I have purified alkaloid from papaya leaf extract. I underwent IR for the same. They are in good accordance of peaks. As a result i performed confirmatory chemical test for all functional group. they were positive. Now I need to make sure for the alkaloid by other means. please suggest
Mr/Miss/Mrs Saraf,
Alkaloids determination by IR is a very complex task, which would be carried out when the molecular structure is relatively simple with characteristic groups. But this is a rare case of alkaloids. Instead IR, there is mass spectrometry (MS / MS modse) for fingerprinting of alkaloids.
Hi, I agree with the comments of Dr. Ivanova. Do you want to know whether your isolate (alkaloid) is already pure? If it is yes, generally at our Lab. we test the purity of our isolate by using TLC (2D-TLC), HPLC-DAD, and DSC. For chromatography we do by using at least 3 chromatography systems; sometime we need more than one method to prove the purity of the isolate.
First of all test purity by TLC, at least in 2 different solvent systems. If the compound gives a single spot, go for melting point. A sharp MP indicate purity of the compound and you can also compare MPs of the reported alkaloids from Papaya in literature and almost confirm your compound. Use your IR data for further confirmation. If it is a common alkaloid available in the market, you can purchase the authentic and compare with TLC and IR. Offcourse H NMR, 13 C NMR, MS, HRMS, MS/MS are also there to authenticate your alkaloid.
I have started with the steps given Rameshkumar K B. This I would be followed by NMR/ GC-MS
Regards
Complete elucidation of the structure of alkaloids is made with IR, UV, and of course nuclear magnetic resonance (1H-NMR and13C-NMR) and mass spectrometer . To be completely sure about the structure you should get good crystals and then get the structure by X-Ray Crystallography.
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