YMC-Triart C18. Multi-layer hybrid material for HPLC ph and temperature stability. Innovative micro-reactor technology

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YMC-Triart C18 Versatile hybrid silica based HPLC Column The Selectivity Company Multi-layer hybrid material for HPLC ph and temperature stability Innovative micro-reactor technology www.ymc.de

2 YMC-Triart Particle Technology... page 4 Physicochemical Characteristics... page 5-6 Surface Modification... page 6-7 Reproducibility Data... page 8-9 Application... page 10-14 Ordering Information... page 15

3 YMC-Triart Chromatographers are seeking versatile HPLC columns which will perform day-to-day with ever-changing ph, buffers and temperature conditions. In order to meet these challenging demands we need HPLC columns that are able to cope with extreme conditions. Where can one find a column suitable for harsh ph conditions in combination with high temperature ranges without sacrificing selectivity? YMC-Triart: the versatile multi-layered hybrid silica material! Multi-layer hybrid material for HPLC ph and temperature stability Innovative micro-reactor technology

4 Particle technology YMC-Triart is a multi-layered material prepared from well-controlled particle formation technology which is adapted from micro-reactor technology. The recently developed production step results in truly narrow particle and pore size distributions! With YMC-Triart, challenging ph and high temperature conditions are no longer a limitation to the day-to-day work in laboratories. Most importantly, due to its unique particle composition, a balanced hydrophobicity and silanol activity are achieved which makes YMC-Triart a First Choice column in method development! YMC-Triart hybrid structure inorganic silane layer organic silane layer

5 Uniform spherical particles YMC-Triart C18 X-Bridge HILIC The new uniform spherical particle support is adopted for YMC-Triart C18. The particle is produced using micro-reactor technology in granulation process. This results in reduction of the back-pressure and leads to more reproducibility in surface modification. Low column back-pressure Column: 5 µm, 150 x 4.6 mm ID Eluent: acetonitrile/water or methanol/water Flow rate: 1.0 ml/min Temperature: 25 C The revolutionary production technique, adapted from micro-reactor flow technology, produces a multi-layered silcia/ organic hybrid stationary phase, with outstanding narrow pore size and particle size distributions which result in low back pressures. YMC-Triart C18 is designed for use under many conditions. Elution with higher viscosity methanol (compared with acetonitrile), YMC-Triart C18 generates lower pressure (approx 30% lower than with conventional phases).

6 Narrow pore distribution 4500 d Vp/ d logd 4000 3500 3000 2500 2000 1500 1000 500 Triart C18 particle Conventional silica-based particle X-Bridge HILIC 0 0 5 10 15 20 Pore size (nm) 25 30 35 This figure shows the pore size distributions of some competitive materials. Compared with other products on the market, YMC-Triart C18 shows a narrower distributon which results in sharper peak shapes. Improved loadability Influence of injection volume on peak shapes 1 2 YMC-Triart C18 solvent for sample acetonitrile injection volume 1.0 µl 1.5 µl 2.0 µl 3.0 µl 1 2 1 Hybrid-silica based C18 2 Silica-based C18 Peak anomaly at 1.5 µl Column: 5 µm, 50 x 2.0 or 2.1 mm ID Eluent: A) water / formic acid (100/0.1) B) acetonitrile / formic acid (100/0.1) 5% B (0-0.5 min), 5-100% B (0.5-2.5 min) Flow rate: 0.4 ml/min Detection: UV at 275 nm 1. OCH2CHCH2NHCH(CH3)2 OH 2. H3CO H3CO CN CH3 C (CH2)3NCH2CH2 OCH3 OCH3 1.5 2.0 2.5 min Propranolol 50 µg/ml CH(CH3)2 Verapamil 50 µg/ml In order to prevent peak errors, there is the limit to the injection volume when the sample is injected in high elution solvents (such as acetonitrile). Compared with traditional columns, more than double the injection volume can injected into YMC-Triart C18 as a result of the extremely narrow particle size distribution.

7 Multi-stage endcapping After bonding the alkyl chain, there are highly reactive and less reactive silanols on the surface. In traditional bonding processes, these reacted with a single capping-compound in one step. However, the highly reactive silanols can be hydrolysed easily which contributes to the poor durability. The less reactive silanols are hard to endcap which results in poor resolution due to peak tailing. Before EC 1 STEP Hydroxyl group End-capping (EC) group 2 STEP 3 STEP YMC-Triart C18 uses a new innovation in end capping called multistage end-capping for its surface modification process. By using a number of compounds with the different reactivities in successive steps, all silanols can be capped to the maximum extent. Ingredients in cough/cold medication: 1. Chlorpheniramine; 2. Detromethorphan; 3. Propyl p-hydroxybenzoate Column: 5 µm, 150 x 3.0 mm ID Eluent: 20 mm KH 2 PO 4 -K 2 HPO 4 (ph 6.9) / acetonitrile (65/35) Flow rate: 0.425 ml/min Detection: UV at 235 nm

8 Batch-to-Batch reproducibility Excellent reproducibility of YMC-Triart C18 is shown even for the analysis for basic and coordination compounds which normally exhibit tailing and adsorption effects. Basic compounds 1 2 3 Coordination compounds 1 2 Lot. E Lot. E Lot. D Lot. D Lot. C Lot. C Lot. B Lot. B Lot. A Lot. A 0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5 min 0.0 2.0 4.0 6.0 8.0 10.0 12.0 14.0 min J091201Y J091201X Column: 5 µm, 150 x 3.0 mm ID Eluent: 20mM KH 2 PO 4 (ph 6.9) / acetonitrile (65/35) Flow rate: 0.425 ml/min Detection: UV at 235 nm Column: 5 µm, 150 x 3.0 mm ID Eluent: acetonitrile / 0.1% H 3 PO 4 (40/60) Flow rate: 0.425 ml/min Detection: UV at 254 nm The reproducibility of packed columns is shown below in terms of theoretical plate number (N) and tailing factor (Tf). YMC Triart C18 packed columns exhibit a very narrow range of variation. 20000 2.5 Theoretical plate number 15000 10000 5000 Theoretical plate number Tailing factor 2.0 1.5 1.0 Tailing factor 0 10 20 30 40 50 Column number 0.5 Column: 5 µm, 150 x 4.6 mm ID Eluent: acetonitrile / water (40/60) Flow rate: 1.0 ml/min Temperature: ambient Sample: butyl benzoate

9 Versatile wide ph stability Phosphate buffer (ph 11.5, 40 C) Rate of theoretical plate number 120% 100% 80% 60% 40% 20% Silica based C18 YMC-Triart C18 Hybrid-silica based C18 0% 0 10 20 30 40 50 Time (hr) Column: 5 µm, 150 x 4.6 mm ID Eluent: 50 mm K 2 HPO 4 -K 3 PO 4 (ph 11.5) / methanol (90/10) Flow rate: 1.0 ml/min Sample: benzyl alcohol Triethylamine (ph 11.5, 40 C) Rate of theoretical plate number 120% 100% 80% 60% 40% 20% Silica based C18 YMC-Triart C18 0% 0 50 100 150 200 250 300 Time (hr) Column: 5 µm, 150 x 4.6 mm ID Eluent: 50 mm triethylamine (ph 11.5) / methanol (90/10) Flow rate: 1.0 ml/min Sample: benzyl alcohol Durability at high temperature ph 6.9, 70 C Rate of theoretical plate number 120% 100% 80% 60% 40% 20% Silica based C18 YMC-Triart C18 Silica based C18 alkaline-resistant type 0% 0 100 200 300 400 500 600 700 Time (hr) Column: 5 µm, 50 x 2.0 mm ID Eluent: 20 mm KH 2 PO 4 -K 2 HPO 4 (ph 6.9) / acetonitrile (90/10) Flow rate: 0.2 ml/min Temperature: 70 C Sample: phenol ph 1, 70 C Rate of theoretical plate number 120% 100% 80% 60% 40% 20% 0% 0 20 40 60 80 100 120 Time (hr) Test conditions Column: 5 µm, 50 x 2.0 mm ID Eluent: acetonitrile / water (60/40) Flow rate: 0.2 ml/min Temperature: 70 C Sample: butyl benzoate YMC-Triart C18 Silica based C18 Hybrid-silica based C18 The columns are stored in acetonitrile/water/tfa (10/90/1) 70 C, and the performance tested every 20 hours. YMC-Triart C18 shows great chemical stability provided by newly developed hybrid-silica. Even in high-ph or high-temperature conditions, the lifetime of YMC-Triart C18 is more than 10x greater than conventional ODS columns.

10 First choice column for method development Hydrogen boning capacity a (caffeine/benzene) Standard ODS The most versatiled columns as first-choice. Hydrophobicity k (amylbenene) Conventional hybrid silica-based ODS columns tend to be less hydrophobic than silica-based columns. YMC-Triart C18 has a higher carbon load, making its hydrophobicity comparable to standard ODS columns, making it a versatile first-choice column for method development. Specification Base organic/inorganic silica Stationary phase C18 (as USP L1) Particle size 3 and 5 µm Pore size 12 nm Carbon load 20% Bonding polymeric type End-capping multi-stage hybrid groups ph range 1 ~ 12 Temperature range ph 1-7: 70 C, ph 7-12: 50 C

11 Basic compounds Column: 5 µm, 150 x 3.0 or 150 x 4.6 mm ID Eluent: 20 mm KH 2 PO 4 -K 2 HPO 4 (ph 6.9) / acetonitrile (65/35) Flow rate: 0.425 ml/min for 3.0 mm ID 1.0 ml/min for 4.6 mm ID Detection: UV at 235 nm The innovative surface modification technology results in excellent peak shapes even for basic compounds that often exhibit peak tailing with conventional silica- and hybrid silicabased ODS columns. Fast LC for conventional HPLC insufficient separation optimisation Baseline separation, but long analysis time Down scaling Eluent: A) water/hfba* (100/0.1) B) acetonitrile/hfba* (100/0.1) Temperature: 37 C Detection: UV at 290 nm Injection: 1 µl (0.1 mg/ml) *heptafluorobutyric acid

12 Coordinating compounds Column: 5 µm, 150 x 3.0 or 150 x 4.6 mm ID Eluent: acetonitrile / 0.1% H 3 PO 4 (40/60) Flow rate: 0.425 ml/min for 3.0 mm ID 1.0 ml/min for 4.6 mm ID Detection: UV at 254 nm YMC-Triart C18 has an extremely low level of metal impurities, much lower than conventional products, enabling to provide excellent peak shape for coordination compounds. Acidic compounds Column: 5 µm, 150 x 3.0 or 150 x 4.6 mm ID Eluent: methanol / 0.1% H 3 PO 4 (5/95) Flow rate: 0.425 ml/min for 3.0 mm ID 1.0 ml/min for 4.6 mm ID Temperature: 37 C Detection: UV at 210 nm YMC-Triart C18 is synthesised using methodology adapted from micro-reactor technology. This technique ensures a reduction of impurities that contribute to peak tailing during the analysis of some types acidic compounds.

13 Comparison of clemastine analysis with conventional columns 10 mm phosphate buffer (ph 6.7)/organic solvent 10 mm CH3COONH4/organic solvent methanol acetonitrile methanol acetonitrile YMC-Triart C18 C18 50 X x 2.0 mmi.d. ID Tf 0.97 Tf 1.11 Tf 1.03 Tf 1.16 F090818H07 F090818I07 F090818F07 F090817G07 Silica based C18 C18 Inertsil Brand ODS-3V C1 50 X x 2.1 mmi.d. ID Tf 1.21 F090727H07 Tf 1.56 F090727I07 Tf 1.13 F090727F07 Tf 1.43 F090803G07 Silica Silica based based C18 C18 Capcell Brand D5 Pak MG-II 50 X x 2.0 mmi.d. ID Tf 0.93 F090717H07 Tf 2.83 F090717I07 Tf 1.05 F090714F07 Tf 2.21 F090714G07 Silica based C18 Silica based C18 Sunfire C18 Brand I6 50 50 x X 2.1 2.1 mmi.d. mm ID Tf 1.03 Tf 3.87 Tf 1.13 Tf 1.88 F090722H07 F090722I07 F090722F07 F090722G07 Hybrid-silica based C18 X-Bridge 50 x 2.1 mm ID Tf 1.14 F090731H07 Tf 1.88 F090731I07 Tf 1.22 F090731F07 10.0 11.0 12.0 min 6.0 7.0 8.0 min 8.0 9.0 10.0 min 6.0 7.0 Tf 1.78 F090731G07 8.0 min Column: 5 µm, 50 x 2.0 or 50 x 2.1 mm ID Eluent: A) 10 mm KH 2 PO 4 -K 2 HPO 4 (ph 6.7) or 10 mm CH 3 COONH 4 B) methanol or acetonitrile 5-90% B (0-10 min), 90% B (10-15 min) Flow rate: 0.2 ml/min Temperature: 25 C Detection: UV at 230 nm Clemastine is a well-known basic compound which readily exhibits peak tailing with conventional ODS columns. YMC-Triart provides sharp separations with many different buffer/ solvent compositions.

14 LC/MS compatibility Column: 5 µm, 50 x 2.0 mm ID Eluent: A) water / formic acid (100/0.1) B) acetonitrile / formic acid (100/0.1) 5% B (0-1 min), 5-100% B (1-5 min), 100% B (5-10 min), 100-5% B (10-10.1 min), 5% B (10.1-12.5 min) Flow rate: 0.4 ml/min Detection: ESI positive, TIC (Mass Range: 50-1000) Column bleeding, caused by the fragments of stationary phase, is the main reason for background noise and limitations on detection limits. No bleed is observed in the test of total ion current (TIC) measured by LC/MS with blank or with YMC-Triart C18. So in terms of the signalnoise ratio (S/N ratio), YMC-Triart C18 can be expect to not only reduce the background noise but to also increase the sensitivity of the analysis. Sure-Fit Connectors Optimise your chromatographic results! Easy column installation without tools Perfect capillary connection every time Optimised resolution and reproducibility Self-adjusting to any port depth regardless of column manufacturer Fingertight to 6000 psi (425 bar) No galling or seizing Available with a range of capillary tubing diameters and lengths for optimal resolution

15 Ordering Information Particle size (µm) Pore size (nm) Dimension (mm) Part number Particle size (µm) Pore size (nm) Dimension (mm) Part number 50 x 2.0 TA12S030502 50 x 2.0 TA12S050502 75 x 2.0 TA12S03L502 75 x 2.0 TA12S05L502 100 x 2.0 TA12S031002 100 x 2.0 TA12S051002 150 x 2.0 TA12S031502 150 x 2.0 TA12S051502 50 x 3.0 TA12S030503 50 x 3.0 TA12S050503 75 x 3.0 TA12S03L503 75 x 3.0 TA12S05L503 3 12 100 x 3.0 TA12S031003 5 12 100 x 3.0 TA12S051003 150 x 3.0 TA12S031503 150 x 3.0 TA12S051503 50 x 4.6 TA12S030546 50 x 4.6 TA12S050546 75 x 4.6 TA12S03L546 75 x 4.6 TA12S05L546 100 x 4.6 TA12S031046 100 x 4.6 TA12S051046 150 x 4.6 TA12S031546 150 x 4.6 TA12S051546 250 x 4.6 TA12S032546 250 x 4.6 TA12S052546 Trademarks SureFit is trademark of Isolation Technologies Capcell Pak MG-II is trademark of Shiseido Co., Ltd. Inertsil ODS-3V is trademark of GL Sciences Sunfire C18, X-Bridge are trademarks of Waters Corp. Gemini NX is trademark of Phenomenex, Inc.

Your local distributor: Hichrom Limited 1 The Markham Centre, Theale, Reading, Berkshire, RG7 4PE Tel: 0118 930 3660, Fax: 0118 932 3484 email: technical@hichrom.co.uk Europe GmbH Schöttmannshof 19 D-46539 Dinslaken Germany TEL. +49(0)2064/427-0, FAX +49(0)2064/427-222 www.ymc.de Co., Ltd. YMC Karasuma-Gojo Bld. 284 Daigo-cho, Karasuma Nisiiru Gojo-dori Shimogyo-ku, Kyoto 600-8106 Japan TEL. +81(0)75-342-4515, FAX +81(0)75-342-4550 www.ymc.co.jp America, Inc. 941 Marcon Boulevard Suite 301 Allentown, PA18109 USA TEL. +1-610-266-8650, FAX +1-610-266-8652 www.ymcamerica.com India Ltd. CX - 07, 3rd Floor, Lobe - 1, Tower - A, The Corenthum, Plot No- A-41, Sector - 62, Noida - 201301 (UP) India. TEL. +91(0)120-4276020 - 25, FAX +91(0)120-4276026 www.ymcindia.com