viridis Columns Bringing a New Dimension to SFC Combining state-of-the-art media manufacturing with industry leading column technology, Viridis

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1 [ viridis columns ]

2 viridis Columns Bringing a New Dimension to SFC Combining state-of-the-art media manufacturing with industry leading column technology, Viridis Supercritical Fluid Chromatography (SFC) Columns bring a new level of reproducibility to the world of laboratory-scale purification.

3 -Ethylpyridine CSH Fluoro- -Ethylpyridine Manufactured for Reliability and Consistency Based upon over 0 years of experience in the separation science industry, Waters chromatographic media sets the standard for quality and batch-to-batch reproducibility. As a primary manufacturer of silica and hybrid chromatographic particles, all stages of product synthesis are meticulously monitored and controlled, ensuring complete characterization of the finished product. This level of manufacturing process understanding delivers chromatographic media with guaranteed consistent performance year after year.

4 Viridis Hybrid Columns Based on the patented Ethylene-Bridged Hybrid () particle technology and Charged-Surface Hybrid (CSH ) particle technology, Viridis Hybrid Columns offer a wide range of selectivities and batch-to-batch reproducibility. The reduction and control of surface silanol activity on Viridis Hybrid particles delivers, under SFC conditions, excellent peak shapes without the use of mobile-phase additives even for well-retained basic achiral compounds. Three available surface chemistries extend the selectivity range of SFC separations: -Ethylpyridine (-EP) Exhibits good retention, peak shape and selectivity properties without the use of additives. Heightens interaction with polar analyte functional groups, when compared to the less polar surface of -EP. CSH Fluoro- Provides good retention for weak bases and alternate elution orders for acidic and neutral compounds, via significantly different selectivity. Chemistry Shape (μm) Pore (Å) Surface Area (m /g) Pore Volume (cc/g) Carbon Load (%) Endcapped -EP Spherical no Spherical n/a n/a CSH Fluoro- Spherical no Columns:. x 0 mm, µm Gradient: 8 minutes Mobile Phase: 0% methanol without additive Flow Rate:.0 ml/min Column Temp.: 0 C Backpressure: 7 psi (0 bar) Injection Volume: µl Compounds:. Coumarin. Flavone. Caffeine. Thymine. Papaverine. Sulfamethoxazole 7. Cytosine 8. Sulfamethizole 7 8 Viridis -EP Part Number: Viridis S = Part Number: Viridis CSH Fluoro- S = 7 Part Number: min Viridis Hybrid SFC Columns are engineered to provide a wider range of selectivity options.

5 Enhanced Selectivity Viridis Hybrid Columns are designed to offer a wide selectivity range. One measure of how different (or similar) the selectivities of two column chemistries are under a given set of SFC conditions, is the Neue Selectivity Value (S).* As shown below, this parameter may be calculated from a plot of retention factors (k) or retention times (t R ) for analytes on a pair of columns under the same chromatographic conditions. A higher selectivity value indicates a higher degree of orthogonality between two columns and/or two sets of chromatographic conditions. t R Viridis CSH Fluoro R = 0.80 S = t R Viridis -Ethylpyridine The Neue Selectivity Values may be used to estimate the degree of orthogonality between the selectivities of a pair of columns under the same LC or SFC conditions. It is calculated as shown from the coefficient of determination (R = 0.80) of a plot of retention times (or retention factors).* As seen below, on a scale of 0 to 00, a higher S value indicates a higher degree of orthogonality. Compounds: caffeine, theophylline, ibuprofen, fenoprofen, hydrocortisone, ketoprofen, sulfamethoxazole, sulfamethizole, coumarin, flavone, adenine, thymine, uracil, prednisone, prednisolone. Selectivity Values -EP CSH Fluoro EP Selectivity Values (S) compared for pairs of Viridis SFC Columns. S may range from 0 (identical selectivity) to 00 (maximum selectivity difference or orthogonality).* Viridis Hybrid SFC columns are shown in light blue. -EP CSH Fluoro- -EP * () U.D. Neue, E.S. Grumbach, J.R. Mazzeo, K. Tran, and D.M. Wagrowski-Diehl Method development in reversed-phase chromatography Chap. in: I.D. Wilson, ed. Bioanalytical Separations, Handbook of Analytical Separations, Vol. Elsevier, Amsterdam (00); () U.D. Neue, J.E. O Gara, and A. Méndez Selectivity in reversed-phase separations: influence of the stationary phase J. Chromatogr. A 7( ): 7 (00); () U.D. Neue and A. Méndez Selectivity in reversed-phase separations: general influence of solvent type and mobile phase ph J. Sep. Sci. 0(7): 99 9 (007).

6 Viridis Columns Based on Waters long history of chromatographic silica production, the Viridis Columns are designed to be highly reproducible and predictable based on tight product specifications and very low metal content. They are available for both analytical screening and in preparative column dimensions for purification. Separation methods can be optimized and scaled up to Viridis Prep OBD Columns. Chemistry Shape (μm) Pore (Å) Surface Area (m /g) Pore Volume (cc/g) Carbon Load (%) Endcapped -EP Spherical no Spherical n/a n/a -Ethylpyridine (-EP) Widely used in achiral SFC separations exhibiting good retention, peak shape and selectivity properties both with and without the use of additives. Unbonded silica heightens interaction with polar compound functional groups, when compared to the less polar surface of -EP column chemistry, resulting in different selectivity. High Mass Loading Viridis Column vs. Viridis -EP Column Column: Viridis -EP OBD, 9 x 0 mm, µm Part Number: 8009 Gradient Separation:. minutes Mobile Phase: % methanol with no additive Flow Rate: 8 g/min at 0 C Sample Conc.: 80 mg/ml Wavelength: 0 nm System Backpressure: 0 bar Compounds:. Coumarin. Flavone. Ibuprofen. Carbamazepine Injection Vol.: 00 µl Total Mass Load: 8 mg Gradient Separation:. minutes Mobile Phase: % methanol with no additive Flow Rate: 8 g/min at 0 C Wavelength: 0 nm System Backpressure: 0 bar.0e-.0e- Compounds:. Flavone. Ibuprofen. Carbamazepine Viridis OBD Column 9 x 0 mm, µm Part Number: e- 0.0 Injection Vol.: 0 µl Total Mass Load: 8 mg.0e- Viridis -EP OBD Column 9 x 0 mm, µm Part Number: e-.0e- Injection Vol.: 70 µl Total Mass Load: 0 mg min min Viridis -EP Columns provide excellent mass loading to ensure maximum throughput. Using the same SFC gradient conditions, different selectivity can be achieved by using a Viridis Column or Viridis -EP Column. The retention time for ibuprofen was reduced and the carbamazepine retention time was increased by switching column chemistries, with little change in the retention of flavone.

7 OBD Column Design Improved Mechanical Stability Laboratory-scale SFC purification presents many challenges to the chromatographer. Inconsistency in column-to-column performance and lifetimes often result in lost samples, repeat purification runs and poor scalability from small- to large-volume columns. Common SFC preparative column failures are due to mechanical instabilities, especially voids in the packed bed. The Viridis Preparative OBD Columns combine the patented* optimized packing process with a unique set of column designs ensuring increased lifetime and greater reproducibility. All Viridis Column characteristics are available in the preparative OBD column format. * US Patent # 7,99,0, UK Patent # GB089 Batch-to-Batch Reproducibility Waters is dedicated to maintain the tightest specifications in the SFC industry. Controlled manufacturing processes and column packing procedures ensure that you receive the best, most reproducible SFC column available. Batch A Batch B µm Column: Viridis -EP,. x 0 mm, µm Part Number: 800 Flow Rate: 0. ml/min Mobile Phase: 90/0 carbon dioxide/methanol Backpressure (BPR): 7 psi (0 bar) Column Temp.: 0 C System: UPSFC (alpha) Injection Vol.:.0 μl Detection: UV at 0 nm (compensated wavelengths 0 0 nm) Compounds:. Toluene. Flavone. Thymine. Papaverine. Prednisolone. Sulfanilamide Batch C Batch D Batch E min This excellent reproducibility is a result of our commitment to maintaining the tightest specifications in the industry. Viridis Hybrid Columns start with high purity raw materials, and are produced using controlled manufacturing processes and column packing procedures that provide today s scientists with the most reproducible SFC column available. Ordering Information Viridis Analytical SFC Columns Dimension -EP CSH Fluoro- -EP. x 0 mm µm x 0 mm μm x 00 mm μm x 0 mm μm x 0 mm μm x 0 mm μm x 00 mm μm x 0 mm μm x 0 mm μm Viridis Preparative SFC Columns Dimension -EP CSH Fluoro- -EP 0 x 0 mm μm x 00 mm μm x 0 mm μm x 0 mm μm OBD 9 x 0 mm μm OBD 9 x 00 mm μm OBD 9 x 0 mm μm OBD 9 x 0 mm μm OBD 0 x 0 mm μm OBD 0 x 7 mm μm OBD 0 x 00 mm μm OBD 0 x 0 mm μm OBD 0 x 0 mm μm OBD 0 x 0 mm μm OBD 0 x 00 mm μm OBD 0 x 0 mm μm OBD 0 x 0 mm μm

8 Sales Offices: Austria and European Export (Central South Eastern Europe, CIS and Middle East) Australia Belgium and Luxembourg Brazil 788 Canada China 8 Czech Republic Denmark Finland France Germany Hong Kong Hungary 0 08 India Ireland 8 00 Italy Japan Korea Mexico The Netherlands Norway Poland Puerto Rico Russia/CIS / Singapore Spain Sweden 8 00 Switzerland Taiwan United Kingdom All other countries: Waters Corporation U.S.A The quality management system of Waters manufacturing facilities in Taunton, Massachusetts and Wexford, Ireland complies with the International Standard ISO 900 Quality Management and Quality Assurance Standards. Waters quality management system is periodically audited by the registering body to ensure compliance. Waters and Viridis are registered trademarks of Waters Corporation. Technology, CSH, OBD, UPSFC, and The Science of What s Possible are trademarks of Waters Corporation. All other trademarks are the property of their respective owners. 0 Waters Corporation. Printed in the U.S.A. July EN SC-FP