Describe the three lines of defense against changes changes in (H+) in terms of their mechanisms and speed of action.

THREE LINES OF DEFENSE

Chemical buffering - in blood, urine, and other fluids onset = instantaneous Respiratory responses - to alter PCO2 as needed to correct pH onset = seconds to minutes Renal responses - to alter [HCO3-] as needed to correct pH onset = hours to weeks

A. 1st Line of Defense: Buffers

The general equation for a buffer system is HA H+ + A-. A represents any anion and HA the undissociated acid. If an acid stronger than HA is added to a solution containing this system, the equilibrium is shifted to the left. Hydrogen ions are "tied up" in the formation of more undissociated HA, so the increase in H+ concentration is much less than it would otherwise be. By the law of mass action, the product of the concentrations of the products in a chemical reaction divided by the product of the concentration of the reactants at K = [H+][A-] [HA] equilibrium is a constant termed K: rearranging K = [H+][A-] [HA]
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[H+] = K [HA] [A-] Log [H+] = log K + log [HA] [A-]

-Log [H+] = -log K - log [HA] [A-]

pH = pK + log [A-] [HA]

When [A-] = [HA], log 1 = 0; pH = pK, and the buffer has its greatest effectiveness. When buffer is more concentrated, it is more effective.

If this equation is solved for H+ and put in pH notation 1, the resulting equation is the one derived by Henderson and Hasselbalch to describe the pH changes resulting from addition of H + or OH to any buffer system (HendersonHasselbalch equation): pH = pK + log [A-] [HA] Recall that when [A-] = [HA], the log of 1 is zero so that pH = pK and that when pK = pH any change in concentration of [A -] or [HA] has the smallest effect on pH; i.e., the buffer is strongest. Note also that the concentration of the components of a buffer system is important in determining the magnitude of a Normal values: 7.4 = 6.1 + log 24 pH change. (See p. 77 for the different (40 x 0.03 ) slopes of the bicarbonate buffer PCO2 = 60 7.4 = 6.1 + log 24 isobar with PCO2 = 20 mm Hg. 1.2 When applied to the bicarbonate buffer system, this equation is: pH = pK + log [HCO3-] [H2CO3] Because H2CO3 is present in very small quantities the clinically relevant form of the equation substitutes [CO2] for [H2CO3] and uses the relationship [CO2] = PCO2 x (Henrys Law) to get a clinically useful form: pH = pK + log [HCO3-] PCO2 x When you have data for HCO3- and PCO2, you can calculate pH. Most of the time a calculator is needed, but if the numbers are such that the ratio of HCO3-/(PCO2 x 0.03) is 1,10 or 100, you can easily calculate that the pH is 6.1, 7.1, or 8.1. 1) Kinds of Buffers Protein Buffer System: HPR H+ + Pr
1

7.4 = 6.1 + log 20 For 7.4 the = bicarbonate 6.1 + 1.3 buffer system

pH = pK + - log [HCO 3 ] pH (if (if ratio ratio of of HCO3 HCO3-/PCO /PCO22 = = 20, 20, pH is is normal) normal) 3] + log 20 7.4 = 6.1 + log 48 [H2CO =6.1
-

80 x 0.03 pH = pK + log [HCO3-] [CO2] pH = pK + log [HCO3-] PCO2 x

Henderson-Hasselbalch Equation

Important in plasma and in cells.

so log [H+] = log K + log [HA] [A ]
-

[H+] = K [HA] [A ]
-

and log [H+] = -log K + log [A-] [HA]

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Ionizable groups on side-chains and terminals. For many proteins, the pK's are close to 7.0 - 7.4, tending to hold pH near these values. Hemoglobin is the major buffer of carbonic acid hydrogen ions in blood H+ + Hb HHb

Phosphate Buffer System: H2PO4- H+ + HPO4-2 Important in cells and in urine. pK about 6.8

Ammonia Buffer System: NH4 H+ + NH3 Ammonia buffering accounts for about 50% of acid excretion and bicarbonate generation. pK of this reaction is 9.2, meaning that there is very little dissociation of NH4+ at normal pH. This is why ammonia is not part of titratable acid and must be measured separately to calculate net acid excretion. Bicarbonate Buffer System: CO2 + H2O H2CO3 H+ + HCO3pK' = 6.1

The bicarbonate buffer system is the most important extracellular buffering system for fixed acids in the body . This is in spite of the fact that its pK is 6.1, far from the regulated pH and that its components are not present in high concentrations. The reason for its relative importance is the fact that two of its components are regulated: HCO3- by the kidneys and CO2 by the lungs. The reaction CO2 + H2O H2CO3 proceeds slowly in either direction unless the enzyme carbonic anhydrase is present. There is no carbonic anhydrase in plasma, but there is a lot in red blood cells. It is also found in high concentration in gastric acid-secreting cells and in renal tubular cells. The catalyzed reaction is essential
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for bicarbonate reabsorption in the kidney. Inhibitors of carbonic anhydrase; e.g., diamox, are consequently potent diuretics (discussed later). An Important Concept: the Isohydric principle All buffer systems are linked, because all the reactions share H. H+ = K1 x [HA 1 ] = K2 x [HA 2 ] = K3 x [HA 3 ] [A-1] [A-2] [A-3] This means: all buffer systems will function together and we need only examine one buffer system in order to assess all buffer systems
+

2) Importance of Buffering
In a classic experiment, shown in this figure, Robert Pitts demonstrated the power of the buffer system of blood to resist pH change due to 8 19 kg dog in vivo addition of fixed acid (HCl in this pH = 7.14 6 1.5 hours case). Much of pH the buffering in 4 11.4 L Distilled Water the dog was pH = 1.84 accomplished by 2 the bicarbonate Robert F. Pitts. Harvey Lect. 48:172. 1953 buffer system. 100 150 50 0 The renal mmol H+ added contribution was minimal due to the short time (1.5 hours).

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B. 2nd Line of Defense: The Control of Respiration

pH = pK + log [HCO3-] PCO2 x As you learned in pulmonary physiology, changes in pH and PCO 2 cause changes in alveolar ventilation. You also learned that changes in alveolar ventilation cause changes in PCO2 and pH. In the next lecture, you will learn how respiratory compensation can compensate for metabolic acid base problems. Control of ventilation is a line of defense or compensation for metabolic acidosis or alkalosis. Chemoreceptors (peripheral and central) controlling respiration are sensitive to changes in hydrogen ion concentration. Metabolic acidosis stimulates ventilation while metabolic alkalosis inhibits ventilation. The response to metabolic alkalosis is less than the response to metabolic acidosis because the decreased alveolar ventilation during alkalosis produces arterial hypoxemia which stimulates the peripheral O2 receptors and increases ventilation. The respiratory response to altered pH from metabolic acidosis or alkalosis is very fast, occurring within seconds.

C. 3rd Line of Defense: Renal Control of Bicarbonate Concentration

pH = pK + log [HCO3-] PCO2

Controls rate of reabsorption of bicarbonate as needed. When necessary, generates new bicarbonate. Both of these processes involve secretion of hydrogen ions into the urine.

II. Role of the liver in AB balance

Produces fixed acids from amino acid metabolism

Produces keto-acids
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Produces HCO3-

20% of total CO2 produced

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