PH AND BUFFER

Acid Base Balance

Prakash Pokhrel
 ACID BASE BALANCE

PH
It is the negative log of the hydrogen ion concentration.
pH = -log [H+]
 pH is a unit of measure which describes the degree of
acidity or alkalinity (basic) of a solution.

 It is measured on a scale of 0 to 14.

 Low pH values correspond to high concentrations of H+

and high pH values correspond to low concentrations of
H+.
PH VALUE
 The pH value of a substance is directly related to the
ratio of the hydrogen ion and hydroxyl ion
concentrations.

 If the H+ concentration is higher than OH- the material

is acidic.

 If the OH- concentration is higher than H+ the material

is basic.

 7 is neutral, < is acidic, >7 is basic

THE PH SCALE
 The pH scale corresponds to the concentration of
hydrogen ions.
 For example pure water H+ ion concentration is 1 x 10^-
7 M, therefore the pH would then be 7.
Acid
Any compound which forms H⁺ ions in solution
(proton donors)
eg: Carbonic acid releases H⁺ ions

Base
Any compound which combines with H⁺ ions in
solution (proton acceptors)
eg:Bicarbonate(HCO3⁻) accepts H+ ions
ACID–BASE BALANCE
 Normal pH : 7.35-7.45
 Acidosis

Physiological state resulting from abnormally low plasma

 Alkalosis

Physiological state resulting from abnormally high plasma

 Acidemia: plasma pH < 7.35
 Alkalemia: plasma pH > 7.45
MEASUREMENT OF PH

The pH can be measured by:

ph strips

Ph indicators

Ph meter
SOME IMPORTANT INDICATORS USED IN A CLINICAL
BIOCHEMISTRY LABORATORY ARE LISTED BELOW:
sr,. INDICATOR Ph range Colour in Colour in
No. acidic ph basic ph
1 Phenophthalein 9.3-10.5 colourless pink

2 Methyl orange 3.1-4.6 red yellow

3 Bromophenol blue 3.0-4.6 yellow blue

4 Methyl red 4.4-6.2 Red yellow

5 Phenol red 6.8 – 8.4 yellow red

6 Litmus 4.5-8.3 red Blue

PH METER
 The pH meter is a laboratory equipment which used to measure
acidity or alkalinity of a solution

 The pH meter measures the concentration of hydrogen ions [H+]

using an ion-sensitive electrode.

 It is the most reliable and convenient method for measuring ph.

 BUFFER
A buffer solution is a solution which resists changes in pH when
a small amount of acid or base is added.

Typically a mixture of a weak acid and a salt of its conjugate

base or weak base and a salt of its conjugate acid.
TYPES OF BUFFERS
Two types :
 ACIDIC BUFFERS –
Solution of a mixture of a weak acid and a salt of this weak
acid with a strong base.
E.g. CH3COOH + CH3COONa
( weak acid ) ( Salt )

 BASIC BUFFERS –
Solution of a mixture of a weak base and a salt of this weak
base with a strong acid.
e.g. NH4OH + NH4Cl
( Weak base) ( Salt)
HOW BUFFERS WORK
 Equilibrium between acid and base.

 Example: ACETATE BUFFER

 CH3COOH  CH3COO- + H+

 If more H+ is added to this solution, it simply shifts the

equilibrium to the left, absorbing H+, so the [H+]
remains unchanged.

 If H+ is removed (e.g. by adding OH-) then the

equilibrium shifts to the right, releasing H+ to keep the
pH constant
•HANDERSON HASSELBALCH
EQUATION
 Lawrence Joseph Henderson wrote an equation, in 1908,
describing the use of carbonic acid as a buffer solution.

 Karl Albert Hasselbalch later re-expressed that formula

in logarithmic terms, resulting in the
Henderson–Hasselbalch equation.
 The Henderson-Hasselbalch Equation derivation

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

[A-]
take the -log on both sides -log Ka = -log [H+] -log
[HA]

[A-]
apply p(x) = -log(x) pKa = pH -log
[HA]

and finally solve for pH…

[A-] [Proton acceptor]

pH = pKa + log = pKa + log
[HA] [Proton donor]
 - The greater the buffer capacity the less the pH
changes upon addition of H+ or OH-

Choose a buffer whose pKa is closest to the desired

pH.

pH should be within pKa ± 1

BUFFER
SYSTEM IN
BODY FLUIDS
 ACIDS
 VOLATILE ACIDS:
 Produced by oxidative metabolism of CHO,Fat,Protein

 Average 15000-20000 mmol of CO₂ per day

 Excreted through LUNGS as CO₂ gas

• FIXED ACIDS (1 mEq/kg/day)

 Acids that do not leave solution ,once produced they
remain in body fluids Until eliminated by KIDNEYS
Eg: Sulfuric acid ,phosphoric acid , Organic acids
Are most important fixed acids in the body
Are generated during catabolism of:
amino acids(oxidation of sulfhydryl gps of cystine,methionine)
Phospholipids(hydrolysis)
nucleic acids
RESPONSE TO ACID BASE CHALLENGE

1. Buffering
2. Compensation
BUFFERS
 First line of defence (> 50 – 100 mEq/day)
 Two most common chemical buffer groups
 Bicarbonate
 Non bicarbonate (Hb,protein,phosphate)
 Blood buffer systems act instantaneously
 Regulate pH by binding or releasing H⁺
CARBONIC ACID–BICARBONATE BUFFER SYSTEM
Carbon Dioxide
 Most body cells constantly generate carbon dioxide
 Most carbon dioxide is converted to carbonic acid, which dissociates into
H+ and a bicarbonate ion
Prevents changes in pH caused by organic acids and fixed acids in ECF

 Cannot protect ECF from changes in pH that result from elevated

or depressed levels of CO2
 Functions only when respiratory system and respiratory control
centers are working normally
 Ability to buffer acids is limited by availability of bicarbonate ions
ACID–BASE BALANCE

The Carbonic Acid–Bicarbonate Buffer System

THE CARBONIC ACID HYDROGENCARBONATE
BUFFER SYSTEM
• The carbonic acid-hydrogen Bicarbonate ion buffer is
the most important buffer system.
• Carbonic acid, H2CO3, acts as the weak acid
• Hydrogen carbonate, HCO3-, acts as the conjugate base
• Increase in H+(aq) ions is removed by HCO3-(aq)
• The equilibrium shifts to the left and most of the H+(aq)
ions are removed
 The small concentration of H+(aq) ions reacts with the
OH-(aq) ions

 H2CO3 dissociates, shifting the equilibrium to the right,

restoring most of the H+(aq) ions

 Any increase in OH-(aq) ions is removed by H2CO3

THE HEMOGLOBIN BUFFER SYSTEM

CO2 diffuses across RBC membrane

 No transport mechanism required

As carbonic acid dissociates

 Bicarbonate ions diffuse into plasma
In exchange for chloride ions (chloride shift)


 Hydrogen ions are buffered by hemoglobin

molecules
 Is the only intracellular buffer system with an immediate effect
on ECF pH
 Helps prevent major changes in pH when plasma PCO is rising
2
or falling
PHOSPHATE BUFFER SYSTEM

 Consists of anion H2PO4- (a weak acid)(pKa-6.8)

 Works like the carbonic acid–bicarbonate buffer system
 Is important in buffering pH of ICF

Limitations of Buffer Systems

 Provide only temporary solution to acid–base
imbalance
 Do not eliminate H+ ions
 Supply of buffer molecules is limited
RESPIRATORY ACID-BASE CONTROL
MECHANISMS
 When chemical buffers alone cannot prevent
changes in blood pH, the respiratory system is the
second line of defense against changes.
 Eliminate or Retain CO₂
 Change in pH are RAPID
 Occuring within minutes
 PCO₂ ∞ VCO₂/VA
PHOSPHATE BUFFER SYSTEM
 The phosphate buffer system (HPO42-/H2PO4-)
plays a role in plasma and erythrocytes.

 H2PO4- + H2O ↔ H3O+ + HPO42-

 Any acid reacts with monohydrogen phosphate

to form dihydrogen phosphate
dihydrogen phosphate monohydrogen phosphate

 H2PO4- + H2O ← HPO42- + H3O+

 The base is neutralized by dihydrogen phosphate
dihydrogen phosphate monohydrogen phosphate
 H2PO4- + OH- → HPO42- + H3O+
29
RENAL ACID-BASE CONTROL
MECHANISMS
 The kidneys are the third line of defence against
wide changes in body fluid pH.
 movement of bicarbonate
 Retention/Excretion of acids
 Generating additional buffers
 Long term regulator of ACID – BASE balance
 May take hours to days for correction
RENAL REGULATION OF ACID BASE BALANCE

 Role of kidneys is preservation of body’s bicarbonate

stores.
 Accomplished by:

 Reabsorption of 99.9% of filtered bicarbonate

 Regeneration of titrated bicarbonate by excretion of:
 Titratable acidity (mainly phosphate)

 Ammonium salts
PROTEINS AS A BUFFER
 Proteins contain – COO- groups, which, like acetate ions
(CH3COO-), can act as proton acceptors.

 Proteins also contain – NH3+ groups, which, like

ammonium ions (NH4+), can donate protons.

 If acid comes into blood, hydronium ions can be

neutralized by the – COO- groups
 - COO- + H3O+ → - COOH + H2O

 If base is added, it can be neutralized by the – NH3+

groups
- NH3+ + OH- → - NH2 + H2O
32
 TITRATABLE ACIDITY
 Occurs when
secreted H+
encounter & titrate
phosphate in tubular
fluid
 Refers to amount of
strong base needed
to titrate urine back to
pH 7.4
 40% (15-30 mEq) of
daily fixed acid load
 Relatively constant
(not highly adaptable)
RENAL REABSORPTION OF
BICARBONATE
Proximal tubule:
70-90%
Loop of Henle:
10-20%
Distal tubule and
collecting ducts:
4-7%
FACTORS AFFECTING RENAL
BICARBONATE REABSORPTION
 Filtered load of
bicarbonate
 Prolonged changes in
pCO2
 Extracellular fluid
volume
 Plasma chloride
concentration
 Plasma potassium
concentration
 Hormones (e.g.,
mineralocorticoids,
glucocorticoids)
 If secreted H+ ions combine with filtered
bicarbonate, bicarbonate is reabsorbed
 If secreted H+ ions combine with
phosphate or ammonia, net acid excretion
and generation of new bicarbonate occur
NET ACID EXCRETION
 Hydrogen Ions
Are secreted into tubular fluid along
 Proximal convoluted tubule (PCT)

 Distal convoluted tubule (DCT)

 Collecting system
AMMONIUM EXCRETION
 Occurs when
secreted H+
combine with NH3
and are trapped as
NH4+ salts in
tubular fluid
 60% (25-50 mEq)
of daily fixed acid
load
 Very adaptable (via
glutaminase
induction)
 AMMONIUM EXCRETION
 Large amounts
of H+ can be
excreted
without
extremely low
urine pH
because pKa
of NH3/NH4+
system is very
high (9.2)
ACID–BASE BALANCE DISTURBANCES

Interactions among the Carbonic Acid–Bicarbonate Buffer

System and Compensatory Mechanisms in the Regulation of
Plasma pH.
FOUR BASIC TYPES OF IMBALANCE

 Metabolic Acidosis
 Metabolic Alkalosis
 Respiratory Acidosis
 Respiratory Alkalosis
 ACID BASE DISORDERS
Disorder pH [H+] Primary Secondary
disturbance response
Metabolic acidosis    [HCO3-]  pCO2

Metabolic alkalosis    [HCO3-]  pCO2

Respiratory acidosis    pCO2  [HCO3-]

Respiratory alkalosis    pCO2  [HCO3-]

METABOLIC ACIDOSIS
 Primary AB disorder
 ↓HCO₃⁻ → ↓ pH

 Gain of strong acid

 Loss of base(HCO₃⁻)
CAUSES OF METABOLIC ACIDOSIS

 LACTIC ACIDOSIS  TOXINS

 KETOACIDOSIS  Ethylene glycol
 Diabetic  Methanol
 Alcoholic  Salicylates
 Starvation  Propylene glycol
 RENAL FAILURE (acute
and chronic)
ACID–BASE BALANCE DISTURBANCES

Responses to Metabolic Acidosis

 METABOLIC ACIDOSIS
 Symptoms are specific and a result of the underlying pathology
 Respiratory effects:

 Hyperventilation

 CVS:

 ↓ myocardial contractility

 Sympathetic over activity

 Resistant to catecholamines

 CNS:

 Lethargy, disorientation,stupor,muscle twitching, COMA,

CN palsies
 Others : hyperkalemia
METABOLIC ALKALOSIS
↑ pH due to ↑HCO₃⁻ or ↓acid
 Initiation process :

 ↑in serum HCO₃⁻

 Excessive secretion of net daily production of fixed

acids
 Maintenance:

 ↓HCO₃⁻ excretion or ↑ HCO₃⁻ reclamation

 Chloride depletion

 Pottasium depletion

 ECF volume depletion

 Magnesium depletion
CAUSES OF METABOLIC ALKALOSIS
I. Exogenous HCO3 − loads
A. Acute alkali administration
B. Milk-alkali syndrome
II. Gastrointestinal origin
1. Vomiting
2. Gastric aspiration
III. Renal origin
1. Diuretics
2. Posthypercapnic state
3. Hypercalcemia/hypoparathyroidism
4. Recovery from lactic acidosis or ketoacidosis
5. Nonreabsorbable anions including penicillin, carbenicillin
6. Mg2+ deficiency
7. K+ depletion
 COMPENSATION FOR METABOLIC ALKALOSIS
 Respiratory compensation: HYPOVENTILATION
 ↑PCO₂=0.6 mm  pCO2 per 1.0 mEq/L ↑HCO3-
 Maximal compensation: PCO₂ 55 – 60 mmHg

 Hypoventilation not always found due to

 Hyperventilation

 due to pain

 due to pulmonary congestion

 due to hypoxemia(PO₂ < 50mmHg)

ACID–BASE BALANCE DISTURBANCES

Metabolic Alkalosis
 METABOLIC ALKALOSIS
 Decreased myocardial contractility
 Arrythmias

 ↓ cerebral blood flow

 Confusion
 Mental obtundation
 Neuromuscular excitability
RESPIRATORY ACIDOSIS
 ↑ PCO₂ → ↓pH
 Acute(< 24 hours)

 Chronic(>24 hours)
 COMPENSATION IN RESPIRATORY ACIDOSIS
Acute resp.acidosis:
 Mainly due to intracellular buffering(Hb,Pr,PO₄)
 HCO₃⁻ ↑ = 1mmol for every 10 mmHg ↑ PCO₂

 Minimal increase in HCO₃⁻

 pH change = 0.008 x (40 - PaCO₂)

Chronic resp.acidosis
 Renal compensation (acidification of urine & bicarbonate retention)
comes into action
 HCO₃⁻ ↑= 3.5 mmol for every 10 mm Hg ↑PCO₂

 pH change = 0.003 x (40 - PaCO₂)

 Maximal response : 3 - 4 days

ACID–BASE BALANCE DISTURBANCES

Respiratory Acid–Base Regulation.

ACID–BASE BALANCE DISTURBANCES

Respiratory Acid–Base Regulation.