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WATER

Chapter 2
Part 2 of 2

started tuesday 9/4


IONIZATION OF WATER
IONIZATION OF WATER

Although many of the proper1es of water can be explained


in terms of the uncharged H2O molecule, the small degree
of ioniza?on of water to hydrogen ions (H+) and hydroxide
ions (OH-) must also be taken into account.

Water has a slight tendency to reversibly ionize:

auto-ioniza1on
HOH
H+ + OH-

Although we commonly show the dissocia1on product of


water as H+ and OH-, free protons (H+ ) do not exist in
solu1on.
IONIZATION OF WATER

Hydrogen ions (H+) formed in the dissocia1on of water are


immediately hydrated to hydronium ions (H3O+).

HOH + HOH H3O+ + OH-


Acid Base

Water acts as BOTH a very weak acid and a very weak base.
IONIZATION OF WATER

For prac1cality, many 1mes is sufficient to describe the


ioniza1on of water as:
HOH
H+ + OH-

as long as you remember that a proton never exists as a free


ion but it always associated with a water molecule.

H3O+ = H+
IONIZATION OF WATER

HOH
H+ + OH-

Because reversible ioniza1on is crucial to the role of water


in cellular func1on, we must have a means of expressing the

EXTENT OF IONIZATION OF WATER

in QUANTITATIVE terms.
IONIZATION OF WATER

aA + bB cC + dD

The posi?on of equilibrium of ANY chemical reac1on is


given by its equilibrium constant, Keq.

[Ceq]c [Deq]d The equilibrium constant is FIXED and


Keq = CHARACTERISTIC for any given chemical
[Aeq]a [Beq]b
reac1on at a specified temperature.

It defines the COMPOSITION OF THE FINAL EQUILIBRIUM


MIXTURE, regardless of the star?ng amounts of reactants
and products.
IONIZATION OF WATER
The degree of ioniza1on of water at equilibrium is small.

H2O H + + OH-

The equilibrium constant, Keq that describes the ioniza1on equilibrium


of water at 25°C is:

Kw = [H+] [OH-] = 10-14

and it is called the ion product of water.


IONIZATION OF WATER

For pure water at 25°C :

Kw = [H+] [OH-] = 10-14

That means:
[H+] = [OH-] = 10-7 M

However, the ion product depends on temperature.

At human body temperature,

[H+] = [OH-] = 1.6 x 10-7 M


but we don’t really use this value, stick with other one
IONIZATION OF WATER

This ([H+][OH-] = 10-14) is true for any aqueous solu?on.

In any aqueous solu1on at 25 °C, no macer what it contains,


the product of [H+] and [OH-] must always equal 1.0 x 10-14.

[H+] [OH-] = 10-14

That means that if the [H+] goes up, the [OH-] must go down
so that the product of the two is s1ll 1.0 x 10-14 .
IONIZATION OF WATER
For instance, if HCl gas is dissolved in water, increasing the [H+],
the [OH-] must decrease.

H+ H+
H+
H+
H+

H2O

H+ OH-

Adding an acid to pure water


•  increases the [H3O+]
•  causes the [H3O+] to exceed 10−7 M
•  decreases the [OH−]
IONIZATION OF WATER
For instance, if NaOH(s) is dissolved in water, increasing the
[OH-], the [H+] must decrease.

OH- OH-
OH-
OH-
OH-

H2O

OH- H+

Adding a base to pure water


•  increases the [OH-]
•  causes the [OH-] to exceed 10−7 M
•  decreases the [H3O+]
IONIZATION OF WATER
The acidity of a solu?on is determined by the amount of
H+ in the solu1on.

[H+] = [OH-] = 10-7

OH-
10-14 10-13 10-12 10-11 10-10 10-9 10-8 10-7 10-6 10-5 10-4 10-3 10-2 10-1 100

100 10-1 10-2 10-3 10-4 10-5 10-6 10-7 10-8 10-9 10-10 10-11 10-12 10-13 10-14
H+
Acidity increases Basicity increases
Neutral zone
IONIZATION OF WATER
To avoid working with such small numbers (nega1ve
powers of ten), the pH scale was invented.
The term pH is defined by the expression:

pH = - log [H+]

The logarithm of a number is the exponent to which another fixed


value, the base, must be raised to produce that number.

log10 102 = 2

if [H+] = 10-7 pH = (-1) log 10-7 = (-1) (-7) = 7


IONIZATION OF WATER

OH-
10-14 10-13 10-12 10-11 10-10 10-9 10-8 10-7 10-6 10-5 10-4 10-3 10-2 10-1 100

100 10-1 10-2 10-3 10-4 10-5 10-6 10-7 10-8 10-9 10-10 10-11 10-12 10-13 10-14
H+
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14

Acidity increases pH Basicity increases

Neutral zone
IONIZATION OF WATER
The pH of a solution describes the acidity of a solution.

pH = 7 NEUTRAL SOLUTION

pH < 7 ACID SOLUTION

pH > 7 BASIC SOLUTION

The higher the [H+] of a solution, the lower the pH, so a low pH
describes an acid solution.
Viceversa, a low [H+] is reflected in a high [OH-], so high pH
describe a basic solution.
IONIZATION OF WATER
In an aqueous solution:
[H+] [OH-] = 10-14

- log {[H+] [OH-]} = - log 10-14



- log [H+] + (- log[OH-]) = - log 10-14

pH + pOH = 14

pH = 14 - pOH

pOH = 14 - pH

IONIZATION OF WATER

What’s the pH of a 0.01 M HCl solution?

pH = - log [H+]

[H+] = 0.01 = 10-2

pH = - log 10-2 = 2

IONIZATION OF WATER
Note that the pH scale is logarithmic, not arithme?c.

That means that an increase or decrease of an integer


value changes the concentra?on by a tenfold.

pH = 3 10 1mes more acidic than pH = 4
pH = 3 100 1mes more acidic than pH = 5
pH = 11 10 1mes more basic than pH = 10
PHYSIOLOGICAL pH RANGE
Most body fluids have a pH range of

6.5 - 8

Most biochemical reaction occur in this range.


In our stomach the pH is much lower (between 1.5 and 3.5).

The LEVEL OF ACIDITY (or pH) of living cells is TIGHTLY


REGULATED and KEPT CONSTANT to allow cells to maintain
their structural integrity, and for cellular processes to function.

An organism, as the human body, needs to be able to


CONTROL THE LEVEL OF ACID within its cells for life to be
possible.
WEAK ACIDS
AND
WEAK BASES
WEAK ACIDS AND BASES
Almost all biochemical reac1ons occur in water.

The BEHAVIOR of all the biochemical substances (molecules


and ions) depends on their:

STATE OF IONIZATION

In other words, on their CHARGE.

Many biochemical molecules are either weak acids or weak


bases.
WEAK ACIDS AND BASES
Acids and bases can be defined by their chemical proper1es
in several different ways.
In biochemistry, the Bronsted–Lowry defini?on of acids and
bases is generally used.

ACID: any substance that donates protons (H+)

BASE: any substance that accepts protons (H+)


WEAK ACIDS AND BASES
For example, when nitrous acid is placed in water, the reac1on that
occurs is:

HNO2 + H 2O H3O+ + NO2-


proton proton hydronium
donor acceptor
(acid) (base)

HNO2 has donated its H+, which was accepted by the water molecule.

No1ce how the H+, once donated by the HNO2 molecule, does not
simply float around in the water freely, but instead associates strongly
with water (the proton acceptor) to form a hydronium ion (H3O+).
Thus, using the Bronsted–Lowry conven1on, the acid proton H+ is
omen wricen as H3O+ to reflect this fact.
WEAK ACIDS AND BASES
Similarly, ammonia (NH3) is a base because it acts as a proton
acceptor:

NH3 + H 2O NH4+ + OH-


conjugate
conjugate
proton proton acid
base

acceptor donor
(base) (acid)

As you can see in the examples above, water can act not only as a
proton acceptor, but also as a proton donor.
WEAK ACIDS AND BASES

STRONG ACIDS AND BASES


They dissociate 100%.

WEAK ACIDS AND BASES


They only partially dissociate.
WEAK ACIDS AND BASES
A molecule change its ionization state by gaining or losing
a proton (H+). net charge +1
net charge -2
????net charge -1

COOH COO- COO- COO-


+H
3N C H
+H
3N C H +H
3N C H H2N C H

CH2 CH2 CH2 CH2


CH2 CH2 CH2 CH2
COOH COOH COO- COO-

fully first second third


protonated deprotona1on deprotona1on deprotona1on

Note that the conjugate base might have or not have a negative
charge, but in all cases it has one less positive charge than the
acid.
WEAK ACIDS AND BASES

For convenience we write the dissocia1on of a weak acid:


-
HA A + H +
weak acid

The equilibrium constant for the dissocia1on of a weak acid


(dissocia?on constant) is:
products

Keq = Ka = [A-] [H+] ___________


[HA] reactants

This CONSTANT describes the intrinsic reac?vity of HA


towards water.
VERRRY important
WEAK ACIDS AND BASES
strong acids have higher Ka’s
Stronger acids, such as phosphoric (H3PO4) and carbonic
(H2CO3) acids, have larger dissocia1on constants; weaker
acids, such as monohydrogen phosphate (HPO42-), have
smaller dissocia1on constants.
weaker acids have smaller Ka’s.

high dissociation low dissociation

high Ka low Ka

Each acid has a characteris1c tendency to lose its proton in


an aqueous solu1on. The stronger the acid, the greater its
tendency to lose its proton.
WEAK ACIDS AND BASES

The STRENGTH OF AN ACID is usually expressed as the pKa


value:

pKa = - log Ka

low pKa high pKa

high ka values low ka values


high dissociation low dissociation
WEAK ACIDS AND BASES

The pKa is a constant for a given acid.

For example:
pKa for ace1c acid = 4.76
pK for formic acid = 3.75
a

Formic acid is therefore a stronger acid than ace1c acid.


WEAK ACIDS AND BASES
losing more than one H+

Some acids are POLYPROTIC: they are capable of loosing more


of one proton.

H2CO3 HCO3- + H+

HCO3- CO32- + H+

Each dissociation involve a separate pKa value


WEAK ACIDS AND BASES
****memorize these!!!!
Examples of weak/strong acids and bases.
TITRATION
OF ACIDS
buret w.
TITRANT

AND
BASES flask with ANALYATE
TITRATION OF ACIDS AND BASES

When acids or bases are added to a solution, the pH changes.

or

H+ H+ OH- OH-
H+ OH-

Solu1on of a
STRONG
OR
WEAK
acid or base
TITRATION OF ACIDS AND BASES

The CONTROLLED addition of an acid or base to a base or acid


solution is called

TITRATION Acid or Base

acid or base
TITRATION OF ACIDS AND BASES
An acid-base 1tra1on is a NEUTRALIZATION reac1on.

HCl + NaOH NaCl + H2O



H+ + OH- H2O

A 1tra1on is an acid-base reac1on, in which the hydrogen
ions (H+) from the acid react with hydroxide ions (OH-) from
the base to form water.
Usually, a acid-base 1tra1on is used as analy1cal technique
where a solu1on of known concentra1on is used to

determine the concentra?on of an unknown solu?on


TITRATION OF ACIDS AND BASES
Titrant and analyte is a pair of acid and base.
Acid-base 1tra1ons are monitored by the change of pH as
1tra1on progresses.
A 1tra1on curve is the plot of the pH of the
analyte solu1on versus the volume of the 1trant
added as the 1tra1on progresses.
TITRATION OF ACIDS AND BASES
The 1tra1on curves contain informa1on about the analyte.

Understanding the 1tra1on curve of weak acids with a


strong base will help us understanding the degree of
ionizion at certain values of pH and the chemical behavior of
amino acids, which can be considered weak polypro1c
acids.
The shape of a 1tra1on curve SHOWS GRAPHYCALLY what
kind of acid or base it has been 1trated.

TITRATION OF ACIDS AND BASES
Titra?on of a strong acid with a strong base
Suppose our analyte is HCl and the 1trant is NaOH.
If we start plosng the pH of the analyte
against the volume of NaOH that we are
adding from the burece, we will get a
1tra1on curve as shown below.
TITRATION OF ACIDS AND BASES
Point 1:
No NaOH added yet, so the pH of the analyte is low (it
predominantly contains H3O+ from the dissocia1on of HCl.

As NaOH is added dropwise, H3O+ slowly starts gesng


consumed by OH- produced by dissocia1on of NaOH.
Analyte is s1ll acidic due to predominance of H3O+.
TITRATION OF ACIDS AND BASES
Point 2:
This is the pH recorded at a 1me point just before complete
neutraliza1on takes place.
TITRATION OF ACIDS AND BASES
Point 3:
This is the EQUIVALENCE POINT (halfway up the steep
curve).
moles of NaOH added =
moles of HCl in the analyte

H3O+ ions are


completely neutralized
by OH- ions
[H3O+] = [OH-]

The solu1on only has salt (NaCl) and water and therefore
the pH is neutral (pH = 7).
TITRATION OF ACIDS AND BASES
Point 3:
The EQUIVALENCE POINT is also called END POINT.

In the case of a 1tra1on of a strong acid with a strong base,


the point is also called NEUTRALIZATION POINT (because it
occurs at pH = 7)
TITRATION OF ACIDS AND BASES
Point 4:
Addi1on of NaOH con1nues, pH starts becoming basic
because HCl has been completely neutralized and now
excess of OH- ions are present in the solu1on (from
dissocia1on of NaOH).
TITRATION OF ACIDS AND BASES

The indicator, if carefully chosen, changes color about the same


time as the equivalence point is reached.
The color change of the indicator is the END POINT.

HCl + NaOH NaCl + H2O

H+ + OH- H 2O

[H+] = [OH-] end point = equivalence point

= neutral point
for a strong acid-base titration
TITRATION OF ACIDS AND BASES
Titra?on of a weak acid with a strong base
Let’s assume our analyte is CH3COOH and the 1trant is
NaOH.
If we start plosng the pH of the analyte
against the volume of NaOH that we are
adding from the burece, we will get a
1tra1on curve as shown below.
TITRATION OF ACIDS AND BASES
Point 1:
No NaOH added yet, so the pH of the analyte is low (it
predominantly contains H3O+ from dissocia1on of
CH3COOH).

But ace1c acid is a weak


acid, so the star1ng pH is
higher than what we
no1ced in the case of a
where we had a strong
acid.
As NaOH is added dropwise, H3O+ slowly starts gesng
consumed by OH- (produced by dissocia1on of NaOH). But
analyte is s1ll acidic due to predominance of H3O+ ions.
TITRATION OF ACIDS AND BASES
Point 2:
This is the pH recorded at a 1me point just before complete
neutraliza1on takes place.
TITRATION OF ACIDS AND BASES
Point 3:
This is the equivalence point (halfway up the steep curve).

At this point, moles of


NaOH added = moles of
CH3COOH in the analyte.

The H3O+ ions are completely neutralized by OH- ions.

The solu1on contains only CH3COONa salt and H2O.


TITRATION OF ACIDS AND BASES
In the case of a weak acid versus a strong base, the pH is
NOT neutral at the equivalence point.

The solu1on is basic (pH ~ 9) at the equivalence point.

HAc + OH- Ac - + H 2O
TITRATION OF ACIDS AND BASES

HAc + OH- Ac - + H 2O

CH3COO- is the conjugate base of the weak acid CH3COOH.

CH3COO- is rela1vely a strong base (weak acid CH3COOH has


a strong conjugate base), and will thus react with H2O to
produce OH-
Ac- + H2O HAc + OH-

thus increasing the pH to ~ 9 at the equivalence point..


TITRATION OF ACIDS AND BASES

HAc + OH- Ac - + H 2O

For any weak acid-strong base titration, the pH at


the equivalence point will be greater than 7
because the conjugate of the weak acid is a base

pH

V of titrant
TITRATION OF ACIDS AND BASES
Point 4:
Beyond the equivalence point (when sodium hydroxide is in
excess) the curve is iden1cal to HCl-NaOH 1tra1on curve.
BUFFERS
BUFFERS
WEAK ACIDS and bases differ from their strong counterparts.

When you put one mole of acetic acid into pure water, only
about 4 in 1000 HAc molecules dissociate into H+ and Ac-.

-
HAc Ac + H+
1000 0 0
996 4 4

Thus, if you start with 1000 HAc, you end up with 996 HAc and 4
each of H+ and Ac-.
BUFFERS
Consider acetic acid in aqueous solution:

-
HAc Ac + H+

Suppose OH- is added (NaOH is added):

1) the added OH- will react with the H+ ions to make water

2) the concentration of H+ ions would go down

3) the pH would go up
BUFFERS

-
HAc Ac + H+

However,
HAc works as a backup source of H+.

As protons are taken away by the added OH- (making water),


they are partly replaced by protons from the HAc.

Weak acids act as a


SOURCE OF PROTONS,
helping to keep the [H+] (and thus the pH) of the
solu?on RELATIVELY CONSTANT
BUFFERS
The weak acid resists changes in pH by releasing H+
to compensate for those “used up” in reac1ng with
the OH- .

pH

The weak acid acts as a


BUFFER

V of titrant
The CURVE FLATTENS.
BUFFERS
At the EQUIVALENCE point, ALL of the weak acid is neutralized
and converted to its conjugate base.

HAc + OH- Ac - + H 2O
acid conjugate
base

(the number of moles of H+ = ADDED number of moles of OH-).

However, the pH at the


equivalence point does not
equal 7.
BUFFERS
Two important points in the titration curve are the:

1) equivalence point
[H+] = [OH-]

2) midpoint of the titration


[proton donor] = [proton acceptor]
BUFFERS
The equivalence point allows us to determine the concentration
of an unknown acid or base.

At the equivalence point, the reactants are in stoichiometric


amounts such that the solution contains [H+] = [OH-].

If you titrate a
monoprotic acid of
Equivalence
unknown
point
concentration, you
can determine its
concentration from
the amount of base
needed to reach the
equivalence point.
BUFFERS
The other important feature is the point of half-neutralization.

HAc + OH- Ac - + H 2O

At the midpoint of the titration (point of half-neutralization), at


which exactly 0.5 equivalent of NaOH has been added, one-half
of the original acetic acid has undergone dissociation, so that
the concentration of the proton donor, [HAc], now equals that of
the proton acceptor, [Ac-].

concentration concentration
of the of the
proton donor proton acceptor
[HAc] [Ac-]
BUFFERS
acid conjugate base [CH3COO-]
[CH3COOH] = [CH3COO-] 100%
50% = 50%

Half-equivalence point
acid:conjugate base
1:1

[CH3COOH]
100%
BUFFERS

[CH3COO-]
[CH3COOH] = [CH3COO-] 100%

The pH at the
half-?tra?on
point is equal
to the pKa of
the weak acid

[CH3COOH]
100%
BUFFERS
Shown here are the titration curves
for CH3COOH, H2PO4-, and NH4+.
Although the titration curves of
these acids have the same shape,
they are displaced along the pH
axis because the three acids have
different strengths.

HAc, (highest Ka, lowest pKa), is


the strongest and is already half
dissociated at pH 4.76. H2PO4-
loses a proton less readily, being
half dissociated at pH 6.86. NH4+
is the weakest acid and does not
become half dissociated until pH
9.25.
BUFFERS
Let’s look again ay the titration curve of acetic acid with a strong
base.
The titration curve of acetic acid has a relatively flat zone
extending about 1 pH unit on either side of its midpoint pH of
4.76.

This relatively flat zone is the


BUFFERING REGION of the
acetic acid–acetate buffer pair.

In this zone, an amount of H+


or OH- added to the system
has much less effect on pH
than the same amount added
outside the buffer range.
BUFFERS
At the midpoint of the buffering region,
where the concentra?on of ace?c acid exactly
equals that of acetate,
the buffering power of the system is maximal

A mixture of equal
concentra?ons of ace1c
acid and acetate ion,
found at the midpoint of
the 1tra1on curve, is a
buffer system.
BUFFERS
The titration curve of a weak acid is that
SHOWS GRAPHYCALLY
that a weak acid and its anion (a conjugate acid-base pair)
can act as a BUFFER.

WEAK ACID CONJUGATE BASE

BUFFER
BUFFERS
so essentially a buffering region is this…
The system slows down a change in pH in the region of
about one pH unit above and one pH unit below the pKa.
its meant to pretty much buffer the solution and regulate the pH

Since the pKa of acetic acid is


4.76, the buffering region of
the acetic acid/acetate buffer
is from about 3.76 to 5.76.

The buffer is maximally strong


at a pH of 4.76.
BUFFERS
The acetic acid/acetate pair work as a buffer because HAc
donate protons when extras are needed (such as when OH- is
added to the solution), and, A- can accept protons when extra H+
are added to the solution (adding HCl, for example).

HAc donate protons A- accepts protons


when OH- are added when H+ are added

The maximum ability to donate or accept protons comes


when
[HAc] = [Ac-]
BUFFERS
The titration curves of
CH3COOH, H2PO4-, and
NH4+ have nearly identical
shapes, suggesting that
these curves reflect a
FUNDAMENTAL LAW or
relationship.

The SHAPE of the


titration curve of
any weak acid is
described by the….
henderson hasselbach eqn
HENDERSON

HASSELBALCH
EQUATION
HENDERSON-HASSELBALCH EQUATION

The Henderson-Hasselbach equa1on relates the pH of an
aqueous solu1on of an acid to the acid dissocia1on constant
of the acid, Ka.

[conjugate base]
pH = pKa + log
[weak acid]
HENDERSON-HASSELBALCH EQUATION

[conjugate base]
pH = pKa + log
[acid]

The Henderson-Hasselbalch equation describes the


relationship between the pH of a solution and
the ratio of the molar concentration of the conjugate base
and its acid in it

The Henderson-Hasselbalch equation


PREDICTS THE RESPONSE of a buffer solution
to a CHANGE IN H+ CONCENTRATION
HENDERSON-HASSELBALCH EQUATION

The H-H shows WHY the pKa of a
weak acid is equal to the pH of the
solu1on at the midpoint of its
1tra1on.

At the midpoint, [HA] equals [A-], and


[A-]
pH = pKa + log

[HA]
pH = pKa + log 1

pH = pKa
HENDERSON-HASSELBALCH EQUATION

According to Henderson-
Hasselbach equa1on, when
the concentra1ons of the
acid and the conjugate base
are the same, i.e, when the
acid is 50% dissociated, pH
of the solu1on is equal to
the pKa of the acid.
EFFECT OF pH ON BIOLOGICAL MOLECULES

Fluids inside biological organisms must be maintained at a

CONSTANT pH

for the health of the organism.


For example, the normal pH (a measure of acidity) of blood
and many other biological fluids is about 7.4. At a blood pH
of 7.0, death in acidic coma results, while at blood pH of 7.8,
death occurs from convulsive contrac1ons (tetany).
valid question…
If the pH of biological fluids is so important, how does an
organism make sure its fluids are all kept at the proper pH
for the correct func1oning of all of its enzymes, even if the
organism ingests material that is acidic or basic?
EFFECT OF pH ON BIOLOGICAL MOLECULES
Weak acids are responsible for keeping the H+
concentra1on, (and thus the pH) of the solu1on they are in
rela1vely constant.
We say that all the biological fluids are

BUFFERED

A solu1on containing a buffering agent tends to

resist changes in pH
so that biochemical reac1ons, which omen produce or use
up acids, can proceed without producing wild fluctua1ons in
cellular pH.
EFFECT OF pH ON BIOLOGICAL MOLECULES
Since pH is so critical to biological molecules, any biochemical
experiment that involves studying proteins or cellular systems
needs to be performed in a buffered solution to keep the
molecules in their natural, or native, conformation.

Thus, an understanding of how buffers can help regulate pH is


essential for any biochemist.
BUFFERS
In cells and tissues, phosphate and bicarbonate buffer systems
maintain intracellular and extracellular fluids at their optimum
(physiological) pH, which is usually close to pH 7.

Enzymes generally work optimally at this pH.


POLYPROTIC ACIDS
Some molecules have more than one ionizable group, so they
have more than one pKa.

The consequence is that a polyprotic acid will ionize at different


pH values. seperate pKa values for each molecule of the polyrpotic acid

Buffers are most effective in a pH range near its pKa.

This is where the titration curve is most shallow, and where the
pH is least affected by added acid or base.
POLYPROTIC ACIDS
For example, in the titration curve of phosphoric acid (blood
buffering system):

pKa1 pKa2 pKa3


H3PO4 H2PO4- HPO42- PO43-

The region in the rectangle is the


buffering region of biological
importance.
Note that the slope of the curve is
shallow within one pH unit of the
pKa value.
This is where the buffering range is
most effective, meaning that the
buffer is able to resist changes in pH.
WATER AS A REACTANT
Water is not just the solvent in which the chemical reactions of
living cells occur; it is very often a direct PARTECIPANT in those
reactions.
Examples:

1) Condensation: water is eliminated

2) Hydrolysis: water is added


(enzymatic depolymerization of proteins, carbohydrates, and
nucleic acids)

3) Combustion: water is eliminated