I am desperate. My column blocked with sample which had LBG. What can i do to remove the LBG (locust bean gum, component with high viscosity) from RP-C18 column?
How about starting from the beginning? What temperature are you conducting your analysis at and what did you dissolve it in and use to analyze it on a "C18" column (IOW: Please provide your detailed HPLC method so we can better understand what you are doing)? It has extremely poor solubility in pure water, esp at room temperature. "Typical" analysis of locust bean gums, processed versions, are run at 50C to 80C using more miscible solvent systems.
Additionally, HPLC columns are consumable items. Consider disposing of the HPLC column if it is clogged and replacing it with a new one. Columns are inexpensive compared to the time and materials used to try and unclog one, then test it to insure it meets specification for re-use. *Save time and money and buy a new column that is appropriate for your desired analysis.
How about starting from the beginning? What temperature are you conducting your analysis at and what did you dissolve it in and use to analyze it on a "C18" column (IOW: Please provide your detailed HPLC method so we can better understand what you are doing)? It has extremely poor solubility in pure water, esp at room temperature. "Typical" analysis of locust bean gums, processed versions, are run at 50C to 80C using more miscible solvent systems.
Additionally, HPLC columns are consumable items. Consider disposing of the HPLC column if it is clogged and replacing it with a new one. Columns are inexpensive compared to the time and materials used to try and unclog one, then test it to insure it meets specification for re-use. *Save time and money and buy a new column that is appropriate for your desired analysis.
The LBG have solubility only in hot water check the temperature of your column then take hot distilled water and fix the column
in column oven, set the temperature according to column manual instruction wash the column for long time with slow floe rate
when you get the desirable back pressure then follow by pure Methanol.
Dear Charia,
if all else fails you could try to hydrolyse the LBG on the column.
This paper provides a protocol for acid hydrolysis of galactomannans
(pH 1.0/70 °C for 15 h)
Article Preparation and characterization of molecular weight fractio...
This should significantly reduce molecular weight and therefore viscosity of the LBG.
As Aqal suggested heating alone may be enough so you should definitely test that first (keep in mind that you should avoid 100 % water as this may cause dewetting of the C18 phase).
Silica based RP-columns are relatively stable towards acidic pH and heat. See for example here:
Article Temperature and pH-stability of commercial stationary phases
However, this depends on the stationary phase of your specific column. Maybe you can find some information in the manual or on the manufacturers home page.
I would only suggest this as a last resort, as prolonged incubation at acidic pH and high T may severely damage your column.
Kind regards,
Lorin
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