THE MAJOR BUFFERING SYSTEMS PAGE!

Buffering System #1: The Bicarbonate Buffering System

Once again, here is the bicarbonate buffering equation. Beginning from the
left, water plus carbon dioxide react reversibly to produce carbonic acid;
continuing to the right, carbonic acid dissociated (reversibly) to form H+ and
HCO3- (bicarb) ions.

H2O + CO2 <---> H2CO3 <---> H+ + HCO3-

Recall that for any acid, the pKa is the pH at which the concentration of intact and ionized acid
is the same: that is, where [HA] = [A-].

At the pKa for carbonic acid (H2CO3), the carbonic acid is 50% ionized. Since carbonic acid is
the source of bicarbonate ions, the pKa for carbonic acid (6.1) is the pKa for the bicarbonate
buffering system!

For the bicarbonate buffering system, then, the pKa is 6.1. This is the pH at which the
bicarbonate buffer exerts its maximum buffering capacity. But wait!!!! The pKa for the bicarb
buffering system is approximately 1.3 pH units below the pH of the blood! With such a wide
disparity, how on earth can the bicarb buffering system be such an effective circulating buffer
in the bloodstream? Even though a pKa of 6.1 is quite far away from the pH of body fluids
(around 7.4), the bicarbonate buffering system remains the major buffering system, on the basis
of elimination of acid over time, used by the body. BUT WHY???

Why is the bicarbonate buffering system such a great buffering system, even
though its pKa is so far below (7.4 - 6.1 = 1.3 pH units) physiological pH?

1) The bicarbonate buffering system is an open system. Using the bicarb buffering system, acid
(H+) can be eliminated as CO2 blown off in the expiratory gas from the lungs... as CO2 is blown
off at the lungs, more carbonic acid can break down to release more CO2 at the lungs; H+ can
also be transported to the kidney where bicarb buffers transfer the acid to various phosphate
buffers in the renal filtrate; the phosphates then carry the H+ out in the urine.

2) the body uses a lot of bicarbonate buffer; the concentration of bicarb in the blood approaches
25 mEq/L

3) carried in circulation, the bicarb buffer reaches every cell in the body

4) and finally, a tubular maxima mechanism in the proximal renal tubule works to efficiently
conserve bicarb... wow... what a buffer!!!

The continuous action of bicarb in removal of CO2 and H+ from the tissues and blood, and the
ability of the bicarb system to eliminate H+ and CO2 at the lung and H+ at the kidney, make the
bicarb system the major buffering system in the body, in terms of acid elimination over time.
This does not mean that the bicarb system has the highest buffering capacity at any instant in
time, it simply means that the bicarb system is actually the buffering system which is handling
the most acid waste over time.

Below, we will learn more about the major "non-bicarbonate" buffering systems, the protein
buffering system, and the phosphate buffering system. Normally, the bicarbonate buffering
system is responsible for elimination of 50% of the acid from the body, while together, the
protein and phosphate buffering systems handle the other 50%. However, as we will learn
below, the protein buffering system actually has the greatest capacity for buffering... and
proteins may account for 75% of the total available "buffering capacity" of the body. In
summary, then, the protein buffering system has the greatest capacity to buffer, but in reality,
the bicarb buffering system is actually eliminating more acid than the other buffering systems.
On this basis, it is not unusual for a pathology instructor to tell their students that the bicarb
buffering system is, functionally, the most important buffering system.

Buffering System #2: Protein (Cellular and Hb) Buffering

"Protein buffering" and "cellular buffering" are the same thing! The cells of the body have lots
of proteins which can buffer against acid and there will be considerable shifting of H+'s back
and forth across cell membranes in acidosis. Recall that cell membranes are approximately 50%
protein by mass. Recall that RBC's are 33%, by weight, Hb. Hb is a globular protein which
circulates in the bloodstream and is capable of buffering tremendous quantities of H+. Because
the buffering capacity of Hb is so great, having 75% of the capacity of the total protein
buffering system, Hb has occasionally been referred to as an entirely separate buffering system,
although we will simply lump Hb into the overall protein buffering system here.

Proteins have amino and carboxyl groups, both of which can donate or accept H+'s; particularly
the a.a., His, with its mitosol group. In Hb, 8% of the a.a.'s are His, such that Hb is a great
buffer. Lots of protein in cells and in plasma can act as buffers. Due to the variety and
complexity of the buffering groups available from protein, proteins have a linear buffering
ability across the physiological range of pH. Despite all the proteins embedded in all the
membranes of all the cells of the body, and while the plasma is rich with protein, Hb still
accounts for the majority (75%) of the overall protein buffering capacity of the body... but we
already know that Hb does a lot more than just transport O2.

In acidosis and alkalosis, protein (cellular) buffering is simple but very important. Because
proteins are spread throughout the body, protein buffering spreads the effects of H+ changes
over the entire body so the overall effect on pH is less, for example, in any one tissue. There are
two small drawbacks to protein buffering. First, while Hb and other circulating proteins
circulate throughout the body and quickly buffer against changes in H+ concentration, most
proteins are locked in place in cells and so act as slow buffers... that is, most of the proteins can
only slowly pull excess H+ out of circulation, which may take hours. Second, while Hb and
plasma proteins carry H+ to the lungs, where much of the H+ can be eliminated in the form of
CO2 + H2O, acid that is buffered by proteins stuck in most of the cells in the body is effectively
"stored" acid. In other words, many cellular proteins buffer against acid by binding acid, but the
cells can slowly become saturated with acid. If the cellular protein system approaches
"saturation" with acid, expect pH to drop quickly.

Buffering System #3: Phosphate Buffers

There are several forms of PO4 buffers (eg. H3PO4 & Na3PO4). Thus, phosphates represent a
complex buffering system with a wide range of pKa's. Phosphate concentrations are low in the
body, plasma and interstitial fluids. Phosphates are not important as buffers in blood but are
important intracellular buffers and are the most important buffers in the urine. Phosphates are
ultimately responsible for holding H+'s in urine so that H+'s can be expelled from the body in
urine. This also allows HCO3- to be reabsorbed from the kidney back into the body, without H+
following the HCO3- back into the body.

If you secrete H+ into the urine you need to buffer against it within the urine in order to
maintain other ionic gradients. Urine buffers (primarily phosphates) allow large amounts of H+
to be secreted without huge changes in pH of the urine; usually the pH of urine is 5-6 but it may
go as low as 4.6. As H+ is secreted into the renal tubule, phosphate salts go from dibasic to
monobasic such that 1 Na+ is released and reabsorbed from the kidney tubule. HCO3- is
reabsorbed along with the Na+ as sodium bicarbonate is generated (NaHCO3). Thus, phosphate
and bicarb may be thought of as working hand in hand; i) the bicarb drags some acid into the
kidney, ii) the phosphate takes the acid from the bicarb and drags it out of the body in the urine,
iii) the bicarb picks up the Na+ released by the phosphate and returns it to the body. Wow...
buffer buddies!

In fact, if phosphates are reduced in the renal filtrate, we lose bicarb in urine at a rapid rate and
the bicarb system quickly becomes ineffective. The bottom line is that, no buffering system in
the body would be effective without the others.

But why does urine sometimes smell like ammonia? (the minor ammonia
buffering system)

Both the PCT and DCT can synthesize ammonia and release it into the renal filtrate. Why do
this? Well, ammonia (NH3) will readily bind H+... and so the fact that cells of the PCT and
DCT can synthesize ammonia gives the kidney a little "buffer booster!" The PCT and DCT
always synthesize some ammonia, which may give off a characteristic ammonia odor when
urine is concentrated. But in an acidosis, NH3 production is increased, and an increasing
amount of NH3 is secreted into the renal filtrate to "boost the buffering" capacity of the renal
filtrate in order to trap additional H+ for elimination in the urine. In an acidosis, then, the
ammonia smell becomes stronger.

Factors Affecting Bicarbonate Reabsorption at the Kidney

The higher the bicarbonate concentration in plasma, beyond a certain point (above a threshold
of 25 mEq/L), the more bicarb will be excreted into the urine. H+ secretion is directly related to
bicarbonate re-absorption. Because much of the H+ secreted in the kidney is secreted in the
form of H2CO3, the more H+ secreted, the more HCO3- that will be reabsorbed! Similarly, when
bicarb levels fall, the PCT will more strongly reabsorb bicarb, although this mechanism is
limited by the fact that bicarb is always strongly reabsorbed.

The bicarbonate buffering system in urine works at an increased rate when acid (H+) levels
become too high for all the lungs to handle further increases by simply blowing off more CO2
at the lung! The bicarbonate concentration in blood is actually about 1 million times higher
than the H+ concentration. However, there are a couple of potential problems with using the
bicarbonate buffering system to actually buffer urine. First of all, if we buffered urine with
bicarb all the time, we would quickly deplete bicarb from the bloodstream... because we would
lose too much bicarb in the urine! In other words, it is preferable to reabsorb bicarb from the
urine, which is exactly what we do in the proximal tubule! (Recall that bicarb is reabsorbed in
the proximal tubule by a tubular maxima mechanism).

David Currie.
Copyright © 2000. All rights reserved. Revised:

From: http://faculty.etsu.edu/currie/buffer.htm