The brief steps are:

First of all you have prepared plant extracts using solvents of various polarity. then isolate the pure fraction from plant extracts using following techniques. First run TLC to know the type of fractions. After that Column Chromatography with HPLC, after isolated pure fraction run FT-IR, LC-MS, HNMR, CNMR for structure elucidation. You can read my papers

Do you have a sensitive but simple test to follow purification steps?   purification?    How much   material/biological activity do you have to start with?

It is that complicated. Do you know what class/type of compound?

i dont know the type of compound, No one has studied the details of this plant. But the crude extract is used to treat for arthritis, it is proven 

Do you have only one extract?(which solvent)

Methanol

For your purpose, bioassay-guided isolation of compounds is suggested. If the crude extract showed activity, then fractionation with selected solvents is needed. From this, you can focus for isolate the active compounds. 

Dear Venugopal

Similar questions were asked by Vinesh Kumar and Danladi Chiroma Husaini and several researchers responded with interesting answers. You may follow these questions for possible leads to your query.

firstly start by the identification of the phytochemical group

but in general either by the use of planner chromatogaphy( not recommended)

column chromatography  ( recommended)

PHPLC  (VERY RECOMMENDED)

Dear Elakkiya Venugopal, You have methanol extract of your plant ok.......Firstly confirm it that it is active against arthritis, if you hv done this then mix the extract with Distilled Water and fractionate with different solvents (Pet. ether, Chloroform, Ethyl acetate, Methanol and Water) then concentrate all the fractions upto a solid mass. Do arthritis activity against all the extracts. from this you will get active extract. when you would successful to find active extract then frctiionate that extract with different combinations of solvent system from low to high polar using column chromatography method. Then repeat the bio-activity and TLC process for all the fractions. After finding active fraction may be you hv isolated compound depending on concentration of compounds, if not repeat the column chromatography process for that active fractions.............Finally u will success to isolated pure compound...........

fisrt of all you have to dissolve plan extract in water and then portioned it into hexane, chloroform, ethylacetate and butanol soluble fraction then perform your experiment on each fraction which fraction has got high active. then perform column chromatography of that fraction which are active.

To isolate an active compound, you may have to first partition the extract using polar solvent like ethanol or even water and then use nonpolar solvent such as hexane or chloroform on the extract from the polar solvent and use a further semipolar solvent such as ethylacetate. You can then test each on the their ability to achieve what you want. the active one can be used further on TLC and followed by column chromatography. the product can be subjected to NMR to determine the structure of the active compound

I would go with response of Carla W Sabandar, orelse it would be exhaustive method usually followed in academics 

The brief steps are:

First of all you have prepared plant extracts using solvents of various polarity. then isolate the pure fraction from plant extracts using following techniques. First run TLC to know the type of fractions. After that Column Chromatography with HPLC, after isolated pure fraction run FT-IR, LC-MS, HNMR, CNMR for structure elucidation. You can read my papers

First prepare the alcoholic extract and the partitioned with polar and non-polar solvents. find the activity of the fractions with TLC, the more active fraction is subjected to column chromatography for the isolation.

prepare the plant extract (dissolve it in water, by sonication you can get a good extract) Then partitioned it with Hexane, ethyl acetate,  butanol. then preform a screening test. If there is any extracts that shows good activity, you can do further purification by CC, HPLC etc to isolate the pure compounds.   

Best of Luck!!!

I think there are good replies for your question previously in this topic. I would like to explore other points aiming to help you.

- If you are going to apply liquid-liquid partition, beware with proportions and volumes. Depends on what solvent system you will use, in wrong proportions, you will probably have trouble to get phases. In addition, some metabolites can favor the formation of emulsions, and this is not good for your separation process. Butanol is very hard to evaporate, try avoid it. I recommend acetonitrile.

- If you are going to apply any chromatography process using silica gel, you should chose the proper solvents for elution. Silica is very reactive with some class of metabolites, so take care about it, or your bioactive compound will be stuck or degradated.

- If you are going to apply HPLC for purification, using DAD as a detector, remember that it only will detect substances with chromophores. We can loose your compound if it doesn't have a portion that absorves "considerably" in UV range (190-800 nm).

- If you are going to use bioguided assay to monitor your fractionation process, you should consider degradation or sinergism just in case of your fractions were not active.

Hope this suggestion may help you.

Good luck.

Dear Venugopal

The attached information may be useful to you. You can also follow the thread on the same subject.

Best wishes.

https://www.researchgate.net/post/Can_someone_list_out_the_methods_of_extraction_crude_extract_compounds_isolation_purification_identification_and_structural_elucidation

Olive leaf harbours antioxidant properties that help protect the body from the continuous activity of free radicals which are highly reactive chemical substances that can cause cellular damage if left unchecked. Some recent research on the olive leaf has shown its antioxidants to be effective in treating some tumors and cancers such as liver, prostate, colon, skin and breast cancer, clinical studies lacking; Olive leaf is especially potent when used in combination with other antioxidants.Somova et al. Antihypertensive, antiatherosclerotic and antioxidant activity of triterpenoids isolated from Olea europaea, subspecies africana leaves, 2003.

• Dr Stevenson, L,. et al. Oxygen Radical Absorbance Capacity (ORAC) Report on Olive Leaf Australia's Olive Leaf Extracts, Southern Cross University, 2005.
• Benavente-Garcia et al. Antioxidant activity of phenols extracted from Olea europaea L. leaves, 2000.
• Saija et al. In vitro evaluation of the antioxidant activity and biomembrane interaction of the plant phenols oleuropein and hydroxytyrosol, 1998.Speroni et al. Oleuropein Evaluated In Vitro and In Vivo as an Antioxidant, 1998.
• Pinelli et al. Quali-quantitative analysis and antioxidant activity of different polyphenolic extracts from Olea europea L. leaves, 2000.
• Hamdi et al. Oleuropein, a non-toxic olive iridoid, is an anti-tumor agent and cytoskeleton disruptor, 2005.
• Dr Stevenson, L,. et al. In vitro Biological Activities of Pure Olive Leaf Extract & High Strength Olive Leaf Extract, 2006.

One suggestion, review all reported studies regarding phytochemistry of your plant sample and others species under same genus to get very valuable information of type of compounds that maybe play significant role for your bioassay. In addition, phytochemicals screening is recommended as a basic guideline for the isolation of compounds.

I hope this article help you to find your answer

You should do activity directed fractionation of the crude extract.

I have a mixture of crude metabolite and i want to find out the concentration of active metabolite from the HPLC chromatogram of known concentration of crude metabolite then how do i go about it? Taking in to account the loading volume (loop volume) for HPLC.

You can use the same investigation method described in the attached search

Article HPLC fractioning to study the synergy and antagonism of rue ...

Thank you @ Naser Jawad Kadhum

https://www.researchgate.net/publication/305769482_HPLC_fractioning_to_study_the_synergy_and_antagonism_of_rue_plant_seeds_alkaloid_to_inhibit_topoisomerase_II_as_antitumor