I have successively extracted leaf powder with Pet. ether, benzene, chloroform, ethanol and water by using soxlet apparatus. Out of this, my water extract showed highest activity. Preliminary phyto-chemical investigation showed presence of flavonoids, phenolics, saponins, and alkaloids. I want to know, what phytochemical is responsible for its activity? Your guidance in this regards, is highly appreciated.
Do you mean to reveal the quality of the ingredients at the level of the groups to which we belong, such as alkaloids and phenols
Or was your question about the names of the compounds?
Hello. You should do an HPLC profile of each extracto; them, you should try a TLC and reveal with different drugs to see which kind of compounds you have. After that you should isolate the different compounds with differents chromatographics techniques and use each single compounds to test their biological activity
In a TOTUM (all compounds) you can't know exactly which phytochemical is responsible for the activity. You have to separate and try again the activity of each compound alone.
Sometimes the activity is due to high dosage compounds. Try to do a dosage of these phytochemicals.
Dear Abusufyan
Little by little. You should be more explicit.
- Which extract show presence of flavonoids, saponins and so on?
- What bio assay did you try?. Why you say the water extract showed the highest activity?.
- You should fraction your positive extract and try the bioassay in each fraction
until get the more active and continue working until isolate the active principle.
Mostly poly-phenols are the water soluble compounds as you mentioned water extract shown higher activity, which may be because of higher quantity of polyphenols presence. So do estimate Total Phenolic and Flavonoid content in water extract
Abusufyan Shaikh, by identify, I suppose you mean to isolate because you can only know the identity of the phytoconstituents after carrying out isolation of the inherent compounds in your water extract. To isolate the compounds, you will carry out fractionation of your water extract using suitable bio-assay guided chromatographic methods till you obtain the compound(s) responsible for the activity.
Let us know if you have specific concern.
Regards.
U should do bioautography first to find which one is the active compound and then by doing prep tlc and then prep hplc u can isolate this active compound and then find the structure by using all the spectroscopic techniques.
You can use LC-MS on the crude extracts. But you have to be explicit
It is impossible to say from what information you have provided. However, if we are talking about antimicrobial assay here it is quite likely the saponin's detergent like effects ruptured bacterial cell walls. The saponin should be very minimal in the ethanol and completely absent in other fractions. To know for sure you need to follow a bio assay guided fractionation. But first you can extract as much as possible from the water using butanol. You shake them together and wait for them to phase separate. Use a separation funnel to get them in different flasks. Eprat a few times. Combine butanol fractions. Completely evaporate butanol from water (there will be some mixing) and also remove butanol from butanol extract under vacuum. Do you have a rotary evaporator? Retest to see if activity persists in water extract. If yes then it must be saponins.
Kindly do chemical test in aqueous extract after that you will go for TLC analysis
All phytochemicals have some kind of bioactivities. It depends on the type of bioassays you're interested in. Phenolic and saponins are shows drag pattern of the peak in LC-MS/HPLC, because of the structure. Generally, flavonoids and alkaloids are a good source of bioactive compounds. Phenolic compounds are a good source of antioxidant properties. Few milligrams of crude water extract can be fractionated using RP-HPLC with fraction collector followed by bioassays and mass spectrometry of fractions. It will give you an idea, phytochemical responsible for bioactivity. Refer to the paper we recently published: Article Screening for bioactive secondary metabolites in Sri Lankan ...
If your water extract is dry, you can redissolve in acetone or methanol. After that, use TLC to identify the presence of the suspected class of compounds by using different derivatizing reagents specific for identification of class of compound of interest. If your extract is aqueous, extract the organic compounds with chloroform or ethyl acetate using solvent-solvent partition. You can then use TLC to analyse the presence of the suspected class of compounds. This is your preliminary step.
Do you mean to reveal the quality of the ingredients at the level of the groups to which we belong, such as alkaloids and phenols
Or was your question about the names of the compounds?
good answer Naser Jawad Kadhum Naser Jawad Kadhum Dhifaf Zeki Harith Rajab Almosawy
This is very similar to my undergraduate project except that i didn´t use water as a solvent. From what i understand in your question the water extract gave the highest bioactivity which your preliminary phytochemical screening indicates the presence of flavonoids, phenolics, saponins, and alkaloids. Each of these metabolites have different levels of activity for either anti-inflammatory, antimicrobial, antidiabetic, etc, so the specific activity will give information on the particular metabolite responsible for it.
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