CAPCELL PAK User's Manual

3-1.Analysis 1. All solvents acceptable for the conventional chemically bonded silica column can be used. 2. The acceptable pH range for capsule-type packing material depends on the bonding group. Table 1 Acceptable pH range for columns Bonding group C 18 C 8 Type ACR UG AQ DD MG Acceptable pH 1~10 2~10 2~9 1.5~10 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 eluate .Prolonged water flow .You may use 100% water containing inorganic salt for AQ (we recommend using a mobile phase containing an organic solvent of 50vol% or more near the acceptable pH limit (around pH9) or 10vol% or more around pH2).

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 resolution 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.

3-3.Notes on Analysis Columns of CAPCELL PAK series generally show similar separation profiles to those of corresponding conventional silica-based columns, although slight selectivity difference may be observed depending on the analyte. When optimizing the conditions for compounds already done with conventional columns, use the same condition as a starting one.

(1) When using C 18, C 8 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. To analyze basic compounds which are protonated under neutral conditions, peak shapes became sharper according to the buffer concentration and amount of organic solvent in mobile phase. 3. If analysis is performed with a neutral or acidic mobile phase after using a basic or strongly acidic mobile phase, retention times and peak shapes of basic compounds may become unstable.

1. Seal the column with the accessory plug and store it in a cold place where there is little temperature fluctuation. 2. Replace the column with solution of organic solvent and water having the same composition as the mobile phase after using solvents containing strong organic acids such as TFA or basic solvents. (Do not use 100% water.) Moreover, for storage of one week or longer, replace the column with acetonitrile. 3. For storage within one month after using, replace the mobile phase with a solution of organic solvent and water having the same composition as the mobile phase and then fill it with the solvent used at the time of shipment. (Refer to the column report.) 4. Avoid using 100% water to rinse a column that is other than AQ.

1. A microcolumn uses a filter-embedded end fitting as shown in Fig. 1. The filter cannot be changed alone. 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

Problems in high performance liquid chromatography are attributable to various causes that cannot all be listed up. The table below describes some comparatively common problems related to the column. Symptom Cause Measures 1. Column pressure rise. Blocking with foreign matter 1. Dust or insoluble matter in the mobile phase or sample solution. 2. Dirt in the tubing. 3. Plunger seal fragment. 4. Precipitation of sample components. • Sonicate the filter or replace it. • Filtrate the mobile phase and sample solution in advance using a membrane filter. • Attach a line filter. • Clean the tubing and replace the plunger seal. • Prepare a sample solution with the mobile phase. 2. Peak splitting, tailing, and broadening. 1. Void in the column head. 2. Dead volume due to inappropriate connections. 3. Inappropriate mobile phase conditions. • Ion suppression method: Inadequate suppression (Too much sample). • Ion-pair method: Inadequate concentration of the ion-pair agent (Too much sample). 4. Column deterioration. * Not repairable in the case of column deterioration or damage to the packing condition. • Reconnect the tubing. • Review the pH, salt concentration, sample amount, and other conditions. • Review the ion pair agent concentration, pH, sample amount, and other conditions. • Check the column performance using standard inspection solution. 3. Retention time too long or unstable. 1. Liquid leak (Indicated on the pressure gage of the pump). 2. Inappropriate mobile phase conditions. 3. Inadequate column equilibration time. • Check the pump and tubing for any leaks. • Secure adequate equilibration time. 4. Retention time too short. 1. Hydrolysis (deterioration) of a bonded groups by strong acid or base. 2. Inappropriate mobile phase conditions. 3. Inadequate column equilibration time. - • Secure adequate equilibration time. CAPCELL PAK 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 Osaka Soda 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 / 10 / 1