Charles Lévesque, M.Sc. Product Manager-Analytical Chemistry Vincent Bédard, B.Sc. HPLC Specialist François Béland, Ph.D. Vice-President of R&D New Generic.

Charles Lévesque, M.Sc.Product Manager-Analytical Chemistry

Vincent Bédard, B.Sc.HPLC Specialist

François Béland, Ph.D.Vice-President of R&D

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

Outlines

HPLC Listing of all C18 SiliaChrom StrategySiliaChrom—Chemistry Review

New Techologies-SB,AQ,XT

Evaluation of SiliaChromRecent SiliaChrom ApplicationsGeneric Approach for HPLC applications

SiliaChrom Focus

Alltima C18Alltima C18 LLAlltima HP C18Alltima HP C18 EPSAlltima HP C18 Hi-LoadAlltima HP C18 AQAlltima HP C18 AmideAlltima WP C18AlphaBond C18Amethyst C18AminoQuantAPEX ODSApollo C18AQUA C18Aquapore OD-300 C18AQUASIL C18Ascentis C18Ascentis Express C18ASI C18ASI microC18Atlantis dC18Atlantis T3Axxi-Chrom C18, ODSBakerbond C18BetaBasic 18BetaMax NeutralBETASIL C18Bio-C18

ACE 300 C18ACE AQACE C18ACE C18-ARAccQTag UltraACQUITY UPLC BEH130ACQUITY UPLC BEH 300ACQUITY UPLC BEH C18ACQUITY UPLC BEH Shield RP18ACQUITY UPLC HSS C18ACQUITY UPLC HSS C18 SBACQUITY UPLC HSS T3ACQUITY UPLC OST C18Adsorbosil C18Adsorbosphere C18Adsorbosphere HS C18Adsorbosphere UHS C18Adsorbosphere XL 300 C18Adsorbosphere XL C18Adsorbosphere XL C18-BAllsphere ODS-1Allsphere ODS-2Allure C18Allure Aqueous C18

BioBasic 18Bio-Sil ODSBio-Sil ODS-10Bondclone C18BR-C18Burdick & Jackson C18Cadenza CD-C18Cadenza 5CD-C18Cadenza CL-C18Cadenza 5CL-C18Cadenza CW-C18Cadenza 5CW-C18Cadenza HS-C18Capcell C18 AG 120Capcell C18 SG 120, 300Capcell C18 UG 80, 120, 300Capcell C18 MGCapcell C18 MGIICapcell C18 MGIIICapcell C18 ACRCapcell C18 AQCapcell C18 IF Chemcosorb 5-ODS-HChromegabond C18Chromegabond WR-C18Chromolith RP-18eClarity Oligo-RPClipeus C18Cogent HPS C18Cogent hQ C18Cogent Simulare C18Cogent MicroBee C18Cogent Bidentate C18Cogent e-Column C18Cosmosil C18 MS-IICosmosil C18 AR-IICosmosil C18 AR-300Cosmosil C18 PAQ

Delta-Pak C18DenaliDevelosil ODSDiazem 2060 C18 60ADiazem 300A C18 4-222Diazem 4-206 C18Diazem 4-230 C18Diazem 4-246 C18Diazem 4-247 C18ECDiazem C18Diazem C18 1000Diazem C18 1120Diazem C18 120ADiazem C18 120A series 43,44,45 and 46Diazem C18 series 200A, 300ADiazem C18 series 2000Diazem C18 series 3000Diazem C18 series 4000Diazem C18 60A

HPLC C18 column listing (L1-USP)

HyPURITY AQUASTARHyPURITY C18IBM OctadecylIB-SIL C18IB-SIL C18 BDInertsil ODSInertsil ODS series 1,2,3ISCO C-18J’Sphere ODS-H80J’Sphere ODS-L80J’Sphere ODS-M80Jupiter C18Kinetex C18Kromasil C18LiChrospher PAHLiChrosorb RP-18LiChrospher 100 RP-18LiChrospher RP18eLuna C18Luna C18(2)

Diazem C18 60A series 1,2,3Diazem C18 series ECDiazem C18 Dual-ZoneDiscovery C18Discovery HS C18Discovery Bo Wide Pore C18Econosil C18Econosphere C18Everest C18Fortis C18Fortis H2oGammabond C18Gemini C18Gemini-NXGenesis C18GP-C18GraceSmart C18Haisil 100, 300 C18Haisil HL C18HALO C18Hi Chrom Reversible ODSHibar II C18Hibar ODSHP-C18HxSil C18HydroBond PS-C18High Strength Silica (HSS) HPLC C18High Strength Silica (HSS) HPLC C18 SBHigh Strength Silica (HSS) HPLC T3Hyperprep HS C18Hypersil 100 C18Hypersil BDS C18Hypersil GOLDHypersil GOLD aQHypersil-ODSHypersil ODS-2

Macrosphere 300 C18MetaSil AQ C18MetaSil ODSMetroSep C18HMetroSep C18AQMetroSep C18 AQ+MetroSep C18SHMetroSep C18EPSMPLC RP-18µBondapak C18µBondapak C18 Radial-PakMicrosorb C18MicroSpher C18 MonoChrom C18nanoAcquity BEH130nanoAcquity BEH300Nova-Pak C18NUCLEODUR C18 GravityNUCLEODUR C18 IsisNUCLEODUR C18 PyramidNUCLEODUR 100 C18 ecNUCLEODUR Sphinx RPNucleosil 100 C18Nucleosil C18Nucleosil C18 ABNucleosil C18 HDNucleosil C18 NautilusNucleosil C18 MPNNucleosil C18 PAHNucleosil C18 PPNODS UltrapakOmniSpher C18Onyx C18Partisil ODSPartisil ODS2Partisil ODS3Partisphere C18

Peco HCODSPecosil C18Pecosphere 3x8CPecosphere 3x8CR C18Pecosphere 5x15CRT C18Pecosphere-3C C18Pecosphere-3CR C18Pecosphere-3x3 C18Peptica C18 serresA,B,CPfarma C18 series A,B,CPinnacle II C18Pinnacle DB C18Pinnacle DB Aqueous C18Platinum C18Platinum EPS C18Platinum 300 C18Polaris C18-APolaris C18-EtherPolygosil 60-7 C18Polygosil C18

HPLC C18 column listing (L1-USP)

Shodex ODSpak F-411Shodex Silica C18MShodex Silica C18PSpectraphysics C18Spheri-(5 and 10) RP-18Spheri-(5 and 10) ODS C18Spherisorb ODS1Spherisorb ODS2Spherisorb ODS-BSTR ODS-IISTR ODS-MSunfire C18 Supelcosil LC-18Supelcosil LC-18-DBSupelcosil LC-318SuperiorexSuperspher 100 RP-18Symmetry C18SymmetryPrep C18Symmetry300 C18SymmetryShield RP18SynChropak RPP

Polygosil C18 60ªPremier C18Prep Nova-Pak HR C18Prevail C18Prevail OAPrevail Select C18PrimeSphere C18-HCProdigy ODS(3)ProntoSIL C18 HProntoSIL C18 SHProntoSIL C18 AQProntoSIL C18 AQ PlusProntoSIL C18 ace-EPSProSphere 300 C18ProSphere 300 PAHProSphere HP C18ProSphere HP C18 AQProSphere HP C18 ZAP!ProteCol-G C18 HQ203ProteCol-P C18 HQ105ProteCol-P C18 HPH125ProteCol-P C18 GP125Purospher RP-18Purospher RP18ePurospher STAR RP18ePursuit C18Pursuit XRs C18Radial-Pak C18Reduced Activity C18Regis ODSResolve C18Resolvex C18Sarasep P C18Scherzo SM-C18SelectaPore 90M, 300P, 300M

Wakosil C18 RS 3 and 5Wakosil 5C18 HG and ARWakopak Wakosil-II 3C18 HGWakopak Wakosil-II 3C18 ARWakopak Wakosil-II 3C18 RSWakopak Wakosil-II 5C18 Wakopak Wakosil-II 5C18-100Wakopak Wakosil-II 5C18 HG Wakopak Wakosil-II 5C18 ARWakopak Wakosil-II 5C18 RSWakopak Wakosil-II 5C18 HG PrepWakopak Wakosil-II 5C18 AR PrepWakopak Wakosil-II 5C18 RS PrepWakopak Handy ODSWakopak Wakosil 5C18Wakopak Wakosil 5C18TWakopak Wakosil 5C18N

SynChropak RPP-1000SynChropak RPP-4000SynChropak RPP-100SynChropak RPS-100Synergi Fusion-RPSynergi Hydro-RPTarga C18TSK-GEL ODS-100VTSK-GEL ODS-100ZTSK-GEL ODS-100STSK-GEL Super-ODSTSK-GEL ODS-80TmTSK-GEL ODS-80TsTSK-GEL ODS-120ATSK-GEL ODS-120TTSK-GEL ODS-140HTPTSK-GEL OligoDNA-RPUltra C18Ultra Aqueous C18Ultra II C18Ultra II Aqueous C18Ultracarb ODS Ultrasphere ODSUltrasphere C-18Ultrasphere C-18-IPUltron VX-ODSUnison UK-C18Unison US-C18Velosep RP-18VisionHT C18VisionHT C18-BVisionHT C18HLVisionHT C18-PViva C18Vydac 201TPVydac 202TPVydac 218MSVydac 218TPVydac 238MSVydac C18

HPLC C18 column listing (L1-USP)

YMC-Pack Hydrosphere C18YMC-Pack ODS-AYMC-Pack ODS-ALYMC-Pack ODS-AMYMC-Pack ODS-AQYMC-Pack Pro C18YMC-Pack Pro C18 RSZodiac 120 C18HZodiac 120 C18AQZodiac 120 C18 EurobondZodiac 120 AC EPSZodiac 120 C18SHZodiac ODSZodiac ODS HZORBAX ODS C18ZORBAX ODS ClassicZORBAX Eclipse Plus C18ZORBAX Eclipse PAHZORBAX Eclipse XDB-C18ZORBAX Extend-C18ZORBAX Rx C18ZORBAX StableBond C18ZORBAX 300 StableBond C18

Wakopak Wakosil 10C18Wakopak Wakosil 5C18 ARWakopak Wakosil 5C18-200Wakopak Wakosil 5C18-200TWakopak Wakosil 5C18-200NWakopak Wakosil 10C18-200Wakopak MS-5C18 GTWakopak Navi C18-5Wakopak Combi ODSWakopak Combi ODS FastXBridge BEH130XBridge BEH300XBridge C18XBridge OST C18XBridge Shield RP18XTerra MS C18XTerra 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...

HPLC C18 column listing (L1-USP)

$$$

A new modern strategy is absolutely necessaryin 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 ActivityPhase compatible with new LC-MS technologyPhase 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

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 High surface area (m2/g) Efficiency pore size (Å) No fines SiliaSphere (SiliCycle) Low metal content High mechanical stability Reproducible loading (critical in reversed-phase)

X

XSupplier A (top)Supplier B (bottom)

(both are real HPLC supports)

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)

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

SiliaChrom-Chemistries Review

Silica-Based HPLC columns for Extremely Acidic Compounds

H3O+

Steric Effect

Note: Not Endcapped silica gel H3O+

SiO

O

Si

OSi

CC

O

O

Si

OH

OO

Si

C

H3C

H3C CH3

CH3

CH3

H3C

C CH3

CH3H3C

CCH3H3C

R

R

CH3

CCH3

CH3

CH3

CH3R=C8 or C18, pH<2.0

New Technology

SiliaChrom-Chemistries ReviewSilica-Based HPLC columns forall type of compounds (pH=2-9)

Note: Partially End capped silica gel

TMS endcapped

Free silianol groups (OH)

R=C18

Ratio of TMS-OH-C18 is well-balanced

New Technology

Polar Surface

SiliaChrom-Chemistries Review

Silica-Based HPLC column forEXTREMELY POLAR compounds

Note: Not Endcapped silica gel

SiO

Si OH

OSi

CH2H3C

CH3

CH2 CH2

HN NH2

O

SiliaChrom HILIC

Normal-Reversed Phases Conditions:

Only SiliaChrom HILIC

SiliaChrom-Chemistries Review

Silica-based HPLC columns for low to high hydrophobic compounds pH=1.5-10.0

Note: Double Endcapped silica gel

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, NH2, CN, 300A, etc…

SiO

Si O

O

Si(CH3)3

Si(CH2)17CH3

H3C

CH3

SiO

Si O

OSi

(CH2)7CH3H3C

CH3

Si(CH3)3

SiO

O

SiO

O

Si

O

O

O

Si

Si

Si

CH3

OO

H3C

O

CH3

O

Si

O

Si

Si

Si

H3C

CH3

CH3

RH3C

CH3

CH3

CH3

CH3

SiliaChrom-Chemistries Review

Silica-Based HPLC columns for EXTREMELY Basic compounds

pH=1.5-12

SiliaChrom XT C18 Series

Replace OH by CH3

(Pseudo-Hybrid tech.)

No bridge(No Backpressure issue as XBridge (Waters))

New Technology

SiliaChrom-Chemistries Review

SiliaChrom SeriesHPLC 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)

(B)(A)

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=7.0Flow rate: 1.0 mL/minDetector: UV scan (PDA), Total scan 200-

600nmTemperature: 23CAll HPLC columns: 4.6x150mm, 5µm

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15Time (min)

-10000

0

10000

20000

30000

40000

50000

60000

70000

80000

90000

100000

110000

120000

130000

140000

150000

uA

U

1.55

7.34

9.73

3.88 4.86

Phenomenex Luna C18(2)

Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline

Low Plate Count (see Results Table)

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Time (min)

-10000

0

10000

20000

30000

40000

50000

60000

70000

80000

90000

100000

110000

120000

130000

140000

150000

160000

170000

180000

uA

U

1.38

6.85

3.294.16 12.99

4.73

Agilent Zorbax SB C18

Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline

Important Tailling

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Time (min)

-10000

0

10000

20000

30000

40000

50000

60000

70000

80000

90000

100000

110000

120000

130000

140000

150000

uA

U

1.49

10.18

8.714.05 5.14

Waters Sunfire C18

Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline

Asymetry

Metal Contamination

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Time (min)

-10000

0

10000

20000

30000

40000

50000

60000

70000

80000

90000

100000

110000

120000

130000

140000

150000

160000

170000

180000

190000

uA

U

1.61

8.75

12.024.15 5.19

SiliaChrom XDB2 C18

Uracil-Toluene-Ethylbenzene-Quinizarin-Amitriptyline

Very good peak shapes

HPLC Column

K’Ethylbenzene

Ethyl/Tol

NEthylbenzene

R AS

(Quinizarine)Tf

(Amitriptyline)

SiliaChrom XDB2 C18 2.22 1.41 28000 3.30 1.10 1.18

Luna C18(2) 2.13 1.42 22000 2.90 1.23 1.20

Zorbax SB C18 2.01 1.45 25000 3.20 1.10 1.55

Sunfire C18 2.45 1.43 35500 3.90 1.80 1.12

SiliaChrom XDB1 C18 3.09 1.44 45000 4.73 1.09 1.65

Summary Results-Case Study

Basic analyte: SiliaChrom XDB2

Less polar analyte: SiliaChrom XDB1

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….

Recent SiliaChrom Applications

Acidic and Basic Compounds (8) with same SiliaChrom HPLC column

1-Maleic Acid

2-Acetaminophen

3-Pseudoephedrine

4-Chlorpheniramine

5-Benzoic Acid

6-Salicylic Acid

7-Dichlorphen

8-Ibuprofen

Recent SiliaChrom ApplicationsAcidic and Basic Compounds (8) with same SiliaChrom HPLC column Phenomenex Luna C18(2) 5µm 4.6x150mm

1

2

34

5

67 8

230 nm

210 nm

230 nm

1

21

2

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

1

2

34

5

67 8

210 nm

230 nm

Recent SiliaChrom ApplicationsFluoroquinolones for Veterinary use by HPLC-Marbofloxacin and Sarafloxacin

SiliaChrom AQ C18 5µm 4.6x150mm

Waters Atlantis dC18 5µm 4.6x150mm Asymmetry

SiliaChrom AQ C18 Atlantis dC18

1.12 1.291.09 1.14

ZwitterionMolecules

Note: SiliCycle has the perfect equivalent (better) with SiliaChrom AQ C18 (or dtC18) than Atlantis dC18 (Waters)

Recent SiliaChrom ApplicationsHalogenated Herbicides by HPLC-Dicamba, Atrazine and 2,4-D

SiliaChrom dtC18 5µm 4.6x150mm

Recent SiliaChrom Applications

Halogenated Herbicides by HPLC-Dicamba, Atrazine and 2,4-D

Supelco Ascentis C18 5µm 4.6x150mm

Asymmetry

SiliaChrom dtC18 Ascentis C18

1.01 1.531.00 0.971.02 1.77

Recent SiliaChrom ApplicationsCatecholamines by HPLC-Polar Compounds

SiliaChrom AQ C18 5µm 4.6x150mmMobile Phase: Water 1.0% AcOH only!

Epinephrine Norepinephrine Dopamine Tyrosine Levodopa

Recent SiliaChrom ApplicationsFaster HPLC with SiliaChrom-Polar and mid-polar Compounds

SiliaChrom AQ C18 5µm 3.0x50mm

Caffeine Chlorthalidone Albendazole Warfarin Tolnaftate Dioctyl Phatalate

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 from AstraZeneca Canada)

Test Mix to evaluate HPLC column from AstraZeneca (Canada)

Recent SiliaChrom Applications

SiliaChrom XDB1 PHENYL 5µm 4.6x150mm

60/40, H2O/MeOH, 1.00 mL/min, 23°C, 254nm : Easy Method!

OH

ClOH

Cl

OH

Cl

Recent SiliaChrom ApplicationsHPLC 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

pH=2.5

50/50, Phosphate Buffer/MeOH, 1.00 mL/min, 23°C, 254nm

pH=7.0

Fronting ?

Assymetry = 0.89

Assymetry = 0.85

Recent SiliaChrom ApplicationsHPLC 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

SiliaChrom AQ C18 (4.6x150mm, 5µm)MeOH:Water, 5:95UV 252nm1mL/min

HPLC Chromatogram Ganciclovir – Polar Compounds (diol-amine)

Recent SiliaChrom Applications

SiliaChrom AQ C18 (4.6x150mm, 5µm)0.4 mM KH2PO4 + 8mM K2HPO4: MeOH, 89:11UV 254nm1mL/min

HPLC Chromatogram of Cefotaxime—Extremely Polar compound (acid-amine-sulfur)

Recent SiliaChrom Applications

SiliaChrom XDB1 C18 (4.6x150mm, 5µm)ACN:Water, 70:30UV 254nm1mL/min

HPLC for mixture of Fenitrothion-Pyriproxyfen—Different Level of Hydrophobicity

Recent SiliaChrom Applications

SiliaChrom XDB2 C18 (4.6x150mm, 5µm)

HPLC Determination for Ephedrine (from Ephedra Sample)

Zorbax SB C18 (4.6x150mm, 5µm)

Recent SiliaChrom Applications

HPLC chromatogram of Berberine-Metal Content Effect

SiliaChrom dtC18 (4.6x250mm, 5µm)ACN: 1%TFA, 35:65UV 346nm1 mL/min

Sunfire C18(4.6x250mm 5µm)

Recent SiliaChrom Applications

SiliaChrom SB C18 (4.6x150mm, 5µm)59:39:2 ACN:Water:TFA (pH = 1.00) UV 270 nm, 1 mL/min

Durability Testing at pH-1.00SiliaChrom SB C18

(Uracil, Phenol, Toluene and Ethylbenzene)

Time/h Retention time(Ethylbenzene)

(min)

Tailing Factor(Ethylbenzene)

(USP)

Theoritical Plates #

(Ethylbenzene) (USP)

0 5.91 1.01 1401424 5.89 1.02 1408548 5.77 1.02 1402372 5.83 1.02 1407696 5.85 1.01 14087

120 5.84 1.02 14050

SD 0.05 0.01 32Mean 5.85 1.02 14056RSD (%) 0.84 0.51 0.23

0 hr

120 hrs

• Poster presented at Pittcon 2010, Orlando FL, USA• LC-GC Application Note—September 2009

Time/h Tailing FactorRetention time(Naphthalene)

(min)

0 1.10 4.857

260 1.14 4.732

300 1.14 4.832

400 1.14 4.873

1 hr

400 hrsSiliaChrom XT C18 (4.6x150mm, 5µm)MeOH: Water/TEA, pH=11.5, 40:60UV 215nm, 1 mL/min

Durability Testing at pH-11.5SiliaChrom XT C18

Naphthalene

1.91

2.25 2.75

1.842.23 2.77

Uracil, Phenol, Nitrobenzene and Naphtalene

SiliaChrom XT C18 (4.6x150mm, 5µm)54.9-44.9-0.2 (ACN-Water-TEA)90C, UV 270 nm 1 mL/min

Durability Testing at pH=11.50, 90CSiliaChrom XT C18 Fidelity

(Uracil, Phenol, Toluene and Ethylbenzene)

0 hr

120 hrs

SiliaChrom HILIC (4.6x150mm, 5µm)ACN 0.1% TFA: Water 0.1%TFA, 10:90UV 280nm1 mL/min

Waters HILIC Silica HPLC column(4.6x150mm, 5µm)

Vitamin B1, Vitamin B6, Vitamin C and Vitamin B2

SiliaChrom HILIC, Vitamins

B1

B6

CB2

Generic Approach with SiliaChrom Series

Final StepPolarityResolutionAcidic, Basic or Neutral

Compounds

SiliaChrom AQ (MeOH/H2O) or

(ACN/H2O), pH=2-9

Not enough retention AcidicSiliaChrom SB

pH=1-2 or HILIC

Basic SiliaChrom XTpH=9-12 Series

or HILIC

SiliaChrom XDB, XDB1 or XDB2

Good Retention, but not enough resolution

SiliaChrom SB, XT or XT Fidelity(low and/or high pH)

Neutral

Note: SiliaChrom AQ is an universal HPLC column (see how particle is designed : TMS-OH-C18)

87% of HPLC success in my Lab!

Conclusion

Low pH High pH High level of Water Hydrophobicity Low Metal Polar Extemely(0.5-2.0) (9-12) Mobile Phase (different Level) Content Analyte Polar Analyte

SiliaChrom SB XT AQXDB, 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!

www.silicycle.com

Thank you!