Water

PH AND BUFFERS
 Cont.
• Total body water is about 50 to 60% of body
weight in adults and about 75% of body weight
in children.
• Approximately 60% of the total body water is
intracellular and 40% extracellular. The
extracellular water includes the fluid in plasma
(blood after the cells have been removed) and
interstitial water (the fluid in the tissue spaces,
lying between cells).
 Cont.
• Because fat has relatively little water associated with
it, obese people tend to have a lower percentage of
body water than thin people, females tend to have a
lower percentage than males, and older people have a
lower percentage than younger people.
Transcellular water is a small, specialized portion of
extracellular water that includes gastrointestinal
secretions, urine, sweat, and fluid that has leaked
through capillary walls as a result of such processes
as increased hydrostatic pressure or inflammation.
 Cont.
• Water is the solvent of life. It bathes our cells, dissolves
and transports compounds in the blood, provides a
medium for movement of molecules into and throughout
cellular compartments, separates charged molecules,
dissipates heat, and participates in chemical reactions.
Most compounds in the body, including proteins, must
interact with an aqueous medium in order to function. In
spite of the variation in the amount of water we ingest
each day and produce from metabolism, our body
maintains a nearly constant amount of water that is about
50 to 60% of our body weight
 Cont.
• Water is the predominant chemical component
of living organisms. Its unique physical
properties, which include the ability to solvate
a wide range of organic and inorganic
molecules, derive from water’s dipolar
structure and exceptional capacity for forming
hydrogen bonds
 Cont.
• An unshielded hydrogen nucleus covalently
bound to an electron-withdrawing oxygen or
nitrogen atom can interact with an unshared
electron pair on another oxygen or nitrogen
atom to form a hydrogen bond.
Cont.
 Cont.
• Metabolic reactions often involve the attack by lone
pairs of electrons of electron-rich molecules termed
nucleophiles on electron-poor atoms called
electrophiles.
• Nucleophilic attack by water generally results in the
cleavage of the amide, glycoside, or ester bonds that
hold biopolymers together.
• Conversely, when monomer units are joined together
to form biopolymers such as proteins or glycogen,
water is a product.
Cont.
 PH and Buffers
• Each day there is always a production of acid
by the body’s metabolic processes and to
maintain balance, these acids need to be
excreted or metabolised.
 Cont.
• Acid is defined as molecule that can cleave off H +
(Arrhenius) or donor of H+ (Brönsted). Base is au
contraire molecule that can cleave off OH - (Arrhenius)
or acceptor of H+ (Brönsted).
• Source of acids in the body is chiefly metabolism,
source of bases is predominantly nutrient.
• Acids and bases undergo either (1) metabolic
conversion (e.g. lactate to glucose in gluconeogenesis,
lactate to pyruvate and oxidation in cardiomyocytes),
or (2) excretion from body.
 Reaction Types
• Proton-productive reactions
• Proton-consumptive reactions
• Proton-neutral reactions
 Proton-productive reactions

a) Anaerobic glycolysis in muscles and erythrocytes

• Glucose → 2 CH3CHOHCOO- + 2 H+
b) Ketogenesis – production of ketone bodies
• Fatty acids → ketone bodies + n H+
c) Lipolysis
• TAG → 3 FA + glycerol + 3 H+
d) Ureagenesis
• CO2 + 2 NH+4 → urea + H2O + 2 H+
 Proton-consumptive reactions

• a) Gluconeogenesis
2 lactate + 2 H+ → Glc
• b) Neutral and dicarboxylic amino acids
oxidation
 Proton-neutral reactions

• a) Complete glucose oxidation

• b) Lipogenesis from glucose
 Cont.
• Human organism (healthy or not) every day produces
great quantities of acids – source of protons. Organism is
acidified by these processes:
• 1) Complete oxidation
• Carbon skeleton → CO2 + H2O → HCO3- + H+
• 2) Incomplete oxidation
• Carbohydrates → glucose → pyruvate, lactate + H+
• Triacylglycerol → fatty acids, ketone bodies + H+
• Phospholipids → phosphate + H+
• Proteins → amino acids→ sulphate, urea + H+
 Cont.
• Acids can be divided into two groups: (1) volatile
acids (respiratory acids), (2) non-volatile acids
(metabolic acids).
• The most important volatile acid is carbonic acid
(H2CO3). H2CO3 is produced by reaction of carbon
dioxide (CO2 is acid-forming oxide) with water. 15
000 – 20 000 mmol CO2 (therefore same amount of
carbonic acid) is produced every day. Respiratory
system however very efficiently eliminates it. This
justifies the term volatile acid.
 Cont.
• Two groups are distinguished among non-volatile acid: (1)
organic, and (2) inorganic. 1 mmol/kg of body weight is
produced every day. Non-volatile acid could be either (1)
metabolised, or (2) excreted (using mainly kidneys).
• Organic non-volatile acids are for example: (1) lactic acid, (2)
fatty acids, (3) ketone bodies (acetoacetic acid, β-
hydroxybutyric acid). They are continually produced by
metabolism (incomplete oxidation of TAG, carbohydrates,
proteins). As organic non-volatile acids are products of
metabolism in normal conditions they are oxidized completely
to CO2 and H2O. Therefore they have no influence on proton
overall balance.
 Cont.
• Inorganic non-volatile acids are: (1) H2SO4
(sulphuric acid is produced by oxidation of
sulfhydryl groups – e.g. in amino acids that
contain sulphur, i.e. cysteine, methionine), (2)
H3PO4 (phosphoric acid is produced by
hydrolysis of phosphoproteins, phospholipids,
nucleic acids). Inorganic non-volatile acids are
predominantly excreted in urine.
 Systems responsible for maintenance of the acid-base balance

• Chemical buffering systems

• Respiratory system
• Kidneys
• Liver
• Myocardium
 Cont.
• Chemical buffering systems
• Buffers react immediately – acute regulation. Capacity of buffers is
not indefinite that is why chemical buffers act only in the short-
term. Chemical buffering systems deal with pH deviations in
common metabolism.
• 2) Respiratory system
• Respiration reacts in 1-3 minutes. Respiratory system regulates
carbon dioxide. Respiration is able to change pCO 2 by its
elimination or retention. Respiratory centre is in brainstem.
• 3) Kidneys
• Kidneys react in hours-days. Their role in acid-base balance is very
complex.
 Cont.
• 5) Myocardium
• Myocardium influences acid-base balance
through lactate and ketone bodies oxidation.
 Buffering systems

• Buffers are substances capable of releasing

and binding H+. Short-term and acute
changes in acid-base balance can be balanced
by buffers. Each buffer keeps its particular
pH. This pH could be calculated by means of
the Henderson-Hasselbalch equation:
• pH = pK + log [conjugated base]/[acid]
 Cont.
• Henderson-Hasselbalch equation for bicarbonate
buffer (HCO3-/CO2):
• pH = pKH2CO3 + log ([HCO3-] / [H2CO3])
• pH = pKH2CO3 + log ([HCO3-] / α x pCO2)
• α is conversion factor, that is used for calculation of
molar concentration (mmol/l) from partial pressure
of CO2 (pCO2). α = 0,226 for pCO2 in kPA, α = 0,03
for pCO2 in mmHg).
• pH = pK ± 1 is range where buffers work optimally.
 Cont.
• In Henderson-Hasselbalch equation above you should notice that for
pH that buffers keep depends primarily on ratio of conjugated base
and acid (Of course concentration of each component is important
but not that much). Therefore it is really important to know the ratio.
Ratio of conjugated base and acid could be calculated from relation
between pH and pK. For example bicarbonate buffer (pH = 7,4; pK =
6,1):
• pH = pKH2CO3 + log ([HCO3-] / [H2CO3])
• 7,4 = 6,1 + log ([HCO3-] / [H2CO3])
• 1,3 = log ([HCO3-] / [H2CO3])
• [HCO3-] / CO2 ≈ 20 / 1
• The ratio in bicarbonate buffer is 20:1 (HCO3- : CO2)
 Cont.
• There are several buffer systems in the body.
The most important include: (1) bicarbonate
buffer (HCO3-/CO2), (2) haemoglobin buffer
(in erythrocytes), (3) phosphate buffer, (4)
proteins, and (5) ammonium buffer. Their
importance differs as it depends on
localization.
Localization Buffer Commentary

Main
ISF
bufferBicarbonate
systems according to body
Buffers metabolic acids
Phosphate Low concentration – limited
compartments significance
Proteins Low concentration – limited
significance
Blood Bicarbonate Buffers metabolic acids
Haemoglobin Buffers CO2 (carbonic acid
production)
Plasma proteins Minor
Phosphate Low concentration – limited
significance
ICF Proteins Significant buffer
Phosphate Significant buffer
Urine Phosphate Responsible for majority of the
titratable urine acidity
Ammonium Significant: elimination of
ammonium nitrogen and
protons; cation
 Blood buffers and their buffer capacity

Buffer Plasma Erythrocytes Together

HCO3- / CO2 35 % 18 % 53 %

Hb / Hb-H+ - 35 % 35 %

Plasma proteins 7% - 7%

Inorganic phosphate 1% 1% 2%

Organic phosphate - 3% 3%

43 % 57 % 100 %
 Cont.
• Because of fact that all buffer systems are in
equilibrium any kind of drift in pH causes
response in all buffer systems. Any
concentration change of any component of any
buffer influences both pH, and all buffer
systems.
 Bicarbonate buffer (HCO3-/CO2)

• Bicarbonate buffer is the most important

buffer system in blood plasma (generally in
the extracellular fluid). This buffer consists of
weak acid H2CO3 (pK1 = 6,1) and conjugated
base HCO3- (bicarbonate).
 Cont.
• HCO3-/CO2 is so called open buffer system.
This means body is capable to actively alter
both bicarbonate, and carbon dioxide. pCO2 is
regulated by respiratory tract (by means of
ventilation – respiratory rate and depth of
breathing). HCO3- levels are altered by the
kidneys and the liver. HCO3- could be both
synthesized, and eliminated.
 Cont.
• pH = pK ± 1 is range where buffers work
optimally. This should mean that bicarbonate
buffer would work best in range 5,1-7,1, but in
pH 7,4 it is very effective because it is open
That is: organism is able to actively change
both components.
 Protein buffers

• Body proteins (plasma proteins and

intracellular) are the most abundant and the
most powerful buffer system in whole
organism. Some amino acids have acid or
basic side chains (His, Lys, Arg, Glu, Asp).
Among blood proteins haemoglobin is the
most important. It provides 35 % of buffering
capacity of blood, remaining proteins provide
only 7 %.
 Role of erythrocytes and haemoglobin in
the acid-base balance
• Intensive change of blood gases occurs in working
tissue. CO2 diffuses to erythrocytes. In the red
blood cell CO2 either (1) binds to haemoglobin
(and carbaminohemoglobin is formed), or (2) reacts
with water. This reaction is catalysed by carbonic
anhydrase (CA, carbonate dehydratase):
• CO2 + H2O ↔ H2CO3
• Produced carbonic acid dissociates:
• H2CO3 ↔ HCO3- + H+
 Cont.
• More than 70% of produced HCO3- leave erythrocyte using special
HCO3-/Cl- antiport. That is bicarbonate is exchanged for Cl -. This
process is called Hamburger´s effect (chloride shift). In carbonic acid
dissociation H+ is produced. Generated protons are buffered by
haemoglobin. Deoxygenated haemoglobin is stronger base than
oxygenated thus deoxygenated is more capable of taking up protons.
• In lungs HCO3- is changed to CO2, using enzyme CA. CO2 is exhaled.
Reaction HCO3- → CO2 + H2O demands H+. Protons for this process are
taken from haemoglobin which affinity to H + has lowered just when it
arrived to lungs where is high pO 2 and haemoglobin become
oxygenated. Reaction catalysed by carbonic anhydrase has reverse
course in lungs in comparison to other tissues:
• HCO3- + H+ → CO2 + H2O
Role of the respiratory tract in maintaining the acid-base balance

• Every day is exhaled approximately 15-20 moles of CO2 by the respiratory

system. CO2 is well soluble in water therefore its concentration in both alveoli
and arterial blood is the same (i.e. pCO2 = 5,33 kPa = 40 mmHg). In venous
blood is pCO2 6,13 kPa = 46 mmHg.
• pCO2 depends – besides other things – on the pulmonary ventilation (=
respiratory minute volume). Pulmonary ventilation is defined as respiratory rate
(RR) multiplied by tidal volume (VT). For understanding following concept you
should recall that pH of buffer depends on ratio of its components (e.g. HCO 3- :
pCO2) and so when ratio changes, pH changes consequently. You can now
easily deduce that:
• 1) increased ventilation leads to drop in pCO2 and that leads to alkalisation
(increased pH)
• 2) decreased ventilation leads to accumulation of CO2 → increased pCO2 and
that leads to acidification (decreased pH)
 Cont.
• There are many ways for controlling
breathing. One of them is chemical control.
Chemoreceptors check both pCO2, and pO2.
Increased pCO2 activates breathing centre.
Sensitivity of chemoreceptors is decreasing
when pCO2 is 8 kPa or higher. Only
remaining stimulus for breathing centre is
decreased pO2.
 Role of the kidneys in maintaining the acid-base balance

• The kidneys take part in maintaining the acid-

base balance by means of:
• 1) Reabsorbing, excreting and producing
bicarbonate
• 2) Excreting or producing H+
 Cont.
• Bicarbonate reabsorption takes place in
proximal tubule cells. In glomerular ultrafiltrate
there is filtered bicarbonate. To the lumen of
the proximal tubule is transported H+. H+ is
transported by Na+/H+ antiport  H+ reacts with
HCO3- and H2CO3 is thus produced. H2CO3 split
up into H2O and CO2. Water and carbon
dioxide get through apical membrane of tubular
cells.
 Cont.
• Inside these cells H2CO3 is again produced. H2CO3
dissociates into HCO3- and H+. Now their fates get
different: (1) H+ becomes again substrate for Na+/H+
antiport and it is transported again to the lumen of the
proximal tubule where it can “catch” another bicarbonate
molecule. (2) Bicarbonate however traverse basolateral
membrane into interstitial fluid (and then to the blood of
the peritubular capillaries). Bicarbonate gets through
basolateral membrane using either Na+/3 HCO3-
cotransport, or anion exchanger (Cl-/HCO3- exchange).
 New bicarbonate production (connected with H + excretion)

• New bicarbonate production takes place in

intercalated cells type A of distal tubule and
collecting duct. These cells absorb CO2 from
the blood and inside the cells carbon dioxide
reacts with water and carbonic acid is thus
produced, catalysed by the enzyme carbonic
anhydrase. Carbonic acid dissociates to H+ and
HCO3-. H+ has totally different fate than
bicarbonate
 Cont.
• (1) H+ is excreted by the H-ATPase to the urine.
This process is active, hence it consumes ATP. In
order to eliminate as much H+ as possible it is
necessary to buffer H+ in the urine. The most
important buffers in the urine are ammonium and
phosphate buffer. (2) Produced bicarbonate is
transported to the blood in peritubular capillaries
exchanged for Cl- (Cl-/HCO3- exchanger in
basolateral membrane). Aldosterone stimulates H+
secretion (and therefore H+ excretion).
Cont.
Ammonium ion excretion
 Compensation and correction of acid-base disturbances

• Compensation is process when organism tries to maintain almost

normal pH. Compensation is performed by system that works
normally, i.e. the acid-base disturbance is caused by the other
system. Compensation thus means metabolic disturbances are
compensated by respiratory system and respiratory disturbances
are compensated by metabolic components of acid-base balance.
• Correction is solving the acid-base problem in the spot where it
started. I.e. metabolic disturbances are solved by metabolic
component of acid-base balance. In the body correction takes
place only in metabolic disorders, i.e. metabolic disorder is
corrected by another component of the metabolic component of
acid-base balance.
 Acidosis
• Acidosis is excessive blood acidity caused by
an overabundance of acid in the blood or a loss
of bicarbonate from the blood (metabolic
acidosis), or by a buildup of carbon dioxide in
the blood that results from poor lung function
or slow breathing (respiratory acidosis)
 Cont.
• Blood acidity increases when people ingest
substances that contain or produce acid or
when the lungs do not expel enough carbon
dioxide.
• People with metabolic acidosis have nausea,
vomiting, and fatigue and may breathe faster
and deeper than normal.
 Cont.
• If an increase in acid overwhelms the body's pH
buffering systems, the blood will become acidic. As
blood pH drops, the parts of the brain that regulate
breathing are stimulated to produce faster and deeper
breathing. Breathing faster and deeper increases the
amount of carbon dioxide exhaled.
• The kidneys also try to compensate by excreting more
acid in the urine. However, both mechanisms can be
overwhelmed if the body continues to produce too much
acid, leading to severe acidosis and eventually coma.
 Cont.
• Metabolic acidosis develops when the amount of acid in the
body is increased through ingestion of a substance that is, or
can be broken down (metabolized) to, an acid—such as wood
alcohol (methanol), antifreeze (ethylene glycol), or large doses
of aspirin (acetylsalicylic acid). Metabolic acidosis can also
occur as a result of abnormal metabolism. The body produces
excess acid in the advanced stages of shock and in poorly
controlled type 1 diabetes mellitus. Even the production of
normal amounts of acid may lead to acidosis when the kidneys
are not functioning normally and are therefore not able to
excrete sufficient amounts of acid in the urine.
 Major Causes of Metabolic Acidosis

• Diabetic ketoacidosis (buildup of ketones)

• Drugs and substances such as acetazolamide, alcohol, aspirin,
and iron
• Lactic acidosis (buildup of lactic acid as occurs as a result of
shock)
• Loss of bases, such as bicarbonate, through the digestive tract
due to diarrhea, an ileostomy, or a colostomy
• Advanced kidney disease
• Poisons such as carbon monoxide, cyanide, ethylene glycol,
and methanol
• Renal tubular acidosis (a form of kidney malfunction)
 Cont.
• Respiratory acidosis develops when the lungs do not
expel carbon dioxide adequately, a problem that can
occur in disorders that severely affect the lungs (such as
emphysema, chronic bronchitis, severe pneumonia,
pulmonary edema, and asthma). Respiratory acidosis can
also develop when disorders of the brain or of the nerves
or muscles of the chest impair breathing. In addition,
people can develop respiratory acidosis when their
breathing is slowed due to oversedation from opioids
(narcotics) or strong drugs that induce sleep (sedatives).
 Major Causes of Respiratory Acidosis

• Lung disorders, such as emphysema, chronic

bronchitis, severe asthma, pneumonia, or
pulmonary edema
• Sleep-disordered breathing
• Disorders of the nerves or muscles of the chest
that impair breathing, such as Guillain-Barré
syndrome or amyotrophic lateral sclerosis
• Overdose of drugs such as alcohol, opioids, and
strong sedatives
 Symptoms
• People with mild metabolic acidosis may have no symptoms but
usually experience nausea, vomiting, and fatigue. Breathing
becomes deeper and slightly faster (as the body tries to correct
the acidosis by expelling more carbon dioxide). As the acidosis
worsens, people begin to feel extremely weak and drowsy and
may feel confused and increasingly nauseated. Eventually, blood
pressure can fall, leading to shock, coma, and death.
• The first symptoms of respiratory acidosis may be headache and
drowsiness. Drowsiness may progress to stupor and coma.
Stupor and coma can develop within moments if breathing stops
or is severely impaired, or over hours if breathing is less
dramatically impaired.
 Diagnosis
• The diagnosis of acidosis generally requires the
measurement of blood pH in a sample of arterial
blood, usually taken from the radial artery in the wrist.
Arterial blood is used because venous blood contains
high levels of bicarbonate and thus is generally not as
accurate a measure of the body’s pH status.
• To learn more about the cause of the acidosis, the
levels of carbon dioxide and bicarbonate in blood is
measured. Additional blood tests may be done to help
determine the cause.
 Treatment
• The treatment of metabolic acidosis depends primarily on the cause. For
instance, treatment may be needed to control diabetes with insulin or to
remove the toxic substance from the blood in cases of poisoning.
• The treatment of respiratory acidosis aims at improving the function of the
lungs. Drugs that open the airways (bronchodilators, such as albuterol)
may help people who have lung diseases such as asthma and emphysema.
People who have severely impaired breathing or lung function, for
whatever reason, may need mechanical ventilation to aid breathing
• Acidosis may also be treated directly. If the acidosis is mild, the
administration of intravenous fluids may be all that is needed. When
acidosis is severe, bicarbonate may be given intravenously. However,
bicarbonate provides only temporary relief and may cause harm—for
instance, by overloading the body with sodium and water.
 Alkalosis
• Alkalosis is excessive blood alkalinity caused
by an overabundance of bicarbonate in the
blood or a loss of acid from the blood
(metabolic alkalosis), or by a low level of
carbon dioxide in the blood that results from
rapid or deep breathing (respiratory alkalosis)
 Cont.
• Metabolic alkalosis develops when the body loses too much
acid or gains too much base. For example, stomach acid is lost
during periods of prolonged vomiting or when stomach acids
are suctioned with a stomach tube (as is sometimes done in
hospitals). In rare cases, metabolic alkalosis develops in a
person who has ingested too much base from substances such
as baking soda (bicarbonate of soda). In addition, metabolic
alkalosis can develop when excessive loss of sodium or
potassium affects the kidneys' ability to control the blood's
acid-base balance. For instance, loss of potassium sufficient to
cause metabolic alkalosis may result from an overactive
adrenal gland or the use of diuretics.
 Cont.
• Respiratory alkalosis develops when rapid,
deep breathing (hyperventilation) causes too
much carbon dioxide to be expelled from the
bloodstream. The most common cause of
hyperventilation, and thus respiratory
alkalosis, is anxiety. Other causes of
hyperventilation and consequent respiratory
alkalosis include pain, low levels of oxygen in
the blood, fever, and aspirin overdose
 Cont.
• Metabolic alkalosis causes
• Loss of acid from vomiting or drainage of the
stomach
• Overactive adrenal gland (Cushing syndrome
and some adrenal tumors)
• Use of diuretics (thiazides, furosemide,
ethacrynic acid)
 Cont.
Respiratory alkalosis causes
• Anxiety
• Aspirin overdose (early stages)
• Fever
• Low levels of oxygen in the blood
• Pain
 Symptoms and Diagnosis
• Alkalosis may cause irritability, muscle
twitching, muscle cramps, or no symptoms at
all. If the alkalosis is severe, prolonged
contraction and spasms of muscles (tetany)
can develop.
• A sample of blood usually taken from an
artery shows that the blood is alkaline.
 Treatment
• Metabolic alkalosis is usually treated by replacing water and
electrolytes (sodium and potassium) while treating the cause.
Occasionally, when metabolic alkalosis is very severe, dilute acid is
given intravenously.
• With respiratory alkalosis, usually the only treatment needed is
slowing down the rate of breathing. When respiratory alkalosis is
caused by anxiety, a conscious effort to slow breathing may make
the condition disappear. If pain is causing the person to breathe
rapidly, relieving the pain usually suffices. When respiratory
alkalosis is due to anxiety alone, breathing into a paper (not a
plastic) bag may help raise the carbon dioxide level in the blood as
the person breathes carbon dioxide back in after breathing it out.