I am working on bioethanol production. How can I quantify the reducing sugars percentage (glucose, xylose, arabinose,galactose, etc.) by using HPLC, after pretreatment, and what are the column conditions for the analysis?
You need to consult relevant literature. I recommend NREL laboratory procedures (LAP) for you. Just Google it and you will see different protocols which would guide you.
I'm not sure if this applies as well for you as i worked on bioethanol production but from starch, we use an aminex HPX-87H column from biorad and its respective H+ cation precolumn (cartridge from biorad), which is useful for the detection of sugars and organic acids, you can look for the attachment on this message as I worked using this protocol from the laboratory analytical procedures (LAP) and it worked great, just follow their instructions.
Best wishes and good luck
For better pretreatment techniques you can follow literature. Recommended columns for sugar analysis are H+ and Pb++ (Make: aminex, phenomex, shodex). For the calculation of percentage you can calibrate HPLC with respective standards.
I had something like this problem during my research, for this purpose you can use HPLC with refractive index detection, with folowing parametes:
Mobile phase - 25 min, from 10 to 20 mg/l Ca-EDTA,
flow rate - 1ml/min,
Column - Ultron PS80-H, with Ultron PS 80G predcolumn or equvalent.
By my opinion it will be work.
Do not hesitate to ask questions. Good luck
In our work by HPLC we use
Column CarboSep CHO 682 (lead), water 0.4 ml/min 80°C. Injection 10µl with RI detection.
For: Cellobiose, Glucose, Xylose, Galactose, Arabinose, Mannose.
I hope it will be usefull for you.
Good work
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