3-1. Mobile Phase 1. All solvents acceptable for the conventional chemically bonded silica columns can be used. 2. The acceptable pH range for Proteonavi is 2 to 10. 3. After full degassing, filtrate the mobile phase using a membrane filter 0.45 µm or smaller to remove dust. A 2-µm filter is used at the column inlet. To prevent foreign matter from clogging the column inlet filter, we recommend using a line filter. 4. The mobile phase stated in the report is sealed in a new column. To change to a mobile phase containing inorganic salt, note the replacement procedure. 5. To prevent column deterioration, avoid the following: •Frequent change of the mobile phase composition or direct change to a mobile phase of low compatibility •Rapid change in column inlet pressure •High column pressure due to the use of a high-viscosity mobile phase •Prolonged water flow 3-2. Preparing a Sample Solution 1. Dissolve the sample in a solvent of the same composition as the eluate wherever possible. 2. Using a solvent with strong dissolving power may lower the separation efficiency or cause the sample to precipitate at the column head. 3. If there is insoluble matter remaining in the sample solution, filtrate the solution using a filter 0.45 µm or smaller. 4. The pH of the sample solution should be set in acceptable pH range for packing material. 3-2. Notes on Analysis 1. Regarding the guard column: Use a column of the same packing material as the main column. If a guard column of different packing material or the chemically bonded silica column of a different manufacturer is used, a separation profile may not be as expected. 2. After gradient analysis is performed, replace the column with eluent containing high content of organic solvent. See 4. for storing.

1. Seal the column with the accessory plug and store it in a cold place where there is little temperature fluctuation. 2. For storage, replace the mobile phase with a solution of acetonitrile and water having the same composition as the mobile phase and then fill it with acetonitrile. Note the replacement procedure to avoid salting out. 3. Avoid using 100% water to rinse a column.

1. An analytical column of up to 6-mm ID uses a filter-embedded end fitting as shown in Fig. 1. The filter cannot be changed alone. If the filter is clogged or the column pressure is high, replace the end fitting. See Table 3 for the replacement parts and repair items. 2. See Fig. 1 for the column connection. If the tubing is inappropriate, especially if a tube for a different type of column is used, the length after the ferrule tip (V in Fig. 1) is often different from the end fitting length L, and a problem may occur. If L is greater than V, dead volume may be generated and cause peak broadening or tailing or deterioration of separation performance. If L is smaller than V, liquid may leak because of inadequate ferrule adhesion. Therefore, we recommend replacing the ferrule together with the column. *If the column is replaced frequently, the male nut may crush the ferrule and liquid may leak. Since tightening the nut too much may cause its head to come off, replace the ferrule at an early stage. 【Fig. 1】 Column connection

Proteonavi is shipped after a strict performance check. However, if you should find any defect, please contact your dealer or Osaka Soda for replacement. Note that ShiseOsaka Sodaido does not warrant the product against column life or deterioration caused by the failure to follow the above handling instructions. Ten or more days after reception by the customer, Osaka Soda will assume that the product was delivered in good condition, and will not accept a later replacement request. 2008 / 9 / 16