lactate quantification - spectroscopy (Nov/23/2005 )
hi any one knows how to measure lactate levels in samples.This is not blood lactate.It is a kind of polymer degradation product in PBS buffer? Preferably any spectroscopic methods???
R-Biopharm has a kit to measure lactic acid. Here is the PDF file of the protocol their kit uses. Their kit has some good and bad points. Good, is that using dual readings and set calculations, you can measure the concentration of your sample without having to make a standard curve everytime. The bad is that you have to do these calculations and I hate math with a passion. Most of these assay kits aren't terribly expensive though.
just a guess, for determination of pyruvate concentration i used lactate-dehydrogenase to reduce pyruvate in a NADH-dependent manner. U can directly follow the reaction using a spectrophotometer and calculate via lambert-beer the initial concentration.
I'm not sure on which side the equilibrium is, but by adapting the concentration of the reactants you might do the reverse reaction and observe the increase in absorption due to the increasing amounts of NADH.
edit: better should have read captain_DNA's reply...