Processing Method for the Quantification of Methanol and Ethanol from Bioreactor Samples Using Gas...

Area of Science:

• Biotechnology
• Analytical Chemistry
• Bioprocess Engineering

Background:

• Methanol is a key carbon source and inducer in methylotrophic yeast bioprocesses, like Pichia pastoris.
• Uncontrolled methanol levels can harm cell cultures and reduce protein production.
• Existing methanol monitoring methods are often costly and complex.

Purpose of the Study:

• To develop a fast, simple, and sensitive method for quantifying methanol and ethanol in biological samples.
• To provide an affordable alternative for monitoring methanol in fermentation processes.
• To aid in optimizing bioprocesses through effective methanol level control.

Main Methods:

• Salting-out assisted liquid-liquid extraction (SO-LLE) technique was employed.
• Potassium carbonate was used for salting out, and ethyl acetate for extraction.
• Quantification was performed using gas chromatography (GC).

Main Results:

• The developed SO-LLE method demonstrated high recovery rates for methanol and ethanol.
• Validation confirmed good linearity, peak resolution, and limit of detection.
• The method proved effective across different sample matrices.

Conclusions:

• The new SO-LLE method offers a cost-effective and efficient approach for methanol and ethanol quantification.
• This technique facilitates better monitoring and control of methanol in fermentation.
• The study contributes to improved bioprocess optimization and recombinant protein production.