Which assay can be used for the quantification of synthetic GSH?
I have tried using the DTNB assay but it over estimates the concentration of the GSH 10x
If you are overestimating by 10-fold, I recommend checking your calculations for an error. The DTNB assay should give the correct result (less any oxidized GSSG in the GSH).
You can follow my paper titiled "In vivo chlorpyrifos induced oxidative stress: Attenuation by antioxidant vitamins" Pesticide Biochemistry and Physiology 88: (2007), 191-196. cited on my researchgate in fulltext form.
There are several methods for estimation of GSH the problem is some of the GSH may oxidized upon exposure to oxygen to form GSSG DTNB is good method however you can use HPLC using C18 column for this method you can look it up in my research gate this is fast can easily distinguish between GSH and GSSG good luck to you
I agree with Abdelrahim, HPLC method will be more precise than DTNB. Use NEM to avoid un specific oxidation
DTNB is not specific for GSH but reacts with all thiols present (e.g. cysteine, protein thiols) in the assay. It largely depends on what kind of sample you have. In case you have a 10x overestimation in pure GSH solutions I agree with Adam i.e. check your calculations for error(s).
- Badges
- Science topic
- Similar topics
- Correction