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The Henderson–Hasselbalch equation can be used to estimate the pH of a buffer solution by approximating the actual concentration ratio as the ratio of the analytical concentrations of the acid and of a salt, MA. The equation can also be applied to bases by specifying the protonated form of the base as the acid.
In biochemistry, the molar absorption coefficient of a protein at 280 nm depends almost exclusively on the number of aromatic residues, particularly tryptophan, and can be predicted from the sequence of amino acids. [6] Similarly, the molar absorption coefficient of nucleic acids at 260 nm can be predicted given the nucleotide sequence.
Absorbance within range of 0.2 to 0.5 is ideal to maintain linearity in the Beer–Lambert law. If the radiation is especially intense, nonlinear optical processes can also cause variances. The main reason, however, is that the concentration dependence is in general non-linear and Beer's law is valid only under certain conditions as shown by ...
Nucleic acid preparations uncontaminated by phenol should have a A 260/280 of around 2. [2] Contamination by phenol can significantly contribute to overestimation of DNA concentration. Absorption at 230 nm can be caused by contamination by phenolate ion, thiocyanates, and other organic compounds.
For example, if the concentration of the conjugate base is 10 times greater than the concentration of the acid, their ratio is 10:1, and consequently the pH is pK a + 1 or pK b + 1. Conversely, if a 10-fold excess of the acid occurs with respect to the base, the ratio is 1:10 and the pH is pK a − 1 or pK b − 1.
Outside the transition range the concentration of acid or conjugate base is less than 10 % and the colour of the major species dominates. Species concentrations calculated with the program HySS for a 10 mM solution of citric acid. pK a1 = 3.13, pK a2 = 4.76, pK a3 = 6.40. A weak acid may be defined as an acid with pK a greater than
Absorbance is defined as "the logarithm of the ratio of incident to transmitted radiant power through a sample (excluding the effects on cell walls)". [1] Alternatively, for samples which scatter light, absorbance may be defined as "the negative logarithm of one minus absorptance, as measured on a uniform sample". [2]
Absorption is a condition in which something takes in another substance. [1] In many processes important in technology, the chemical absorption is used in place of the physical process, e.g., absorption of carbon dioxide by sodium hydroxide – such acid-base processes do not follow the Nernst partition law (see: solubility).