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Home > Maintenance
Videos FAQ Instrument Opetating Guide Maintenance
Proper Cleaning and Use of SepaFlash Columns
How To Use and Clean Chromatography Column
Usage: The sample should be kept clean. It is best to remove the insoluble and strongly retained impurities in advance. The sample can be pretreated for a certain amount of time, such as: SPE, filter membrane. At the same time, solid sample loading is used for samples with poor solubility.
Rinsing method: 1) For contaminants, first wash with alcohol (methanol, ethanol, isopropanol, recommended isopropanol) 5-10 column volumes, if it can not be removed, strong elution solvent (tetrahydrofuran, chloroform) Wait 5) column volume, and finally rush out the strong elution solvent with 5-10 column volumes of alcohol. Finally, it is stored in a normal phase system such as n-hexane/isopropanol, or dried by blowing.
2) If the normal phase column is contaminated with water, remove 10 times column volume with isopropyl alcohol.
Usage and Preservation Method of C4 Rapid Separation Column
1) The C4 rapid separation column is packed by dry packing, and at least 20 column volumes must be washed with 100% methanol before the first use, and then 10 column volumes are equilibrated with the mobile phase;
2) The pH of the mobile phase should be maintained between 2.5-7.0;
3) Properly store the C4 rapid separation column for multiple use, plug the end cap and store it in ACN/H2O (80:20) or MeOH/H2O (80:20). If there is buffer salt in the system, do not contain the same proportion of the buffered phase of the buffer salt is flushed 5-10 column volumes.
C8 Rapid Separation Column Usage and Preservation Method
1) The C8 rapid separation column is packed by dry packing, and at least 20 column volumes must be washed with 100% methanol before the first use, and then 10 column volumes are equilibrated with the mobile phase;
2) The pH of the mobile phase should be maintained between 2.5-7.0;
3) Properly store the C8 Rapid Separation Column for multiple use, plug the end cap and store it in ACN/H2O (80:20) or MeOH/H2O (80:20). If there is buffer salt in the system, do not contain the same proportion of the buffered phase of the buffer salt is flushed 5-10 column volumes.
C18 Rapid Separation Column Usage and Preservation Method
1) The C18 rapid separation column is packed by dry packing. Before using it for the first time, it must be flushed with 100% methanol for at least 20 column volumes, and then the mobile phase is equilibrated with 10 column volumes;
2) The pH of the mobile phase should be maintained between 1.5 and 7.5;
3) Properly store the C18 Rapid Separation Column for multiple use, plug the end cap and store it in ACN/H2O (80:20) or MeOH/H2O (80:20). If there is buffer salt in the system, do not contain The same proportion of the buffered phase of the buffer salt is flushed 5-10 column volumes.
Usage and Preservation Method of NH2 Rapid Separation Column
1) The NH2 rapid separation column can be used either in normal phase or in reverse phase. It should be noted that the normal phase solvent and the reverse phase solvent are immiscible. When replacing the mobile phase, ensure that the new mobile phase is miscible with the original preservation solution;
2) It is recommended to use isopropyl alcohol to rinse 30 times column volume at low flow rate and 10 times column volume using mobile phase; it can not be used to separate aldehyde group, carbonyl compound and reducing sugar in normal phase; Pay attention to the PH range of the mobile phase and the range of the water phase. The lower the pH, the higher the water ratio, the more the risk of hydrolysis, the recommended pH is 3.0-7.0;
3) Preservation of NH2 rapid separation column: When using the normal phase, the column is rinsed and stored with n-hexane-acetonitrile (99:1). When used in reverse phase, if it is not used for a short period of time, it can be stored in methanol or acetonitrile. When it is not used for a long time, the methanol should be replaced with isopropyl alcohol and chloroform, and finally stored in n-hexane-acetonitrile (99:1).
ARG Rapid Separation Column Usage and Preservation Method
1) Before using the ARG Rapid Separation Column, equilibrate the column with 50 times column volume with a 50/50 acetonitrile/water buffer system, then equilibrate 20 times column volume with the initial mobile phase. The injection interval requires 10 column volumes to initiate the mobile phase. Balance the column;
2) The mobile phase maintains not less than 5% of the aqueous phase solution, the organic ratio is not less than 40%, the buffer salt is ammonium formate or ammonium acetate, and the phosphate buffer salt cannot be used to prevent the buffer salt from being precipitated;
3) Methanol and acetonitrile are recommended for the sample preparation solvent. It is not recommended to use pure water or DMSO to dissolve;
4) ARG rapid separation column is recommended to be stored in 95% acetonitrile if not used for a long time; if there is buffer salt in the system, rinse 5-10 column volumes with the same proportion of mobile phase without buffer salt;
5) If reused, rinse 20-50 column volumes with low flow rate of isopropanol before use again (flow rate is referenced to pressure, rinse column volume is used as a reference for previous use conditions) and then rinsed according to the first use method.
Usage and Preservation Method of Diol Rapid Separation Column
If the Diol separation column is used for normal phase separation, the method of use is as follows:
Keep the sample clean, it is best to remove insoluble and strong retained impurities in advance, and the sample can be pre-treated, such as: SPE, filter membrane. At the same time, solid sample loading method is adopted for samples with poor solubility;

Rinsing method: a. For contaminants, first wash with alcohol (methanol, ethanol, isopropanol, recommended isopropanol) 5-10 column volumes, if it can not be removed, strong elution solvent (tetrahydrofuran, chloroform) Wait 5) column volume, and finally rush out the strong elution solvent with 5-10 column volumes of alcohol. Finally, it is stored in a normal phase system such as n-hexane/isopropanol, or dried and stored;
b. If the normal phase column is contaminated with water, transfer 10 times column volume with isopropanol.
Usage and Storage Method of CN Rapid Separation Column
1) CN rapid separation column can be used either in normal phase or in reverse phase. It should be noted that the normal phase solvent and the reverse phase solvent are immiscible. When replacing the mobile phase, ensure that the new mobile phase is miscible with the original preservation solution;
2) It is recommended to use isopropyl alcohol to rinse 30 times column volume at low flow rate and 10 times column volume with mobile phase. Pay attention to the PH range of mobile phase and the range of water comparison. The lower the pH, the lower the PH. The higher the water ratio, the more the risk of hydrolysis;
3) Preservation of CN Rapid Separation Column: When using the normal phase, rinse the column and store it with n-hexane-ethanol (95:5). When used in reverse phase, if it is not used for a short period of time, it can be stored in methanol or acetonitrile. When it is not used for a long time, the methanol should be replaced with isopropanol and chloroform in sequence, and finally stored in n-hexane-ethanol (95:5).
Maintenance of SepaBean Machine
Mobile Phase Replacement
Fill in sample information, TLC information (normal phase), HPLC information (inversion), separation parameters, gradients can be generated automatically or manually.
• Positive phase inversion
When A and B flow are the same, change to isopropanol, rinse with Flow Rate=30ml/min for 10 minutes, change to methanol when A and B flow are the same, rinse with Flow Rate=30ml/min for 10 minutes, A mobile phase change Pure water, also flow rate = 30ml / min, rinse for 10 minutes, this will ensure that the pipeline has been rinsed. While flushing the tubing, ensure that the collection needle is also cleaned.
• Reverse phase positive phase
When the flow of A and B is the same, change to methanol, rinse with Flow Rate=30ml/min for 10 minutes, change the flow of A and B to isopropanol, rinse with Flow Rate=30ml/min for 10 minutes, and change the A mobile phase. Into petroleum ether, B mobile phase is changed to ethyl acetate, and also rinsed for 10 minutes with Flow Rate=30ml/min, which ensures that the pipeline has been rinsed clean. While flushing the tubing, ensure that the collection needle is also cleaned.
Fuse Replacement
The fuse replacement process is as follows:
1. Turn off the power switch.
2. Remove the power cable from the power outlet.
3. Open the fuse back cover with a flat-blade screwdriver.
4. After replacing the new fuse, push the fuse cover
Hardware Maintenance
1) Cleaning solvent filter
Remove the clogged solvent filter from the bottle head assembly, place the filter in a beaker containing isopropyl alcohol, and rinse the filter thoroughly with water.
2) Pump
The pump is a maintenance-free, valveless metering pump. It is not recommended that the customer disassemble the pump assembly and only operate the solvent connection to the pump.
3) Replace the xenon lamp and tungsten lamp
1. Turn off the power of the instrument and wait for 15-30 minutes to cool the light source.
2. Use the hexagon socket to unscrew the two fixing nuts of the side panel to remove the panel. You can see the entire detector component, loosen the tray of the fixture detector counterclockwise, carefully pull out the tray to the appropriate position, and remove the xenon lamp. For the power cord of the tungsten lamp, use a Phillips screwdriver to loosen the nut that fixes the xenon lamp and the tungsten lamp. Carefully pull out the xenon lamp and the tungsten lamp (be careful to wear gloves, do not touch the lamp glass window) and place it on clean white paper.
3. Install new xenon lamp and tungsten lamp, pay attention to the position of the fixed lamp source and the position of the lamp source window (see picture), tighten the upper and lower fixing nuts, plug in and put down the power cord. Push the tray back to its original position, tighten the retaining nut and install the side panel.
4. Plug in the instrument power and network cable and turn on the instrument.
4) Cleaning the flow cell
If the flow cell is contaminated or clogged, set the pump parameters to be flushed with a small flow rate solvent, or unscrew the flow cell inlet and outlet lines, connect the two-way joint, use a syringe to absorb the solvent that is good for blocking the sample, and slowly rinse until it is flushed. open.
5) Maintenance plan
Maintenance project cycle
Check the tubing and fittings to see if the tubing is wrinkled every week, or if the flow rate is not accurate, the fitting has leaks and there are air bubbles in the tubing
Replace piping and fittings annually
Check the xenon lamp energy every quarter, or when the detector noise is high and the sensitivity is reduced
Replace the xenon lamp When the xenon lamp is not lit or the xenon lamp energy is not enough.
Cleaning the flow cell quarterly
Confirm that the solvent bottle is clean daily
Troubleshooting for SepaBean machine
Detector Failure and Elimination
Problems Possible Causes Solutions
Baseline noise Detection pool window contamination 1、Flush the detection cell with 1 mol/L nitric acid, water and new solvent
2、Remove the detection pool, open the cleaning or replace the pool window quartz
Bubbles in the sample cell 1、Suddenly increase the flow to drive out the bubbles.
2、Add a BPR at the exit end of the test cell to increase the pressure inside the test cell.
Detector light source failure 1、Check the xenon lamp setting status
2、Check the lamp usage time, lamp energy, and number of starts
3、Replace the xenon lamp
Solvent leakage Tighten or replace the connector
Small bubbles pass through the detection cell 1、The mobile phase is carefully degassed
2、Increase the back pressure of the test cell
3、System leak detection
Particles passing through the detection pool 1、Cleaning the detection pool
2、Check the separation column outlet sieve
Baseline drift Detection pool window contamination 1、Flush the detection cell with 1 mol/L nitric acid, water and new solvent
2、Remove the detection pool, open the cleaning or replace the pool window quartz
Detector temperature change System constant temperature
Detector light source failure Replace xenon lamp
The original mobile phase is not completely removed. Rinse the system with a new mobile phase to replace the solvent, or replace it with a compatible solvent.
Solvent storage bottle contamination Clean solvent bottles with new mobile phase balancing system
Strongly adsorbed components are eluted from the separation column 1. Wash the separation column with a solvent with strong elution ability before the next separation.
2. Using solvent gradient
Large spikes appear in the curve Bubbles pass through the test cell. 1. Degas the solvent and thoroughly flush the system.
2. Check if the connection system leaks
Negative peak Injection failure Use the injection valve to verify that there are no air bubbles in the sample loop during injection
The mobile phase used is impure. Use a chromatographically pure mobile phase or purify the solvent.
The curve panel signal rises stepwise; the flat head peak; the baseline cannot be returned to zero. The output range of the detector is not set properly. Reset the output range of the detector.
As the pump reciprocates, there is a spike in the detection tank. Bubbles in the detection cell Remove the inlet tube of the detection tank from the joint of the separation column, and use a syringe to push methanol from the outlet end to remove air bubbles.
Pump Failure and Elimination
Problems Possible Causes Solutions
The pump is running, but no solvent is output. The solvent reservoir is empty. Fill the reservoir.
The actual flow rate is lower than the set value. The filter head is dirty. Clean or replace the filter head.
The gradient valve has dirt. Clean or replace the gradient valve.
The pipe has wrinkles. Replace the pipe.
No solvent is delivered. Pump is not running, power switch is not turned on. Turn on the Power switch.
The solvent is placed lower than the pump head Move the solvent to a position higher than the pump head or horizontal position with the pump head
The pressure rises too high. The pipe is blocked. Find the blockage and handle it.
The inner diameter of the pipe is too small. Replace the appropriate inner diameter line
Online filter blocking Stainless steel screens for cleaning or replacing inline filters
Column blocking Replace column
Flow cell blockage Cleaning flow cell
Pump stops during operation The pressure exceeds the high pressure limit. 1、Reset the maximum pressure limit.
2、Replace the column
3、Replace the appropriate inner diameter pipeline
4、 Cleaning the flow cell
Pump overheat protection View fan and circuit
The pump flow rate becomes smaller The air bubbles in the pump gather. Open the pump head and the pipeline, let the pump run at high flow rate, and eliminate the air bubbles.
The solvent filter is clogged. Open the pump head inlet pressure cap. If the solvent does not flow out of the infusion tube very quickly, the filter is clogged and needs to be cleaned or replaced.
The two solutions in the pump are not mutually soluble. A solvent is used between the two solvents to dissolve the two mutually insoluble solvents.
Plunger seal leak Replace the plunger seal
Strongly adsorbed components are eluted from the separation column Bubbles accumulate in the pump head. Let the pump run at high flow rates, eliminate bubbles.
Solvent stratification in the pump Using a transition solvent to dissolve the two
The pump is not tightly sealed. Tighten the fixing screws.
The infusion line is leaking or partially blocked Check the pipeline one by one to eliminate
The pump has a click and cannot start normally Motor failure Stop the pump and check
Line voltage is too low Increase line voltage
Excessive pressure See "Pressure is too high"
The column pressure is too high The stigma is blocked by impurities Disassemble the column head and clean the column head filter. If the impurity particles have entered the bed , carefully remove the sediment and the contaminated packing, and then fill in the same packing, do not leave a gap in the column head; The method is to add a filter in front of the column.
There is column pressure after pumping, but no mobile phase flows out of the detector. Serious leakage in the system. Repair the injection valve or the tubing and fasteners between the pump and the detector
Flow path blockage Clear the inlet of the injector, the injection line or the connecting line between the column and the detector or the particles of the detector
The inlet end of the column is blocked by particles Clean or replace the column inlet filter; replace one column as needed; filter all samples and solvent
Increased column pressure and reduced flow The inlet of the detector or detector is partially blocked Disassemble and clean the test cell and tubing
Contact Us
Order Email: ca_order@santaitech.com
Support Email: ca_support@santaitech.com
+1 418-580-0437
MON - FRI 9 AM - 5 PM
North America Production and assembling site:
214 Brunswick, Pointe-Claire,Quebec, H9R 1A6 Montréal
Canada
Administration site:
375,Labbe Street, Thetford Mines,Québec,Canada