Chapter 4 Reagents
Chapter 4 Reagents
Chapter 4 Reagents
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Acknowledgment
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Objective.
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Out line
4.1. Definitions
4.2. Types of solution
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4 : Reagents (solution) used in
clinical laboratories
4.1. Definitions:
Solution
A solution: - is a homogenous mixture of two or more
components that can be varied in composition within
certain limit.
Every solution consists of two parts, the solvent
and solute
Solvent: - is the substance that dissolves the solute.
The solvent in most cases are liquid.
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Reagents (solution) ……………..
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Reagents (solution) ……………..
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4.2. Types of solution
1.Solid solution: - are mixtures of two or more solids and they are
dispersed or mixed randomly throughout one another resulting a
heterogeneous mixture. Eg. Alloys
2.Gaseous solution :-is a homogeneous mixture of two or more gases
E.g. Air is a solution of O2, N2, H2, CO2 & other gases.
3. Liquid solution: - is a solution where the solvent liquid & solute are
also liquid but it can be solid or gas.
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4.3 Expression of the strength of solution
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Expression of the strength of …….
c. Concentrated solution
Large amount of solute dissolved in a small amount of solvents
Example, 98% H2SO4.
c. Saturated
It is one in which a given volume of solvent has dissolved
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2. Quantitative expressions of
concentration of solution
The concentration of solutes in a solution can be
expressed quantitatively in physical units or chemical
units.
A. Physical unit:
Percentages
Density
Specific gravity
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Quantitative expressions…..
1. Percentage
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Quantitative expressions…..
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Exercises -1
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Quantitative expressions…..
2. Parts per unit.
This is to express the number of parts by volume or by
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Quantitative expressions…..
B. Chemical units.
Commonly used chemical unit to express the quantity of solutes in a
given solution are:-
Mole
Equivalents
Molarity
Normality
Molality
1. Mole.
1 mole of a give substance is its atomic weight or molecular weight
expressed in gram.
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Quantitative expressions…..
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Quantitative expressions…..
Solution
1 mole C6H12O6= its molecular weight
1mole C6H12O6 = 180gm
2mole = ?
180gm x 2moles = 360gm
1 mole
* 360 gm glucose is dissolved in one liter of solution to give
2 moles of glucose.
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Quantitative expressions…..
Exercises
How can you prepare 1.5 mole of HCL in litter?
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Quantitative expressions…..
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Quantitative expressions…..
Solution
Actual mass = 20gm
Molecular wt = 40gm
Volume = 0.2 lit (200ml)
M =Actual Mass
Mol.wt x vol in liter
M= 20gm
40x 0.2 = 2.5M Na OH
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Quantitative expressions…..
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Quantitative expressions…..
Example-1 what is the normality of KOH. If 5.6gm KOH is
dissolved in 1 liter
of solution?
Solution
Actual mass = 5.6gm
Volume = 1 lit
N= Actual Mass in gram
Equ. Wt x vol in lit
= 5.6gm = 0.1NKOH
56 x 1
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Quantitative expressions…..
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Quantitative expressions…..
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Quantitative expressions…..
Assume 1 L of solution:
5.86 moles ethanol = 270 g ethanol
927 g of solution (1000 mL x 0.927 g/mL)
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Dilution of solution
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Dilution …
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Dilution …
Example: to make 45 ml of 30% solution from 70% solution
C2= 30%
V2= 45ml
C1= 70%
V1= C2 x V2
C1
V1= 30 x 45 = 19.3ml
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There fore 19.3ml of 70% solution must be diluted with
25.7ml of distilled to obtain 45ml of 30%solution.
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Methods of dilution
A. Simple dilution
A general process of preparing less concentration solutions from
a solution of greater concentration.
E.g.,
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Methods of dilution …
B. Serial dilution
It is defined as multiple progressive dilutions ranging
from more concentrated solutions to less concentrated
solutions.
Example ???
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Dilution…..
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4.4. Reagent solutions
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Reagent …
Reagent
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4.4.1. Types of reagent solutions
1. Stock reagent solution
Is a concentrated reagent solution which is diluted to
prepare a working solution.
Has a longer shelf life and occupies less space in
storage than the working solution.
2. Working –reagent solution
Can be diluted from the stock solution or prepared
directly from the reagent chemical following the
recommended procedures.
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Types of reagent …
3. Saturated solution
Is a solution in which further solute can not be
dissolved
A saturated solution must have the deposits of the
solute at the bottom of the reagent bottle.
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4.4.2. REAGENT HANDLING
1. Personal safety precautions
Assume all chemicals are potentially harmful to a certain degree.
Before picking
After picking
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Reagent …
container
Always add a reagent slowly and observe the reaction
Always pour more concentrated solutions slowly into
water or less concentrated solutions
3. Labeling of reagents and solutions
Every reagents and solution in the laboratory should be
labled appropriately .
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4.5. Grades of chemicals
Chemicals are produced in various purities or grades.
Some of these includes:
1. Reagent grade or Analytical grade(AR) grade
Are of a high degree of purity.
Often used in the preparation of reagents in
clinical laboratory.
2. American chemical society(ACS)
Are reagent grade chemicals that meet the
standards of ACS.
Labeled as ACS
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Grade of …
3. Chemically pure grade(CP) grade:
Sufficient for different analysis in the clinical laboratory
Since their designation does not reveal the limits of
available.
4. USP and NF grade
Are reagents that meet the specifications of united states
pharmacopeia(USP) or the National formulary(NF)
Are less pure than the CP grade.
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Grade of …
5.Purified, practical or pure grade
Are not generally used in the clinical laboratory but
may be used as starting materials for synthesis of
other chemicals of better purity.
6.Technical or Commercial grade
Are used only for industrial purposes and are not
generally used in the preparation of reagents for the
clinical laboratory.
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4.6. Standard solutions
These are solutions in which the concentration of a given
chemical is precisely known and are used to determine the
value of an identical chemical with unknown concentration of
a given solution.
Chemicals that are used to prepare these solutions should be
of analytical grade.
Since poor standard solutions cause errors in the estimation
of the intended substances.
their accurate preparation is of paramount importance in
order to obtain accurate and precise laboratory findings in
medical laboratories.
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Standard……
highest purity and can be used directly for the exact measurement
of substances of unknown concentration in a given solution.
Primary standard solution should be made of substances that are:
Free of impurities,
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Standard……
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Standard….
In the preparation of secondary standard solutions the
following points should be taken into consideration:
Using analytical balance for weighing.
exact measurements.
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4.7. Staining solutions
Staining solutions are solutions that contain colored
dyes.
These solutions can contain basic, acidic or neutral
dyes.
Different stains are used in medical laboratories to give
an artificial color for the substances to be identified from
a given biological specimen (whole blood, body fluids,
urine, etc.).
The substances may be identified by their characteristic
reaction with the staining solutions.
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Staining …
1. Basic stains
2. Acidic stains
3. Neutral stains
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Staining …
1. Basic Stains
Are stains in which the colouring substance is contained
in the base part of the stain and the acidic part is
colourless.
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Staining …
2. Acidic Stains
Are stains in which the colouring substance is contained
in the acidic part of the stain and the base part is colourless. E.g.
eosin.
3. Neutral Stains
Are stains in which the acidic and basic components of stains
are coloured.
Neutral dyes stain both nucleic acid and cytoplasm. e.g
Giemsa’s stain, Wright’s stain.
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Staining………
Different types of blood cells, bacteria, parasites, and
tissues together with their cellular elements can be
stained by using appropriate types of stains
(differential stains) such as Giemsa stain, Wright
stain, Gram stain, Leishman stain, Acid Fast Stain,
etc.
Simple stains are used to reveal the morphology
(shape, size and content) of an organism(s) and
single dye is utilized for the procedure.
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Summary questions
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References
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Next chapter will be on : STERILIZATION AND
DISINFECTION
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