i used protocols described by Yoshida et al.1976. any other suggestion?
10/19/17
Dear Mohd,
If you want a simple method for measuring carbohydrate (CHO), you can use the procedure of DuBois et al. (See below.). The reagents are 5% w/v phenol in water and concentrated H2SO4. It works very well, is quick & easy to perform and sensitive. I modified the procedure a bit to improve sensitivity and save on reagents.
Procedure: Sample + DI-water = 0.50 mL. Add 0.50 mL 5% phenol, followed by 2.5 mL concentrated H2SO4. Mix well. (Caution! Addition of H2SO4 makes the final solution very hot!) Allow ~20-30 min to cool, then read Abs @ 490 nm. You can use glucose as a standard. With the above procedure, you can generate a calibration curve from 5- to 90 uG CHO. The article lists the various sugars, etc. with which the method works. It works for simple sugars (mono- and oligosaccharides) as well as polysaccharides. It does not work with amino sugars.
M. Dubois, K.A. Gilles, J.K. Hamilton, P.A. Rebers, F. Smith, Colorimetric method for determination of sugars and related substances, Anal. Chem. 28 (1956) 350–356.
Another assay procedure you can try is attached. It uses anthrone and H2SO4. I've never used it, but it may work for you.
Good luck!
Bill Colonna Center for Crops Utilization Research, Iowa State University, Ames, IA USA
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