1.7μm Fortis UHPLC Columns - dichrom

Strength in Technology1.7μm FortisUHPLC ColumnsUltra High PressureChromatographyImproveyourUHPLCPerformance8 Chemistry ChoicesIncrease EfficiencyIncrease SpeedImprove ResolutionGreater SensitivityLower Backpressure

UHPLC Columns– Optimised for Ultra High Pressure LC (UHPLC)– Available in 8 different Selectivities– Operate at 18,000psi– Fully Scalable analytical to prepSpeedResolutionSelectivitySensitivity

1.7μm Phase Chemistry Selectivity1.7μm Fortis C18– General UHPLC use– Method Development from pH 1-12AcidsBasesNeutralsUHPLC1.7μm Fortis H2o– Polar endcapped– Increased polar retentionHydrophilic analytesOrganic acidsCatecholamines1.7μm Fortis Diphenyl– Unique di-phenyl structure– Metabolite profiling– Separate positional isomersMetabolitesPositional IsomersHydrophilic / Hydrophobic analytes1.7μm Fortis C8– General UHPLC use– Method DevelopmentLipidsSteroidsHighly Hydrophobic analytes1.7μm Fortis HILIC– High polar retention– Homogenous silanol concentration– Improve MS sensitivityCarboxylic acidsNucleotidesVitamins1.7μm Fortis HILIC Diol– Alternate selectivity to bare silica– Stable bonding– HILIC or Normal phase modeSteroidsProteinsMetabolites1.7μm Fortis Cyano– Cyano functionality– Reversed phase or Normal phaseExplosivesPesticidesSteroids1.7μm Fortis Amino– Reproducible, Robust bonding– Reversed phase, Normal phase orIon exchange modeSaccaridesOligonucleotidesSteroids

UHPLC1.7μm UHPLC Columns- 380m 2 /g Surface Area Provides Increased Peak Capacity– Available in 8 Phase Chemistries– Operate to 18,000psi– Fully Scalable to Analytical and Prep Size1.7μm Fortis particles are designed to provide characteristics, which will aid in increased productivity within ultra high pressurechromatography (UHPLC). Designed to be robust, reproducible and fully scalable with 3μm, 5μm and 10μm particles. 1.7μm Fortis particleswill operate upto 1200bar providing increased efficiency at high linear velocities, whilst allowing speed and sensitivity to be achieved on allthe latest UHPLC systems. By choosing a high surface area UHPLC phase the analyst can increase peak capacity using their existing columndimension, or maintain existing capacity whilst lowering backpressure on a shorter column.High Efficiency with Lower Backpressure1.7μm Fortis C18 provides increased efficiencyover 3μm and 5μm particles. This provides theopportunity to increase resolution or speed ofanalysis.– Higher EfficiencyCompare 1.7μm Fortis C18 with your existingcolumn to see high retention, high efficiency.4-EthylanilineN,N-DimethylanilineNapthalene1.7μm Fortis C1830x2.1mm134 bar– Lower BackpressureUse the high surface area to lower yourcolumn length and reduce the backpressurein the system.4-EthylanilineN,N-DimethylanilineNapthaleneAcquity ® BEH C18 1.7μm50x2.1mm315 bar0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0MinutesComparison of Hydrophobicity and Peak Shape1.7μm Fortis C18 50x2.1mmSurface Area 380m 2 /g1.7μm Fortis C18380m 2 /gEfficiency 191,670Peak Shape(N,N-Dimethylaniline)1.03Psi - 0.4ml/min(60:40 ACN:Water)225bar1.7μm BEH C18185m 2 /gAcquity ® BEH 1.7μm C18 50x2.1mmSurface Area 185m 2 /gEfficiency 167,400Peak Shape(N,N-Dimethylaniline)1.28 Psi - 0.4ml/min(60:40 ACN:Water)292barXbridge ® , Acquity BEH ® and UPLC ® , are registered trademarks of Waters Corporation. Fortis is not associated with these companies. Comparative separations/resultsmay not be representative of all applications. All columns are original manufacturers own.

ComparisonPressure Comparison 1.7μm Fortis C18 vs 1.7μm Acquity ® BEH C18UHPLCHigh surface area of the silica 1.7μmFortis C18 gives you the choice to lowerbackpressure:– Lower Backpressure1.7μm Fortis C18 provides less backpressurethen many of the other UHPLC columnsavailable– Shorten ColumnIf you can then use a shorter column and stillhave the same retention due to the highersurface area, you will reduce the pressureeven further with no loss of separation.Pressure (bar)400350300250200150100501.7μm BEH C1850x2.1mm1.7μm Fortis C1850x2.1mm1.7μm Fortis C1830x2.1mm05 20 40 60 80 100% OrganicComparison of UHPLC Columns - Sharper Peak Shapes1.7μm Acquity ® BEH C18 50x2.1mm1.8μm Epic ® C18 50x2.1mm1. Sulfanilamide 2. Sulfamerazine 3. Sulfamethoxaxole 4. Hydrocortisone 5. KetoprofenData Courtesy of: GNF, San Diego1.7μm Fortis ® C18 30x2.1mmBest Peak Shape andXbridge ® , Acquity BEH ® and UPLC ® , are registered trademarks of Waters Corporation. Epic C18 is a registered trademark of ES Industries. Fortis is not associatedwith these companies. Comparative separations/results may not be representative of all applications. All columns are original manufacturers own.

UHPLCHigh Performance– Use 1.7μm Fortis particles as a traditional UHPLC column– Use 1.7μm Fortis particles in place of core-shell– Fully pH stable 1-12- Fully Scalable to analytical and Preparative scale- Can be used on traditional HPLC Instruments**1.7μm Fortis UHPLC columns can be used in UHPLC systems or in ‘standard 400-600bar systems’ to produce ultra-high pressure or ultrahighperformance chromatography. If you use a high surface area stationary phase (Fortis = 380m 2 /g) then in comparison with smallersurface area phases, such as hybrid’s and core-shell, you will gain distinct advantages:+ 380m 2 /g +30x2.1mm65 bar= N 182kpH 1-121.7μm Fortis 1.7um HybridUHPLC particle+ 185m 2 /g +50x2.1mm160 bar= N 167kpH 1-12+ 200m 2 /g +50x2.1mm80 bar= N 178kpH 2-8 **Core-Shell* pH range for gradient** Column length and optimisation required

0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00MinutesMinutes0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.000.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00 0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00MinutesMinutesComparison1.7μm Fortis C18 vs 1.7μm Acquity ® BEH C18UHPLCHigh surface area of the silica 1.7μmFortis C18 gives you the choice to lowerbackpressure or increase Retention/Resolution:OPTION 1– Increase Peak Capacity1.7μm Fortis C18 high surface area willincrease retention over the same lengthAcquity ® BEH column leading to moreavailable resolution.OPTION 11.7μm Waters BEH ® C181.145mins315bar50x2.1mmOPTION 2OPTION 2– Reduce Backpressure1.7μm Fortis C18 high surface area meansthat you can use a shorter column tomaintain the same retention as an Acquity ®BEH column but reduce backpressure evenfurther1.7μm Fortis C181.95mins230bar50x2.1mmFor longer retention, use same column length= High peak capacity / resolution1.7μm Fortis C181.15mins139bar30x2.1mmFor same retention, use shorter column length= Less backpressure1.7μm Fortis C18 vs 2.6μm Kinetex ® C18High surface area of the silica 1.7μmFortis C18 gives you the choice to lowerbackpressure or increase Retention/Resolutionover core-shell particles:2.6μm Kinetex ® C1850x3.0mm0.8ml/minOPTION 1– Increase Peak Capacity1.7μm Fortis C18 high surface area willincrease retention over the equivalent lengthKinetex ® C18 column, leading to moreresolution.OPTION 2– Reduce Backpressure1.7μm Fortis C18 high surface area meansthat you can use a shorter column to maintainthe same retention as a Kinetex ® C18 columnbut reduce backpressureOPTION 11.7μm Fortis C18227barN=179,000175barN=169,00050x3.0mm0.8ml/minOPTION 21.7μm Fortis C18145barN=166,00030x3.0mm0.8ml/min– Other Gains- 1.7μm Fortis C18 will operate at extendedpH ranges over core-shell.- 1.7μm Fortis C18 can be scaled to analyticaland preparative scale unlike core-shell.For longer retention, use same column length= High peak capacity / resolutionFor same retention and performance, useshorter column length= Less backpressureXbridge ® , Acquity BEH ® and UPLC ® , are registered trademarks of Waters Corporation. Kinetex ® is a registered trademark of Phenomenex. Fortis is not associatedwith these companies. Comparative separations/results may not be representative of all applications. All columns are original manufacturers own.

UHPLC1.7μm Fortis C18 pH options– pH selectivity for method development– pH stable 1-12– Gives high speed of equilibration1.7μm Fortis C18 will operate across the pH spectrum giving the analyst the ability to optimise the correct pH region for their separation.Quickly equilibrating from formic acid to ammonium acetate through to ammonia allows pH, as a method variable, to be rapidly evaluated.Resolution of compounds can be changed radically by altering pH to optimise separation between compound classes.34Column: 1.7μm Fortis C18 30x2.1mmp/n: F18-020201Gradient: 10 - 50% in 5minFlow:0.4ml/minTemp: 20°CWavelength: 254nm1. Uracil2. Procaine3. Fenuron4. 3-Nitrobenzoic acid12pH 2.2431AcidicBasic2NeutralpH 7.2432pH 11.21

Critical Considerations in UHPLCFully ScalableAll Fortis phases can be scaled from 1.7μmall the way through analytical 3μm and 5μmparticles to prep size without any change inretention profile.1.7μm Fortis C18N = 191,670UHPLC– Improve transfer of methodsBy combining the same surface area, pore sizecharacteristics with the identical bonding youhave the ability to scale up or down, also theconfidence to transfer to another laboratorywithout change in selectivity– IssuesIf a small particle used in UHPLC is not thesame as its larger 3μm and 5μm particle thenchanges in resolution and retention can occur,neither of which are acceptable in methodvalidation.1.7um Fortis C18 will alleviate allthese potential issues, leaving theanalyst confident in method transfer.3μm Fortis C18N = 130,1915μm Fortis C18N = 88,7080 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0MinutesHPLC_Acquity_21nov_011AU1.751.51.251.07.5e-15.0e-12.5e-10.0-2.5e-1-5.0e-10.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40HPLC_X Brige_21nov_0112: Diode Array2.90Range: 1.781.25Competitor1.7μm Hybrid C180.240.731.07Competitor3.5μm Hybrid C181.481.622.022.122.242.332.572.762.802: Diode ArrayRange: 2.1541.0AU7.5e-15.0e-12.5e-10.250.811.191.602.362.252.452.192.692.940.01.73-2.5e-1-5.0e-1Major Pharmaceutical Company DataTime0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40Sensitivity GainsPeak height increases in UHPLC due to therise in efficiency (N) from the smaller particle,but it is also inversely proportional to peakwidth, so symmetrical peaks will lead toincreased sensitivity.– Sharp Peak ShapesAll Fortis phases are designed to give thesharpest possible peak shapes.– High EfficiencyMoving from 3μm to 1.7μm Fortis C18gives a peak height increase of 27% in thisexample. The increase from 5um particles is0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0Minutes- All columns 50x2.1mm

UHPLCUHPLC Method Development1.7μm Fortis columns will allow the transfer of methods from traditional HPLC to UHPLC, saving both time and solvent. In order to performmethod transfer there are several ‘method development’ calculators available to help in making appropriate changes to column dimension,flow rate and gradient conditions. If done properly the overall method time will reduce but resolution and selectivity of solutes will remainconstant or indeed improve. Download at : www.uhplccolumns.com/UHPLC_CalculatorEquivalent UHPLC Column - ‘Separating Power’First consideration is the ability to scale themethod down in column dimension, lengthand diameter:– Equivalent UHPLC columnIf you can retain equivalent column platecount or ‘separating power’ then it is mucheasier to scale down effectively.– ExampleIf you move from a 5μm 150x4.6mm to a1.7μm 50x2.1mm the equivalent separationshould be achieved but a several foldimprovement in analysis time will beachievedColumnLengthEfficiency of5μm250 22,000Efficiency of3μmEfficiency of1.7μm150 12,700 16,800 26,460100 8,300 10,700 21,00050 4,000 6,000 11,20030 3,200 7,00020 3,000Method Development CalculatorDOWNLOAD AT:www.uhplccolumns.com/UHPLC_Calculator

UHPLC Method DevelopmentScaling a Method - IsocraticTo scale to a UHPLC column first we changeflow rate and injection volume in order tomaintain the linear velocity across the methodand not overload the column– Change Flow rateRs = 12.745μm 150x4.6mm1.0 ml/minUHPLCF 2= F 1x (Dc 2/ Dc 1) 2HPLC 20mins– Change Injection VolumeRs=6.732V 2= V 1x (Dc 2x L 2)2( Dc 1x L 1)F 2= New flow rateF 1= Original flow rateDc 2= New column DiameterDc 1= Original column DiameterL 2= Length of new columnL 1= Length of original columnV 2= New injection VolumeV 1= Original injection Volume0.0010.00 20.003.4 6.8UHPLC 2minsRs = 12.133.4 6.81.7μm 50x2.1mm0.2 ml/minUHPLC 6.8mins1.7μm 50x2.1mm0.4 ml/minScaling a Method - GradientIn order to change our gradient we must aimto keep the slope and the start point the samebut lower the time the gradient runs in.Altering the gradient time, retains the samelinear gradient and slope, but reduces the runtime:5μm 150x4.6mm Fortis C181.0 ml/minHPLC 20mins– Change Gradienttg 2= tg 1x (F 1/ F 2) x (Dc 22/ Dc 12) x (L 1/ L 2)10.0020.001.7μm 50x2.1mm Fortis C180.2 ml/minUHPLC 6.8minstg 2= New Gradient timetg 1= Original Gradient time0.003.4 6.8

100.0095.0090.0085.0080.0075.0070.0065.0060.0055.0050.000.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00Minutes88.0086.0084.0082.0080.0078.0076.0074.0072.0070.0068.0066.0064.0062.0060.0058.0056.0054.0052.0050.000.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00Minu tesSATIN120.00115.00110.00105.00100.0095.0090.0085.0080.0075.0070.0065.0060.0055.0050.0045.0040.0035.0030.00SATIN0.00 0.50 1.00 1.50 2. 00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00MinutesSATINUHPLCUHPLC Method DevelopmentResolution vs Efficiency vs Selectivity1.7μm Fortis C18 will provide hydrophobicselectivity which is suitable for manycompounds. However as the resolutionequation shows us having multiple phasechemistries available is a definite advantageeven in UHPLC. Selectivity can then be usedin conjunction with higher efficiency.1.7μm Fortis UHPLC columns are alsoavailable as:– 1.7μm Fortis Diphenyl– 1.7μm Fortis H2o (Polar C18)– 1.7μm Fortis C8– 1.7μm Fortis Cyano– 1.7μm Fortis HILIC– 1.7μm Fortis HILIC DIOL– 1.7μm Fortis AminoN • 3.02.52.01.51.00.5Nk0.01.00 1.05 1.10 1.15 1.20 1.250 5000 10000 15000 20000 250000 5 10 15 20 25NkImprove SelectivityIf we are scaling a method and hoping that anincrease in efficiency alone will provide thenecessary resolution we can be disappointed.3μm Fortis C18150x2.1mm– Efficiency AloneScaling from 3μm to 1.7μm C18 has notprovided baseline resolution between thecompounds.mV– Efficiency & SelectivityAdding selectivity by choosing an alternativephase chemistry has allowed us to go fasteron a shorter column and now achieve fullbaseline separation.EfficiencyAlone1.7μm Fortis C18150x2.1mmEfficiency +Selectivity1.7μm Fortis Diphenyl100x2.1mm– ConclusionIn this instance 1.7μm Fortis Diphenyl providesmore resolution than C18. This then leads tothe ability to increase speed by use of shortercolumns.mV4minsmV3.5minsIsonicotinamide and Nicotinamide

UHPLC Sample Filter– Low volume in-line filter for all UHPLC columns– No backpressure increase– Increase lifetime of UHPLC columns– Change over time seconds not minutesUHPLCColumn Protection - No lose in performanceFortis UHPLC in-line filters are direct connectdesign, fitting in between the UHPLC columnand the conventional system fitting to filter outparticulate matter.They contain low dead volume and pressure.In-line filters are ideal for 1.7μm Fortis UHPLCcolumns where extra packed bed from a guardwould be detrimental.Without In-line FilterRt - 3.81 minN - 174,000 plates/meterWith In-line FilterRt - 3.83 minN - 172,000 plates/meterUHPLC in-line filters are manufactured towithstand 20,000psi.– In-line Filter or Guard cartridgeIn-line filters are more suitable to many instances0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00MinutesColumn Protection - No loss in performanceUHPLC column fitting is of crucial importance,since the addition of the smallest “dead” or voidvolume to these new low volume UHPLC systemswill severely impact upon the performance of thecolumn.UHPLC GuardUHPLC trFortis UHPLC Fitr– In-line Filter or Guard cartridgeIn-line filters are more suitable to many instancesof UHPLC since with very short run times guardcolumns will add retention that is not required.Guards can also reduce the efficiency of thesystem.– In-line Filter comparisonIn-line filters are not all the same, both efficiencyand peak shape can be affected by a filter thatis not optimal.Fortis In-line filters are optimised forUHPLC.UHPLC In-line FiltersUHPSAV2 UHPLC In-line filter pk 2UHPSAV4 UHPLC In-line filter pk 4

UHPLCUHPLC Fittings– Perfect fit every time– No dead volume– No tools required– Change over time seconds not minutesFortis UHPLC fittings are designed to offer the perfect fit for all UHPLC columns. Quickly change the ferruledepth to adapt to any column. Hand-tight fitting requires no tools. Fitting is ideal for 1.7μm Fortis UHPLCcolumns as they are manufactured to withstand 20,000psi.Correct FittingUHPLC column fitting is of crucial importance,since the addition of the smallest “dead” or voidvolume to these new low volume UHPLC systemswill severely impact upon the performance of thecolumn. Even the smallest 1mm void producedfrom fitting the column can lead to a sharpdecrease in efficiency and peak shape from whatshould be achieved.420.00400.00380.00360.00340.00320.00300.001-2mm Void180.00170.00160.00150.00140.00No Void - Perfect Fit- Adjustable fittingsStainless steel fittings have been widely adapteddue to the high pressures involved, however ifthe ferrule is also stainless steel and immovableonce in place then this can create a void whenswitching between different manufacturerscolumns.A fully adjustable UHPLC fittingshould always be adapted in order toensure that the fitting of the columnis perfect every single time regardlessof UHPLC hardware.280.00260.00240.00220.00200.00180.00160.00140.00120.00100.0080.0060.0040.00Asy = 1.32N = 165,960600.00 1.00 2.00 3.00 4.00130.00120.00110.00100.0090.0080.0070.0060.0050.00Asy = 1.06N = 1 9, 000.00 1.00 2.00 3.00 4.00UHPLC FittingsUHPFIT-2 UHPLC Fitting pk 2UHPFIT-4 UHPLC Fitting pk 4

1.7μm UHPLC part numbersFortis C18Column Length20 30 50 100 1502.1 F18-020101 F18-020201 F18-020301 F18-020501 F18-020701UHPLCColumn Diameter 3.0 - F18-030201 F18-030301 F18-030501 -4.6 - F18-050201 F18-050301 F18-050501 -Fortis DiphenylColumn Length20 30 50 100 1502.1 FPH-020101 FPH-020201 FPH-020301 FPH-020501 FPH-020701Column Diameter 3.0 - FPH-030201 FPH-030301 FPH-030501 -4.6 - FPH-050201 FPH-050301 FPH-050501 -Fortis H2o(Polar Endcapped C18)Column Length20 30 50 100 1502.1 FHO-020101 FHO-020201 FHO-020301 FHO-020501 FHO-020701Column Diameter 3.0 - FHO-030201 FHO-030301 FHO-030501 -4.6 - FHO-050201 FHO-050301 FHO-050501 -Fortis C8Column Length20 30 50 100 1502.1 F08-020101 F08-020201 F08-020301 F08-020501 F08-020701Column Diameter 3.0 - F08-030201 F08-030301 F08-030501 -4.6 - F08-050201 F08-050301 F08-050501 -Fortis CyanoColumn Length20 30 50 100 1502.1 FCN-020101 FCN-020201 FCN-020301 FCN-020501 FCN-020701Column Diameter 3.0 - FCN-030201 FCN-030301 FCN-030501 -4.6 - FCN-050201 FCN-050301 FCN-050501 -Fortis HILICColumn Length20 30 50 100 1502.1 FHI-020101 FHI-020201 FHI-020301 FHI-020501 FHI-020701Column Diameter 3.0 - FHI-030201 FHI-030301 FHI-030501 -4.6 - FHI-050201 FHI-050301 FHI-050501 -Fortis HILIC DIOLColumn Length20 30 50 100 1502.1 FDI-020101 FDI-020201 FDI-020301 FDI-020501 FDI-020701Column Diameter 3.0 - FDI-030201 FDI-030301 FDI-030501 -4.6 - FDI-050201 FDI-050301 FDI-050501 -Fortis AminoColumn Length20 30 50 100 1502.1 FNH-020101 FNH-020201 FNH-020301 FNH-020501 FNH-020701Column Diameter 3.0 - FNH-030201 FNH-030301 FNH-030501 -4.6 - FNH-050201 FNH-050301 FNH-050501 -

WORLDWIDE AVAILABILITYFortis products are available worldwide. For the distributorin your country, contact Fortis international Sales Office, UKby telephone, fax or email: info@fortis-technologies.com45 Coalbrookdale RoadClayhill Industrial ParkNestonCheshire, UKCH64 3UGt: +44 151 336 2266f: +44 151 336 2669www.fortis-technologies.come: info@fortis-technologies.com Austria Bangladesh Brazil Canada China Columbia Czech Republic Ecuador Egypt France Germany Greece Holland Hong Kong Hungary India Ireland Israel Italy Japan Korea Malaysia Mexico Netherlands Norway Puerto Rico Poland Portugal Romania Russia Singapore South Africa Spain Sweden Switzerland Taiwan Thailand Turkey USAFor technical support or applications contact :technicalsupport@fortis-technologies.comFor more information VISIT :www.fortis-technologies.comFortis and Fortis Pace are trademarks of Fortis Technologies Ltd.Xbridge ® , Acquity BEH ® and UPLC ® , are registered trademarks of Waters Corporation. Kinetex ® is a registered trademark of Phenomenex. Epic is a trademarkof ES Industries. Fortis is not associated with these companies. Comparative separations/results may not be representative of all applications. All columns areoriginal manufacturers own. ©2012 Fortis Technologies Ltd. All rights reserved.