New Generic Approach for HPLC analysis: The Modern Silica Based HPLC Columns; SiliaChrom Series

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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 Analytical Chemistry 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) 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 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 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 Delta-Pak C18 Denali Develosil ODS Diazem 2060 C18 60A Diazem 300A C Diazem C18 Diazem C18 Diazem C18 Diazem C18EC Diazem C18 Diazem C Diazem C 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

4 HPLC C18 column listing (L1-USP) 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) 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 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) Shodex ODSpak 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 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 Wakosil C18 RS 3 and 5 Wakosil 5C18 HG and AR Wakopak Wakosil-II 3C18 HG Wakopak Wakosil-II 3C18 AR Wakopak Wakosil-II 3C18 RS Wakopak Wakosil-II 5C18 Wakopak Wakosil-II 5C Wakopak Wakosil-II 5C18 HG Wakopak Wakosil-II 5C18 AR Wakopak Wakosil-II 5C18 RS Wakopak Wakosil-II 5C18 HG Prep Wakopak Wakosil-II 5C18 AR Prep Wakopak Wakosil-II 5C18 RS Prep Wakopak Handy ODS Wakopak Wakosil 5C18 Wakopak Wakosil 5C18T Wakopak Wakosil 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 Wakopak Wakosil 10C18 Wakopak Wakosil 5C18 AR Wakopak Wakosil 5C Wakopak Wakosil 5C18-200T Wakopak Wakosil 5C18-200N Wakopak Wakosil 10C Wakopak MS-5C18 GT Wakopak Navi C18-5 Wakopak Combi ODS Wakopak Combi ODS Fast XBridge BEH130 XBridge BEH300 XBridge C18 XBridge OST C18 XBridge Shield RP18 XTerra MS C18 XTerra RP 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 improve today silica based HPLC columns? Better bonding chemistry/ improved control for better reproducibility. What about the matrix? (support silica) High Quality Level of the Support (Silica) narrowest particule size distribution perfectly spherical high level of purity X High surface area (m 2 /g) Efficiency pore size (Å) No fines SiliaSphere (SiliCycle) Low metal content X High mechanical stability Reproducible loading (critical in reversed phase) Supplier A (top) Supplier B (bottom) (both are real HPLC supports)

9 Generic Approach/Most popular applications Now better support (Silica) Similar chemistries than other HPLC suppliers Improved quality and efficiency HPLC column for low ph (not hybrid, pure silica based) HPLC column for high 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, better sensitivity and peak shape for quaternary ammonium compounds) HPLC column for HILIC technology (extremely polar compounds)

10 Portofolio of SiliaChrom Columns A column for every need: ONLY 7 Chemistries! SiliaChrom SB C18 and SB C8 SiliaChrom XDB1 C18,C8, C4, CN, AMINO, PHENYL, DIOL, Silica SiliaChrom XDB and XDB2 (C8&C18) SiliaChrom AQ C18 (dtc18) SiliaChrom HILIC SiliaChrom XT C18 (XT Fidelity C18) NO MORE CHEMISTRIES

11 New Technology SiliaChrom Chemistries Review Silica Based HPLC columns for H Extremely Acidic Compounds 3 C H 3 C C CH R=C8 or C18, ph<2.0 3 CH 3 Si C O Si O CH C 3 O CH3 H3 C CH 3 O Si OH CH 3 H C CH O 3 C 3 CH 3 Si O Si C C CH O 3 CH 3 H 3 C CH 3 H 3 O + Steric Effect Note: Not Endcapped silica gel H 3 O + R R

12 New Technology SiliaChrom Chemistries Review Silica Based HPLC columns for all type of compounds (ph=2 9) TMS endcapped 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 O SiliaChrom HILIC Normal Reversed Phases Conditions: Only SiliaChrom HILIC HN NH 2 H 3 C CH Si 2 CH 2 CH 2 Si O CH 3 O Si OH Note: Not Endcapped silica gel

14 SiliaChrom Chemistries Review Silica based HPLC columns for low to high hydrophobic compounds ph= SiliaChrom XDB 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, NH 2, CN, 300A, etc H 3 C (CH Si 2 ) 7 CH 3 Si O CH 3 O Si O Si(CH 3 ) 3 H 3 C (CH Si 2 ) 17 CH 3 Si O CH 3 O Si O Si(CH 3 ) 3 Note: Double Endcapped silica gel

15 New Technology SiliaChrom Chemistries Review Silica Based HPLC columns for EXTREMELY Basic compounds ph= SiliaChrom XT C18 Series O O O O Si Si Si Replace OH by CH 3 (Pseudo Hybrid tech.) H 3 C O CH 3 Si CH 3 Si O CH 3 O O CH 3 Si O Si H 3 C R H 3 C O CH 3 CH 3 Si Si O CH 3 O CH 3 Si No bridge (No Backpressure issue as XBridge (Waters))

16 SiliaChrom Chemistries Review

17 SiliaChrom Series HPLC Classification-USP/NIST Approaches All SiliaChrom HPLC columns are evaluated by USP and NIST approaches: Uracil: void volume marker Toluene: hydrophobic retention Ethylbenzene: methylene selectivity and plate count determination Amitriptyline: activity towards bases (silianol activity evaluation) (A) Quinizarin: activity towards chelating reagents (metal content evaluation) (B) (Pharmacopeial Forum, Vol. 31(2) March-Apr. 2005, p.637) (NIST, SRM 870, Certificat Issue Date:December 19th 2002, Valid until September 30th 2010) (A) (B)

18 SiliaChrom Evaluation 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 nm Temperature: 23 C All HPLC columns: 4.6x150mm, 5µm

19 Phenomenex Luna C18(2) uau Low Plate Count (see Results Table) Time (min) Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline

20 Agilent Zorbax SB C Important Tailling uau Time (min) Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline

21 Waters Sunfire C18 Asymetry Metal Contamination uau Time (min) Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline

22 SiliaChrom XDB2 C18 uau Very good peak shapes Time (min) Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline

23 Summary Results-Case Study HPLC Column K Ethylbenzene α Ethyl/Tol N Ethylbenzene R AS (Quinizarine) Tf (Amitriptyline) SiliaChrom XDB2 C Luna C18(2) Zorbax SB C Sunfire C SiliaChrom XDB1 C Basic analyte: SiliaChrom XDB2 Less polar analyte: SiliaChrom XDB1

24 Recent SiliaChrom Applications 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 with SiliaChrom-Polar and mid-polar Compounds Isomer separation for Chlorophenol series 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 SiliaChrom Applications Acidic and Basic Compounds (8) with same SiliaChrom HPLC column 1 Maleic Acid 4 Chlorpheniramine 2 Acetaminophen 7 Dichlorphen 5 Benzoic Acid 3 Pseudoephedrine 8 Ibuprofen 6 Salicylic Acid

26 Recent SiliaChrom Applications Acidic and Basic Compounds (8) with same SiliaChrom HPLC column Phenomenex Luna C18(2) 5µm 4.6x150mm 210 nm nm

27 Recent SiliaChrom Applications Acidic and Basic Compounds (8) with same SiliaChrom HPLC column (same HPLC conditions than Luna C18(2)) SiliaChrom AQ C18 5µm 4.6x150mm 210 nm nm

28 Recent SiliaChrom Applications Fluoroquinolones for Veterinary use by HPLC Marbofloxacin and Sarafloxacin SiliaChrom AQ C18 5µm 4.6x150mm Zwitterion Molecules Waters Atlantis dc18 5µm 4.6x150mm Asymmetry SiliaChrom AQ C18 Atlantis dc Note: SiliCycle has the perfect equivalent (better) with SiliaChrom AQ C18 (or dtc18) than Atlantis dc18 (Waters)

29 Recent SiliaChrom Applications Halogenated Herbicides by HPLC-Dicamba, Atrazine and 2,4-D SiliaChrom dtc18 5µm 4.6x150mm

30 Recent SiliaChrom Applications Halogenated Herbicides by HPLC-Dicamba, Atrazine and 2,4-D Supelco Ascentis C18 5µm 4.6x150mm Asymmetry SiliaChrom dtc18 Ascentis C

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

32 Recent SiliaChrom Applications Faster HPLC with SiliaChrom-Polar and mid-polar Compounds SiliaChrom AQ C18 5µm 3.0x50mm 95/5 0.1%TFA in H 2 O/ACN 5/95 0.1% TFA in H 2 O/ACN in 2.25 min ml/min, 23 C, 254 nm (LC MS conditions from AstraZeneca Canada) Test Mix to evaluate HPLC column from AstraZeneca (Canada) Caffeine Chlorthalidone Albendazole Warfarin Tolnaftate Dioctyl Phatalate

33 Recent SiliaChrom Applications SiliaChrom XDB1 PHENYL 5µm 4.6x150mm 60/40, H 2 O/MeOH, 1.00 ml/min, 23 C, 254nm : Easy Method! OH OH OH Cl Cl Cl

34 Recent SiliaChrom Applications 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? Assymetry = 0.85 ph=2.5 50/50, Phosphate Buffer/MeOH, 1.00 ml/min, 23 C, 254nm Assymetry = 0.89 ph=7.0

35 Recent SiliaChrom Applications HPLC study for Naloxone-pH=2.5, 7.0 and 10.5 SiliaChrom XT C18, 5µm, 4.6x150mm 50/50, H 2 O/MeOH 0.1% NH 4,OH, 1.00 ml/min, 23 C, 254nm Assymetry = 0.99 ph=10.5 Remember, ph range for XT is Good Chromatography Tool

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

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

40 Recent SiliaChrom Applications HPLC chromatogram of Berberine-Metal Content Effect Sunfire C18 (4.6x250mm 5µm) SiliaChrom dtc18 (4.6x250mm, 5µm) ACN: 1%TFA, 35:65 UV 346nm 1 ml/min

41 Durability Testing at ph-1.00 SiliaChrom SB C18 Time/h Retention time (Ethylbenzene) (min) Tailing Factor (Ethylbenzene) (USP) Theoritical Plates # (Ethylbenzene) (USP) SD Mean RSD (%) hr 120 hrs SiliaChrom SB 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 SiliaChrom XT C18 Time/h Naphthalene Tailing Factor Retention time (Naphthalene) (min) hr SiliaChrom XT C18 (4.6x150mm, 5µm) MeOH: Water/TEA, ph=11.5, 40:60 UV 215nm, 1 ml/min hrs Uracil, Phenol, Nitrobenzene and Naphtalene

43 Durability Testing at ph=11.50, 90 C SiliaChrom XT C18 Fidelity 0 hr SiliaChrom XT C18 (4.6x150mm, 5µm) (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 B1 C B2 Vitamin B1, Vitamin B6, Vitamin C and Vitamin B2

45 Generic Approach with SiliaChrom Series Neutral Note: SiliaChrom AQ is an universal HPLC column (see how particle is designed : TMS-OH-C18)

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

47 Conclusion Low ph High ph High level of Water Hydrophobicity Low Metal Polar Extemely ( ) (9 12) Mobile Phase (different Level) Content Analyte Polar Analyte SiliaChrom SB XT AQ XDB, XDB1 XDB2 dt AQ HILIC 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!

48 Thank you!

49 Si Acid Base Neutral Ionic Hydrophobic Hydrophilic SiliCycle Inc. manufactures a variety of HPLC columns for reversed phase and normal phase applications. The SiliaChrom series contain more than 30 different phases, and we continue to develop additional unique and powerful HPLC column. Most of the SiliaChrom product line are based on silica we produce in our own laboratories since In fact, you can be completely assured of the quality, from raw materiel synthesis through packing, testing and customer s applications. ID (mm) Analytical 2.1, 3.0, 4.6 Semi-Preparative 10.0, 20.0 Preparative 30.0, 50.0 SiliaChrom HPLC columns for the most popular HPLC applications presenting superior performance for any type of compounds. Broad ph range ( ) Compatible with 100% aqueous mobile phase High surface coverage Minimum buffer concentration needed Low bleed Wide variety of chemistries available Excellent column efficiency Long column lifetime Tightest specification SiliaChrom SB C18 SiliaChrom SB C8 SiliaChrom AQ C18 SiliaChrom HILIC SiliaChrom dtc18 SiliaChrom XDB C8 & C18 SiliaChrom XDB1 C8 & C18 SiliaChrom XDB2 C18 SiliaChrom AQ C18 SiliaChrom HILIC SiliaChrom dtc18 SiliaChrom XDB1 CN SiliaChrom XDB1 AMINO SiliaChrom XT C18 SiliaChrom XDB C8 & C18 SiliaChrom XDB1 C8&C18 SiliaChrom XDB2 C18 SiliaChrom dtc18 SiliaChrom XDB1 C4 SiliaChrom XDB1 PHENYL SiliaChrom XDB1 Si SiliaChrom RPC SiliaChrom AQ C18 SiliaChrom HILIC SiliaChrom XDB1 DIOL SiliaChrom dtc18 SiliaChrom SCX & SAX SiliaChrom IEC SC & WC SiliaChrom IEC SA & WA SiliCycle HPLC Columns Pore Size Particule Size Surface Area Carbon Loading ph Range USP Code Applications Chromatographic Properties Similar Phases SiliaChrom SB C18 SiliaChrom SB C8 SiliaChrom AQ C18 SiliaChrom HILIC SiliaChrom XT C18 SiliaChrom XDB C18 SiliaChrom XDB C8 SiliaChrom XDB1 C18 SiliaChrom XDB1 C8 SiliaChrom XDB1 CN SiliaChrom XDB1 Si SiliaChrom XDB1 PHENYL SiliaChrom XDB1 DIOL SiliaChrom XDB1 AMINO SiliaChrom XDB1 C4 SiliaChrom SCX SiliaChrom SAX SiliaChrom XDB2 C18 SiliaChrom dtc18 150Å 150Å 100Å 100Å 150Å 150Å 150Å 100Å 100Å 100Å 100Å 100Å 100Å 100Å 100Å 100Å 100Å 100Å 100Å 3.0, 5.0µm 3.0, 5.0µm 3.0,5.0µm 3.0,5.0µm 3.0,5.0µm 3.0,5.0µm 3.0,5.0µm 3.0,5.0µm 3.0,5.0µm 5.0µm 3.0, 5.0µm 5.0µm 5.0µm 5.0µm 5.0µm 3.0,5.0µm 3.0,5.0µm 3.0,5.0µm 2.5,3.0,5.0µm m 2 /g m 2 /g 380m 2 /g m 2 /g 200m 2 /g 200m 2 /g 200m 2 /g m 2 /g m 2 /g m 2 /g m 2 /g m 2 /g m 2 /g m 2 /g m 2 /g 380m 2 /g 380m 2 /g 380m 2 /g m 2 /g 12% 7% 18% 8% 15% 15% 8% 22% 14% 5% Not Applicable 12% 5% 6% 7% 10% 6% 18% 18% Not Applicable Not Applicable L01 L07 L01 Please Call L01 L01 L07 L01 L07 L10 L03 L11 L20 L08 L26 L09 L14 L01 L01 Ideal for MS and ELSD of neutral to slightly polar analytes Selectivity and peak shape similar to SiliaChrom SB C8 Ideal for analyses that require > 90% water (buffer) Ideal for MedChem laboratories-isolation of very polar analytes Excellent durability for high ph-ideal for basic compounds Ideal for barbiturates, fat-soluble vitamins, fatty acids, steroids Selectivity and peak shape similar to SiliaChrom XDB C18 Hydrophobic C18 phase suitable for analyses of a wide range of compounds Applications similar to SiliaChrom XDB1 C18 Excellent for basic pharmaceuticals, steroid and other basic compounds Ideal for normal phase applications Ideal for polynuclear aromatic hydrocarbons, purines and polar aromatics Excellent for normal phase applications with more hydrophobic activity Superior general purpose amino phase. Idel for carbohydrates Ideal for peptides and small proteins Ideal for charged analytes Ideal for charged analytes Perfect peak symmetry for acidic, basic and neutral compounds Universal C18 for most popular applications (highest purity of silica gel) Very stable at low ph (1.0), low bleeding for LC-MS Very stable at low ph (1.0), low bleeding for LC-MS Accept very aqueous (or buffer) mobile phases Accept normal and reversed phases conditions Very stable at high ph (up to 12), low bleeding for LC-MS Designed for larger molecules and highly hydrophobic compounds Less hydrophobic than SiliaChrom XDB C18 Designed for maxiumum hydrophobicity and efficiency for dirty sample Less hydrophobic than SiliaChrom XDB1 C18-good C8 for high ph Maximum hydrophobicity and accept normal and reverse phases conditions Destined for normal phase conditions--high surface area-very low metal content Highly retentive phase for aromatic an unsaturated compounds Highly cyano phase with reduced silianol activity Recommanded for normal phase analysis Exceptionnally stable C4 with high bonding coverage at low silianol activity Highly retentive phase for strong cation exchanger chromatograpy Highly retentive phase for stong anion exchanger chromatography A very retentive, phase that exibits excellent peak shape for a wide range of compounds Highly retentive and selective for reversed phase separation of neutral and polar analyte Zorbax SB C18, Lichrospher RP C18 Zorbax SB C8 Zorbax SB Aq, Atlantis dc18, Inertsil ODS-3 Unique Unique, Gemini, Gemini NX Symmetry C18, Zorbax XDB C18, Discovery C18 Symmetry C8, Zorbax XDB C8 Sunfire C18, Luna C18, Ascentics C18, Altima HP C18 Sunfire C8, Luna C8, Ascentis C8 Luna CN, Zorbax SB CN Luna HILIC, Luna Silica Zorbax SB Phenyl Nucleosil Diol, Luna Diol Luna NH2 Jupiter C4 Luna SCX Agilent SB-SAX Sunfire C18, Luna C18(2) Unique YMC ODS AQ C18, Atlantis T3, Atlantis dtc18 SiliaChrom XDB, XDB1 and XDB2 C18 SiliaChrom XDB1 PHENYL SiliaChrom XDB1 DIOL SiliaChrom SB C8 R=(CH 2 ) 7 CH 3 SiliaChrom AQ C18 R=(CH 2 ) 17 CH 3 CHIRAL SiliaChrom SB C18 R=(CH 2 ) 17 CH 3 SiliaChrom dtc18 R=(CH 2 ) 17 CH 3 (very pure silica gel) SiliaChrom XT C18 R=(CH 2 ) 17 CH 3 SiliaChrom OD CHIRAL SiliaChrom AD SiliaChrom XDB, XDB1 C8 SiliaChrom SAX SiliaChrom HILIC SiliaChrom XDB1 CN SiliaChrom SCX SiliaChrom XDB1 C4 SiliaChrom XDB1 AMINO SiliaChrom Si SiliaChrom IEC WA Size Exclusion Columns (Silica Based) SiliaChrom GF-100 (100 A) SiliaChrom GF-300 Amide (300 A) SiliaChrom GF-300 (300 A) [email protected], Tel.: , Toll-free: SILICYCLE (North America only) SiliaChrom IEC SA SiliaChrom IEC SC PSDVB Polystyrene - Divinylbenzene Polymer SiliaChrom RPC Copyright 2009 SiliCycle Inc. All rights reserved. SiliaChrom IEC WC