Unit -2

Sample preparation for analysis

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 2.1. Introduction
 Pharmaceutical analysis is a science which deals with identification and/or
quantification of drug in raw-materials and dosage forms.
In order to identify and quantify the drugs that is contained in a product, the
drug may need to be separated from its matrix.
Some times components which can interfere with the analysis, must be
removed.
To do this majority of samples require a specific pretreatment.

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 2.2. Steps in chemical analysis
 The substance can be a solid, a liquid, a gas, or a biological material.
 An analysis involves several steps and operations depend on:
• the particular problem
• your skill and expertise
• the apparatus or equipment.

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Fig. 1.1. Steps in an analysis
I. Plan the Analysis
A. Define the problem
B. Gather Analytical Information and Choose the best suited techniques
Factors need to be considered in method selection process are:
1. Sample type and size required
2. Accuracy required
3. Sample preparation needed
4. Cost of analysis (total investment)
5. Number of sample to be analyzed
6. Time required for analysis
7. Skill required
8. Complexity of the sample
9. Number of components in the sample 5
II. Sampling
•
Snowball Sampling
To produce meaningful information, an analysis must be performed on a
representative sample.
• Sometimes proper sampling is the most difficult step in an analysis and
the source of greatest error.
Types of sampling techniques
• Random sampling: d/t fractions are represented by chance.
• Systematic sampling: taking increments at predetermined intervals.
• Stratified sampling: is a random sampling from each group.
• Convenience sampling: selecting samples on the basis of availability.

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III. Sample preservation
 It is an important step, because there is usually a delay between sample
collection and analysis.
 It ensures that the sample retains its physical and chemical characteristics
so that the analysis truly represents the object under study

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 2.3. Preliminary Treatment of Sample
(sample preparation)
 Sample preparation is a process required for the transformation of a sample to
make it amenable for chemical analysis or to improve the analysis.
This is because the samples cannot be analyzed directly without some form of
pretreatment in many analytical measurements
Sample preparation is an essential step in analysis because it greatly
influence the measurement:
−Reliability, -Accuracy and -Cost
This step may be the most time consuming in analysis and affects significantly
the analytical information. 8
The sample preparation depends on the analytical techniques to be employed
and their capabilities.
are three key issues that necessitate pre-treatment of a sample:
1. the sample is in the wrong physical state for the analysis method (e.g.,
the method requires a liquid but you have a solid sample),
2. the sample has interfering matrix components that may give either a
false positive or negative reading in the measurement, and
3. the sample has too low analyte concentration to be detected by the
instrument.
Methods like extraction, distillation or precipitation can be applied for this
purpose.
 Extraction
Is the most common methods to separate active ingredient from the matrix.
There are different types of extractions:
Solid– liquid extraction
The sample is placed in a vessel, and the solvent is added to dissolve the
sample components.
The container is shaken for a given period of time, and then the insoluble
components are removed by filtering or centrifugation.
This process works well if the analytes are readily soluble in the solvent and
not tightly bound to the matrix.
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Solution– solvent extraction
A two-phase liquid–liquid extraction system is used.
 Separate layers will be formed after shaking the components
In general more efficient extraction is achieved with several extractions than
with a single utilizing the same total volume.

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 Example: in analyzing phytochemical constituents of a plant, processes within
sample preparation include:
 Cutting and drying the plant part needed

Homogeniza Clean-up
tion by Size (isolation
Extraction Concentration Analysis
reduction and
(grinding purification)
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 Distillation
Distillation is the process by which a liquid is heated to create a vapor and
then condensed back into a liquid again.
It is used to separate a components of a mixture based on different boiling
points.
Precipitation
Chemical precipitation is the process of conversion of a material dissolved in
solvent (solution) into solid by converting the substance into insoluble form
or by making the solution a super saturated one.
 Analysis
 Measurement of different parameters of the analyte using selected
method.
 Calculating the results: Analyte concentrations from experimental data
need to be calculated.
 Estimate reliability of results: Some measure of the uncertainties
associated with computed results need to be provided.
 Compiling data
− All existing data have to properly compiled and maintained

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 Types of Chemical Analysis
Four basic types with respect to the information which is furnished
1. Proximate Analysis—the amount of each component in a sample is
determined with no concern as to the actual compounds present
2. Partial analysis—determination of selected components in a sample
3. Trace constituent Analysis—determination of specified components
present in very minute quantity (less than 1ppm)
4. Complete Analysis—proportion of each component of the sample is
determined
 Areas of Chemical Analysis and Questions They Answer
Quantitation:
− How much of substance X is in the sample?

Detection:
− Does the sample contain substance X?

Identification:
− What is the identity of the substance in the sample?

Separation:
− How can the species of interest be separated from the sample matrix for better
quantitation and identification?