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BIOCHEMISTRY

1.1. Introduction to Water, Acids, Bases, and Buffers


Dr. Alba
2018A
Alvarez, E.M. A., Alvarez, L., Alvero, G., Amadore, R., Amog, P., Ang, D.
Outline
WATER
A. Biochemistry of Water
B. Properties of Water
C. Water as Solvent
D. Water and Thermal Regulation
E. Total Body Water
II. OSMOLALITY
III. pH
A. Acid
B. Base
C. Henderson-Hasselbach Equation
IV. BUFFER

o
o

I.

Objectives
To be able to differentiate among individuals
its distribution and factors affecting it
To be able to apply the properties of water
and its uses in our daily life
To recognize the role of water and its
biochemical properties
To be able to determine acids, bases, and
buffers in everyday living
To be able to apply Hendeson-Hasselbach
equation
To familiarize with cases concerning water
balance and basic acid-base problems
I. WATER
Predominant chemical component of a living
organism
Forms an essential part of body cells and
fluids
Matrix of many living reactions
Universal solvent
Medium in intracellular and extracellular
processes
Transport system
Cushion
High surface tension
Liquid at room temperature
o High boiling point
o High freezing point

Alvarez|Alvarez|Alvero|Amadore|Amog|Ang

June 18, 2014


1st Semester, A.Y. 2014-2015

Temperature regulation
High latent heat of vaporization

A. Biochemistry of Water

Consists of 3 atoms: 2 hydrogen atoms and


1 oxygen atom
Dipolar:
o Allows it to form hydrogen which is
responsible for the solvent property
o Both with positive and negative ends
Has 2 unshared electrons that form an
electron dense cloud
Has strong tendency to form hydrogen
bonds
o Acts as hydrogen donor (acid) and acts
as
hydrogen
acceptor
(base)
simultaneously
Example: Its ionization may mean
an intermolecular proton transfer to form
hydronium ion (H3O+) and hydroxide
ion (OH-)
o Water is a reactant or product in many
metabolic processes
Bent molecule with a bond angle of 105

Figure 1. Water Molecule

Type of covalent bond is called a polar bond


o Due to the separation of charge
Partial negative on the oxygen is twice the
strength of a hydrogen since it shares
electrons with 2 hydrogen atoms.
Both the hydrogen and oxygen atoms form
hydrogen bonds and participate in the
hydration shell
Oxygen binds with 2 hydrogen atoms so each
water molecule is hydrogen bonded to 4 close
water molecule in fluid

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molecules: stay in hydration shell for 2.4


nanoseconds
D. Water and Thermal Regulation

Figure 2. Hydrogen Bonding

Hydrogen Bond
o Enable water to dissolve many organic
biomolecules with functional groups which
can participate in binding
1. Oxygen atoms of aldehydes
2. Ketones
3. Amides
4. Alcohol and Amines
B. Properties of Water
1. Cohesion
- attraction between particles of same
substance
surface tension
- strength of waters surface
- enables insects to walk on water
2. Adhesion
- attraction between two different
substances
3. High conductivity
4. High latent heat of vaporization
C. Water as Solvent

Dissolution: water forms bonds and


electrostatic interactions
Hydrogen bonds: strong enough to dissolve
polar molecules in water and to separate
charges; weak enough to allow movement of
water and solutes
H-bond between two water molecules: 4kcal
(1/20th of strength of covalent bond)
Waters strong dipole and high dielectric
constant enable water to dissolve large
quantities of charged compounds
Average H-bond between water molecules:
lasts only 10 picoseconds; each water

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Structure of water: allows it to resist


temperature change
High heat of fusion = large dropp of
temperature as needed to convert liquid water
to solid state of ice
High thermal conductivity = facilitates
dissipation of heat
High heat capacity and heat of vaporization

E. Total Body Water

Total amount of body water: CONSTANT


Tissue concentration: VARIES
Average lean adult male:
60% fluids, 40% solids
Average lean adult female:
55% fluids, 45% solids
Table 1. Amount of Total Body Water Among
Sexes, Period of Development and Based on
Weight
Infant

Male Adult

Female Adult

Thin

80

65

60

Average

70

60

55

Obese

65

55

45

Factors affecting amount of body water:


Body weight - heavier you are the less total
body water
Age - older you are the less total body water
Sex - men are leaner, therefore have more
total body water than females
Amount of fat - fatter you are the less total
body water

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Computations for total body water and amount of


water in different body compartments:
Total body water = 0.6 x body weight
Extracellular fluid (ECF)= 0.2 x body weight
o Interstitial fluid = 0.75 x ECF
o Plasma fluid = 0.25 x ECF
Intercellular fluid (ICF) = 0.4 x body weight
Facts on water and its distribution
Half-life of water: 9.3 days
Fluctuation: less than 1% of body weight
per day
Intake and output: 2000mL at normal
conditions
Affected by:
o Climate
o Activity
o Diet
o State of health
Table 2. Typical Daily Water Balance (average
build, normal, adult, male)
Water Intake
Drinking Water
600
Water in other beverages
400
Performed Water
700
Metabolic Water
300
Total
2000
Water Output
Skin
500
Lungs
400
Urine
1000
Stool
100
Total
2000

II. OSMOLALITY
Water is distributed dependent on the
concentration of solutes in the body
It is proportional to total concentration of all
dissolved molecules
Osmotic force attraction of particles in a
solution
o Dependent on number of particles or ions
in a solution
o Water flows from region of low osmotic
pressure to a region of high osmotic
pressure

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Osmolality - measure of osmotic pressure,


total concentration of dissolved particles in a
solution
Unit of osmotic pressure: osmole (OSM)
= 10milliosmole per kg solvent
Equation of total osmolality:
[

where:
2[Na] = [Na] x [Cl]
GLU = glucose (FBS fasting blood sugar;
FBG fasting blood glucose)
BUN = urea; blood urea nitrogen

Osmolality is regulated by changing the total


body water and not by changing body solute
Measurement of osmolality (water balance) in
one compartment will reflect osmolality in all
compartments
Serum sodium concentration - measure of
body osmolality; used to diagnose disorders of
water balance

Fluid Compartments: (Chief osmotic particles)


Extracellular: Na, HCO3, Cl
Intracellular: K, Mg, PO4
Table 3. Distribution of Cations and Anions
Between ECF & ICF
In
ECF
ICF
mEq/L
Na
145
12
K
4
150
Ca
5
0.001
Cl
105
5
HCO3
25
12
P
2
100
pH
7.4
7.1
Control state:
Hypertonic NaCl intracellular K will move out
to extracellular fluid; crenate
Water extracellular Na will move in to the
intracellular fluid; lyse
Isotonic saline equal movement; controlled
state
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III. pH
Dissociation of water
Forms H+ and OH- and extent is 0.0000001 M
or 10-7 mol/l the concentration of hydrogen
ions in a solution
It is the negative log of hydrogen ion
expressed in moles/liter (mol/L)
Expresses the relationship between H ion
concentration and the hydroxide ion
concentration and the concentration of water at
equilibrium
Sorensen first introduced the term pH in
1909
pH of pure water = 7
pH 7.0 neutral pH; H and OH are equal
Higher pH = low concentrations of H

Dissociation constant of water (Kd or Keq)


Expresses the relationship between H ion
concentration and the hydroxide ion
concentration and the concentration of water at
equilibrium
Because water dissociates to small extent, it is
constant at 55.5M: multiplication of Kd of water
by 55.5 gives the ion product of water (Kw)
=([+][]) / [2]
=(+)()=11014
* Water Molecules exhibit a slight tendency to
dissociate.
Act both as acid and base -> its ionization
may mean an intermolecular proton transfer to
form hydronium ion (H3O) and hydroxide ion
A. Acid

Higher H ion concentration and lower OH ion


concentration than pure water
Donate a hydrogen ion to a solution
Bronsted Lowry
o Proton or H donor in a reaction
o Usually tastes sour
o Turns blue litmus paper to red
Lewis
o Potential electron pair acceptor in a
reaction to form a covalent bond
Arrhenius
o Compounds that dissociates in water to
form H+
Strong and Weak acids

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During metabolism, several acids increases


H in the body thus decreases the pH
o Weak or strong dependent on the degree
of dissociation
o A strong acid is the one that completely
dissociates or 100% ionizes in water
solution
Metabolic acids: metabolized at the level of
the liver and lungs
Non-metabolic acids: excreted by the
kidneys

Equilibrium constant for the dissociation of a


weak acid:
The tendency of an acid to dissociate and donate
a hydrogen ion to a solution is a Ka
()+()+()
=([+][]) / []
=log10

Acids produced by the Body:


o Volatile acids
CO2
o from decarboxylations in citric acid cycle
o 13,000-20,000 mmol/day
o Eliminated through the lungs
CO2 + H2O = H2CO3 = H+ + HCO3carbonic anhydrase
o

Non-volatile acids (Non-carbonic/Fixed


acids)
50-80 mol/day
Organic (such as lactic acid and ketone
bodies) or inorganic
Due to catabolism of food:
1. Diet:
a) Sulfuric acid H2SO4 (contains
methionine, cysteine and cystine)
from sulfur-containing amino acids
b) Phosphoric acid (H3PO4) from
phosphoproteins/phospholipids
c) Uric acid from purine metabolism
2. Intermediary metabolism:
a) Lactic acid from carbohydrates
when O2 is low
b) Ketoacids from fats (acetoacetic
(acetone) and Beta-hydroxybutyric
acid when DM is uncontrolled)
3. Bicarbonate losses in the stool:
a) Diarrhea addition of non-volatile
acids

B. Base
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Basic solutions have a higher OH ion


concentration and lower H ion concentration
than pure water
Bronsted Lowry
o Proton or H acceptor in a reaction
o Turns bitter and feels slipper or soapy
o Turns red litmus paper to blue
Lewis
o Potential electron pair donor in a
reaction to form a covalent bond
Arrhenius
o Compounds that dissociates in water to
form OH (hydroxide)
Strong and Weak bases
o A strong base is the one that completely
dissociates

C. Henderson-Hasselbach Equation

Equation relating the pH to the composition of


the buffer solutions

IV. BUFFER
Consists of a weak acid and it conjugate base
or a weak base and it conjugate acid
Resists changes in pH
Buffer Capacity
o Equivalents of (H) or (OH) required to
change 1L of buffer by 1.0 pH unit
o Measure as the effectivity of a solution to
resist pH changes
o Maximum buffer capacity occurs at +/- 1
pH unit on either side of the pKa

Acid-Base Buffer Systems:

Conjugate acid-base pairs that bind or release


H+ when an acid or base is added to he body
so that the change in pH is minimized
1st line immediate defense against pH changes
Bicarbonate-carbonic acid: major buffer
system of the body

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A. Blood pH

7.36 and 7.44 (7.35 to 7.45)


Intracellular pH at 7.1
Extracellular pH must be maintained to allow
metabolic functions of the liver, beating of the
heart, and conduction of neural impulses is
6.8-7.8

B. Buffers in the Body


1. Bicarbonate Buffer System (ECF)
Major source of metabolic acid in the
body: CO2 (produced principally from fuel
oxidation from the TCA or tricarbocylic
acid cycle)
Normal: 13 mol of CO2/day
Regulation of CO2 availability by
respiration

Increased HCO3, increased pH alkaline


Increased pCO2, decreased pH acidic
2. Hemoglobin Buffer System (RBC)
Bicarbonate buffer system +
haemoglobin: buffering the blood,
transporting CO2 in the lungs
3. Phosphate Buffer (ICF)
Non-volatile acids that is produced from
body metabolism cannot be excreted as
CO2 is excreted in the urine (usually from
sulfuric acids, phosphates, and ammonia)
C. Acid-Base Balance
---------------

Regulated by:
1. Liver
o Metabolizes proteins that produces
hydrogen ions
2. Lungs
o Removal or retention of CO2
Net gain of CO2: Hypoventilation
Net loss of CO2: Hyperventlation
o Excess CO2 combine with water to
form carbonic acid (H2CO3)
3. Kidneys
o Generate new HCO3
o Regulate plasma bicarbonate by
reabsorbing filtered bicarbonate

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It affects hydrogen ion excretion thru


the formation of titrable acids and
ammonium (synthsis)
Excrete acid in the form of dihydrogen
phosphate (H2PO4), bicarbonate
(HCO3), ammonia (NH3)

D. Acid-Base Disorders

Change in Extracellular pH
o conditions that initially affect either HCO3(base) or pCO2 (acid-carbonic acid)
leading to a shift in pH from normal;
(homeostasis is lost)
o May be caused by:
Renal or respiratory function is
abnormal
Acid or base overload overwhelms the
capacity to excrete
Changes in plasma H concentration and pH
can be induced by alterations of pCO2 and
plasma bicarbonate (HCO3-) concentration
pCO2: regulated by respiration -> primary
disturbance: respiratory acidosis/alkalosis
primary
disturbance
in
bicarbonate
concentration(HCO3-)
:metabolic
acidosis/alkalosis
Evokes a compensatory response that is the
same direction as the primary disturbance

Arterial Blood Gases (ABGs)


pH=7.35-7.45
pCO2=35-45 mmHg
HCO3=21-28 mmHg
pO2=80-100 mmHg
Anion gap
Means
of
approximating
the
total
concentrations of anions other than Cl- and
HCO3- in the plasma
Sum of the major cations in the plasma minus
the sum of the major anions
=([Na+] + [K+]) ([Cl-] + [HCO3-]
=12mEq/L

1. Metabolic acidosis fall in the plasma


bicarbonate and a low pH (pH<7.35)
Compensatory response:
a. Increase in ventilation
b. Renal excretion of excess acid (takes
several days) total restoration
Caused by:
o Loss of alkali (diarrhea, GIT fistulas and
drainages)
o Accumulation of fixed acids (increase in
lactic acid due to shock/hypoxemia,
uncontrolled diabetes mellitus, renal
failure)
2. Metabolic alkalosis elevation of plasma
bicarbonate and a high pH (pH>7.45)
Caused by:
o Bicarbonate administration loss of acid
(relative increase in alkali): hydrogen loss
like vomiting, use of diuretics, ingestion of
alkali compounds (Maalox)
3. Respiratory acidosis increase in pCO2
(pH<7.35); decrease effective alveolar ventilation;
increased CO2 leads to decreased pH (or increase
in production of carbonic acid)
Caused by:
o neuromuscular trouble, depression of
airway regulation, lung disease or airway
obstruction

At risk: abnormal surgery px, analgesia


4. Respiratory alkalosis decreased in pCO2
(pH>7.45)
Caused by:
o hyperventilation (CNS disorders,
medication), conditions that increases the
excretion of carbon dioxide,
hypermetabolic states (overstimulation of
the respiratory center)

*Recommended video:
https://www.youtube.com/watch?v=eK2dBdBRvCU

ACIDOSIS: pH less than 7.35


ALKALOSIS: pH more than 7.45
GUIDE QUESTIONS
Alvarez|Alvarez|Alvero|Amadore|Amog|Ang

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Who among the follow has the highest total body


water?
a. average male- 60% fluids
b. obese women- fatter you are the lower the
total body fluids
c. thin infant- younger therefore has
highest total body water compared to
other options
d. thin male- has high total body water but is
older so less body water compared to thin
infant
Which of the following statements about water id
true?
a. It is the universal solute
b. It forms a minor part of all body cells and
fluids
c. The total amount of body water varies
d. Water has a high latent heat of
vaporization
SOURCES
Devlin, T. (2011). Textbook of Biochemistry with
Clinical Correlations, Wiley-Liss 7th edition.
New York: A John Wiley & Sons, Inc.,
Publication
Murray, R.K., et al., (2009) Harpers Illustrated
Biochemistry, 28th ed, Prentice Hall
International
Oxford Dictionary of Biochemistry and Molecular
Biology, OKxford University Press Inc., New
York

Alvarez|Alvarez|Alvero|Amadore|Amog|Ang

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