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(HPLC: Amino Acid Analysis) Shutting Down the System
(HPLC: Amino Acid Analysis) Shutting Down the System
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Answer
• For 1 to 2 Days
When a series of analyses is complete and similar analyses are scheduled in 1 to 2 days time, replace the solution inside the reaction reagent flow line with purified water, as follows.
When a series of analyses is complete and similar analyses are scheduled in 1 to 2 days time, replace the solution inside the reaction reagent flow line with purified water, as follows.
1. Turn OFF the detector lamp.
2. Set the parameters as follows:
2. Set the parameters as follows:
Pump A T.Flow (LC-20AB total flow rate): 0.3 mL/min
Pump A B.Conc（LC-20AB-B ratio(%)): 0 %
SV (Pump A) (FCV-11ALS port): A (Mobile Phase A (AA-MA))
Pump A B.Conc（LC-20AB-B ratio(%)): 0 %
SV (Pump A) (FCV-11ALS port): A (Mobile Phase A (AA-MA))
B.Flow (Flow rate of LC-40D 1st): 0 mL/min
C.Flow (Flow rate of LC-40D 2nd): 0 mL/min
Oven T: 0 ºC(No temperature control)
C.Flow (Flow rate of LC-40D 2nd): 0 mL/min
Oven T: 0 ºC(No temperature control)
3. With this status, pump for 10 to 20 minutes until the separation column cools down to room temperature.
You can open the column oven door.
4. Stop the operation of all pumps (including reaction reagent delivery pumps) and column oven.
5. Set purified water bottles to two LC-40D (reaction reagent delivery pumps).
6. Open the drain valve, and purge the pumps with purified water for about two minutes. Close the drain valve, and pump purified water at 0.2 mL/min for about 10 minutes.
Do not purge the LC-20AB (mobile phase delivery pump), but rather leave it without operation.
You can open the column oven door.
4. Stop the operation of all pumps (including reaction reagent delivery pumps) and column oven.
5. Set purified water bottles to two LC-40D (reaction reagent delivery pumps).
6. Open the drain valve, and purge the pumps with purified water for about two minutes. Close the drain valve, and pump purified water at 0.2 mL/min for about 10 minutes.
Do not purge the LC-20AB (mobile phase delivery pump), but rather leave it without operation.
NOTE:
• Be sure to purge the reaction reagent delivery pumps with purified water, and further pump purified water for about 10 minutes. This allows for washing away all salt in reaction coils and fluorescence detector flow line. Otherwise, reaction reagent left in flow lines may cause salt crystallization or clogs, since it contains high-concentration inorganic salt.
• To leave purified water bottles until next analysis, place them higher than the waste container. If they are placed lower than the waste container, waste liquid may flow back. To avoid backflow, be sure to dispose of waste liquid after analysis.
• To leave purified water bottles until next analysis, place them higher than the waste container. If they are placed lower than the waste container, waste liquid may flow back. To avoid backflow, be sure to dispose of waste liquid after analysis.
7. Stop the reaction reagent delivery pumps. Turn off the system unit power.
8. Dispose of waste liquid.
8. Dispose of waste liquid.
• For 3 Days or More
When shutting down the system for 3 days or more, (1) rinse the separation column and ammonia trap column with mobile phase C, (2) remove the columns from the system, and (3) rinse the entire flow line with purified water. To do so, follow the procedure below:
When shutting down the system for 3 days or more, (1) rinse the separation column and ammonia trap column with mobile phase C, (2) remove the columns from the system, and (3) rinse the entire flow line with purified water. To do so, follow the procedure below:
1. Turn off the detector lamp.
2. Set the parameters as follows:
2. Set the parameters as follows:
Pump A T.Flow (LC-20AB total flow rate): 0.3 mL/min
Pump A B.Conc（LC-20AB-B ratio(%)): 0 %
SV (Pump A) (FCV-11ALS port): A (Mobile Phase A (AA-MA))
Pump A B.Conc（LC-20AB-B ratio(%)): 0 %
SV (Pump A) (FCV-11ALS port): A (Mobile Phase A (AA-MA))
B.Flow (Flow rate of LC-40D 1st): 0 mL/min
C.Flow (Flow rate of LC-40D 2nd): 0 mL/min
Oven T: 0 ºC(No temperature control)
C.Flow (Flow rate of LC-40D 2nd): 0 mL/min
Oven T: 0 ºC(No temperature control)
3. With this status, pump for 10 to 20 minutes until the separation column cools down to room temperature.
You can open the column oven door.
4. Stop the operation of all pumps (including reaction reagent delivery pumps) and column oven.
5. When load pressure to the columns drops down to almost zero, remove the separation column and ammonia trap column.
You can open the column oven door.
4. Stop the operation of all pumps (including reaction reagent delivery pumps) and column oven.
5. When load pressure to the columns drops down to almost zero, remove the separation column and ammonia trap column.
NOTE: Be sure to seal the columns after removed. Once packing materials dry out, column performance will significantly decrease.
6. Attach coupling joints in the places of the separation column and ammonia trap column.
7. Replace all mobile phase/reaction reagent containers with purified water bottles, and purge all HPLC pumps with purified water.。
8. Close the drain valve and pump purified water from all pumps for about 10 minutes.
Use the same flow rates for mobile phase/reaction reagent delivery pumps as those in analysis.
9. Stop all pumps and turn off the system unit power.
10. Dispose of waste liquid.
7. Replace all mobile phase/reaction reagent containers with purified water bottles, and purge all HPLC pumps with purified water.。
8. Close the drain valve and pump purified water from all pumps for about 10 minutes.
Use the same flow rates for mobile phase/reaction reagent delivery pumps as those in analysis.
9. Stop all pumps and turn off the system unit power.
10. Dispose of waste liquid.
NOTE: To leave purified water bottles until next analysis, place them higher than the waste container. If they are placed lower than the waste container, waste liquid may flow back. To avoid backflow, be sure to dispose of waste liquid after analysis.
• For long-term storage
Purified water will putrefy as time elapses. When not using the system for long time (a few weeks), first purge the entire flow line with purified water, and replace the purified water with methanol or ethanol.
NOTE: Pumping high-concentration alcohol through separation column or ammonia trap column decreases column performance. Be sure to rinse the entire flow line with purified water before installing the columns.
NOTE: Pumping high-concentration alcohol through separation column or ammonia trap column decreases column performance. Be sure to rinse the entire flow line with purified water before installing the columns.
If the column is to be long out of service (for six months or longer), click here for store it.
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