INLINE HPLC DEGASSING MODULES

    INLINE HPLC VACUUM DEGASSER / DEGASSING MODULES:

Degassing your mobile phase is critical to maintaining a stable baseline without fluctuations. Loss of HPLC pump prime or cavitation, baseline instability and poor reproducibility often result when the mobile phase is not degassed. Modern electronic vacuum degassing systems are a great convenience (just plug them in). Changing heavy Helium gas cylinders, monitoring gas pressures and levels over time have been replaced in many labs with the sound of small gurgling vacuum pumps. While continuous helium sparging provides the best type of degassing for HPLC and benchtop ultrasonic baths are poorly suited to degassing HPLC mobile phases, vacuum degassers are widely used. 

• Since sonicators and in-line vacuum degassers do not specifically remove oxygen from the gas inside the liquid, continuous helium sparging should still be used for applications where low mobile phase oxygen levels are needed to maintain high-sensitivity (i.e. Fluorescence detection or very low UV detection). While the convenience gained through the use of these in-line vacuum devices can be great, there are some important downsides to this technology. One of them will be mentioned here.

Total Channel Volume: The volume of liquid contained in the tubing and vacuum chamber of EACH vacuum degassing channel can be enormous compared to the tiny volume often found in systems which utilized Helium sparging alone. When the system is turned off, this solvent often sits stagnant over night, allowing air to bleed in, and must be flushed out when the system is re-initialized for use the following day. *If you just switched bottles of mobile phase, then the old mobile phase is still inside the system and needs to be completely flushed out before use or you will have some very strange chromatographic results in the coming weeks! This process takes time and should be performed on every channel in your system, even if you are not going to use them all. How much volume you ask ?

Let’s look at the chamber volumes found in a few commonly used vacuum degassing units (You can look up the spec’s or measure it on your own).

• Agilent/HP Brand Degassing Module, Model G1322A; Each vacuum channel chamber has a volume of about 10 to 12 ml. The interconnecting tubing (solvent frit to bottle head, bottle head to degassing unit and degassing unit to pump inlet) can add another 10 mls more. In this example, we recommend that you flow at least 30 mls of the appropriate solvent through the line before use. This will insure that the channel is primed with fresh mobile phase. That step is for EACH channel so it is best to flush them one-at-a-time, in sequential order, to keep track of them.

• Agilent/HP Brand Micro Degassing Module, Model G1379A, G1379B or G4225A:  Each vacuum channel chamber has a volume of about 1 ml. The interconnecting tubing (solvent frit to bottle head, bottle head to degassing unit and degassing unit to pump inlet) can add another 10 mls more. In this example, we recommend that you flow at least 20 mls of the appropriate solvent through the line before use. This will insure that the channel is primed with fresh mobile phase (it is best to flush them one-at-a-time, in sequential order,100% A, 100% B ... to reduce miscibility concerns).

• Shimadzu DGU-20A3, DGU-20A5, DGU-20A3R & DGU-20A5R. Each vacuum channel in these modules has a volume of between 0.4 and 0.5 mL. We recommend that you flow at least 15 mls of the appropriate solvent through each line before use.

• Waters Alliance 2690, 2695, 2790, 2795, 1525, DG2 Systems and Acquity BSM/QSM. Each vacuum channel in these modules has a 0.5 ml volume. Because these are integrated degassers (exc for the DG2 stand-alone), the interconnecting tubing volumes are also low ~ 5 ml each. For this example, we recommend you flush each channel with at least 10 mls of mobile phase before use. Remember to flush them one-at-a-time, in sequential order, for best results.

NOTE: Some early models of HPLC vacuum degassing systems had individual chamber volumes of 30 to 50 mls each ! When you add the total tubing volume to that you end up with a flush volume of 40 to 60 mls per channel. These systems took a very long time to flush out.

To save time performing these initial “flush” steps, you should take advantage of the highest available flow rate setting of your pump. With the priming valve open (so the solvent goes to waste, not to the column), set the pump flow rate to 5 or 10 ml/min at 100% for the first channel (i.e. 'A') and you will be able to flush out a single channel in two or three minutes. Repeat for each of the other channels and then adjust the flow rate slowly back down to your normal equilibration flow rate before closing the prime purge valve again. Our lab performs this flushing procedure at the start of each day to insure the HPLC columns receive fresh solvent. This procedure should also be performed any time a mobile phase bottle is changed out to a different bottle or the liquid is "topped off" too.

Please keep in mind that running liquid at 10 ml/min during the flush phase in most analytical degassing modules will not result in very efficient degassing as degassing efficiency is a function of flow rate (surface area of liquid exposed over time. The lower the flow rate, the better the degassing). You will still need to wait a period of time until the new solvent is fully degassed once again, but at least you will know it is free of any older mobile phase. *Remember, everything you do before you start an analysis contributes to the final result.

•  Make sure that your degassing module is 100% fully functional. These modules usually wear out after between three to five years of normal use. We have found that some models like the Agilent G1379A and G1379B's may wear out sooner (~ 3 years). Problems with leaks, liquid exiting the vacuum pump exhaust line, bubbles, contamination and failed vacuum pumps are very common. These are maintenance items.  If the unit is repaired early, the cost may be much less than delaying repair so have the system properly tested and diagnosed at the first sign of trouble. Anytime the degasser's performance drops below the manufacturer's specification, your hear unusual noises and/or liquid is observed exiting from the vacuum pump's exhaust line, your entire HPLC system may fall out of compliance. You must repair the degasser module to use it. The degasser is an important component of your entire HPLC system so it must be maintained and serviced just like the other modules. RED or Yellow Error lights (on Agilent/HP models) or "Degasser hardware fault" error (Waters) on these modules usually indicates a need for professional service. 

  References:

    TROUBLESHOOTING INFO PAGES:  

• Waters Brand HPLC Vacuum Degasser Modules: 
  

• Agilent / HP Brand HPLC Vacuum Degasser Modules:
  

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