CHAP TER 4

BASIC REQUIREMENTS
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PART 1

SAMPLING
Sampling is the most critical aspect of an analysis.
The significance and accuracy of the measurements
can be limited by the sampling process.

SAMPLING

The ease or complexity of sampling will depend on

the nature of the sample.

Is a process to get
homogeneous sample.

Representative means that content of analytical

sample reflects content of bulk sample.

Homogeneous means that the analytical sample has

the same content throughout.

representative

and

CLASSIFICATION OF ANALYSIS
Method

Sample Weight (mg)

Sample Volume (L)

i) Meso

>100

>100

ii) Semimicro

10 100

50 - 100

iii) Micro

1 10

< 50

iv) Ultramicro

<1

Classification of the constituents in a sample

i)
Major
>1%

ii)

Minor

0.1 1 %

iii)

Trace

< 0.1 %

iv)

Ultratrace in the range of few parts per million or less.

SAMPLING
Deciding

how to obtain a sample for analysis

depend on:

1)

The size of the bulk to be sampled.

2)

The physical state of the fraction to be analyzed

(solid, liquid, gas)

3)

The chemistry of the material to be assayed.

(Nothing

can be done that would destroy or alter the

identity or quantity of the analyte)

Obtaining a representative sample is the first step of an analysis.

The gross sample is several small portions of the
sample. This is reduced to provide a laboratory

sample.
An aliquot of this sample is taken for the analysis sample.

In the clinical laboratory, gross

sample is usually satisfactory for
use as a sample because it is not
large and homogeneous
(blood and urine)

STEPS INVOLVED IN SAMPLING BULK

MATERIAL
Identify the population from which the
sample is to be obtained.

Collect a gross sample that is truly

representative of the population being
sampled.
Reduce the gross sample to a laboratory
sample that is suitable for analysis.
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SAMPLING SOLID
Inhomogeneity of the material, make sampling of
solids more difficult.

The easiest way to sample a material is:

GRAB SAMPLE: the sample taken at random and
assumed to be representative.
For reliable results, it is best to take 1/50 to 1/100 of
the total bulk. The larger the particle size, the larger
the gross sample should be.

The gross sample must be reduced in size to obtain a

laboratory sample.

SAMPLING SOLID
CONING AND QUARTERING
This process is continued until the gross sample is
small enough to be transported to the laboratory.

SAMPLING SOLID
CONING AND QUARTERING
There is no specific technique that can be used for taking the
samples. Using an example, explain how to sample either a solid,
liquid, or gas sample. (5 marks)

Sampling solid
Using the method cone and quarter.
Divide a pile of material into quarter.
Take a sample from each quarter of the pile and crush these
sample and form into a smaller conical pile.
Flatten the conical pile and cut into equal quarters. Two
opposite quarters are chosen at random.
Crush the quarter further.
The whole steps are repeated until a laboratory samples obtain.
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SAMPLING SOLID
TABLET SAMPLER
Designed to take a sample of
tablets and capsules as
containers are filled.

SCOOPER
A stainless steel scoop
with a capacity of 5 oz,
148 ml. Perfect for
sampling powder, soil,
gravel, and other
materials.
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SAMPLING LIQUID
Liquid

samples are homogeneous and are much easier

to sample.

The gross sample

If

can be relatively small.

liquid samples are not homogeneous, and have only

small quantity, they can be shaken and sampled
immediately.

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SAMPLING LIQUID
Sampling depends on the types of liquids:

LARGE VOLUME OF LIQUIDS (IMPOSSIBLE TO MIX)

- If in a pipe, after passing through a pump when they
have undergone the most thorough mixing

LARGE STATIONARY LIQUIDS (LAKES, RIVERS)

- thief sampler, which is a device for obtaining
aliquots at different levels

BIOLOGICAL FLUIDS
The timing of sampling biological fluids is very
important (blood composition varies before and after
meals)
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Aliquot: sample or a portion of the total amount of a solution

SAMPLING LIQUID

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SAMPLING GASES
Tend to be homogeneous.
Large volume of samples is required because of their
low density.

Air analysis: Use a `Hi-Vol sampler that is containing

filters to collect particulates.
Liquid displacement method: The sample must has
little solubility in the liquid and DOES NOT REACT with
the liquid

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Breath sample: The subject could blow into evacuated

bag.

SAMPLING GASES

Hi-Vol sampler

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SAMPLE STORAGE & PRESERVATION

1.

Samples storage purpose:

There is a time gap between when the sample is taken and

the actual analysis is being carried out.
2.

For LIQUIDS samples, make sure that it is kept in bottles

with stoppers.

3.

ACIDIC LIQUID samples can be stored in glass container.

Whereas BASIC LIQUID samples in plastic container.
SOLID SAMPLES is easier to keep and have less chance to
be adulterated by foreign matters. Sometimes it can also
get absorbed or adsorbed to the wall of the container.

4.
5.

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SAMPLE STORAGE & PRESERVATION

An important aspect of the sampling process
Samples are preserved to prevent from:
Decomposition

Precipitation
Loss

Loss

of metals from water samples

of water from hygroscopic material

of volatile analytes from water samples (volatile

organic compounds)

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SAMPLE STORAGE & PRESERVATION

Preparing a laboratory sample

Converting the sample to a useful form:
Solids

are usually ground to a suitable particulate

size to get a homogeneous sample.

Dry

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the samples to get rid of absorption water.

Purpose of drying solid samples :

To ensure that the exact weight is obtained during
the QUANTITATIVE chemical analysis.
How it is done ?:
Solid samples dried in oven at 105 - 110oC for
1-2 hours.
Plant and tissue samples dried by heating
Problems associated with drying of samples :
1. Samples might decompose at high temperature.
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2. Some samples are sensitive to heat, therefore drying can

be carried out in a desiccators.

DEFINING:
REPLICATE SAMPLES
Replicate

samples are always performed unless the

quantity of the analyte, expense or other factors
prohibit.

Replicate

samples are portion of a material of

approximately the same size that is carried through an
analytical procedure at the same time and the same
way.

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PREPARING SOLUTIONS
OF THE SAMPLE
A solvent is chosen that dissolves the whole
sample without decomposing the analytes
SOURCES OF ERROR
i) Incomplete dissolution of the analyte.
ii)

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Losses of analyte by the volatilization.

iii) Introduction of analyte as a

contamination.
iv) Contamination from the reaction of the solvent
with vessel walls.

SAMPLE PREPARATION &

DISSOLUTION METHODS
SAMPLE DISSOLUTION is the digestion or
mineralization of a sample to render it soluble and to
destroy organic matter that may interfere with the
recovery of the analyte.

Sample dissolution procedures can be divided into

1. Dry ashing
2. Oxidative ashing
3. Wet digestion
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1. DRY ASHING
The sample is slowly combusted at a high
temperature (400 700oC) in a muffle furnace.
Atmospheric O2 serves as the oxidant, that is
organic matter is burned off, leaving behind
inorganic residue that is soluble in dilute acid.
Oxidizing aids may be employed.

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1. DRY ASHING
SIMPLE DRY ASHING
no chemical aids.
Pb, Zn, Co, Cr, Mo, Sr, Fe traces can be recovered with little loss by
retention and volatilization.

Usually a porcelain crucible can be used.

Example: Lead is volatilized at temperature more than 500 oC,
especially if chlorine is present (blood and urine samples).
- Pt crucible are preferred for lead for minimal retention losses.
-If an oxidizing material (Mg(NO3)2) is added to sample, the
ashing efficiency is enhanced.

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1. DRY ASHING
IF THE SAMPLE ARE LIQUIDS AND WET TISSUES:

The sample are dried on a stream bath or by gentle heat

before they are placed in a muffle furnace.

The heat from the furnace should be applied GRADUALLY

UP to full temperature to prevent rapid combustion and
foaming.

After dry ashing is complete, the residue is usually leached

from the vessel with 1 or 2 mL concentrated or 6 M HCl and
transfer to a flask or beaker for further treatment.
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DRY
ASHING
ADVANTAGES

Simpilicity
Free from
contaminations since
few or no reagents are
added

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DISADVANTAGES
Volatilization of elements

and
losses by retention on the walls
of the vessels.
Adsorbed metals on the
vessel may in turn
contaminate future samples.
2-4 hour are needed for dry
ashing

2. WET DIGESTION

A method for the decomposition of an organic

material, such as resins or fibers, into an ASH by
treatment with a boiling oxidizing acid or mixture of
acids.
- The acids oxidize organic matter to CO2, H2O and
other volatile products, which are driven off, leaving
behind salts or acids of the inorganic constituents.

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2. WET DIGESTION
Principle:

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1. Usually use combination of ACIDS to achieve a complete

dissolution
2. Mixture of HNO3 and H2SO4 is often used
3. A small amount (5 mL) of H2SO4 is used with larger volumes
of HNO3 (20 to 30 mL).
4. Usually performed in a Kjeldahl flask.
5. HNO3 destroys the bulk of organic matter, but it does not get
hot enough to destroy the last traces.
6. It is boiled off during the digestion process until only H2SO4
remains and dense, white SO3 fumes are evolved and begin
to reflux in the flask.
7. At this point, the solution gets very hot, H2SO4 acts on the
remaining organic material.

2. WET DIGESTION
Principle:
8. If the organic matter persists, more HNO3 may be
added.
9. Digestion is continued until the solution clears.
10. All digestion procedures must be performed in a
FUME HOOD.

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WET
DIGESTION

ADVANTAGES

Superior in term of
RAPIDITY
Freedom from loss by
retention

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DISADVANTAGES

Introduction of
impurities from the
reagent necessary for
the reaction.

ELIMINATING
INTERFERENCES
Interferences are substances that prevent
direct measurement of the analyte and MUST BE
REMOVED.
May included separation steps:

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i)

Precipitation

ii)

Chromatography

iii)

Distillation

iv)

Dialysis

v)

Extraction into an immiscible solvent