Does any one have any idea on how to remove fat from plants without reducing the phytate concentration needed for hydrolysis by phytase?
All I can provide is that oil extraction protocol I have, but it removes the lipids in general:
The sample is treated with 20 mL of n-hexane in a Soxhlet extractor overnight to obtain oils, which are sealed under N2 and stored in a freezer (-5°C) until HPLC analysis.
You can also check the oil extraction official method published by the association of analytical communities (AOAC) in 1965. I hope this helps.
Weight 1 g of your sample (fresh or dry), ground it in presence of liquid nitrogen until to obtain fine tissue particles or powder , place the powder in a glass flask and add it with 10 mL of n-hexane. Cover the flask from light and mix it gently by 10 h at room temperature. Stop mix, then place the flask on bench without disturbing, after 20 min, remove the supernatant (n-hexane), repeat three more times the n-hexane extraction as described above. Dry your powder in hood or at room temperature by 4 hour or if you prefer, dry the powder at 45 C by 2 h. Then extract your tissues to recover proteins or phytates using the current methodology in your hands.
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