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Charles L évesque , M.Sc . Product Manager- Analytical Chemistry Vincent B édard , B.Sc . HPLC Specialist François Bé PowerPoint Presentation
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  1. New Generic Approach for HPLC analysis: The Modern Silica-Based HPLC Columns; SiliaChrom®Series Charles Lévesque, M.Sc. Product Manager-AnalyticalChemistry Vincent Bédard, B.Sc. HPLC Specialist François Béland, Ph.D. Vice-President of R&D

  2. Outlines • HPLC Listing of all C18 • SiliaChrom Strategy • SiliaChrom—Chemistry Review • New Techologies-SB,AQ,XT • Evaluation of SiliaChrom • Recent SiliaChrom Applications • Generic Approach for HPLC applications • SiliaChrom Focus

  3. HPLC C18 column listing (L1-USP) • BioBasic 18 • Bio-Sil ODS • Bio-Sil ODS-10 • Bondclone C18 • BR-C18 • Burdick & Jackson C18 • Cadenza CD-C18 • Cadenza 5CD-C18 • Cadenza CL-C18 • Cadenza 5CL-C18 • Cadenza CW-C18 • Cadenza 5CW-C18 • Cadenza HS-C18 • Capcell C18 AG 120 • Capcell C18 SG 120, 300 • Capcell C18 UG 80, 120, 300 • Capcell C18 MG • Capcell C18 MGII • Capcell C18 MGIII • Capcell C18 ACR • Capcell C18 AQ • Capcell C18 IF • Chemcosorb 5-ODS-H • Chromegabond C18 • Chromegabond WR-C18 • Chromolith RP-18e • Clarity Oligo-RP • Clipeus C18 • Cogent HPS C18 • Cogent hQ C18 • Cogent Simulare C18 • Cogent MicroBee C18 • Cogent Bidentate C18 • Cogent e-Column C18 • Cosmosil C18 MS-II • Cosmosil C18 AR-II • Cosmosil C18 AR-300 • Cosmosil C18 PAQ • Alltima C18 • Alltima C18 LL • Alltima HP C18 • Alltima HP C18 EPS • Alltima HP C18 Hi-Load • Alltima HP C18 AQ • Alltima HP C18 Amide • Alltima WP C18 • AlphaBond C18 • Amethyst C18 • AminoQuant • APEX ODS • Apollo C18 • AQUA C18 • Aquapore OD-300 C18 • AQUASIL C18 • Ascentis C18 • Ascentis Express C18 • ASI C18 • ASI microC18 • Atlantis dC18 • Atlantis T3 • Axxi-Chrom C18, ODS • Bakerbond C18 • BetaBasic 18 • BetaMax Neutral • BETASIL C18 • Bio-C18 • Delta-Pak C18 • Denali • Develosil ODS • Diazem 2060 C18 60A • Diazem 300A C18 4-222 • Diazem 4-206 C18 • Diazem 4-230 C18 • Diazem 4-246 C18 • Diazem 4-247 C18EC • Diazem C18 • Diazem C18 1000 • Diazem C18 1120 • Diazem C18 120A • Diazem C18 120A series 43,44,45 and 46 • Diazem C18 series 200A, 300A • Diazem C18 series 2000 • Diazem C18 series 3000 • Diazem C18 series 4000 • Diazem C18 60A • ACE 300 C18 • ACE AQ • ACE C18 • ACE C18-AR • AccQTag Ultra • ACQUITY UPLC BEH130 • ACQUITY UPLC BEH 300 • ACQUITY UPLC BEH C18 • ACQUITY UPLC BEH Shield RP18 • ACQUITY UPLC HSS C18 • ACQUITY UPLC HSS C18 SB • ACQUITY UPLC HSS T3 • ACQUITY UPLC OST C18 • Adsorbosil C18 • Adsorbosphere C18 • Adsorbosphere HS C18 • Adsorbosphere UHS C18 • Adsorbosphere XL 300 C18 • Adsorbosphere XL C18 • Adsorbosphere XL C18-B • Allsphere ODS-1 • Allsphere ODS-2 • Allure C18 • Allure Aqueous C18

  4. HPLC C18 column listing (L1-USP) • Diazem C18 60A series 1,2,3 • Diazem C18 series EC • Diazem C18 Dual-Zone • Discovery C18 • Discovery HS C18 • Discovery Bo Wide Pore C18 • Econosil C18 • Econosphere C18 • Everest C18 • Fortis C18 • Fortis H2o • Gammabond C18 • Gemini C18 • Gemini-NX • Genesis C18 • GP-C18 • GraceSmart C18 • Haisil 100, 300 C18 • Haisil HL C18 • HALO C18 • Hi Chrom Reversible ODS • Hibar II C18 • Hibar ODS • HP-C18 • HxSil C18 • HydroBond PS-C18 • High Strength Silica (HSS) HPLC C18 • High Strength Silica (HSS) HPLC C18 SB • High Strength Silica (HSS) HPLC T3 • Hyperprep HS C18 • Hypersil 100 C18 • Hypersil BDS C18 • Hypersil GOLD • Hypersil GOLD aQ • Hypersil-ODS • Hypersil ODS-2 • Macrosphere 300 C18 • MetaSil AQ C18 • MetaSil ODS • MetroSep C18H • MetroSep C18AQ • MetroSep C18 AQ+ • MetroSep C18SH • MetroSep C18EPS • MPLC RP-18 • µBondapak C18 • µBondapak C18 Radial-Pak • Microsorb C18 • MicroSpher C18 • MonoChrom C18 • nanoAcquity BEH130 • nanoAcquity BEH300 • Nova-Pak C18 • NUCLEODUR C18 Gravity • NUCLEODUR C18 Isis • NUCLEODUR C18 Pyramid • NUCLEODUR 100 C18 ec • NUCLEODUR Sphinx RP • Nucleosil 100 C18 • Nucleosil C18 • Nucleosil C18 AB • Nucleosil C18 HD • Nucleosil C18 Nautilus • Nucleosil C18 MPN • Nucleosil C18 PAH • Nucleosil C18 PPN • ODS Ultrapak • OmniSpher C18 • Onyx C18 • Partisil ODS • Partisil ODS2 • Partisil ODS3 • Partisphere C18 HyPURITY AQUASTAR HyPURITY C18 IBM Octadecyl IB-SIL C18 IB-SIL C18 BD Inertsil ODS Inertsil ODS series 1,2,3 ISCO C-18 J’Sphere ODS-H80 J’Sphere ODS-L80 J’Sphere ODS-M80 Jupiter C18 Kinetex C18 Kromasil C18 LiChrospher PAH LiChrosorb RP-18 LiChrospher 100 RP-18 LiChrospher RP18e Luna C18 Luna C18(2) • Peco HCODS • Pecosil C18 • Pecosphere 3x8C • Pecosphere 3x8CR C18 • Pecosphere 5x15CRT C18 • Pecosphere-3C C18 • Pecosphere-3CR C18 • Pecosphere-3x3 C18 • Peptica C18 serresA,B,C • Pfarma C18 series A,B,C • Pinnacle II C18 • Pinnacle DB C18 • Pinnacle DB Aqueous C18 • Platinum C18 • Platinum EPS C18 • Platinum 300 C18 • Polaris C18-A • Polaris C18-Ether • Polygosil 60-7 C18 • Polygosil C18

  5. HPLC C18 column listing (L1-USP) • Polygosil C18 60ª • Premier C18 • Prep Nova-Pak HR C18 • Prevail C18 • Prevail OA • Prevail Select C18 • PrimeSphere C18-HC • Prodigy ODS(3) • ProntoSIL C18 H • ProntoSIL C18 SH • ProntoSIL C18 AQ • ProntoSIL C18 AQ Plus • ProntoSIL C18 ace-EPS • ProSphere 300 C18 • ProSphere 300 PAH • ProSphere HP C18 • ProSphere HP C18 AQ • ProSphere HP C18 ZAP! • ProteCol-G C18 HQ203 • ProteCol-P C18 HQ105 • ProteCol-P C18 HPH125 • ProteCol-P C18 GP125 • Purospher RP-18 • Purospher RP18e • Purospher STAR RP18e • Pursuit C18 • Pursuit XRs C18 • Radial-Pak C18 • Reduced Activity C18 • Regis ODS • Resolve C18 • Resolvex C18 • Sarasep P C18 • Scherzo SM-C18 • SelectaPore 90M, 300P, 300M • SynChropak RPP-1000 • SynChropak RPP-4000 • SynChropak RPP-100 • SynChropak RPS-100 • Synergi Fusion-RP • Synergi Hydro-RP • Targa C18 • TSK-GEL ODS-100V • TSK-GEL ODS-100Z • TSK-GEL ODS-100S • TSK-GEL Super-ODS • TSK-GEL ODS-80Tm • TSK-GEL ODS-80Ts • TSK-GEL ODS-120A • TSK-GEL ODS-120T • TSK-GEL ODS-140HTP • TSK-GEL OligoDNA-RP • Ultra C18 • Ultra Aqueous C18 • Ultra II C18 • Ultra II Aqueous C18 • Ultracarb ODS • Ultrasphere ODS • Ultrasphere C-18 • Ultrasphere C-18-IP • Ultron VX-ODS • Unison UK-C18 • Unison US-C18 • Velosep RP-18 • VisionHT C18 • VisionHT C18-B • VisionHT C18HL • VisionHT C18-P • Viva C18 • Vydac 201TP • Vydac 202TP • Vydac 218MS • Vydac 218TP • Vydac 238MS • Vydac C18 ShodexODSpak F-411 Shodex Silica C18M Shodex Silica C18P Spectraphysics C18 Spheri-(5 and 10) RP-18 Spheri-(5 and 10) ODS C18 Spherisorb ODS1 Spherisorb ODS2 Spherisorb ODS-B STR ODS-II STR ODS-M Sunfire C18 Supelcosil LC-18 Supelcosil LC-18-DB Supelcosil LC-318 Superiorex Superspher 100 RP-18 Symmetry C18 SymmetryPrep C18 Symmetry300 C18 SymmetryShield RP18 SynChropak RPP Wakosil C18 RS 3 and 5 Wakosil 5C18 HG and AR WakopakWakosil-II 3C18 HG WakopakWakosil-II 3C18 AR WakopakWakosil-II 3C18 RS WakopakWakosil-II 5C18 WakopakWakosil-II 5C18-100 WakopakWakosil-II 5C18 HG WakopakWakosil-II 5C18 AR WakopakWakosil-II 5C18 RS WakopakWakosil-II 5C18 HG Prep Wakopak Wakosil-II 5C18 AR Prep WakopakWakosil-II 5C18 RS Prep Wakopak Handy ODS WakopakWakosil 5C18 WakopakWakosil 5C18T WakopakWakosil 5C18N

  6. HPLC C18 column listing (L1-USP) YMC-Pack Hydrosphere C18 YMC-Pack ODS-A YMC-Pack ODS-AL YMC-Pack ODS-AM YMC-Pack ODS-AQ YMC-Pack Pro C18 YMC-Pack Pro C18 RS Zodiac 120 C18H Zodiac 120 C18AQ Zodiac 120 C18 Eurobond Zodiac 120 AC EPS Zodiac 120 C18SH Zodiac ODS Zodiac ODS H ZORBAX ODS C18 ZORBAX ODS Classic ZORBAX Eclipse Plus C18 ZORBAX Eclipse PAH ZORBAX Eclipse XDB-C18 ZORBAX Extend-C18 ZORBAX Rx C18 ZORBAX StableBond C18 ZORBAX 300 StableBond C18 • WakopakWakosil 10C18 • WakopakWakosil 5C18 AR • WakopakWakosil 5C18-200 • WakopakWakosil 5C18-200T • WakopakWakosil 5C18-200N • WakopakWakosil 10C18-200 • Wakopak MS-5C18 GT • WakopakNavi C18-5 • WakopakCombi ODS • WakopakCombi ODS Fast • XBridge BEH130 • XBridge BEH300 • XBridge C18 • XBridge OST C18 • XBridge Shield RP18 • XTerra MS C18 • XTerra RP18 • 378 HPLC column as C18 in 2010 • Which is the best one! • Do I need to buy all! • What is my C18 strategy? • ...etc... $$$

  7. A new modern strategy is absolutely necessary in 2010 for HPLC end-users New Generic Approach to Find Appropriate HPLC column (Total Solution for HPLC end-users) Better matrix (silica-gel) -Stationary Phase Specification (manufacturers) Phase according to Polarity (acidic and basic analytes) Phase according to Hydrophobicity(% carbon loading) Phase according to Low Metal Content and Silianol Activity Phase compatible with new LC-MS technology Phase for very aqueous mobile phase (100% buffer or water) Etc... Note: Every Pittcon and Analytica Events present new C18, more and more HPLC C18/year

  8. New Modern HPLC Strategy • How to improvetodaysilica-based HPLC columns? • Betterbondingchemistry/ improved control for betterreproducibility. • What about the matrix? (support—silica) • High QualityLevel of the Support (Silica) • narrowest particule size distribution • perfectlyspherical • highlevel of purity • High surface area (m2/g) • Efficiency pore size (Å) • No fines SiliaSphere (SiliCycle) • Lowmetal content • High mechanicalstability • Reproducibleloading (critical in reversed-phase) X X Supplier A (top) Supplier B (bottom) (both are real HPLC supports)

  9. GenericApproach/Most popular applications • Nowbetter support (Silica) • Similarchemistriesthanother HPLC suppliers • Improvedquality and efficiency • HPLC column forlow pH (not hybrid, pure silicabased) • HPLC column forhigh pH (pseudo-hybrid) • HPLC column for 100% aqueous mobile phase (or buffer) • HPLC column for LC-MS (no bleeding) • HPLC column for QC/QA • validation and method development (easy method) • No tailling, bettersensitivity and peakshape for quaternary ammonium compounds) • HPLC column for HILIC technology(extremely polar compounds)

  10. Portofolio of SiliaChromColumns A column for everyneed: ONLY 7 Chemistries! SiliaChromSB C18 and SB C8 SiliaChrom XDB1 C18 ,C8, C4, CN, AMINO, PHENYL, DIOL, Silica… SiliaChromXDB and XDB2 (C8&C18) SiliaChrom AQ C18 (dtC18) SiliaChromHILIC SiliaChrom XT C18 (XT Fidelity C18) NO MORE CHEMISTRIES

  11. SiliaChrom-Chemistries Review New Technology Silica-Based HPLC columns for Extremely Acidic Compounds R=C8 or C18, pH<2.0 H3O+ Steric Effect H3O+ Note: Not Endcapped silica gel

  12. SiliaChrom-Chemistries Review New Technology TMS endcapped Silica-Based HPLC columns for all type of compounds (pH=2-9) Free silianol groups (OH) R=C18 Polar Surface Ratio of TMS-OH-C18 is well-balanced Note: Partially End capped silica gel

  13. SiliaChrom-Chemistries Review Silica-Based HPLC column for EXTREMELY POLAR compounds SiliaChromHILIC Normal-Reversed Phases Conditions: Only SiliaChromHILIC Note: Not Endcapped silica gel

  14. SiliaChrom-Chemistries Review Silica-based HPLC columns for low to high hydrophobic compounds pH=1.5-10.0 SiliaChromXDB C8 (8%) and C18 (15%) SiliaChrom XDB1 C8 (14%) and C18 (22%) SiliaChrom XDB2 C18 (18%) 1) Different carbon loadings (8-22%C) 2) XDB2=Luna C18(2) 3) XDB1= Phenyl, NH2, CN, 300A, etc… Note: Double Endcapped silica gel

  15. SiliaChrom-Chemistries Review New Technology Replace OH by CH3 (Pseudo-Hybrid tech.) SiliaChromXT C18 Series Silica-Based HPLC columns for EXTREMELY Basic compounds pH=1.5-12 No bridge (No Backpressure issue as XBridge (Waters))

  16. SiliaChrom-Chemistries Review

  17. SiliaChromSeriesHPLC Classification-USP/NISTApproaches • All SiliaChrom HPLC columns are evaluated by USP and NIST approaches: • Uracil: void volume marker • Toluene: hydrophobicretention • Ethylbenzene: methyleneselectivity and plate count determination • Amitriptyline: activitytowards bases (silianolactivityevaluation) (A) • Quinizarin: activitytowardschelatingreagents (metal content evaluation) (B) (Pharmacopeial Forum, Vol. 31(2) March-Apr. 2005, p.637) (NIST, SRM 870, Certificat Issue Date:December 19th 2002, ValiduntilSeptember 30th 2010) (A) (B)

  18. SiliaChromEvaluation • HPLC System: Thermo Surveyor with PDA • (ChromQuest-Data handling version 5.0) • Mobile Phase: MeOH/buffer (80/20, v/v) • Buffer: 20mM of phosphate buffer adjusted at pH=7.0 • Flow rate: 1.0 mL/min • Detector: UV scan (PDA), Total scan 200-600nm • Temperature: 23C • All HPLC columns: 4.6x150mm, 5µm

  19. 150000 140000 130000 120000 110000 1.55 100000 90000 7.34 80000 uAU 9.73 70000 60000 3.88 4.86 50000 40000 30000 20000 10000 0 -10000 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Time (min) Phenomenex Luna C18(2) Low Plate Count (seeResults Table) Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline

  20. 180000 170000 160000 150000 140000 1.38 130000 120000 110000 100000 90000 6.85 uAU 80000 70000 3.29 60000 4.16 50000 12.99 40000 30000 20000 10000 4.73 0 -10000 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Time (min) Agilent ZorbaxSB C18 Important Tailling Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline

  21. 150000 140000 130000 120000 1.49 110000 100000 90000 80000 uAU 70000 10.18 60000 8.71 4.05 50000 5.14 40000 30000 20000 10000 0 -10000 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Time (min) Waters SunfireC18 Asymetry Metal Contamination Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline

  22. 190000 180000 170000 160000 150000 1.61 140000 130000 120000 110000 100000 uAU 90000 80000 8.75 70000 12.02 60000 4.15 5.19 50000 40000 30000 20000 10000 0 -10000 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Time (min) SiliaChrom XDB2 C18 Very good peak shapes Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline

  23. Summary Results-Case Study Basic analyte: SiliaChrom XDB2 Less polar analyte: SiliaChromXDB1

  24. Recent SiliaChromApplications • Acidic and Basic Compounds (8) with one SiliaChrom HPLC column (AQ C18) • Fluoroquinolones for Veterinary use by HPLC-Marbofloxacin and Sarafloxacin • Halogenated Herbicides by HPLC-Dicamba, Atrazine and 2,4-D • Catecholamines by HPLC- Polar Compounds • Faster HPLC withSiliaChrom-Polar and mid-polar Compounds • Isomerseparation for Chlorophenolseries by simple HPLC method • HPLC study for Naloxone-pH=2.5, 7.0 and 10.5 • HPLC Chromatogram of Ganciclovir– Polar Compound (diol-amine) • HPLC Chromtogram of Cefotaxime—Extremely Polar Compound (acid-amine-sulfur) • And more….

  25. Recent SiliaChromApplications Acidic and Basic Compounds (8) withsameSiliaChrom HPLC column 1-MaleicAcid 4-Chlorpheniramine 7-Dichlorphen 2-Acetaminophen 5-BenzoicAcid 3-Pseudoephedrine 8-Ibuprofen 6-SalicylicAcid

  26. Recent SiliaChromApplications Acidic and Basic Compounds (8) withsameSiliaChrom HPLC column Phenomenex Luna C18(2) 5µm 4.6x150mm 210 nm 1 2 2 5 2 230 nm 230 nm 4 6 7 1 3 8 1

  27. Recent SiliaChromApplications Acidic and Basic Compounds (8) withsameSiliaChrom HPLC column (same HPLC conditions than Luna C18(2)) SiliaChrom AQ C18 5µm 4.6x150mm 210 nm 2 5 230 nm 4 6 7 3 8 1

  28. Recent SiliaChromApplications Fluoroquinolones for Veterinary use by HPLC-Marbofloxacin and Sarafloxacin SiliaChrom AQ C18 5µm 4.6x150mm Zwitterion Molecules Waters Atlantis dC18 5µm 4.6x150mm Note: SiliCycle has the perfect equivalent (better) with SiliaChromAQ C18 (or dtC18) than Atlantis dC18 (Waters)

  29. Recent SiliaChromApplications Halogenated Herbicides by HPLC-Dicamba, Atrazine and 2,4-D SiliaChrom dtC18 5µm 4.6x150mm

  30. Recent SiliaChromApplications Halogenated Herbicides by HPLC-Dicamba, Atrazine and 2,4-D SupelcoAscentis C18 5µm 4.6x150mm

  31. Recent SiliaChromApplications Catecholamines by HPLC-Polar Compounds SiliaChrom AQ C18 5µm 4.6x150mm Mobile Phase: Water 1.0% AcOH only! EpinephrineNorepinephrine Dopamine Tyrosine Levodopa

  32. Recent SiliaChromApplications Faster HPLC withSiliaChrom-Polar and mid-polar Compounds SiliaChrom AQ C18 5µm 3.0x50mm 95/5 0.1%TFA in H2O/ACN – 5/95 0.1% TFA in H2O/ACN in 2.25 min. 1.75 mL/min, 23°C, 254 nm (LC-MS conditions fromAstraZeneca Canada) Test Mix to evaluate HPLC column from AstraZeneca (Canada) CaffeineChlorthalidoneAlbendazoleWarfarinTolnaftateDioctylPhatalate

  33. Recent SiliaChromApplications SiliaChrom XDB1 PHENYL 5µm 4.6x150mm 60/40, H2O/MeOH, 1.00 mL/min, 23°C, 254nm : EasyMethod!

  34. Recent SiliaChromApplications • HPLC study for Naloxone-pH=2.5, 7.0 and 10.5 SiliaChrom XT C18, 5µm, 4.6x150mm 80/20, Phosphate Buffer/MeOH, 1.00 mL/min, 23°C, 254nm Fronting ? pH=2.5 Assymetry = 0.85 50/50, Phosphate Buffer/MeOH, 1.00 mL/min, 23°C, 254nm pH=7.0 Assymetry = 0.89

  35. Recent SiliaChromApplications • HPLC study for Naloxone-pH=2.5, 7.0 and 10.5 SiliaChrom XT C18, 5µm, 4.6x150mm 50/50, H2O/MeOH 0.1% NH4,OH, 1.00 mL/min, 23°C, 254nm pH=10.5 Assymetry = 0.99 Remember, pH range for XT is 1.5-12—Good Chromatography Tool

  36. Recent SiliaChromApplications HPLC Chromatogram Ganciclovir – Polar Compounds (diol-amine) SiliaChrom AQ C18 (4.6x150mm, 5µm) MeOH:Water, 5:95 UV 252nm 1mL/min

  37. Recent SiliaChromApplications HPLC Chromatogram of Cefotaxime—Extremely Polar compound (acid-amine-sulfur) SiliaChrom AQ C18 (4.6x150mm, 5µm) 0.4 mM KH2PO4 + 8mM K2HPO4: MeOH, 89:11 UV 254nm 1mL/min

  38. Recent SiliaChromApplications HPLC for mixture of Fenitrothion-Pyriproxyfen—Different Level of Hydrophobicity SiliaChrom XDB1 C18 (4.6x150mm, 5µm) ACN:Water, 70:30 UV 254nm 1mL/min

  39. Recent SiliaChromApplications HPLC Determination for Ephedrine (from Ephedra Sample) Zorbax SB C18 (4.6x150mm, 5µm) SiliaChrom XDB2 C18 (4.6x150mm, 5µm)

  40. Recent SiliaChromApplications HPLC chromatogram of Berberine-Metal Content Effect SunfireC18 (4.6x250mm 5µm) SiliaChromdtC18 (4.6x250mm, 5µm) ACN: 1%TFA, 35:65 UV 346nm 1 mL/min

  41. Durability Testing at pH-1.00 SiliaChrom SB C18 0 hr 120 hrs SiliaChromSB C18 (4.6x150mm, 5µm) 59:39:2 ACN:Water:TFA (pH = 1.00) UV 270 nm, 1 mL/min (Uracil, Phenol, Toluene and Ethylbenzene) • Poster presented at Pittcon 2010, Orlando FL, USA • LC-GC Application Note—September 2009

  42. Durability Testing at pH-11.5 SiliaChromXT C18 Naphthalene 2.75 2.25 1 hr 1.91 2.23 2.77 SiliaChromXT C18 (4.6x150mm, 5µm) MeOH: Water/TEA, pH=11.5, 40:60 UV 215nm, 1 mL/min 1.84 400 hrs Uracil, Phenol, Nitrobenzene and Naphtalene

  43. Durability Testing at pH=11.50, 90C SiliaChrom XT C18 Fidelity 0 hr SiliaChromXT C18 (4.6x150mm, 5µm) 54.9-44.9-0.2 (ACN-Water-TEA) 90C, UV 270 nm 1 mL/min 120 hrs (Uracil, Phenol, Toluene and Ethylbenzene)

  44. SiliaChrom HILIC, Vitamins SiliaChrom HILIC (4.6x150mm, 5µm) ACN 0.1% TFA: Water 0.1%TFA, 10:90 UV 280nm 1 mL/min Waters HILIC Silica HPLC column (4.6x150mm, 5µm) B6 C B2 B1 Vitamin B1, Vitamin B6, Vitamin C and Vitamin B2

  45. Generic Approach with SiliaChrom Series 87% of HPLC success in my Lab! Neutral Note: SiliaChrom AQ is an universal HPLC column (see how particle is designed : TMS-OH-C18)

  46. Conclusion ONLY 8 Chemistries to cover more than 85% of Regular HPLC Application using Silica-Based as support • Be aware for your HPLC column strategy, Marketing is not real SCIENCE!

  47. www.silicycle.com Thank you!