I am working on one plant for that I require to prepare methanolic and aqueous extract of leaf. Which method I have to use for that? I have to do cell line testing as well as the characterization of bioactive compound.
Hello Khushali Upadhyay! for the methanolic extraction you can try with the soxhlet method, maybe coupled with ultrasound waves for decreased the solvent volume that you will use and, the extraction time. After that, you must eliminate the solvent with a rotatory evaporator, so then you could use that extract on biological evaluations. For water extraction, maybe the decoction method is better.
After that, for the isolation of chemical compounds, you can use Column chromatographic, TLC, paper chromatography techniques, or preparative HPLC.
Best Regards!
Hi khushali,
First of all, you have to prepare the powder of your plant leaf by freeze- drying method following by milling step.
1- For methanolic extraction:
* you need to test many different mixtures of water-methanol 10, 20, 30...100 % methanol (v/v).
* Then, you need to choice the temperature and time of soaking step.
* You need to evaporate the solvent (methanol) by rotatory evaporator.
2- For aqueous extraction:
* You prepare the freeze-dried powder of plant leaf.
* You need to choice the temperature and the time of soaking step.
* You need to perform a separation step by using filtration or/and centrifugation method.
3- For isolation the bioactive compounds of your extract, you need a preparative HPLC.
Good lucks
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