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The Henderson–Hasselbalch equation can be used to model these equilibria. It is important to maintain this pH of 7.4 to ensure enzymes are able to work optimally. [10] Life threatening Acidosis (a low blood pH resulting in nausea, headaches, and even coma, and convulsions) is due to a lack of functioning of enzymes at a low pH. [10]
Conversely, when pH = pK a, the concentration of HA is equal to the concentration of A −. The buffer region extends over the approximate range pK a ± 2. Buffering is weak outside the range pK a ± 1. At pH ≤ pK a − 2 the substance is said to be fully protonated and at pH ≥ pK a + 2 it is fully dissociated (deprotonated).
Buffer capacity falls to 33% of the maximum value at pH = pK a ± 1, to 10% at pH = pK a ± 1.5 and to 1% at pH = pK a ± 2. For this reason the most useful range is approximately pK a ± 1. When choosing a buffer for use at a specific pH, it should have a pK a value as close as possible to that pH. [2]
The concentration of water, [H 2 O], is omitted by convention, which means that the value of K w differs from the value of K eq that would be computed using that concentration. The value of K w varies with temperature, as shown in the table below. This variation must be taken into account when making precise measurements of quantities such as pH.
It consists of a mixture of 0.04 M boric acid, 0.04 M phosphoric acid and 0.04 M acetic acid that has been titrated to the desired pH with 0.2 M sodium hydroxide. Britton and Robinson also proposed a second formulation that gave an essentially linear pH response to added alkali from pH 2.5 to pH 9.2 (and buffers to pH 12).
Add 2.84 mM of HCl to shift the buffer to 7.3 mM HPO 4 2− and 4.6 mM H 2 PO 4 − for a final pH of 7.4 and a Cl − concentration of 142 mM. The pH of PBS is ~7.4. When making buffer solutions, it is good practice to always measure the pH directly using a pH meter. If necessary, pH can be adjusted using hydrochloric acid or sodium hydroxide.
The useful buffer range for tris (pH 7–9) coincides with the physiological pH typical of most living organisms. This, and its low cost, make tris one of the most common buffers in the biology/biochemistry laboratory. Tris is also used as a primary standard to standardize acid solutions for chemical analysis.
The pKa of KHP is 5.4, so its pH buffering range would be 4.4 to 6.4; however, due to the presence of the second acidic group that bears the potassium ion, the first pKa also contributes to the buffering range well below pH 4.0, which is why KHP is a good choice for use as a reference standard for pH 4.00. [8] [9]