Viridis SFC Columns Contents Thank you for choosing a Viridis® SFC column. The Viridis SFC packing materials are designed to provide excellent stability, minimal column bleed, and high mass loading. The Viridis SFC packing materials are GETTING STARTED manufactured in an ISO 9000 certified plant using ultra-pure reagents. a. Safety Considerations b. Column Installation c. Column Equilibration d. Initial Column Efficiency Determination Each batch of Viridis SFC material is tested and the results are held to narrow specification ranges to assure excellent, reproducible performance. Every column is individually tested and a Performance Chromatogram is provided with each column along with the Certificate of Batch Analysis. Table 1. Physical Characteristics V. CONNECTING THE COLUMN TO THE SFC SYSTEM a. Column Connectors and System Tubing Considerations b. Band Spreading Minimization VI. TROUBLESHOOTING VII. COLUMN CLEANING, REGENERATING and STORAGE a. Cleaning and Regeneration b. Storage Carbon Load (%) iV. SCALING UP/DOWN ISOCRATIC METHODS Particle Shape a. Sample Preparation b. Solvents c. Pressure

Each Viridis SFC column comes with a Certificate of Analysis and a Viridis SFC columns are shipped dry. Equilibrate the column with a mini- Performance Test Chromatogram. The Certificate of Analysis is specific mum of 10 column volumes of the mobile phase prior to use. (Refer to to each batch of packing material and includes the gel batch number, Table 2 for a listing of empty column volumes.) analysis of unbonded particle and analysis of bonded particles. The Performance Test Chromatogram is specific to each individual column d. Initial Column Efficiency Determination and contains the gel batch...

3. If the sample is not dissolved in the mobile-phase modifier, Table 3 may be helpful in scaling-up the flow rate to maintain sepa- ensure that the sample, solvent and mobile phases are miscible rations in larger column internal diameters. in order to avoid sample precipitation. Filter sample with 0.2 μm Table 3. Flow Rate Scaling Chart membranes to remove particulates. If the sample is dissolved in a solvent, ensure that the membrane material does not dissolve in the solvent. Contact the membrane manufacturer with solvent Columns and Supplies Catalog for additional information.) filters are...

3. If a leak occurs between the stainless steel compression screw The presence of a void in the flow stream reduces column performance. fitting and the column endfitting, a new compression screw This can occur if a Parker ferrule is connected to a Waters style end- fitting, tubing and ferrule must be assembled. 4. An arrow on the column identification label indicates correct Figure 3: Parker Ferrule in a Waters Style Endfitting direction of solvent flow. Correct connection of 1/16 inch outer diameter stainless steel tubing leading to and from the column is essential for high-quality chromatographic...

ViI. COLUMN CLEANING, REGENERATING AND STORAGE Figure 5 shows the influence of tubing internal diameter on system band-spreading and peak shape. As can be seen, the larger tubing diameter causes excessive peak broadening and lower sensitivity. Changes in peak shape, peak splitting, shoulders on the peak, shifts Figure 5: Effect of Connecting Tubing on System in retention, change in resolution or increasing backpressure may indicate contamination of the column. Flushing with a neat organic solvent, taking care not to precipitate additives, is usually suffi- cient to remove the contaminant. If...

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