Dear Sujata, try chloroform fraction, i think most of the alkaloids come in that 

Dear Sujata

Please find some method in attachments. Although basifying plant material with ammonia water and extracting it with chloroform or methanol is mostly used and validated method.

Good luck

thank  you for the anwers . But separation of the individual alkaloids from the fraction how it will be carried out

If your alkaloid is UV active then try it with in preparative TLC plate. If not, then pass the fraction with column using isocratic elution. First try to search literature, whether there is any report on the same plant for the isolation of alkaloids or not, if yes then first try their method.

i got in methanol but its not uv active and only soluble in dmso

Dear Sujata

Just make partition studies with your Methanol fraction using various pH with various solvent. You definitely get the pure compound. 

I can think of 2 easy ways of doing it. In both cases extract with methanol.

The first method involves multiple extractions. First partition between dichloromethane and acidic water. I like to use formic or acetic acid in case the water soluble compounds are also of interest- these acids can be evaporated. Next, make the aqueous  solution basic with ammonia (using the acids I mentioned earlier, the ammonium salts are also volatile) and partition again with dichloromethane.  After this step, the alkaloids will be in the dichloromethane as free bases.

The other way is using an strong cation exchange (SCX) column. The advantage of this method is that one can start separating different alkaloids. The links below describe the methods I use:

http://www.isco.com/WebProductFiles/Applications/101/Application_Notes/AN40_Purification_of_Alkaloids_with_RediSep_Rf_SCX_Columns.pdf

http://www.isco.com/WebProductFiles/Applications/101/Application_Notes/AN98_Hints%20for%20Strong%20Ion%20Exchange%20Resins.pdf

This link also shows some methods for subsequent purification:

http://www.isco.com/WebProductFiles/Applications/101/Poster_and_Paper_Reprints/Alkaloid_Purification_Strategies_Using_Flash_Chromatography.pdf

thank you @jack silver

Great Discussion on this thread..

I m trying to purify alkaloids too.

I m performing TLC for my crude extract. But not sure which band is Alkaloid.. 

Whats the UV range for Alkaloids in general, if it is UV Active...

@ Lokesh: Alcaloids have a wide variety of structures, and absorb at many different wavelengths. Some ( notably quinine, harmine) will even flouresce.

I suggest taking a small amount of the crude extract (20-50 milligrams will be sufficient)  and doing the solvent extract described above. Run TLC on the final extract, the bands you see on your TLC will be alkaloids.

Dragendorff reagent consists of (Nitrooxy)oxobismuthine (BiNO4xH2O), tartaric acid and KI and it is used for the DC detection of alkaloids and - more general - tertiary amines (and also some secundary ones). The Tertiary amine is protonated due to the tartaric acid and an ion pair is formed that consists of [BiI4]- and [HNR3]+. In dependence of the nature of the tert. amine (or alkaloid) this ion pair has a yellow...orange...red...brown collor. Secondary amines will create less colored spots.

use it