MLAB 2401:

Clinical Chemistry

Basic Principles and Practice of Clinical

Chemistry
Part One

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UNITS OF MEASURE

■ Measurement requires a numerical value and a unit

❑ Laboratory results almost always have units of measurement associated
with them

❑ SI units:
❑ length ( meter )

❑ mass ( gram )

❑ quantity ( mole )

❑ Volume ( liter )

❑ Time ( second )

■ Basic units describe unrelated physical quantities

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Unit of Measure: Prefixes
■ Common prefixes and abbreviations that are added to units of measure:
■ deci (d) 10-1
■ centi (c) 10-2
■ milli (m) 10-3
■ micro ( μ) 10-6
■ nano (n) 10-9
■ pico (p) 10-12
■ femto (f) 10-15

■ Example: A common unit of liquid measurement is a deciliter( dl ), or one –

tenth of a liter

■ Combine a prefix with a basic unit results in a statement of a specific length,

weight or volume
❑ Reporting clinical chemistry results may be in units such as :
■ mg / dL
■ g / dL
■ mEq / L

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Scientific Notation

■ True scientific notation format:

❑ 1.22 X 104

❑ BUT in hemo, for example a hemoglobin result

would look like = 12.2 X 103

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Water Specifications
■ Tap water is unsuitable for lab use (too many impurities)
■ Types of water purification techniques
■ Distillation – removes most organic matter
■ Reverse osmosis-removes organic, ionic, microbial, and viral
contaminants
■ Ultrafiltration – removes particulate matter, bacteria, emulsified solids
■ Deionization – ions removed

■ Reagent Grades of water

■ Type I Purest – Required for sensitive tests
■ Type II Acceptable for most uses
■ Type III OK for washing glassware
■ CAP - QC of water : pH, electrical resistance, bacterial culture

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Water filtration system for
Automated chemistry analyzer.

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 Solutions
■ The clinical lab almost always uses solutions. A solution means that
something has been dissolved in a liquid. In the clinical laboratory the
solvent we measure most of the time is human plasma. The solute is
whatever the substance is we want to measure.
❑ Solute + Solvent = Solution

■ Solvent- The substance in which the solute is dissolved in

■ Solute- substance dissolved in a liquid

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Concentration
■ Amount of one substance relative to the amounts of the other substances in
the solution.

■ Concentration can be measured in many different units

❑ Percent Solutions:
■ % w/w- weight per weight
■ %v/v – volume per volume
■ % w/v – weight per volume (parts of solute / 100 totals parts )

❑ Molarity: Moles / Liter

❑ Molality: Moles / 1000 grams solvent

❑ Normality: equivalent weight/ liter

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Expressing Concentration:
Percent Solution

■ % w/w – percentage weight per weight

❑ Most accurate method of expressing concentration
❑ Can be cumbersome (especially with liquids), not often used in
clinical labs.

❑ % w/w = gram of solute OR gram of solute per 100.0 g of solution

100.0 g of solution

❑ How many grams of NaOH are needed to make a 25.0% w/w

solution using deionized water as the solvent?

25.0% w/w = X g of solute in 100 g of solution

X= 25.0 g NaOH

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Expressing Concentration:
Percent Solution
■ % w/v – percentage weight per volume
❑ Easiest & most commonly used
❑ Accurate if temperature controlled

■ %w/v= g of solute OR g of solute per 100.0 mL of solution

100 mL of solution

What is the %w/v of a solution that has 15.0 g of NaCl dissolved into a total
volume of 100 mL deionized water?

X% w/v = 15.0 g NaCl

100 mL of solution
X= 15.0 %
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Expressing Concentration:
Percent Solution
■ % v/v –percentage volume per volume
❑ Least accurate
❑ Used when both substances are liquids
❑ Note: volumes of liquids are not necessarily additive

❑ %v/v= mL of solute OR milliliter of solute per 100 mL of solution

100 mL of solution

■ How many milliliters of ethanol are needed to make a 75.0% v/v

solution using deionized water as the solvent?
75.0% v/v EtOH = X mL EtOH in 100 mL of solution
= 75.0 mL EtOH

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Expressing Concentration:
Molarity
■ Three components of Molarity
❑ Gram weight of solute
❑ Solute’s gram molecular weight
❑ Solvent quantity

■ Number of moles per one liter of solution

■ Mole = 6.022 X 1023 number of atoms or
molecules OR
■ Mole= Molecular weight in grams
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Determinig Molarity: First step

■ Molecular Weight
❑ Sum of the atomic weights of each element in the compound
❑ What is the molecular weight of Na3PO4?
■ Step 1: Sodium has an atomic weight of 22.99, but there are
3 molecules so 22.99*3= 68.97
■ Step 2: Phosphorus has an atomic weight of 30.97, and only 1
molecule, so 30.97 *1= 30.97
■ Step 3: Oxygen has an atomic weight of 16, but there are 4
molecules ,so 16*4= 64.00
■ Step 4: Add 68.97+ 30.97+ 64.00= 163.94 gram molecular
weight

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Determining Molarity: Next Step
❑ How many grams are contained in one mole of Na3PO4?
❑ Use the formula for mole calculations
Number grams of solute
Gram molecular weight of solute

1 mole Na3PO4 = X g Na3PO4

gram molecular weight(gmw)
X= 163.94 g Na3PO4

So, 163.94 grams of trisodium phosphate are contained in 1 mole of

trisodium phosphate or 6.022 X 1023 trisodium phosphate molecules
weigh 163.94 grams

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Determinig Molarity: Final Step
■ Molarity (M) = 1 mole of solute
1L of solution
We are asked to make a 1.00 L volume of a 0.100 molar solution of trisodium phosphate.
How many grams would we need?
M= grams
gmw
1.00 L of solution

0.100 molar= X grams of Na3PO4

163.94 gmw of Na3PO4

1.00 L of solution

(0.100M)(1.ooL) = X g

163.94 gmw

0.100= X
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163.94
Expressing Concentration:
Molality
■ Amount of solute per one kg of solvent
■ Expressed in terms of weight per weight or
moles per 1000 grams of solvent
■ Used to measure the physical properties of
solutions
■ Molality = 1 mole of solute
1 kg of solvent

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 Expressing Concentration:
Normality

■ Number of gram equivalent weights per liter

of solution
■ Requires gram equivalent weight and valence
■ Formula
❑ N= Number of grams of solute
Gram equivalent weight of solute
1.00L of solution

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Expressing Concentration:
Normality
❑ First Step
❑ Gram equivalent Weight / Liter
■ Gram equivalent weight is equal to the gram molecular
weight of a substance divided by its valence
■ Valence = the electrical charge of an ion, or the number
of moles that react with 1 Mole H+

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Determining Equivalent Weights

■ Example

▪ The MW of calcium = 40 grams

▪ Calcium ions carry a +2 electrical charge ( valence = 2 )
▪ Equivalent Weight of calcium = 40 / 2 = 20 gram equivalent
weight

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Solution Properties
■ Titration – Method of measuring concentration of one solution by comparing
it with a measured volume of a solution whose concentration is known

■ General formula: when you have a volume and concentration of one,

and either the volume or the concentration of the other: V1 C1 = V2 C2

❑ For Example:
How many mls of 1.0 N HCl is required to prepare 25 mls of 0.5 N HCl ?
( 1.0 N ) ( ? mls ) = ( 0.5 N ) ( 25 mls)
? mls = 12.5 mls

You would need to add 12.5 mls of 1.0 N HCl to 12.5 mls of deionized water
( a total volume of 25 mls) to prepare 25 mls of 0.5 N HCl

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Solution Properties
■ Density – An expression in terms (usually) of
a mass per unit of volume

❑ Many examples - including specific gravity,

osmolality

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pH and Buffers

■ Buffers resist change in acidity

■ Buffers are usually weak acids ( or bases) and their salts

■ pH is the unit used to measure acidity ( Hydrogen ion concentration )

■ “p” = “negative log” of the concentration of a substance in solution.
■ Example: pH = - log [H+]

■ The Hydrogen ion concentration of deionized H2O is 1 x 10-7 M

■ The negative log of 10-7 = 7. The pH of H2O is 7.0

■ The pH scale ranges from 0 - 14

■ pH 7 = neutral
■ pH > 7 = alkaline (basic)
■ pH < 7 = acid

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Temperature
■ Measurement of temperature is an important component of
the clinical lab. Instruments, refrigerators and incubators are
required to operate within specific temperatures that must be
maintained and monitored daily.
❑ Examples
■ Heat blocks, water baths, and incubators shall be
maintained at least +/- 1 degree C. of the desired
temperature
■ Refrigerators shall be maintained at 2 -8 degrees C.

■ Each laboratory must have a NIST calibrated thermometer in

order to ensure the accuracy of other thermometers in the
laboratory

■ Out-of-range temperatures should be addressed asap

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Temperature
■ Scientific measurement of temperature is always expressed in the
Celsius ( C) scale , not Fahrenheit ( F )

❑ Celsius scale: 0 degrees = freezing point of water

100 degrees = boiling point of water

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Conversion: Temperature
■ Conversion of Celsius to Fahrenheit and Fahrenheit to
Celsius
❑ F° = ( C ° x 1.8 ) + 32

❑ C° = ( F ° - 32 )
1.8
■ For example:
❑ Your refrigerator at home is probably around 40 ° F. What is that in
Celsius?
■ Celsius= 40-32 = 4.4
1.8

❑ Water boils at 100 ° C. What is that expressed in Fahrenheit?

■ (1.8)(100) +32 = 212

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Examples of conversions
How many mls are there in 2.5 liters?

The question you have to ask yourself is, what is the relationship between
liters and mls? The answer : 1 liter = 1000 ml
But now what?

We want to get rid of the “liters’ units and end up with “mls” … Right ?

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1.25 liters = _____ mls ? Remember, write a fraction that does two things:

1. Equals 1
2. Gets rid of unwanted units and / or adds needed units

100 mg = _________ ug ?

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Dilutions
■ A ratio of the concentrate to the total (final) volume.
❑ A 1:4 dilution has a 1 volume of sample and 3 volumes of diluent
mixed together.
■ Any volume can be used to create this dilution, but it must be the
same unit of volume
■ Keep in mind the sample size when making your dilution
❑ For example: a 2:3 dilution could contain:
■ 2 mL serum: 1 mL pure water
■ 20 µL of serum: 10 µL of pure water
■ 0.2 mL of serum: 0.1 mL of pure water

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Dilutions for the Clinical Laboratory

Example:

A technician performed a laboratory analysis of

patient’s serum for a serum glucose determination.
The patient’s serum glucose was too high to read on
the glucose instrument.
The technician diluted the patient’s serum 1:2 and
reran the diluted specimen, obtaining a result of 210
g/dl. To correct for the dilution, it is necessary to
multiply the result by the dilution factor (in this case
x 2).
The final result is 210 g/dl x 2 = 420 g/dl.
Examples of dilutions and dilution factors

Parts Parts Total Dilution Dilution

Specimen Diluent Volume Factor

1.0 1.0 2.0 1:2 2

1.0 2.0 3.0 1:3 3

1.0 3.0 4.0 1:4 4

1.0 9.0 10.0 1 : 10 10

0.5 4.5 5.0 1 : 10 10

0.2 1.8 2.0 1 : 10 10

0.2 9.8 10.0 1 : 50 50

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Serial Dilutions
■ In these types of questions, you are given a series of tubes.
■ Each tube having a measured amount of a diluent.
■ You are instructed to add a specified amount of specimen into the first
tube, mix well and transfer a specified amount of the mixture to the next
tube, etc.

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Serial Dilutions
■ Example:
❑ 6 tubes, each with 0.5 mL DI water
❑ Add 0.2 mL serum to first tube and serially dilute
❑ Find the dilution in tube # 6

■ Find the dilution factor (will be the same in each of these tubes)

■ 1/dil factor x 1/dil factor x 1/dil factor (etc. 6 times)

■ Result multiplying the numerator 1x1x1x1x1x1x1x = 1
■ Multiplying the denominators

■ Will give the result as 1 / 1838

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 Resources

■ Serial dilution
❑ http://tinyurl.com/cw7e3ok

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References
■ Bishop, M., Fody, E., & Schoeff, l. (2010). Clinical Chemistry:
Techniques, principles, Correlations. Baltimore: Wolters
Kluwer Lippincott Williams & Wilkins.
■ Doucette, L. (2011). Mathematics for the Clinical Laboratory
(2nd ed.). Maryland Heights, MO: Saunders.
■ Sunheimer, R., & Graves, L. (2010). Clinical Laboratory
Chemistry. Upper Saddle River: Pearson .

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