CAPCELL PAK ST instruction manual

• 1. Column Handling
• 2. Column Installation
• 3. Analysis
• 4. Storage of columns
• 5. About the column connection
• 6. Troubleshooting

Thank you for purchasing the Specialty CAPCELL PAK ST for Streptromycin high.

To ensure the stability and long life of CAPCELL PAK ST column, please read and follow this manual carefully for the correct of use products.

1. Column Handling

1. Handle the column with great care. Strong shock may cause damage.
2. Attach or detach the column when the pressure gage indicates zero.
3. Maximum column operating pressure is 20 MPa.

2. Column Installation

1. The column joint is a male-nut type for 1/16 inch OD tubing. Make sure that the tubing joints of the system fit correctly and that the ferrule tips are inserted completely into the joints. (See Fig. 1.)
2. Before attaching the column, displace the liquid in the system with the mobile phase to be used.
   * Verify the solvent supplied at the time of column shipment (mentioned in the enclosed column report) and note the replacement procedure to avoid salting-out.
3. Install column in the direction of the arrow on the column label.

3. Analysis

3-1. Mobile phase

1. A mobile phase indicated in the column report is enclosed with the new column. In the case of displacement with other mobile phase containing a mineral salt, note the displacement procedures to prevent precipitation.
2. 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.
3. Avoid the following practices and factors to prevent column deterioration.
   • Frequent change of the mobile phase composition
   • Rapid change in pressure (some Mp at column inlet
   • Continued using at pressure exceeding 15 MPa
   • Prolonged water flow

3-2. Preparation of sample solutions

1. Preferably, dissolve the sample in the same solvent of as the mobile phase.
2. Be aware of the fact that if you dissolve sample in solvent with a high water content, resolution will be reduced. Solvent with organic solvent content of higher than 50% is recommended.
3. If the dissolved sample is poor solubility in mobile phase, it will be separated out and cause suddently increase of the column pressue.
4. Set the pH of the sample solution within the acceptable range of pH for the column (pH 2-7).

4. Storage of columns

1. Hermetically seal with accessory plugs and store in a cold place where temperature fluctuation is negligible.
2. If the storage period is shorter than one month, displace the mobile phase with a mixed solution of water and organic solvent of the same composition as the mobile phase used (without acid or inorganic salt)(avoid the displacement with water alone).
3. If the storage period is longer than one month, after the procedure in 2, displace the mobile phase with the solvent used in shipment (see the column report) and then place in storage.

5. About the column connection

For tubing, follow the instructions in Fig. 1. Inappropriate tubing, particularly when a tube used for a different type of column is used as is, the tip length beyond the ferrule (V in Fig. 1) may not match the length of the end fitting bore (L in Fig. 1), which leads to troubles.

Where L>V, dead volume occurs and may cause the broadening or tailing of peak, or limit separation performance.
Where L<V, inadequate sealing of the ferrule causes liquid leakage. Therefore, it is recommended that the ferrule be changed concurrently with the column change.

* After frequent column replacement the male-nut may present a crushed ferrule and cause liquid leakage. In such a case, tightening too hard may cause the head portion of the nut to break off. Therefore, replace the ferrule in good time.

Fig. 1

6. Troubleshooting

Since it is impossible to enumerate all of the troubles associated with HPLC analysis work have diverse causes, only those which occur relatively often in and around the column are cited:

Symptom	Causes	Remedies
1. Rise in column pressure	Clogging with foreign matter 1. Foreign particles, insolubles in eluants and sample solutions 2. Water deposits in the tubing 3. Fragments of plunger seal 4. Precipitation of sample constituents	• Clean the filter with an ultrasonic device or replace. • Filter the solvent and sample solution through a membrane filter beforehand. • Install a liquid line filter. • Clean the tubing and replace the plunger seal • Prepare the sample solution with the mobile phase
2. Splitting, tailing or broadening of peak	1. Occurrence of dead volume due to faulty tubing 2. Inappropriate conditions of mobile phase 3. Deterioration of column * Restoration is impossible when deterioration of column or void in packing material bed has occurred.	• Dismantle the tubing once and reconnect. • Ascertain sample solvent and injection volume. • Verify column performance using the standard testing solution.
3. Retention time too long or unstable	1. Liquid leakage (this can be judged by the movement of pressure gauge on the pump) 2. Inappropriate conditions of the mobile phase 3. Insufficient time for column stabilization	• Inspect the liquid leakage from pump and tubing system. • Take sufficient time for stabilization.
4. Retention time too short	Insufficient time for column equilibration	• Take sufficient time for stabilization.

While CAPCELL PAK ST columns undergo strict verification of performance before shipping, if any defect is identified, please inform us or our distributor of it.
However, please take note that we are not responsible for matters related to column service life or problems caused by the failure to follow the above mentioned instructions for care and use. Please note also that we assume the product is assumed to have been delivered in good condition if no claim is made within ten days from receipt by the customer, and requests for replacement submitted thereafter will not be accepted.

June 1st 2009