Uptisphere CS Evolution Core Shell columns for fast & highly efficient identification & quantification of small molecules

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Uptisphere CS Evolution Core Shell columns for fast & highly efficient identification & quantification of small molecules www.interchim.com www.blog.interchim.com www.forum.interchim.com www.facebook.com/interchim

Uptisphere CS Evolution superficially porous & mono-dispersed silica Uptisphere CS Evolution consists of superficially porous & mono dispersed 2,6μm particles of high purity silica. They are formed by a solid core of 1.6 microns diameter and welded to its surface, a totally porous silica layer of 0,50 μm thickness. Benefit of its technology: Thin diffusion zone which allows rapid mass transfer. Mono-dispersed and "heavy" particles ensure optimum columns packing. Efficiency & reproducibility are superior to equivalent totally porous columns, h <1.8. Low back pressure Uptisphere CS Evolution guarantee high efficiencies and very fast separations with lower back pressures for (U)HPLC applications.

Core-Shell technology boots productivity without compromising with resolution.

CS Evolution silica characterization Particle size Superposition of three consecutive measurements: very good homogeneity One Gaussian, narrow, ranging from 1.7 to 3.1 microns Analyzes done with Mastersizer 2000 wet process (suspended in acetonitrile) Laboratoire SMS - université de Rouen

CS Evolution silica characterization Surface Area BET = 126 m 2 /g Laboratoire SMS - université de Rouen 5

Uptisphere CS Evolution superficially porous & mono-dispersed silica Maximum Operational Surface Technology (MOST) Interchim technology advantages & benefits Before bonding, Uptisphere CS Evolution particles undergo a proprietary treatment similar to a recondensation of the porous silica layer, which has the effect of maximizing the operational surface density. Enhance surface with selected silanol ready for bonding for both hilic & reverse phase mode High temperature stability Allows direct transfer from Uptisphere CS Evolution to Interchim purification stationary phases

29 Si RMN after ITM proprietary treatment & C18 bonding Red = micro wave Blue = reflux φ 3 - no bonding detected (M) Step 2 - reflux + MW φ 2 - reflux + MW Step φ 1-1 MW - reflux + MW φ 1 - reflux + MW Laboratoire SMS - université de Rouen

Elementary analysis after ITM proprietary treatment & C18 bonding Synthesis Bonding type n c M w (g. mol -1 ) Treatment ( C) %C Grafting rate * (µmol/m 2 ) Microvawe -Si( ) 2 C 18 H 37 20 311,65 Regular 9,54 3,60 Microvawe -Si( ) 2 C 18 H 37 20 311,65 φ 1 9,06 3,39 Microvawe -Si( ) 2 C 18 H 37 20 311,65 φ 2 7,68 2,82 Microvawe -Si( ) 2 C 18 H 37 20 311,65 φ 3 0,50 0,17 * Based on a surface area of 126 m²/g Impact on chromatographic results Regular Step 1 Step 2 Qty of alkyl chains 5,24 4,53 5,34 Hydrophobicity 1,51 1,53 1,51 Steric Selectivity 1,50 1,74 1,55 Potential of hydrogen links 0,43 0,37 0,52 Exchange potential @ ph: 2.7 0,06 0,05 0,05 Exchange potential @ ph: 7.6 0,30 0,30 0,68 Laboratoire SMS - université de Rouen

Thermogravimetric studies High temperature stability C18 bonded & end-capped material Empty crucible (%) Normalized C18 silica (%) 20 to 300 C 0-0,1 300 to 600 C - 0,5-9,5 600 to 900 C - 1,75-0,5 Total loss 20 to 1000 C - 3-10,5 9

C18-HB Selectivity Uptisphere CS Evolution Capacity Productivity O SI O SI C 18 H 37 Uptisphere CS Evolution C18-HB 85Å - 130m 2 /g 2.6 µm Bonding: C18 monofonctionnal %C: 8.0 End-capping: one-step Maximum Operational Surface Technology ph stability: 1.5 to 8.0 USP code: L1 Suitable for non polar compounds separation. Exhibits a very hydrophobic surface. HB shows excellent mechanical stability under high temperature. Application: non-polar organic compounds

C18 Selectivity Uptisphere CS Evolution Capacity Productivity O SI OH O SI C 18 H 37 Uptisphere CS Evolution C18 85Å - 130m 2 /g 2.6 µm Bonding: C18 monofonctionnal %C: 9,0 End-capping: one-step Maximum Operational Surface Technology ph stability: 1.5 to 7.5 USP code: L1 Serves a broad-ship of analytical & prep LC requirements for separating non polar compounds Application: non-polar organic compounds

C18-RP Selectivity Uptisphere CS Evolution Capacity Productivity O R O O SI C 18 H 37 Uptisphere CS Evolution C18-RP 85Å - 130m 2 /g 2.6 µm Bonding: C18 monofonctionnal %C: 6,0 End-capping: multi step mixte ph stability: 1.5 to 8.0 USP code: L1 Application: mid-polar organic compounds Suitable for mid & non polar compounds separation. RP shows excellent mechanical stability that provides long-life & make it an excellent tool for analysis under acidic or basic conditions.

Hydrophobic selectivity (Tanaka Méthylène) CS competitor 2.7µm C18-50 x 4.6mm CS Evolution 2.6µm C18-RP - 50 x 4.6mm Uptisphere 3.0µm C18-RP - 150 x 4.6mm 1) Uracil, 2) Ethylbenzene, 3) n-propylebenzene, 4) n-butylbenzene, 5) n-pentylbenzene MeOH-H2O @ 1 ml/min - UV: 254 nm Similar selectivity for Hydrophobic probes CS competitor C18 2.7µm 50 x 4.6mm CS Evolution C18-RP 2.6µm 50 x 4.6mm Uptisphere C18-RP 3µm 150 x 4.6mm α butylbenzene/pentylbenzene 1.56 1.54 1.50

Polar selectivity CS competitor 2.7µm C18-50 x 4.6mm 1) Uracil, 2) Benzoic acid, 3) p-toluic acid, 4) p-ethylbenzoic acid, 5) Methyl benzoate ACN-formic acid buffer ph: 2.8 @ 1 ml/min - UV: 254 nm Better selectivity & resolution for Polar probes CS concurrente C18 2.7µm 50 x 4.6mm CS Evolution C18-RP 2.6µm 50 x 4.6mm Ethylbenzoic /methyl benzoate 1.36 1.39 Rs Ethylbenzoic /methyl benzoate 6.63 7.19 Uptisphere CS Evolution 2.6µm C18-RP - 50 x 4.6mm

Basic compounds CS competitor 2.7µm C18-50 x 4.6mm Uptisphere CS Evolution 2.6µm C18-RP - 50 x 4.6mm 1) Uracil, 2) p-hydroxybenzoic acid, 3) Pyridine, 4) Dimethylpyridine ACN-buffer ph: 5.7 @ 1 ml/min - UV: 254 nm Good resolution & symmetry with Basic probes CS concurrente C18 2.7µm 50 x 4.6mm CS Evolution C18-RP 2.6µm 50 x 4.6mm pyridine/dimepyridine - 1.19 Rs pyridine/dimepyridine - 2.39

C18-AQ Selectivity Uptisphere CS Evolution Capacity Productivity O P OH O SI C 18 H 37 Uptisphere CS Evolution C18-AQ 85Å - 130m 2 /g 2.6 µm Bonding: C18 monofonctionnal %C: 6.5% End-capping: Mixte ph stability: 1.5 to 7.0 USP code: L1 Suitable for mid & non polar compounds separation. RP shows excellent mechanical stability under 100% aqueous mobile phase conditions. Application: mid-polar organic compounds 100% water compatible

RP/SCX Selectivity Uptisphere CS Evolution Capacity Productivity O SI C n H 2n+1 OH O O SI (CH 2 ) 3 O S H + USP code: L44 O O - Uptisphere CS Evolution RP/SCX 85Å - 130m 2 /g 2.6 µm Bonding: mixed mode RP/SCX monofonctionnal %C: 8.5 End-capping: one step ph stability: 2.0 to 7.0 Ion exchange & hydrophobic chains are bonded onto the surface of silica providing unique selectivity. Compounds that possess basic functionality are retained by ion exchange functionality. Application: Mid-polar & non polar organic compounds under cationic form

MVP Selectivity Uptisphere CS Evolution Capacity Productivity O SI (CH 2 ) n PEG OH O SI (CH 2 ) n PEG USP code: Uptisphere CS Evolution MVP 85Å - 130m 2 /g 2.6 µm Bonding: monofonctionnal %C: 7.0 End-capping: propriétaire ph stability: 2.0 to 7.5 Suitable for Alkylbenznes, PAHs applications Application: Planar & PI compounds. Peptides

Linear regressions of log k in function of log P o/w for an homologous serie of ( ) Alkylbenzenes & ( ) PAHs log k log k log k log k 1,50 1,00 0,50 competitor C18 R² = 0,9966 0,80 0,70 0,60 0,50 0,40 0,30 CS Evolution MVP R² = 0,9884 R² = 0,9968 0,20 0,00-0,50 0 2 4 6 8 10 R² = 0,9863 0,10 0,00-0,10 0 2 4 6 8 10-1,00 CN-H 2 O 85:15, v/v log P o/w -0,20-0,30 CN-H 2 O 60:40, v/v log P O/W 2,50 2,00 competitor Amide R² = 0,994 1,80 1,60 1,40 competitor Phenyl-Hexyl R² = 0,9966 1,50 1,20 1,00 1,00 0,80 0,60 0,50 0,00 R² = 0,985 CN-H 2 O 60:40, v/v 0 2 4 6 8 10 0,40 0,20 0,00 R² = 0,9916 CN-H 2 O 60:40, v/v 0 2 4 log P 6 8 10 o/w log P o/w

Hilic-HIT Selectivity Uptisphere CS Evolution Capacity Productivity O O H H Uptisphere CS Evolution Hilic-HIT 85Å - 130m 2 /g 2.6 µm Bonding: proprietary End-capping: proprietary ph stability: 1.5 to 7.0 Maximum Operational Surface Technology USP code: L3 Application: water-soluble compounds Aqueous normal phase separation (ANP) of water-soluble compounds. Typical mobile phase: water / ACN (> 70%) ANP is an excellent alternative to RP purification for highly polar compounds

Selectivity, Efficiency & Resolution CS competitor 2.7µm Hilic - 50 x 4.6mm Uptisphere CS Evolution 2.6µm HIT - 50 x 4.6mm

Symmetry for basic compounds As pyridine : 1,86 As 2.6-dymethylpyridine : 3,43 As pyridine : 1,70 As 2.6-dymethylpyridine : 2,82 CS competitor 2.7µm Hilic - 50 x 4.6mm Uptisphere CS Evolution 2.6µm HIT - 50 x 4.6mm

Stimulant compounds (90/10) - ACN/Acétate d ammonium 40 mm Flow rate = 1,5 ml/min T = 35 C UV : 254 nm Sample: 1 - norephedrin 2 - ephedrin 3 - hydroxyephedrin 4 - synephrin Uptisphere CS Evolution 2.6µm HIT

Aromatic Amino-acids (85/15) - ACN/Acétate d ammonium 100 mm ph: 4 Flow rate = 0,5 ml/min T = 25 C UV : 254 nm Samples : 1 - tryptophan 2 - phenylalanin 3 - tyrosin Uptisphere CS Evolution 2.6µm HIT

Silica Selectivity Uptisphere CS Evolution Capacity Productivity O SI OH Uptisphere CS Evolution Silica 85Å - 130m 2 /g 2.6 µm Bonding: none End-capping: none ph stability: 1.5 to 7.0 USP code: L3 Specific of non-ionic, polar organic compounds analysis Application: Non-ionic, polar organic compounds

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