I have the article "Colorimetric Analysis of Sugars" from 1957 (cited below) and it proposes several methods. I have read recent papers where the anthrone reaction is used, but the article proposes several other methods and I am wondering whether some of the others might be better.
As of right now, I haven't tried any yet. I culture my bacteria with glucose and use a commercial enzymatic kit to measure it. The kit was designed to measure blood glucose.
My goal is to escalate the process to industrial proportions and use sucrose instead of glucose. I have been told that I can hydrolyse sucrose with acid and use my same glucose commercial kit, but this is also expensive.
Does anyone know or currently use a better method for sucrose? All the help is much appreciated, thank you!!
http://dx.doi.org/10.1016/S0076-6879(57)03350-9
Veronica,
Gilbert Ashwell published an updated set of procedures to distinguish between aldo- and keto- sugars in 1966, you may wish to read it:
New Colorimetric Methods of Sugar Analysis
http://ac.els-cdn.com/0076687966080108/1-s2.0-0076687966080108-main.pdf?_tid=36b863b0-1b73-11e5-976a-00000aab0f27&acdnat=1435261915_8c0b3a16f0db67b960c5b9fcba51519e
You probably will not find kits for these reagents but they require very few things, they are simple and straightforward. You do not need even a spectrophotometer, an old-fashioned colorimeter does a fine job.
In case of sucrose, probably would want to acid hydrolyze the disaccharide to monomers first. I have successfully used this method decades ago and it proved quite reliable., with the caveat that it is more time consuming.
Another very old method is the Luff Schoorl method to determine glucose.
http://www.sigmaaldrich.com/catalog/product/fluka/34400?lang=en®ion=N
A few years back, we were using a diabetic strip meter to measure excess glucose in our yeast bioreactor fermentations. It worked very nicely and was fast, accurate and reproducible. So, after hydrolysis of sucrose with either acid or treatment with the enzyme invertase, you could try employing this type of meter. The good thing is this is easy to test out with some standard curves with known quantities of glucose, then hydrolyzed sucrose to see how well they agree.
I hope this is helpful in some way.
Dear Veronica,
cheap depends on what you already have, e.g. if you ahave access to an an HPLC System with an refractive index detector and an "amino-phase" this would be cheap in Terms of running costs if you optimize runtime etc.
Challenge of your question is that you won't have that much analyte, since you work with cell cultures -however this might not be aproblem when you swithc to a bigger scale.
When do you not have access to HPLC, then I second the suggestion of Kees (Luff Schoorl) - I have made hundreds of that and it really works fine.
In addition, you may want to have a look at quantitative TLC (Thin Layer Chromatography). You can even prepare the plates yourself if bying is to expensive. When you do not own a Densitometer you can visually compare spots to references. Advantage would be very limited sample preparation (depending on your Matrix, of course).
Good luck!
Holger
Thank you all so much for answers!
My request for "cheap" was aimed at a method that didn't require to buy kits or expensive reactants. Tausif posted the article and there was the sulfuric acid-phenol method, I think I will try that.
Thank you all!!
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology