Systems buffer

Na20 20 4H20 (60). Mixtures of B(OH)2 and B(OH) 4 appear to form classical buffer systems where the solution pH is governed primarily by the acid salt ratio, ie, [B(OH)3]/[B(OH) ]. This relationship is nearly correct for solutions of sodium or potassium (1 2) borates, eg, borax, where... 

Aqueous solutions of citric acid make excellent buffer systems when partially neutralized because citric acid is a weak acid and has three carboxyl groups, hence three p-K s. At 20°C pifj = 3.14, pi 2 4.77, and = 6.39 (2). The buffer range for citrate solutions is pH 2.5 to 6.5. Buffer systems can be made using a solution of citric acid and sodium citrate or by neutralizing a solution of citric acid with a base such as sodium hydroxide. In Table 4 stock solutions of 0.1 Af (0.33 N) citric acid are combined with 0.1 Af (0.33 N) sodium citrate to make a typical buffer solution. 

Flue Ga.s Desulfuriza.tion. Citric acid can be used to buffer systems that can scmb sulfur dioxide from flue gas produced by large coal and gas-fired boilers generating steam for electrical power (134—143). The optimum pH for sulfur dioxide absorption is pH 4.5, which is where citrate has buffer capacity. Sulfur dioxide is the primary contributor to acid rain, which can cause environmental damage. 

Disc electrophoresis was first iatroduced ia the early 1960s (11—13) as various techniques using polyacrylamide gels were being explored and designed. Original work employed several buffer systems and different polyacrylamide gels in order to first concentrate and then separate compounds (14). 

Seals, particularly the restricted leakage type, require auxiliary piping, control valves, and other items. If more than a suggested schematic is to be included in the vendor s scope of supply, a statement to that effect should be made. For the oil buffered systems, if a separate oil system is required, this must be stated, and it can be detailed in the accessory section with the lube system. For dry gas seals, the type of support system must be specified. The vendor s proposal should be requested to outline materials and to... 

The role that acid and base catalysts play can be quantitatively studied by kinetic techniques. It is possible to recognize several distinct types of catalysis by acids and bases. The term specie acid catalysis is used when the reaction rate is dependent on the equilibrium for protonation of the reactant. This type of catalysis is independent of the concentration and specific structure of the various proton donors present in solution. Specific acid catalysis is governed by the hydrogen-ion concentration (pH) of the solution. For example, for a series of reactions in an aqueous buffer system, flie rate of flie reaction would be a fimetion of the pH, but not of the concentration or identity of the acidic and basic components of the buffer. The kinetic expression for any such reaction will include a term for hydrogen-ion concentration, [H+]. The term general acid catalysis is used when the nature and concentration of proton donors present in solution affect the reaction rate. The kinetic expression for such a reaction will include a term for each of the potential proton donors that acts as a catalyst. The terms specific base catalysis and general base catalysis apply in the same way to base-catalyzed reactions. 

The experimental detection of general acid catafysis is done by rate measurements at constant pH but differing buffer concentration. Because under these circumstances [H+] is constant but the weak acid component(s) of the buffer (HA, HA, etc.) changes, the observation of a change in rate is evidence of general acid catalysis. If the rate remains constant, the reaction exhibits specific acid catalysis. Similarly, general base-catalyzed reactions show a dependence of the rate on the concentration and identity of the basic constituents of the buffer system. 

Mount the upper end piece of the filling tube and immediately connect it to the pump, open the column outlet, and start the pump at a flow rate of 10 ml/min (delivering water or any desired buffer system of low ionic strength). Optionally, the column outlet can be additionally connected to a water jet pump, which has to be operated during the first 2 min of packing. The water... 

Equilibrate the packed gel with at least 3 bed volumes at 1 ml/min of the desired buffer system. It is recommended to include 50-300 mM of salt in the buffer. 

FIGURE 7.14 A Fractogel EMD BioSEC Superformance column (600-16) was loaded with 500 /il of BSA, ovalbumin, and cytochrome c (5/5/3 mg/ml) at I ml/min. The test covered 100 individual runs with the standard proteins as samples. The buffer system used was 20 m/VI sodium dihydrogen phosphate, 300 m/VI NaCI, pH 7.2. After each individual run the column was rinsed with I /VI NaOH (60 min with I /VI NaOH at 2 ml/min). No significant change in retention times and resolution was observed after 100 cycles. 

Target protein is not stable in the chosen buffer system for SEC... 

In the development of a SE-HPLC method the variables that may be manipulated and optimized are the column (matrix type, particle and pore size, and physical dimension), buffer system (type and ionic strength), pH, and solubility additives (e.g., organic solvents, detergents). Once a column and mobile phase system have been selected the system parameters of protein load (amount of material and volume) and flow rate should also be optimized. A beneficial approach to the development of a SE-HPLC method is to optimize the multiple variables by the use of statistical experimental design. Also, information about the physical and chemical properties such as pH or ionic strength, solubility, and especially conditions that promote aggregation can be applied to the development of a SE-HPLC assay. Typical problems encountered during the development of a SE-HPLC assay are protein insolubility and column stationary phase... 

A second way to achieve constancy of a reactant is to make use of a buffer system. If the reaction medium is water and B is either the hydronium ion or the hydroxide ion, use of a pH buffer can hold Cb reasonably constant, provided the buffer capacity is high enough to cope with acids or bases generated in the reaction. The constancy of the pH required depends upon the sensitivity of the analytical method, the extent of reaction followed, and the accuracy desired in the rate constant determination. 

The sensitivity of the equilibrium constant to temperature, therefore, depends upon the enthalpy change AH . This is usually not a serious limitation, because most reaction enthalpies are sufficiently large and because we commonly require that the perturbation be a small one so that the linearization condition is valid. If AH is so small that the T-jump is ineffective, it may be possible to make use of an auxiliary reaction in the following way Suppose the reaction under study is an acid-base reaction with a small AH . We can add a buffer system having a large AH and apply the T-jump to the combined system. The T-jump will alter the Ka of the buffer reaction, resulting in a pH jump. The pH jump then acts as the forcing function on the reaction of interest. 

FIGURE 2.15 A buffer system consists of a weak acid, HA, and its conjugate base, A. The pH varies only slightly in the region of the titration curve where [HA] = [A ]. The unshaded box denotes this area of greatest buffering capacity. Buffer action when HA and A are both available in sufficient concentration, the solution can absorb input of either H or OH, and pH is maintained essentially constant. 

Buffers are solutions that tend to resist changes in their pH as acid or base is added. Typically, a buffer system is composed of a weak acid and its conjugate base. A solution of a weak acid that has a pH nearly equal to its by definition contains an amount of the conjugate base nearly equivalent to the weak acid. Note that in this region, the titration curve is relatively flat (Figure 2.15). Addition of H then has little effect because it is absorbed by the following reaction ... 

The pH then remains relatively constant. The components of a buffer system are chosen such that the of the weak acid is close to the pH of interest. It is at the that the buffer system shows its greatest buffering capacity. At pH values more than one pH unit from the buffer systems become ineffec-... 

The important buffer system of blood plasma is the bicarbonate/carbonic acid couple ... 

X 10 M), and an equivalent amount of OH (its usual concentration in plasma) would swamp the buffer system, causing a dangerous rise in the plasma pH. How, then, can this bicarbonate system function effectively The bicarbonate buffer system works well because the critical concentration of H2CO3 is maintained relatively constant through equilibrium with dissolved CO2 produced in the tissues and available as a gaseous CO2 reservoir in the lungs. ... 

Under the conditions of temperature and ionic strength prevailing in mammalian body fluids, the equilibrium for this reaction lies far to the left, such that about 500 CO2 molecules are present in solution for every molecule of H2CO3. Because dissolved CO2 and H2CO3 are in equilibrium, the proper expression for H2CO3 availability is [C02(d)] + [H2CO3], the so-called total carbonic acid pool, consisting primarily of C02(d). The overall equilibrium for the bicarbonate buffer system then is... 

For most free amino acids and small peptides, a mixture of alcohol with water is a typical mobile phase composition in the reversed-phase mode for glycopeptide CSPs. For some bifunctional amino acids and most other compounds, however, aqueous buffer is usually necessary to enhance resolution. The types of buffers dictate the retention, efficiency and - to a lesser effect - selectivity of analytes. Tri-ethylammonium acetate and ammonium nitrate are the most effective buffer systems, while sodium citrate is also effective for the separation of profens on vancomycin CSP, and ammonium acetate is the most appropriate for LC/MS applications. 

Those which maintain certain conditions but do not take part in electrode reactions examples are electrolytes added to improve conductivity, pH buffer systems and ions which maintain complex ion equilibria. 

By taking the logarithm of both sides of Equation 14.1 and multiplying through by — 1, you can show (Problem 79) that, for any buffer system,... 

Equation 141 is a completely general equation, applicable to all buffer systems. The calculation of [H+]—and hence pH—can be simplified if you keep two points in mind. 

A buffer has a limited capacity to react with H+ or OH- ions without undergoing a drastic change in pH. To see why this is die case, consider Figure 14.3, which applies to the H2C03-HC03- buffer system (IQH2CO3 = 4 X 10-7). 

Consider the buffer system shown in the box below. The symbol 9 represents a mole of the weak acid the symbol 9 represents a mole of its conjugate base. The pH of the buffer is 6.0. What is Kz of the weak acid ... 

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