The excess iron is added to the serum to bind all the ferritin in the serum, and the excess iron is adsorbed by adding the iron adsorbent. The iron bind with the ferritin is separated from the protein by the action of acid solution and reductant. Fe3+ in serum is reduced to Fe2+, Fe2+ binds with bipyridine to form pink complex. In a certain range, the amount of TIBC is positively correlated with the depth of color. The iron content measured is, minus serum iron value, which is called unsaturated iron binding force. Total iron binding capacity minus serum iron value is unsaturated iron binding capacity (UIBC).
Under the action of reductant, iron ions in samples can be reduced into ferrous ions (Fe2+). The latter then bind to probe and form complexes, which has a maximum absorption peak at 593 nm. The concentration of iron can be calculated by measuring the OD value at 593 nm indirectly.
Under acid conditions, the oxidizing material in the sample can oxidize Fe2+ to Fe3+, which binds highly with xylenol orange to produce a blue-purple complex. When the pH of solution is in the range of 2-3, its maximum absorption wavelength is around 590 nm, and the color depth is proportional to the content of oxidation substances in a certain concentration and a certain time, so as to indirectly calculate the total oxidation state of the sample.
Plant total phenol is a common secondary natural metabolite in plants. There are several kinds of phenolic compounds, such as hydroxybenzoic acid, hydroxy cinnamic acid, flavonoids, chalcone, flavonoids, lignin, coumarin and astragalus. Phenolic compounds are antioxidants that delay or prevent oxidation and oxygen radical reactions.
Under alkaline conditions, tungsten-molybdenum acid can be reduced by phenols and produce blue compounds, which has a characteristic absorption peak at 760 nm. The content of total phenols in sample can be calculated indirectly by measuring the absorbance at 760 nm.
Phosphodiesterase (PDEs) hydrolyzes the intracellular second messenger (cyclic adenosine phosphate or cyclic guanosine phosphate), thereby ending the biochemical effects of these second messengers. The principle of this kit is that by hydrolyzing the substrate with total PDEs, the resulting substance has a maximum absorption peak at 405 nm. The OD value at 405 nm and the standard curve were measured to calculate the total PDEs activity in the sample.
Cell and tissue samples are treated with lysates containing protease inhibitors and phosphatase inhibitors to prevent the enzymes in the sample from being released to hydrolyze or dephpsphprylate protein due to the disruption of the membrane system.
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