What do you thing about this method using Agilent ZORBAX Carbohydrate column? Do you think its better for me to use longer column?
Hi,
Rebaudioside can be analysed using Amide column and ELSD (Evoparation Light scattering detector.
I have attached application note for the same. Hope this will help you.
Regards,
Agilent has an excellent app note on steviol glycoside analysis on their Web site: www/agilent.com
Reversed phase HPLC with UV detection at 210nm has been used to determine constituents in commercial Stevia products, but the methodology is limited in its ability to separate all components of interest, especially when higher order oligosaccharides are present. Another problem is that any single UV wavelength is a detection compromise as the constituents exhibit a wide range of absorbance maxima (193, 204, 236, 238, 284nm). Some are only weakly chromophoric.
The Evaporative Light Scattering Detector (ELSD) is a valuable complement to spectroscopic detectors for HPLC. The ELSD makes a measurement of photons scattered from semi- and non-volatile particles that have been dried of mobile phase through evaporation. Because its response is independent of the light absorbing properties of molecules, it can reveal sample components that UV detectors miss and provide a more accurate profile of relative component abundance than is possible with a spectroscopic detector.
Here i had attach an application note of Shimadzu. Hope it will we helpful to you
Thank you guys..rebaudioside a is my interest compound..in the paper using 150mm of column but i think in my extraction i need longer column maybe 250mm bcoz diterpene glycosides contain many compound to avoid problems later. I'll try soon, hope for the best.
Fast and isocratic HPLC-method for steviol glycosides analysis from Stevia rebaudiana leaves
Bergs, D., Burghoff, B., Joehnck, M., Martin, G., Schembecker, G., Journal fur Verbraucherschutz und Lebensmittelsicherheit, Volume 7, Issue 2, June 2012, Pages 147-154
Kolb, N. , Herrera, J.L., Ferreyra, D.J., Uliana, R.F., Analysis of sweet diterpene glycosides from Stevia rebaudiana: Improved HPLC method, Journal of Agricultural and Food Chemistry, Volume 49, Issue 10, 2001, Pages 4538-4541
Liu, J., Li, S.F.Y., Separation and determination of Stevia sweeteners by capillary electrophoresis and high performance liquid chromatography, Journal of Liquid Chromatography, Volume 18, Issue 9, 1995, Pages 1703-1719
Mauri, P, Catalano, G., Gardana, C., Pietta, P., Analysis of Stevia glycosides by capillary electrophoresis, Electrophoresis, Volume 17, Issue 2, February 1996, Pages 367-371
Pieri, V., Belancic, A., Morales, S., Stuppner, H., Identification and quantification of major steviol glycosides in stevia rebaudiana purified extracts by 1H NMR spectroscopy, Journal of Agricultural and Food Chemistry, Volume 59, Issue 9, 11 May 2011, Pages 4378-4384
Rajasekaran, T., Ramakrishna, A., Udaya Sankar, K., Giridhar, P., Ravishankar, G.A.,
Analysis of predominant steviosides in Stevia rebaudiana Bertoni by liquid chromatography/electrospray ionization-mass spectrometry, Food Biotechnology,
Volume 22, Issue 2, April 2008, Pages 179-188
Sakamaki, N., Matsumoto, H., Hagino, K., Nakazato, M., Yasuda, K., Simultaneous determination of stevioside, rebaudioside A and glycyrrhizic acid in foods by HPLC,
Journal of the Food Hygienic Society of Japan, Volume 45, Issue 2, 2004, Pages 81-86
Woelwer-Rieck, U., Lankes, C., Wawrzun, A., Wüst, M., Improved HPLC method for the evaluation of the major steviol glycosides in leaves of Stevia rebaudiana, European Food Research and Technology, Volume 231, Issue 4, 2010, Pages 581-588
- Badges
- Science method