Thank you so much. I have been searching this information for more than 1 week. You explained it in an understandable way. Can we use this calculation for NADPH?
Yes - NADPH absorbs light directly at 340nm so you can use Beer's law directly. Note that NADH also absorbs at this wavelength, so you will need to design your experiment carefully to ensure it is NADPH you are measuring specifically. Hope that helps!
I want to know the molar absorption coefficient of phenylhydrazone. Where can i find. Because instead of NADH, i have glyoxylate phenylhydrazone formation as product
No - in this case if there is a 1:1 conversion of reactant -> product and both absorb at the same wavelength you won't be able to measure the change in absorbance. You'd need to extract the product specfically
Great tutorial! And about the volume of the reaction? Do you think we should consider it?
Most helpful answer I've found online, thank you!
This was great helped A LOT Thanks!
Thank you so much. I have been searching this information for more than 1 week. You explained it in an understandable way. Can we use this calculation for NADPH?
Yes - NADPH absorbs light directly at 340nm so you can use Beer's law directly. Note that NADH also absorbs at this wavelength, so you will need to design your experiment carefully to ensure it is NADPH you are measuring specifically. Hope that helps!
I want to know the molar absorption coefficient of phenylhydrazone. Where can i find. Because instead of NADH, i have glyoxylate phenylhydrazone formation as product
I don’t know as have never worked on that reaction but looks like this paper might be useful? pubmed.ncbi.nlm.nih.gov/10731695/
If the reactant and product absorb same wavelength then it is possible to use spectrophotometer to find the absorbance
No - in this case if there is a 1:1 conversion of reactant -> product and both absorb at the same wavelength you won't be able to measure the change in absorbance. You'd need to extract the product specfically
Thanks