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BIOCHEMISTRY (Lab)

 Measurement if PH-one of the most PH scale


important and frequently used
0-1.99 Strongly acidic
procedures in Biochemistry.
2-4.99 Moderate acidic
 Confirmation and functions o 5-6.99 Weakly acidic
biomolecules are affected by: 7 Neutral
7.01-9.00 Weakly Basic
Hydrogen, hydroxyl ion and
9.01-12.00 Moderate Basic
aqueous environment 12.01-14.00 Strongly Basic
 Intracellular and extracellular fluids
of living organisms=different pH Saliva 6.80 Weakly
 pH (puissance ‘ hydrogen )(power of acidic
hydrogen)
by Sorenen Aspirin 2.95 Moderate
 convinent way of expressing the acidic
Baking 8.75 Weakly
exact acidity and basicity of a
Soda Basic
solution Vinegar 2.73 Moderate
 In aqueous solution Acidic acidic
properties are due to Milk of
hydrogen ions H+ or Magnesia
hydronium ions 1M 7.06 Weakly
Albumin Basic
Blood 7.35- Weakly
 Indicators are usually complex
7.45 Basic
organic compound that are of one Urine 5.85
color if the hydrogen ion
concentration is above a certain
value and of a diff color if it lower PREPARATION Of Buffer
 Eg. pH paper indicators
1.) 100 ml of acetate buffer Solution
 pH meter - electrical instrument ( for from 0.1 M acetic acid and Sodium
precise results) Acetate
 25°C pH= 7 (neutral) pH=<7 acidic pKa 4.73
pH=>7 basic pH 5.22
 Buffer solution = mixture of weak 2.) Measure the volume of th reagents
acid and its conjugate base, or a in Graduated Cylinder
weak base and its conjugate acid. 3.) Mix in a 100ml Volumetric Flask
Ability to resist sudden changes in 4.) If the measured pH deviates from
pH the assigned value more than 0.10 ,
BIOCHEMISTRY (Lab)

add drop wise either 0.1M HCL or


0.1 M NaOH to bring the buffer -/+
0.05 of the desired pH( 5.22) in the first method, prepare a solution
with an acid and its conjugate base by
How does such pH remain constant? dissolving the acid form of the buffer in
about 60% of the volume of water
 ph remains constant because of required to obtain the final solution
buffer. Buffered maintains the pH of volume. Then, measure the pH of the
solution using a pH probe. The pH can
solution by adjusting the direction of be adjusted up to the desired value
their chemical reaction. using a strong base like NaOH. If the
buffer is made with a base and its
Why is it important that the pH body conjugate acid, the pH can be adjusted
fluids like blood remain virtually using a strong acid like HCl. Once the
constant? pH is correct, dilute the solution to the
final desired volume.
 impairment of these transporters
causes dysfunction of cells, diseases, Alternatively, you can prepare solutions
of both the acid form and base form of
and a decrease in physical the solution. Both solutions must contain
performance associated with the same buffer concentration as the
abnormal pH. Additionally, it is concentration of the buffer in the final
known that fluid pH in the solution. To get the final buffer, add one
solution to the other while monitoring the
interstitial space of metabolic
pH.
tissues is easily changed due to little
pH buffering In a third method, you can determine the
exact amount of acid and conjugate
COMPUTATION base needed to make a buffer of a
certain pH, using the Henderson-
pKa: A quantitative measure of the Hasselbach equation:
strength of an acid in solution; a
weak acid has a pKa value in the pH=pKa+log([A−][HA])
approximate range −2 to 12 in water
and a strong acid has a pKa value of Sodium acetate,
less than about −2.
CH₃COONa, also abbreviated NaOAc, is
the sodium salt of acetic acid. This colorless
Buffer solutions are resistant to pH
deliquescent salt has a wide range of uses.
change because of the presence of an Wikipedia
equilibrium between the acid (HA) and
its conjugate base (A–). The balanced
equation for this reaction is:
Formula: C2H3NaO2
HA⇌H++A−
BIOCHEMISTRY (Lab)

Molar mass: 82.0343 g/mol

IUPAC ID: Sodium acetate

Boiling point: 881.4 °C

Density: 1.53 g/cm³

Soluble in: Water

Acetic acid

Chemical compound

DescriptionAcetic acid, systematically


named ethanoic acid, is a colourless liquid
organic compound with the chemical
formula CH₃COOH. When undiluted, it is
sometimes called glacial acetic acid.
Vinegar is no less than 4% acetic acid by
volume, making acetic acid the main
component of vinegar apart from water.
Wikipedia

Formula: CH3COOH

IUPAC ID: Acetic acid

Molar mass: 60.052 g/mol

Density: 1.05 g/cm³

Boiling point: 118.1 °C

Melting point: 16.6 °C

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