3 Replies Latest reply on Jan 24, 2013 9:27 AM by An-Phong Le

    Hydrogen Bonding and UV Absorbance

    mahdi rabiei

      Hi

      Does hydrogen bonding of acrylamide with another molecule affect its absolute contribution in UV absorbance?

      in detail if X concentration of acrylamide has AX absorbance at 200 nm, by adding another molecule which not absorb at this wavelength and interact with acrylamide only by hydrogen bonding, does the absorbance value (AX) affected? Why ? if yes, increases or decreases? why?

      (the UV absorbance measured around 200 nm because of C=O bonds)

      Thanks in advance for your prompt response

        • Re: Hydrogen Bonding and UV Absorbance
          Deborah Fox Walsh

          Hi Mahdi,

            If you don't see the expected response at the predetermined wavelength for UV absorbance measurement, think about where you may be losing your molecule of interest during the experimental process.  Is it soluble?  Did you attempt to isolate the molecule? Was the recovery poor, was it lost during the extraction process?  Consider the HPLC system, is it performing properly? Are you using the correct column, or could it be your mobile phase or gradient? Is your molecule bonding irreversibly to the column or vials?

          Although I don't have any specific experience with acrylamide, these are some of the things I'd consider.....

            • Re: Hydrogen Bonding and UV Absorbance
              mahdi rabiei

              Hi Deborah

              thank you for your response

              actually I measured the UV absorbance of a certain Concentration of acrylamide (which i dissolved in water), and in parallel I measured the UV absorbance of a certain Concentration of a protein which i dissolved in water at the same wavelength.

              and subsequently when I mixed the solutions, the absorbance of the mixture was significantly less than sum of two absorbance values.

              all I know about the mixture is that there are only hydrogen bonds between acrylamide and the protein.