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# MLAB 2401:

Clinical Chemistry
Chapter 3: Basic Principles and Practice
of Clinical Chemistry, part 1

UNITS OF MEASURE

## Measurement requires a numerical value and a unit

SI units:
length ( meter )
mass ( gram )
quantity ( mole )
Volume ( liter )
Time ( second )

with them

deci (d)
10-1

centi (c)
10-2

milli (m)
10-3

micro ( )
10-6

nano (n)
10-9

pico (p)
10-12

femto (f)
10-15

tenth of a liter

weight or volume

mg / dL
g / dL
mEq / L
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Conversions

## Most conversions within the metric system occur in units of TEN

where changing a unit of measure to a higher or lower designation
requires moving the decimal one place either to the left or to the
right.

## When converting measures in either the high end of the scale

(example kilo to mega) or the low end of the scale (examples milli to
micro, micro to nano, etc.) the decimal must be moved three places
right or left as the prefix designations are assigned only to every
third unit in the extreme ends.

Example of a conversion

## 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 ?

1000 mls
2.5 Liter
2500 mls
1 Liter

## Remember, write a fraction that does two things:

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

1 0 0 0 m ls
1 . 2 5 L i t e r s 1 L i t e r 1 2 5 0 m l s

100 mg =

_________ ug ?

1000 ug
1 0 0 m g 1 m g 1 0 , 0 0 0 0 u g
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## Physiological Saline is used in Blood Banks and Hematology to prepare

Red Blood Cell suspensions.
Physiological Saline is usually listed as being 0.9 % NaCl
0.9 grams of NaCl is added to 100 mls deionized water to make
physiological saline
What is the Normality (N) of physiological saline?

0 .9 g r a m s N a C l 1 E q W t N a C l 1 0 0 0 m l s

0 .1 5 N
1 0 0 m ls w a te r 5 8 g r a m s 1 L ite r
Unwanted units cancel out
leaving EqWt / Liter = N

Fraction = 1

Fraction = 1

## Conversions are manipulations of the units not the values !!!

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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

Temperature

## Scientific measurement of temperature is always expressed in the Celsius ( C) scale ,

not Fahrenheit ( F )

## 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.

Each laboratory must have a NIST calibrated thermometer in order to ensure the
accuracy of other thermometers in the laboratory

Celsius scale:

## Conversion of Celsius to Fahrenheit and Fahrenheit to Celsius

F = ( C x 1.8 ) + 32

C = ( F - 32 )

1.8

Conversion: Temperature

## Conversion of Celsius to Fahrenheit and Fahrenheit to

Celsius

F = ( C x 1.8 ) + 32

C = ( F - 32 )

1.8

For example:

1.8

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

F a h r e n h e i t 1 .8 1 0 0 3 2 2 1 2
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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.

## Mixtures of substances the substances in a solution are not in

chemical combination with one another.

## Dispersed phase - the substance is dissolved (the solute)

The substance in which the solute is dissolved is the solvent.
Solute + Solvent = Solution

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Concentration

## Concentration refers to the amount of one substance relative to the

amounts of the other substances in the solution.
Expressing Concentration

Example : mg/dL

## % v/v percentage volume per volume

Most accurate method of expressing concentration, but can be cumbersome (especially with
liquids), not often used in clinical labs.
Example :mg/gm

Example : mL/L

## Note: volumes of liquids are not necessarily additive.

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Solution Properties

## % Solutions: w/w, v/v , w/v (parts of solute / 100 totals parts )

Note: liquids + liquids and solids + solids alters the total parts,
but solutes + solvents does not

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What is a Mole?

molecules

Molecular Weight

## The molecular weight( MW ) of hydrogen = 1.0

That means that 6.022 X 1023 hydrogen atoms weighs 1.0 gram

## 1 mole of water weighs 18 grams

That means that 6.022 X 1023 H2O molecules weigh 18.0 grams

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Solution Properties

/ Liter

## Working with normality, is most important when

dealing with acid or bases in neutralization reactions.

## Equivalent Weight = MW / Valence

Valence = the electrical charge of an ion, or the
number of moles that react with 1 Mole H+

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Equivalent Weight

## The valence is the electrical charge of the substance

1 Equivalent weight of any substance reacts with 1 Equivalent Weight
of hydrogen ions

Example

## The MW of calcium = 40 grams

Calcium ions carry a +2 electrical charge ( valence = 2 )
Equivalent Weight of calcium = 40 / 2 = 20 grams

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Solution Properties

Normality

N = M x valence
M = N / valence
M is always < N

Calculation tips

## Use ratio and proportion when NOT changing concentration.

For calculations changing concentrations (as in titrations), use:V1C1
= V2C2
Important to remember that you cannot make a solution more
concentrated.

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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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pH and Buffers

1
H

## 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 H 2O 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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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

## 2 mL serum: 1 mL pure water

20 L of serum: 10 L of pure water
0.2 mL of serum: 0.1 mL of pure water

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Dilutions

## Find the dilution factor:

0.1 mL serum
2.9 mL DI water
1.0 mL reagent A
1.0 mL reagent B
5.0 mL total volume

0.1 mL serum
5.0 mL total

=1
X

Dilution is 1/50

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Parts
Specimen

Parts
Diluent

Total
Volume

Dilution

Dilution
Factor

1.0

1.0

2.0

1:2

1.0

2.0

3.0

1:3

1.0

3.0

4.0

1:4

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 / 838

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Resources

http://www.youtube.com/watch?
v=ZqdU3VfQ_Tc

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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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Water Specifications

## Distillation removes most organic matter

Reverse osmosis
Filtration
Deionization ions removed

## Type I Purest Required for sensitive tests

Type II Acceptable for most uses
Type III OK for washing glassware

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## Water filtration system for

Automated chemistry analyzer.

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