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Chemistry Forums for Students => Analytical Chemistry Forum => Topic started by: Carli11235 on February 20, 2021, 09:02:11 AM

Title: Ferrous Lactate Determination by HPLC
Post by: Carli11235 on February 20, 2021, 09:02:11 AM
I am trying to figure out if using HPLC/UV to quantify iron (II) lactate in an aqueous solution is possible. Would the ferrous lactate dissociate when diluted to the low concentration needed for analysis? I cannot quantify the lactate ion alone due to interference of other molecules at that wavelength which cannot be adequately separated.
Title: Re: Ferrous Lactate Determination by HPLC
Post by: Babcock_Hall on February 20, 2021, 10:22:33 AM
This is not a direct answer to your question, but lactate might in principle be determined spectrophotometrically by enzymatic oxidation to pyruvate (although the equilibrium constant for this process with NAD is not favorable).  There is a series of books called Methods of Enzymatic Analysis by Bergmeyer that could be consulted.
Title: Re: Ferrous Lactate Determination by HPLC
Post by: Carli11235 on February 20, 2021, 10:53:32 AM
Thank you for the suggestion! Though, are too many additional compounds in the mixture that absorb at the same wavelength for that to work. Hence, the desire for a chromatographic method.
Title: Re: Ferrous Lactate Determination by HPLC
Post by: Babcock_Hall on February 20, 2021, 11:31:13 AM
Just to be clear, NADH absorbs at 340 nm.  From your answer I assume that you have interferences there.