The total saponin content is determined by colorimetry methods, using gin- senoside Rb1 as standard. Measurements is carried out at 550 nm, and the results give a total saponin content of ginsenoside equivalent saponin content per gram of the plant extract. For structural elucidation NMR and mass spectrometry methods are used.
Dear Ghasem,
Please go through this article. I hope this article may help you.
Makker, H. P. S.; Becker, K. Nutrients and antiquality factors in different morphological parts of the Moringa oleifera tree. J. Agric. Sci. Cambridge 1997, 128, 311–322.
Isolation of Saponin-Rich Guar Extract
A total of 5 samples from guar meal were used in this study. Guar meal was extracted by refluxing 25 g of guar meal with 250 ml of ethanol: water, 1:1 (v: v) for 3 h in a simple reflux apparatus. Refluxed extracts were cooled and filtered through 150 µm Hassan et al. 25
(Watman No. 2) then 125 µm pore size (Watman No. 114) filter papers. Ethanol was removed from the filtered extract by evaporating under reduced pressure in a roto-evaporator (Buchi,
Rinco Instrument Co., Inc., Greenville, IL, Switzerland, model 310391) until two-thirds of the initial volume was removed. The remaining aqueous extract was partitioned with n-butanol, 1:1 (v: v) overnight at room temperature using a separatory funnel. The upper n-butanol extract was collected in a glass flask and the lower aqueous extract was collected and further partitioned with n-butanol two more times to increase the yield of crude saponin extract. The
butanol extracts were pooled and evaporated to remove the butanol or dryness under vacuum at temperature below 50°C using the roto-evaporator procedure described above. A minimum volume of distilled water was added to the dry n-butanol extract and the resulting material was freeze-dried and weighed.
Taken from the paper link
http://netjournals.org/pdf/AMPR/2013/1/13-013.pdf
I agree with Mr Mudhir . But One change at the final stage After removal of Butanol under vacuum ,weight of residue taken to constant weight and calculated.
Please go to the article:
VIGO, Cléverson Luiz dos Santos ; NARITA, Emília ; MARQUES LC . Validação da metodologia de quantificação espectrofotométrica das saponinas de Pfaffia glomerata (Spreng.) Pedersen - Amaranthaceae. Revista Brasileira de Farmacognosia, Maringá, PR, v. 13, n.supl. 2, p. 46-49, 2003.
http://dx.doi.org/10.1590/S0102-695X2003000400006
One of the best methods to determine saponins in plants is using HPLC ou UPLC determination. Look in the literature for the papers of Dr. Chigen Tsukamoto he has a great experience in saponins determination. Dr. Tsukamoto belongs to The Department of Applied Biological Chemistry, Faculty of Agronomy Iwate University, Japan.
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