Group-2 biochemistry practical:

Subject : biochemistry

Topic : pH & buffers

Lecturer: Alexander N. Tan Jr., M.D.

Group members :

1. Arjun kataria
2. Parth faldu
3. Nayan gadhiya
4. Roshani chaudhari
5. Pallak gautam
6. Krishna godhani
7. Sophia givertz
8. Anurag kamlakar
9. Sandesh gamot
 CHAPTER ONE: pH AND BUFFERS

pH :

the pH of solution is the common logarithm of the reciprocal of the

hydrogen ion concentration expressed as:

ph=log 1/H+

pH is detected and measured by using a pH meter, but the most common

detectors are the color changes of acid-base indicators. an acid-base
indicator is a weak acid. the ionized and unionized forms of the indicator
have different colors:

Hn + H2O <====> H+ + in-

When the indicator is placed in an solution the equilibrium is shifted

to the Hin form predominants. In an alkaline solution depends upon the
ratio of Hn and is related to pH in the following equaton:

Log10 [in-]/ [HIn] = pH-pKa

(comes from Handerson-Hasselbalch equation)

The color of the indicator solution is therefore a measure of the pH.

BUFFERS :

A buffer solution is one of that resits a change solution in pH when a

small amount of acid or base is added. A buffer solution contain a weak
acid, HA and its conjugate base. An explain is the mixture of acetic acid
and sodium acetate.
 Buffers are significant in biochemistry. An excellent example is its in
blood. blood contains phosphates, carbohydrates, and proteins as buffering
components and has a content pH between 7.35 and 7.45 even though
there are large amount of acids and bases contantly introduced by
designation of foods, absorption, muscular activity and respiration.

Procedure:

pH determination

1. prepare the following samples:

a. defibrinated blood

b. fresh milk

c. freshly volded urine

2. Determine the approximate pH of the sample using the pH paper by

following this steps:

a. dip a piece of paper in the sample for about 10 seconds. remove the
paper and place it on a watch glass.

b.match the color produced in the pH paper with the color chart to
determine the pH of the sample.

c.record this results.

3.determine the accurate pH of the samples using the pH meter.

QUESTIONS AND PROBLEMS:

1. What is the H+ concentration of a phosphate buffer solution if its

pH is 7.4?
Ans.

pH=7.4
[H+]=?
Now pH = log1/[H+]
7.4 = log1/[H+]
[H+] = log1/
[H+] = 3.98 X 10-8 M
So like this we can find H+ concentration of any solution by
using this equation

2. What is the pH of a 0.125 M NaOH solution? (clue: pH + pOH =

14)
Ans.

Same as above with some twist …..

pH= ??
[OH-]=0.125 M

So pOH = -log[OH-]
pOH = invlog

3. What is the significant of buffer systems in humans?

 Ans. There are three buffer systems in the human body. The protein
buffer system is the most important and widely operating buffer in the
bodies fluid. An example of a protein buffer is hemoglobin. The
phosphate buffer system is made up of two ions, hydrogen phosphate
ion, and dihydrogen phosphate ion. The bicarbonate buffer system
function is to maintain the pH level in blood mammals. The
bicarbonate buffer system neutralize the pH enzymes in the stomach
and intestines.

4. What is the Henderson-hasselbalch equation?explain.

Ans.

In chemistry, the Henderson–Hasselbach equation describes the
derivation of pH as a measure of acidity (using pKa, the negative log
of the acid dissociation constant) in biological and chemical systems.
The equation is also useful for estimating the pH of a buffer solution
and finding the equilibrium pH in acid-base reactions (it is widely used
to calculate the isoelectric point of proteins).
The equation is given by:

pH = pKa + log [A-]/[HA]

5. what is the normal pH of blood?urine?gastric juice? why is it

necessary to know the pH of these physiologic fluids?

Ans. Normal ph of the …

lood ph

Urine ph

Gastric juice ph–

6.what is the biochemical relevance of pH?

Ans. pH is a measure of acidity or alkalinity and is measured on a scale of

0-14, with 0 being 100% acidic and 14 being 100% basic. A pH of 7 is
neutral, neither acidic or basic. pH has relevance to biochemical systems,
because most biological systems require a mainly neutral pH environment
to survive. For example, the pH of blood is 7.4 with a very narrow range.

7.calculate the ph of 0.01 M HCL