GL Sciences Inc.

Inertsil 3 Series

New Inertsil ODS Sprint Inertsil Ph-3New Inertsil ODS-3 4um New Inertsil Hilic

Inertsil ODS-3 Inertsil DiolInertsil ODS-3V Inertsil CN-3 Inertsil ODS-EP Inertsil NH2Inertsil ODS-P Inertsil AX

New Inertsil Sulpha C18 Inertsil CXInertsil C8-3 Inertsil SIL-100A

Column Category

GL Sciences Inc.

Inertsil 2 Series

Inertsil ODS-2Inertsil C8Inertsil C4Inertsil PhInertsil SIL-150A

Column Category

GL Sciences Inc.

Inertsil for Proteins and Peptides

Inertsil WP300-C18Inertsil WP300-C8Inertsil WP300-CInertsil WP300-DiolInertsil Peptides-C18Inertsil SIL-150A

Column Category

GL Sciences Inc.

Controlling FactorsBase Silica-Gel Chemical Bonding Selectivity of Packings Inertness of Packings Durability of Packings Column Performance

Inertsil Quality Control

GL Sciences Inc.

QC Tests for Lots

Inertsil Quality Control

Category Method DescriptionBase Silica-Gel SEM Sphericality and Surface Smoothness by Scanning Electron Microscopy.

Ph Particle Size Uniformity by Nitrogen Adsorption Laser Ray Particle Analyzer.M Metals by Atomic Emission and ICPPh Particle Size, Surface Area, Pore Diameter and Pore Volume. by Nitrogen Adsorption, Laser Ray Particle Analyzer

Chemical Bonding C Carbon Loading by Elementary AnalysisSi NMR Residual Silanol by Si-NMR Schro Residual Silanol by Chromatographic test

Selectivity of Packings T Hydrophobicity Tanaka Test MixtureT Stereoselectivity Tanaka Test MixtureT Hydrogen-bonding capability Tanaka Test Mixture

Inertness of Packings P Inertness to Basic compounds PyridineAm Inertness to Basic compounds Ethylaniline, Benzylamine, AminopyridineAc Inertness to Acidic compounds Formic Acid, Acetic AcidCu Inertness to Complexes Oxine-Copper

Durability of Packing D Durability to Low pH mobile phase at pH 2.0D Durability to High pH mobile phase at pH 9.0

GL Sciences Inc.

Inertsil Quality Control

Sphericality and Surface Smoothness by Scanning Electron Microscopy

GL Sciences Inc.

Inertsil Quality Control

Particle Size, Surface Area, Pore Diameter and Pore Volume are Controled by Using Nitrogen Adsorption and Laser Ray Particle Analyzer.

0.000

200.000

400.000

600.000

800.000

1000.000

1200.000

FSQ5-

138

FSQ5-

139

FSQ5-

140

FSQ5-

141

FSQ5-

142

FSQ5-

143

FSQ5-

144

FSQ5-

145

FSQ5-

146

FSQ5-

147

Pore

Volu

me &

Surf

ace A

rea

0.00

20.00

40.00

60.00

80.00

100.00

120.00

Pore

Siz

e

Part

icle

Siz

e

Surface Area (m2/g)

Pore Volume (uL/g)

Pore Size ()

Particle Size (m)

GL Sciences Inc.

Inertsil Quality Control

Packings with an Uniform Size and Smooth SurfaceShow Very Low Column Back Pressure

Column back pressure

0

50

100

150

200

250

0 20 40 60 80 100 120

Methanol concentration in H2O

e

PRODIGY 5 ODS-2 (5um) S/NO 143250

TSKgel ODS-80Ts QA (5um) C-No.W0004

YMC-Pack Pro C18 (5um) No.042527052(W)

J'sphere ODS-M80 (4um) No.042522712(W)

PRODIGY 5u ODS(3) 100A (5um) S/NO 145865

COSMOSIL 5C18-AR (5um) Manf.No.H11279

L-column ODS (5um) No.101097

ZORBAX SB-C18 (5um) Serial No.CL3106

Mightysil RP-18 (5um) Lot.No.960134

Inertsil ODS-3 (5um) TQ5-1219

GL Sciences Inc.

Inertsil Quality Control

High Base Silica-Gel PurityAmount of Metals Are Inpected by

Atomic Emission and ICP

GL Sciences Inc.

Inertsil Quality Control

High Base Silica-Gel Purity ( Amount of Metals Are Inpected by Atomic Emission and ICP)

GL Sciences Inc.

Inertsil Quality Control

Chemicaly Bonded Carbon Loading AreInspected by Elementary Analysis

GL Sciences Inc.

Inertsil Quality Control

Chemical Bonding - Residual Silanol Groups areinspected by 29Si-NMR

Silanol Si-OH

Slica gel structure Si-O-Si

Silanol Si-OH

GL Sciences Inc.

Inertsil Quality Control

Chemical Bonding - Residual Silanol Groups areinspected by 29Si-NMR

remaining silanol

ODS groups

C18H37- Si-O-

GL Sciences Inc.

Inertsil Quality Control

Chemical Bonding - Residual Silanol Groups areinspected by 29Si-NMR

End-capped groups

CH3)3- Si-O-

GL Sciences Inc.

Inertsil Quality Control

29Si-NMR spectra of Inertsil ODS-3 and commercially available ODS packing materials

Inertsil ODS-3Commercially available ODS column A

residual silanol

endcapping

residual silanol

endcappingendcapping

residual silanol

Commercially available ODS column B Commercially available ODS column C

End-capping groupCH3)3- Si-O-

GL Sciences Inc.

Inertsil Quality Control

-Residual Silanol Groups-by Chromatographic Tests (Pyridine/Phenol)

Column : 250x4.6mmI.D. Eluent:CH3OH/H2O=30/70Detection : UV at 254 nmColumn Temperature : 40Peak identification : 1) Pyridine 2) Phenol

0 2 4 6 8 10 12 14 16 18

Time (min)0 2 4 6 8 10 12 14 16 18

Time (min)0 2 4 6 8 10 12 14 16 18

Time (min)0 2 4 6 8 10 12 14 16 18

Time (min)

GL Sciences Inc.

Inertsil Quality Control

GL Sciences is using Prof. Tanakas selectivity test for QC

(n-amylbenzene) / ( n-butylbenzene) : Hydrophobicity, (CH2) (triphenylene) / ( o-terphenyl) : Stereo selectivity, (t/o) (caffeine ) / (phenol) : Hydrogen-bonding capacity, (c/p) According to Prof. Tanakas group: The (CH2) value is a better measure of the surface coverage than the carbon content. The (t/o) values are determined by the functionality of silylating

reagents and the surface coverage. The (c/p) divided by (CH2) illustrates the effect of hydrogen bonding by canceling the difference in the hydrophobic property.

GL Sciences Inc.

Inertsil Quality Control

Separation selectivity test for ODS columnsk (n-Amylbenzene) / k (n-Butylbenzene) : the hydrophobicity of the ODSk (Triphenylene) / k (o-Terphenyl) : the stereo-selectivity of the ODSk (Caffeine) / k (Phenol) : the quantity of unreacted silanol groups in the ODS

0 10 20Time (min)

1)Uracil

2) Caffeine

3) Phenol4) Butylbenzne5)o-Terphenyl6) Amylbenzene

7) Triphenylene H3C N

CH3

C

GL Sciences Inc.

Inertsil Quality Control

-Inertness to Basic Compounds-Pyridine/Phenol

GL Sciences Inc.

Inertsil Quality Control

-Inertness to Acidic Compounds-Formic acid/ Acetic acid

GL Sciences Inc.

Inertsil Quality Control

-Inertness to Chelating Compounds-Oxine Copper

GL Sciences Inc.

Inertsil Quality Control

-Durability to Acidic Condition -0.1% Trifluoroacetic acid in H2O pH2.0

GL Sciences Inc.

Inertsil Quality Control

-Durability to Alkaline Condition CH3OH/0.01M Na2HPO4(pH9)=40/60

GL Sciences Inc.

Inertsil Quality Control

-Column Performance Measurement-

CalucalationN : Number of theoreticalplatesRt : Retention timeW/2 : Half-height width

150x4.6mmI.D. 13,000 more250x4.6mmI.D. 22,000 more

S : peak asymmetry 5%peakheight

1leading,fronting1TailingPeak asymmetry 0.95

1.10

w/2

GL Sciences Inc.

Inertsil Quality Control

GL Sciences Inc.

Inertsil Quality Control

Slide Number 1Slide Number 2Slide Number 3Slide Number 4Slide Number 5 Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control Inertsil Quality Control