Osmometry

Quiz
• Which of the following is not a colligative
property of solutions?

A. pH
B. Freezing point
C. Osmotic pressure
D. Vapor pressure
 Quiz
• Which of the following describes the basis for the freezing
point osmometer?

A. The freezing point depression is directly proportional to the

amount of solvent present.
B. The freezing point depression varies as the logarithm of the
concentration of solute.
C. The freezing point is raised by an amount that is inversely
proportional to the concentration of dissolved particles in the
solution.
D. The freezing point is lowered by an amount that is directly
proportional to the concentration of dissolved particles in the
solution
 Quiz
• Which of the following may be associated with the
colloid osmotic pressure (COP) osmometer?

A. Utilizes a cooling bath set at -7°C

B. Measures total serum osmolality
C. Negative pressure on reference (saline) side
equivalent to COP of sample
D. Measures contribution of electrolytes to osmolality
 Quiz
• Given the following information, calculate the
plasma osmolality in milliosmoles per kilogram:
sodium—142 mmol/L; glucose—130 mg/dL; urea
nitrogen— 18mg/dL.

A. 290
B. 291
C. 295
D. 298
 Introduction
• Osmometry measures the total solute particles
concentration in a liquid that contribute to the
osmotic pressure of a solution
• Solutes added to a solvent alter the solvent in
several ways
• These changes, collectively referred to as
colligative properties, are dependent on the
total number of articles present in the solution
• The actual mass of the particles is irrelevant; a
small molecule will exert the same effect as a
large molecule
Colligative properties

Properties determined by the number of

particles in solution rather than the type of
particles.

Vapour pressure lowering

Freezing point depression
Boiling point elevation
Osmotic pressure elevation
 How Vapor Pressure Lowering Occurs
 Solute particles take up space in a solution.
 Solute particles on surface decrease number of
solvent particles on the surface.
 Less solvent particles can evaporate which
lowers the vapor pressure of a liquid.
 Vapor Pressures of Pure Water and a Water Solution
The vapor pressure of water over pure water is greater than the vapor
pressure of water over an aqueous solution containing a nonvolatile
solute.

Solute particles take up

surface area and lower
 Boiling Point Elevation
When a non volatile solute is added to solvent:
• Vapor pressure of solvent is lowered

• solution formed must be heated to higher

temperature than boiling point of pure solvent to
reach a vapor pressure of 1 atm.

• This means that non volatile solute elevates the

boiling point of the solvent which we call boiling
point elevation
 Freezing Point Depression

•Addition of a non-volatile solute to a solution

lowers the freezing point of the solution relative
to the pure solvent.
•The change in freezing point is equal to the
molal freezing-point constant multiplied by
the molality of the solution.
ΔTf = kf m (eqn 1)
•The molal freezing-point constant used is the
constant for the solvent, not the solute.
 Osmosis
There are many times in nature when a solvent will pass
spontaneously through a semipermeable membrane,
which is a membrane permeable to solvent, but not
solute

The osmotic pressure, Π, is the pressure that must be

applied to stop the influx of solvent
 Osmolality
• What is the best way of measuring this?
• Osmotic Pressure, the pressure generated across
a semi-permeable membrane with water on the
one side and the solution you are measuring on
the other, would seem to offer the best choice
with such high pressures being produced from
relatively low solution strengths.
• In practice it is impossible to measure due to the
problem of getting a perfect semi-permeable
membrane, small ions will always get through
 Osmolality
• Even if the perfect membrane did exist the pressures
exerted are so immense, 23 atmospheres for
1000mOsmols, that it would be sure to burst!
• Of course, if you want know about only the larger
molecules, such as proteins, you can measure Osmotic
Pressure with a Colloid Osmometer and select an
appropriate membrane with a cut-off of 10,000 or
20,000 Daltons which will hold back the protein
molecules but allow everything else to get through.
• But this is no use if you need to know the total
Osmolality.
 Osmolality
• Measuring Boiling Point Elevation is messy and
the change in BP is only small, clearly this is not
an option for blood samples, unless you want a
blood curdling experience,
• Vapour Pressure works, but there is a drawback
when measuring solutions containing other
liquids which exert a vapour pressure of their
own, for example a blood sample containing
alcohol will give a misleadingly high result.
Freezing point depression osmometer
Components
• 1. A thermostatically controlled cooling bath or
block maintained at -7oC
• 2. A rapid stir mechanism to initiate (seed)
freezing of the sample
• 3. A thermistor probe connected to a circuit to
measure the temperature of the sample
• Thermistor is a glass bead attached to a metal
stem whose resistance varies rapidly and
predictably with temperature
Freezing point depression osmometer
• 4. A galvanometer that displays the freezing
curve and that is used as a guide when the
measuring potentiometer is used
• 5. A measuring potentiometer (variable
resistor) is used to null the current in the
circuit
• 4 & 5 can be replaced by a LED display that
indicates the time course of the freezing curve
and the final result
Freezing point depression osmometer
Freezing point depression osmometer
 Analysis
• Sample with thermistor probe and stirring wire
lowered into the bath
• With gentle stirring , sample is super-cooled* to a
temperature several degrees below its freezing
point (-7oC)
• When sufficient super-cooling has occurred , the
sample is raised to a point above the liquid in the
cooling bath
• Vigorous stirring of the sample initiates freezing
of the super-cooled solution
 Analysis
• The freezing is only to the slush stage, with about 2-3%
of solvent solidifying
• The released heat of infusion initially warms the
solution and the temp plateaus (equilibrium temp)
• At the end of equilibrium temp plateau, the
galvanometer again indicates decreasing temp as the
sample freezes further toward a complete solid
(eqn 1)
Freezing curve
 Clinical uses
Serum osmolality
• Screening for Toxin ingestion
• Alcohols (methanol, isopropanol)
• Glycols
• Monitoring conc of osmotically active agents
• Mannitol
• Evaluation of hyponatremia
• Exclusion of pseudohyponatremia
• Presence of other osmotic agents (glycine,
glucose)
 Urine osmolality

• Evaluation of increased urine output

• Primary polydipsia (low serum, low urine osm)
• Diabetes insipidus (high serum, low urine osm)
• Diabetes mellitus (high serum, high urine osm)
• Evaluation of decreased urine output
• Dehydration (high urine osmolality, negative free
water clearance)
• Acute tubular injury (0 free water clearance)
• Evaluation of renal acidification defects
• Urine osmolar gap=NH4+ excretion
 Stool osmolality
• Separation of secretory from osmotic
diarhoea
Osmotic diarrhoea
• Presence of unabsorbed solutes
• Due to laxative abuse, malabsorption
Secretory diarrhoea
• Due to agents damaging mucosa
(inflammation, drugs, infections)
 Homework
• A 48 year-old female presented with severe diarrhoea on several occasions over a six month
period. Prior to her final admission she had diarrhoea for four days consisting of 10-12 watery
motions a day. She also complained of three periods of hot flushes.
• The plasma and faecal electrolytes were as follows:
•
• Plasma
• Na 142 mmol/L
• K 2.4 mmol/L
• Cl 110 mmol/L
• HCO3 18 mmol/L
• Urea 6.1 mmol/L
• Creat 90 umol
•
• Faeces
• Na 76 mmol/L
• K 75 mmol/L
• Osmol 284 mmol/L
•
•
a) Comment on the results
b) What is the differential diagnosis?
c) Upon further investigations urinary 5HIA was found to be largely elevated. Explain why the clinical
features tend to be delayed in this condition.
THE END