18 Gravity, 1.8 μm

Electron microscopic image of NUCLEOSHELL particlesUtilizingaproprietaryprocessofsyn-thesis, NUCLEOSHELL particles exhibit a distinct narrow particle size dis-tribution (d90/d10 ~1.1). Columnspacked with NUCLEOSHELL core shell particles feature exceptional separa-tionefficiencieswith theoreticalplatenumbers easily comparable to totallyporous sub 2 micron particles.

Indirectcomparisonwiththe“con-ventional” sub 2 micron phases, NUCLEOSHELL columns only gen-erateabout60%ofthebackpres-sure and can be operated with the majorityofconventionalHPLCsys-tems.Inordertodevelopthemaxi-mum performance of NUCLEOSHELL columns, we recommend reducing extra column voidsbyusing suit-able capillaries (< 0.15mm innerdiameter) and specially adapteddetector cells. Moreover detector settings should be optimized byincreasingthemeasuringrateorbydecrease of the time constant.

Pressure drop

0

100

200

300

400

500

600

0 2 4 6 8 10 12

Kinetex™ C18

Ascentis® Express C18NUCLEOSHELL RP18

Poroshell 120 EC-C18

NUCLEODUR® C18 Gravity, 3 μm

dp2

Φ · LC · η · uΔp =

Columns: 50 x 4.6 mmEluent: CH3CN – H2O (70:30, v/v)Temperature: 25 °C

Pres

sure

[bar

]

Linear velocity [mm/s]

= pressure drop= flow resistance (non-dimensional)= column length= viscosity= linear velocity = particle diameter

ΔpΦLCηu dp

NUCLEODUR® C18 Gravity, 1.8 μm

www.mn-net.com4

Features of core-shell silica particlesStability under acidic and basic

conditionsColumn: 50 x 4.6 mm NUCLEOSHELL RP 18, 2.7 μmEluent: acetonitrile – 1 % TFA in H2O, pH 1 (50:50, v/v)Flow rate: 1.3 mL/minTemperature: 80 °CDetection: UV, 254 nmSample: anthracene

0

20

40

60

80

100

0 5000 10000 15000 20000 25000 30000

pH = 1

Column volumes

% in

ital r

eten

tion

(ant

hrac

ene)

Columns: 50 x 4.6 mm NUCLEOSHELL RP 18, 2.7 μm 50 x 4.6 mm Ascentis® Express C18, 2.7 μm 50 x 4.6 mm Poroshell 120 EC-C18

Eluent: 20 mM Na borate – 10 mM NaOH – methanol, pH 10 (21:49:30, v/v))

Flow rate: 1.5 mL/minTemperature: 40 °CDetection: UV, 220 nmSample: toluidine

0

2000

4000

6000

8000

10000

12000

0 5000 10000 15000 20000

Ascentis ExpressNUCLEOSHELL

Poroshell 120

Plat

es (t

olui

dine

)

Column volumes

pH = 10

TheabovefigureshowsacolumnstabilitytestofNUCLEOSHELLRP18atmobilephaselevelspH1andpH10comparedwithtwocompetingphases.

Acriterionforthelong-termstabilityofthecol-umn at pH extremes is the percentage decrease ofinitialretentionandinitialplates,respectively.Thecolumncanalsobeoperatedatelevatedtem-peratures without loss in retention behavior, ef-ficiencyorpeaksymmetry.

Temperature stabilityStability test:Column: 50 x 2 mm NUCLEOSHELL RP 18, 2.7 μmEluent: A) 10 mM ammonium formate – methanol

(9:1, v/v) + 120 μL formic acid, ~ pH 4 B) 10 mM ammonium formate – methanol (1:9, v/v) + 120 μL formic acid, ~ pH 4 0–100 % B in 7 min

Flow rate: 0.5 mL/minTemperature: 100 °CDetection: UV, 220 nmPeaks:1. Phenol2. Naphthalene

0 2 4 6 8 min

38 h34 h30 h26 h22 h

MN Appl. No. 125400Efficiency test:Eluent: acetonitrile – water (60:40, v/v)Flow rate: 0.33 mL/minTemperature: 25 °CDetection: UV, 254 nmSample: anthracene

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

Batch-to-batch reproducibilityColumn: 50 x 4 mm NUCLEOSHELL RP 18, 2.7 μmEluent: methanol – 25 mM KH2PO4 pH 7

(70:30, v/v)Flow rate: 1 mL/minTemperature: 40 °CDetection: UV, 254 nmPeaks: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

Uniformly shaped NUCLEOSHELL particles com-bined with optimized bonding technology safe-guardtightlypackedcolumnsfor100%reproduc-ible results.

5www.mn-net.com

NUCLEOSHELLPeak capacity

Columns: 100 x 4.6 mm NUCLEOSHELL RP 18, 2.7 μm NUCLEODUR® C18 Gravity, 1.8 μm NUCLEODUR® C18 Gravity, 3 μm NUCLEODUR® C18 Gravity, 5 μm

Eluent: A) acetonitrile, B) water, 40–100 % A in 4 minFlow rate: 1.5 mL/minTemperature: 25 °CDetection: UV, 230 nmPeaks:1. Acetophenone2. Benzoin3. Propiophenone

4. Butyrophenone5. Benzophenone6. Valerophenone

0 1 2 3 4 min

3 4

56

2

1

Max. pressure [bar]

Resolution (4,5)

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

Peak capacity

100125

163

216

0

50

100

150

200

250

nc: peak capacitytg: gradient timeW: peak width (at baseline)

NUCLEODUR®5 μm 3 μm 1.8 μm 2.7 μm

NUCLEOSHELL

n c (n

orm

aliz

ed)

Wtgnc = 1 + ( )

Thepeakcapacityisameasureofthenumberofsample analytes that can be separated on HPLCcolumns per time unit. Narrow peaks increase thepeakcapacityandefficiencyofanalyticalcol-umns. The example shows, that in comparisonwith totally porous NUCLEODUR® silica (1.8 µm)NUCLEOSHELLprovides33%higherpeakcapacity.

Loading capacityNUCLEOSHELL columns allow reliable quantifica-tioninawideanalyticaldetectionrange.Retentiontimeandpeakwidthat50%height remaincon-stant with increasing column load although core-shellparticlesaresuspectedofshowingaslightlylower loading capacity compared to fully poroussilica materials.

Loading capacityColumn: 50 x 3 mm NUCLEOSHELL RP 18, 2.7 μmEluent: acetonitrile – 25 mM KH2PO4, pH 3

(70:30, v/v)Flow rate: 0.66 mL/minTemperature: 30 °CDetection: UV, 285 nmPeaks:1. Valerophenone

0

200

400

600

800

0 2 4 6 8 10

R2 = 0.9991

Area

Load on column [μg]

9 μg

0 1 2 3 min

3 μg

0.9 μg0.3 μg

Normalized column parameters

0

0.2

0.4

0.6

0.8

1

1.2

0 2 4 6 8 10

Peak width(at 50 % peak height)

Retention time

Efficiency(at 10 % peak height)

Load on column [μg]

www.mn-net.com6

NUCLEOSHELL RP 18 Key features:

· Based on core-shell particle technol-ogyforfastandefficientHPLC

· Suitable for LC/MS and HPLC at pH extremes(pH1–11)

· Superior base deactivation, ideal for method development

Technical characteristics: Octadecylmodification,multi-endcapped;poresize90Å,particlesize2.7μm,carboncontent7.5%

Recommended application: Overallsophisticatedanalyticalseparations, e.g., analgesics, anti-inflammatorydrugs,antidepressants;herbicides;phytopharmaceuticals;immunosuppressants

USP L1

NUCLEOSHELL RP 18NUCLEOSHELLRP18 isbasedoncore-shellpar-ticle technology silica. A unique derivatizationprocess generates a homogeneous surface with ahighdensityofbondedsilanes(carboncontent~7.5%).Thefollowingthoroughendcappingsup-pressesanyunwantedpolarinteractionsbetweenthe silica surface and the sample, which makes NUCLEOSHELLRP18particularly suitable for theseparationofbasicandotherionizableanalytes.Theextremelyreducedsilanolactivityofthephasecanbedemonstratedbyapplyingbasicanalytes,suchas tricyclic antidepressants.The chromato-gram on page 7 shows a sharp elution profile(superior resolution!)of thesehighlypolar com-pounds with an excellent asymmetry value foramitriptylineof1.12.

Tanaka plot of NUCLEOSHELL RP 18 Thediagrambelowunderlinesthedistincthydro-phobiccharacteristicsandthelowsilanolactivityofthestationaryphase.

0.5

1hydrogen bonding capacity

5

10capacity

0.5

1

ion exchange capacity pH 7.6

1

2

hydrophobicity

0.1

0.2

ion exchange capacity pH 2.7

1

2

stericselectivity

Parameters of the Tanaka diagramCapacity=k’(pentylbenzene)Hydrophobicity=α(pentylbenzene,butylbenzene)Stericselectivity=α(triphenylene,o-terphenyl)Hydrogenbondingcapacity(silanolcapacity)=α(caffeine,phenol)Ionexchangecapacityat2differentpHvalues(2.7and7.6)=α(benzylamine,phenol)

The separation of 13 β-lactam antibiotics illus-trateshowtimeofanalysiscanbeshortenedtoafractionalpartbyusingcore-shellparticleswith-out loss of resolution at moderate back pressure.

13 β-lactam antibiotics in less than 3 minColumns: 50 x 4 mm NUCLEOSHELL RP 18, 2.7 μm

150 x 4 mm NUCLEODUR® C18 Gravity, 5 μmEluent: A) acetonitrile; B) 20 mM KH2PO4 pH 3.5

10 % A (0.5 min) → 50 % A in 1.5 min (0.5 min 50 % A) 10 % A (3 min) → 50 % A in 9 min (3 min 50 % A)

Flow rate: 2 mL/min, 1 mL/minPressure: 270 bar, 110 barTemperature: 25 °C Detection: UV, 220 nmPeaks:1. Amoxicillin2. Ampicillin3. Cephalexin4. Cefotaxime5. Cefoxitin

6. Cefamandole7. Cephalothin8. Piperacillin9. Penicillin V10. Oxacillin

11. Cloxacillin12. Nafcillin13. Dicloxacillin

0.0 0.4 0.8 1.2 1.6 2.0 min

0 2 4 4 8 10 12 min

1

23

46

5

7 8 9

1011

1213

1

23 4

6

5

7 89

10 11

12

13

2.5 min270 bar

MN Appl. No. 124940

7www.mn-net.com

NUCLEOSHELLTricyclic antidepressants · comparison of selectivity and resolution

Columns: 50 x 4.6 mm each NUCLEOSHELL RP 18, 2.7 μm Ascentis® Express C18 Kinetex™ 2.6 μm C18 Poroshell 120 EC-C18

Eluent: methanol – acetonitrile – 25 mM KH2PO4 pH 7 (22.5:22.5:55, v/v/v)

Flow rate: 2 mL/min Pressure: 224 bar, 239 bar, 248 bar, 212 barTemperature: 40 °C Detection: UV, 220 nm

Asymmetry (amitriptyline)

Resolution (8, 9)

NUCLEOSHELL 1.12 3.35Ascentis® Express 2.07 1.91Kinetex™ 1.33 n.a.Poroshell 1.05 1.95

1

2

3

7

89

NUCLEOSHELL

Ascentis

Kinetex

Poroshell

6

5

4

0 2 4 6 8 10 12 14 16 18 min

1

2

3

4

0 1 32 min

Peaks:1. Protriptyline2. Desipramine3. Maprotiline4. Nortriptyline5. Doxepin6. Imipramine7. Amitriptyline8. Clomipramine9. Trimipramine

MN Appl. No. 124960

NUCLEOSHELL RP 18 combines innovative silica technology and excellent surface de-activation, that outperforms conventional C18 silicas in terms of efficiency, resolution and speed. Due to the applied core-shell particle design the back pressure at elevated flow rates remains at a moderate level and permits the use of existing HPLC equipment in many cases. NUCLEOSHELL RP 18 with ex-tended pH stability, low bleed characteristics in LC/MS applications and overall robust-ness is an ideal tool for method development and routine analysis in modern HPLC.

www.mn-net.com8

NUCLEOSHELL HILIC

NPC IC

HILIC

RPC

eluent

analyte

adso

rben

t

Key features: · Based on core-shell particle

technologyforfastandefficientHPLC ·Idealforreproducibleandstable

chromatographyofhighlypolaranalytes

·Veryshortcolumnequilibrationtimes Technical characteristics:

Ammonium-sulfonicacidmodifiedsilica;poresize90Å, particlesize2.7μm; carboncontent1.3%; pHstability2–8.5;suitableforLC/MS

Recommended application: Hydrophiliccompoundssuchas

organic polar acids and bases, polar natural compounds, nucleosides, oligonucleotides, amino acids, peptides, water soluble vitamins

NUCLEOSHELL HILICHydrophilic interaction chromatography (HILIC)is a separation technique using polar stationaryphases and organic-aqueous mobile phases. Aminimumwater content of at least 2% is indis-pensabletoprovideapermanentwaterlayerbe-tween the adsorbent surface and the organic frac-tionof themobile phase. The samplemoleculesbecomeseparatedinapartitionchromatography,inwhichpolaranalytesaremorestronglyretainedthanneutral,lesshydrophiliccompounds.Conse-quently,increasingtheaqueouspartinthemobilephase will diminish retention of the polar sample constituents.InthiswayHILICbehavesinversetoclassicalRPchromatography.Theparticularreten-tionprofileofHILICenablesthechromatographyof very polar and often small molecules, whichwon’tshowanyretentiononC8orC18reversedphases.

Ultra-fast separations at moderate back pressureNUCLEOSHELL HILIC is a core-shell technologybasedstationaryphasewithacovalentlybonded3-N,N-dimethylaminopropanesulfonicacidligand(pat.pend.).Thebetaine characterof the strongion-exchanger results in full charge balancing and facilitatesfastequilibrationtimes.

+ N

CH3

CH3

SiO

2

SO3 –

Good separation of polar compounds like the physiologicallyimportantsubstancescreatineandcreatininecanbeachievedonNUCLEOSHELLHILICaswellasonNUCLEODUR®HILIC,1.8µmatsimi-lar retention, but much lower back pressure.

Separation of creatine and creatinineColumns: 50 x 4 mm NUCLEOSHELL HILIC, 2.7 μm

50 x 4 mm NUCLEODUR® HILIC, 1.8 μmEluent: acetonitrile – 10 mM ammonium acetate

pH 4.0 (90:10, v/v)Flow rate: 1.7 mL/minPressure: 129 bar

180 barTemperature: 25 °CDetection: UV, 210 nm Peaks:1. Creatinine2. Creatine

1

2

0.0 0.2 0.4 0.6 0.8 1.0min

MN Appl. No. 124990

9www.mn-net.com

NUCLEOSHELLSeparation of catecholamines

Columns: 100 x 4 mm NUCLEOSHELL HILIC, 2.7 μm 100 x 4 mm NUCLEOSHELL HILIC, 2.7 μm 250 x 4 mm NUCLEODUR® HILIC, 3 μm

Eluent: acetonitrile – 100 mM ammonium formate pH 3.2 (80:20, v/v)Flow rate: 4 mL/min, 1 mL/min, 1 mL/minPressure: 395 bar, 95 bar, 116 barTemperature: 25 °CDetection: UV, 280 nmPeaks:1. DOPAC2. Serotonin3. Dopamine4. Epinephrine5. Norepinephrine6. DOPA7. DOPS

0 5 10 min

1

2

3

4

5

6

7

12 min12 mL (solvent

consumptionper run)

1

2

3

4

56

7

4.5 min4.5 mL

1

1 min4 mL

core-shell {fully porous

2

3

4

56

7

MN Appl. No. 125440

Thechromatogramsshowthemethodtransferfromafullyporous3µmHILICphaseto2.7µmcore-shellsilicawithequalselectivityfeatures.Runtimehasbeencutdownto1min.Columnbackpressureremainsmodest<400bar,whilesolventdemandisreducedtolessthan35%.

NUCLEOSHELL HILIC provides stable and reproducible chromatogra-phy, comprising all the benefits of a state-of-the-art core-shell silica.

Core-shell silica:

separation in 1 min

pressure < 400 bar

www.mn-net.com10

ApplicationsSeparation of ketones

Columns: 50 x 3 mm NUCLEOSHELL RP 18, 2.7 μm 50 x 2 mm NUCLEODUR® C18 Gravity, 1.8 μm 125 x 2 mm NUCLEODUR® C18 Gravity, 5 μm

Eluent: acetonitrile – water (60:40, v/v)Flow rate: 4 mL/min, 1.25 mL/min, 0.33 mL/min Pressure: 540 bar, 774 bar, 89 barTemperature: 25 °CDetection: UV, 254 nm Injection: 1 μL, 1 mg/mL of each compound in eluentPeaks:1. Acetophenone2. Eugenol3. Propiophenone4. Butyrophenone5. Benzophenone6. Valerophenone

0 1 2 3 min

1

2

3

45 6

12

3

4

56

12

3

45

6

1

2

3

4

56

1

2

3

45

6

NUCLEOSHELL

0.0 0.2 0.4 min

NUCLEODUR®

1,8 μm

0,0 0,2 0,4 min

NUCLEODUR®

1.8 μm

MN Appl. No. 124920

Tricyclic antidepressantsColumns: 50 x 4.6 mm NUCLEOSHELL RP 18, 2.7 μm

250 x 4.6 mm fully porous C18, 5 μmEluent: methanol – acetonitrile – 25 mM KH2PO4 pH 7

(22.5:22.5:55, v/v)Flow rate: 2 mL/min, 1.3 mL/minPressure: 224 bar, 190 barTemp.: 40 °CDetection: UV, 220 nm

0 50 100 150 min

1

2

3

4 5

6 78

9

0 5 10 15 min

1

18 min36 mL

180 min234 mL

2

3

4

5

6 78

9

20

Peaks:1. Protriptyline2. Desipramine3. Maprotiline4. Nortriptyline5. Doxepin6. Imipramine7. Amitriptyline8. Clomipramine9. Trimipramine

MN Appl. No. 125420

time sav

ing

90 %

eluent s

aving

85 %

11www.mn-net.com

NUCLEOSHELLAcidic pharmaceuticals

Columns: 50 x 4 mm NUCLEOSHELL RP 18, 2.7 μm 150 x 4 mm fully porous C18, 5 μm

Eluent: A) acetonitrile, B) 25 mM KH2PO4 pH 7, 25–40 % A in 2.2 min, 25–40 % A in 20 min

Flow rate: 1.5 mL/min, 0.5 mL/minPressure: 219 bar, 92 barTemp.: 20 °CDetection: UV, 215 nm Peaks:1. Ketoprofen2. Fenoprop3. Fenoprofen4. Flurbiprofen

5. Ibuprofen7. Carprofen8. Diclofenac9. Meclofenamic acid

0.0 0.5 1.0 1.5 2.0 min

12

34

5

6

7

8

2.2 min3.3 mLRs (2, 3) = 1.69

20 min10 mLRs (2, 3) = 1.70

0 5 10 15 min

1

2

34

5

6

7

8

MN Appl. No. 125430

Separation of steroidsColumns: 50 x 3 mm NUCLEOSHELL RP 18, 2.7 μm

125 x 3 mm NUCLEODUR® C18 Gravity, 3 μmEluent: A) acetonitrile, B) water

30–80 % A in 1 min (0.4 min 80 % A) 30–80 % A in 5 min (2 min 80 % A)

up to 90 % time saving

66 % solvent saving

Flow rate: 2 mL/min 1 mL/min

Pressure: 350 bar 280 bar

Temperature: 25 °C Detection: UV, 240 nmInjection: 1 μL, 1 mg/mL of each compound in eluent

0 21 3 4 5 min

32

1 4

5

6

7

8 93

4

5

61.4 min2.8 mL

7 min7 mL

2

1 7

8 9

Peaks:1. Estriol2. Prednisolone3. Cortisone4. Estradiol5. Testosterone6. Estrone7. 6a-Methyl-11b-hydroxyprogesterone8. 6a-Methyl-17a-hydroxyprogesterone9. Progesterone

MN Appl. No. 124930

www.mn-net.com12

ApplicationsNon-steroidal anti-inflammatory drugs

Columns: 50 x 4.6 mm NUCLEOSHELL RP 18, 2.7 μm Ascentis® Express C18

Eluent: acetonitrile – 20 mM KH2PO4 pH 2.5 (40:60, v/v)

Flow rate: 2.5 mL/minPressure: 268 bar, 281 barTemperature: 22 °CDetection: UV, 230 nmInjection: 1 μL, 1 mg/mL of each compound in eluent

0 1 2 3 min

3

1

2

6

45 8

Rs = 1.85 Rs = 1.59

9 10 1112

7

Rs = 1.25 Rs = 1.14

Peaks:1. Acetylsalicylic acid2. Sulindac3. Piroxicam4. Suprofen5. Tolmetin6. Naproxen7. Diflunisal8. Fenoprofen9. Flurbiprofen10. Niflumic acid11. Indomethacin12. Ibuprofen

MN Appl. No. 124970

Water-soluble vitaminsColumn: 100 x 4 mm NUCLEOSHELL HILIC, 2.7 μmEluent: A) acetonitrile – 100 mM ammonium acetate

pH 3.2 (90:10, v/v), B) water; 4 % B (1 min) → 20 % B in 1.6 min (0.7 min 20 % B)

Flow rate: 2 mL/minPressure: 218 barTemperature: 25 °CDetection: UV, 260 nm Peaks:1. PABA (p-aminobenzoic acid)2. Nicotinamide3. Vitamin B6 (pyridoxine)4. Riboflavin5. Nicotinic acid6. Vitamin C (ascorbic acid)7. Vitamin B1 (thiamine)8. Folic acid9. Vitamin B12 (cyanocobalamine)

0 1 2 3 min

12

3 4

5

67

8

9

MN Appl. No. 125450

Anions and cationsColumn: 100 x 4 mm NUCLEOSHELL HILIC, 2.7 μmEluent: A) 30 mM ammonium formate pH 3,

B) acetonitrile 80 % B (3 min) → 20 % B in 7 min

Flow rate: 1.5 mL/minPressure: 200 barTemperature: 40 °CDetection: CAD (Nebulizer: 35 °C) Peaks:1. Nitrate2. Chloride3. Potassium4. Sodium5. Phosphate6. Sulfate

0 1 2 3 4 5 6 min

1

3

4

5

6

MN Appl. No. 125460

good

selectiv

ity

and re

solution

13www.mn-net.com

NUCLEOSHELLMetformin

Columns: 50 x 4 mm NUCLEOSHELL HILIC, 2.7 μm 50 x 4 mm NUCLEODUR® HILIC, 1.8 μm

Eluent: acetonitrile – 10 mM ammonium acetate pH 3.2 (75:25, v/v)

Flow rate: 3 mL/min 1.5 mL/min

Pressure: 202 bar 167 bar

Temperature: 25 °CDetection: UV, 218 nm

0.0 0.5 1.0 min

1

1

2

2

NUCLEOSHELL

NUCLEODUR®

Peaks:1. Dicyandiamide2. Metformin

MN Appl. No. 125470

5-FluorouracilColumn: 50 x 4 mm NUCLEOSHELL HILIC, 2.7 μmEluent: acetonitrile – 10 mM ammonium acetate

(95:5, v/v)Flow rate: 2.5 mL/minPressure: 119 barTemperature: 25 °CDetection: UV, 254 nmPeaks:1. 5-Fluorouracil2. Uracil

0.0 0.2 0.4 min

1

2

MN Appl. No. 125480

Analysis of an energy drinkColumn: 100 x 4 mm NUCLEOSHELL HILIC, 2.7 μmEluent: acetonitrile – 100 mM ammonium acetate

pH 5.0 (90:10, v/v)Flow rate: 1.7 mL/minPressure: 126 barTemperature: 35 °CDetection: UV, 254 nm

1

2

34

5

6

0 1 min

Peaks:1. Caffeine2. Niacinamide3. Pyridoxine4. Benzoic acid5. Sorbic acid6. Riboflavin

MN Appl. No. 125010

Amino acidsColumns: 50 x 4 mm NUCLEOSHELL HILIC, 2.7 μmEluent: acetonitrile – 100 mM ammonium acetate pH 4.0 (80:20, v/v)Flow rate: 1.5 mL/minPressure: 105 barTemperature: 25 °CDetection: UV, 215 nm Peaks:1. Phenylalanine2. Phenylglycine3. Tyrosine4. Histamine

MN Appl. No. 125000

4

3

2

1

0.0 0.4 0.8 1.2 1.6 2.0 min

www.mn-net.com14

Packed columns · Ordering informationEC standard columns for analytical HPLC

Analyticalcolumnsystemmadeofstainless steel

M8outerthreadsonbothends

Combination of sealing element and veryfine-meshedstainlesssteelscreen,PTFEringandfittingadapter

ColumnheadsSW12,withinnerthreadsM8x0.75andUNF10-32(=1/16”fitting)

Asscrew-onguardcolumnsystemwerec-ommend the Column Protection System used with EC guard column cartridges with 4mmlength(seenextpage).

NUCLEOSHELL RP 18 Poresize90Å;octadecylmodification,multi-endcapped,7.5%C;eluentincolumnCH3CN – H2O

Length→ 50 mm 100mm 150mm EC guard columns*

NUCLEOSHELL RP 18, 2.7 µm particlesize2.7µmECanalyticalcolumns2mmID 763132.20 763134.20 763136.20 4x2mm:763138.203mmID 763132.30 763134.30 763136.30 4x3mm:763138.304mmID 763132.40 763134.40 763136.40 4x3mm:763138.304.6mmID 763132.46 763134.46 763136.46 4x3mm:763138.30

ECcolumnsinpacksof1,guardcolumnsinpacksof3

NUCLEOSHELL HILIC Poresize90Å;ammonium–sulfonicacidmodification,endcapped,1.3%C;eluentincolumnCH3CN – H2O

Length→ 50 mm 100mm 150mm EC guard columns*

NUCLEOSHELL HILIC, 2.7 µm particlesize2.7µmECanalyticalcolumns2mmID 763332.20 763334.20 763336.20 4x2mm:763338.203mmID 763332.30 763334.30 763336.30 4x3mm:763338.304mmID 763332.40 763334.40 763336.40 4x3mm:763338.304.6mmID 763332.46 763334.46 763336.46 4x3mm:763338.30

ECcolumnsinpacksof1,guardcolumnsinpacksof3

*ECguardcolumnsrequiretheColumnProtectionSystemCartridgeHolder(seeright)

15www.mn-net.com

NUCLEOSHELLColumn Protection SystemInnovative and universal screw-on guard column holder systemSuitable for all analytical HPLC columns with 1/16” fittings

CartridgesfilledwithspecifiedNUCLEOSHELL,NUCLEODUR®, and NUCLEOSIL® HPLC adsorbents

Idealprotectionforyouranalyticalmaincolumn →significantincreaseincolumnlifetime

Minimized void volume → suitable also for ultra fast HPLC

Special ferrules →pressurestabilityupto1034bar(15000psi)

Visualcontaminationcheck→ in-time changing of the guard column

Guardcolumnlength4mm,ID2mm(formaincolumnswith2mmID)orID3mm(formaincolumnswith3,4and4.6mmID)

UNIVERSALRPguardcolumnsavailableforall HPLC columns under RP conditions

Content of the Column Protection System

Description REF

Column Protection System 718966

Details Content

Cartridge Holder 1

Capillaries 2

Ferrules 3

Wrenches 2

Manual 1

Replacement parts for the Column Protection System • Ordering information

Description Pack of REF

Ferrules 5 718967Replacement connector including O-ring 1 718968Capillarytubes,nutsandmetalferrules 3 718969Wrench(size12and14mm) 1 718970EC4/2UNIVERSALRPguardcolumn(formaincolumnswith2mmID) 3 728777.20EC4/3UNIVERSALRPguardcolumn(formaincolumnswith3,4and4.6mmID) 3 728777.30

Visual Contamination Check

The cartridge is assembled with a white filter membrane.

Adiscolorationofthefiltermembraneusuallyindicatesthatthecartridgeshouldbe replaced.

Incaseofcolorlesscontaminants,arisingbackpressureand/orlossofchro-matographic performance advise to change the guard column.

Trademarks:NUCLEODUR®(MACHEREY-NAGELGmbH&Co.KG,Germany),Ascentis®(Sigma-AldrichCo.,USA),Kinetex™(PhenomenexInc.,USA),Poroshell(AgilentTechnologiesInc.,USA) Imagecredits:©by-studio/Jaeeho(Fotolia.com)

www.mn-net.com

www.mn-net.comMACHEREY-NAGEL GmbH & Co. KG · Neumann-Neander-Str. 6–8 · 52355 Düren · GermanyFrance:MACHEREY-NAGEL EURLTel.: +33 388 68 22 68Fax: +33 388 51 76 88E-mail: sales-fr@mn-net.com

Switzerland:MACHEREY-NAGEL AGTel.: +41 62 388 55 00Fax: +41 62 388 55 05E-mail: sales-ch@mn-net.com

Germanyand international:Tel.: +49 24 21 969-0Fax: +49 24 21 969-199E-mail: info@mn-net.com

MACHEREY-NAGEL

EN ISO 9001: 2008CERTIFIED

USA:MACHEREY-NAGEL Inc.Tel.: +1 484 821 0984Fax: +1 484 821 1272E-mail: sales-us@mn-net.com

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