Richard Lab:Dilute-acid pretreatment
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Procedure for 1.5 g of dry mass
Dilute-acid pretreatment is a commonly used method of increasing the digestibility of lignocellulosic biomass. While it is difficult for the average lab to achieve the prescibed conditions for standard pretreatment, this protocol is a reduced severity treatment which is achievable using a standard autoclave.
- Drying Oven
- Filtration aparatus
- Air lock (optional)
- Aluminum weigh trays
- 0.8% Sulfuric Acid (w/w)
- Fiber Filters
- Prepare biomass samples as necessary (grinding, blending, etc.)
- Weigh out ~1g of biomass into a preweighed aluminum tray (do replicates of this).
- Be sure to record the weight of the biomass as well as the weight of the tray.
- Place the trays in a 105°C oven overnight.
- Prepare 0.8% (w/w) sulfuric acid by adding 89 parts water and 1 part 72%.
- Once the moisture content is determined, weigh the amount of wet biomass correlating to 1.5g dry mass to a 125ml flask.
- Record this weight and repeat for all samples.
- Apply 45ml of 0.8% sulfuric acid to each sample.
- Put the flasks (with biomass and acid inside) in the airlock and apply vacuum until bubbles stop rising. Repeat 2X.
- If an airlock is unavailable just let the flasks (with biomass) sit at on the bench for 1-2 hours.
- Cap all flasks with foil.
- Autoclave at 121°C for 30min on the liquid cycle.
- Immediately after the cycle is finished, remove the samples and filter using a sidearm flask and a fiber filter
- Wash the biomass with 200ml water using the filter.