Instrumental

Methods of Analysis

Danni Ramdhani

 Chapter 1: Drug registration and assessment

1.1. Summary of particulars
1.2. Chemical and pharmaceutical documentation
1.3. Reports of Experimental and biological studies
1.4. Report on Clinical Studies
1.5. Company profile
Chapter 2: The Analytical Problem

2.1.
2.2.
2.3.
2.4.
2.5.

Sampling/Sample Handling
Experimental errors
Choice of methods of an analysis
Statistics of data analysis
Validation of analytical procedure

 Chapter 3: Drug stability and degradation product

3.1. Chemical purity and its control

3.2. International pharmacopoeia and official monograph

3.3. Radiopharmaceutical
3.4. Stability indicating assay

Chapter 4: Function group analysis

4.1. Classical analysis

4.2. Instrumental analysis

4.2.1. Spectral methods

4.2.2. Electro analytical methods

Chapter 5: Automation in pharmaceutical analysis

5.1. Flow injection analysis

5.2. HPLC

5.3. Gas chromatography.

5.4. Mass spectroscopy

 Chapter

6: Determination of active ingredients in

different dosage forms and in biological fluids

6.1. Determination of active ingredients in tablets

6.2. Determination of active ingredients in capsules
.
6.3. Determination of active ingredients in semisolid
6.4. Determination of active ingredients in eye drops
6 5. Determination of active ingredients in injection
6.6. Determination of active ingredients in Suppositories.

6.7. Determination of active ingredients in aerosols

inhalation

6.8. Radioimmunoassay
Chapter 7: Quality assurance of pharmaceuticals

7.1. Good Manufacturing Practice G.M.P

7.2. National and international organization

7.3. Departments in pharmaceutical companies

7.4. ISO and BSI

Instrumental Methods of
Analysis
Goal detection of analyte concentration and/or
structure
Method design an instrument that furnishes this
information based upon some particular response
of the analyte.
USP Categories :
Category 1: Quantitation of major components or
active

ingredients

Category 2: Determination of impurities or

degradation products
Category 3: Determination of performance
characteristics

Stimuli for Various Instrumental

Methods

Light

Optics

Electricity

Fragmentation
Equilibrium

Common Instrumental
Methods

Optical Spectroscopic Techniques

Ultraviolet and Visible (UV-Vis) Spectroscopy
Fluorescence and Phosphorescence

Atomic Spectroscopy - both emission (ICP-AES) and absorption (AAS)

Xray Spectroscopy
Nuclear Magnetic Resonance (NMR) Spectroscopy

Chromatographic Techniques
Gas Chromatography (GC)
High Performance Liquid Chromatography (HPLC)
Capillary Electrophoresis

Electrochemical Techniques
Voltammetry
Polarography
Coulometry

Miscellaneous Techniques
Mass Spectrometry

Accuracy vs. Precision

Accuracy
How well a
measurement
agrees with an
accepted value

Precision
How well a series
of measurements
agree with each
other

Accuracy vs. Precision

Examples of Precision and Accuracy:

Low Accuracy
High Precision

High
Accuracy
Low Precision

High Accuracy
High Precision

Hasil dikatakan tepat apabila penyebaran (dispersi) hasil

dalam satu seri penetapan kecil.

Ukuran Ketepatan
Kisaran (Range).
Deviasi rata-rata (mean deviation) = d = MD

Standar Deviasi = s = SD

Standar Deviasi Relatif = RSD = CV (dalam %)

Systematic vs Random
Errors
Systematic
Error
Avoidable error
due to
controllable
variables in a
measurement.

Random
Errors
Unavoidable
errors that are
always present in
any
measurement.
Impossible to
eliminate

 Experimental Errors

Random (in determinant) and Systematic

(determinant) errors

The term error as used here refers to the numerical

difference between a measured value and the true value.

For example, the % composition of a standard sample
certified by the national Bureau of standards may be
treated as correct in evaluating a new analytical method;
differences between the standard values and the results
obtained by the new method are then treated as error in
the latter.
One can attempt to minimize errors but cannot eliminate
them completely

Random errors
small fluctuations introduced in nearly all analyses. Arise
from variation of external conditions over which the
observer has no control
For example, repeated
measurements of the same property often differ even if
they are performed on a single instrument that is calibrated
and operated properly.

 Systematic errors
Systematic errors cause the results to vary from the correct value
in a predictable manner and can often be identified and corrected.
An example of a systematic error is improper calibration of an
instrument. The measurement lacks accuracy. It is even possible that
repeated measurements with this broken instrument will give
reproducible results (high precision), but every one of them will deviate
from the true value (low accuracy).
Systematic errors have been classified as personal, instrumental and

methodic

1. personal errors:

Are those due to the carelessness of the observer and include such
things as misreading a burette or overshooting the end point, or loss in
weight during washing and filtration, and mathematic error in
calculations.
2. Instrumental error:

Error due to the instruments themselves and include the use of

uncalibrated equipments e.g. Weights, burettes, pipette, flasks.
3. Methodic:

Errors are those inherent in the method itself, incomplete

Minimization of the errors

1. Calibration of the equipment and application of correction factor.
All of the equipments should be corrected and appropriate
correction factor applied.
2. Use of blank experiments
Carrying out a separate determination without the sample under
the same experimental condition to overcome the effect of
impurities introduced through reagents, solvents and vessels.
3. Standard addition method
A known amount of the constituent being determined (standard) is
added to the sample which is then analyzed for the total amount
of the constituent present. The difference between the analytical
results for the sample with and without the added standard gives
the recovery of the amount of added constituent.
4. Use of internal standard
It is used in HPLC and GC; fixed amount of the suitable standard
is added with different amount of the sample to minimize the error
during introduction of the sample.

 Choice of methods of an analysis

The pharmacist who has need for analytical data usually finds himself faced
with an array of methods which could be used to provide the desired information.
Unfortunately, there are no generally applicable rules that can be applied; the
choice of method is thus a matter of judgment. Such judgment is difficult, and the
ability to make it will come only with experience.

Factors plays an important role in the selection of the

analytical methods of analysis.

1.

Concentration range of the sample to be determined.

if for example the element present to the extent of a few ppm
as impurities or degradation products it is not suitable to use volumetric
or gravimetric methods and use other sensitive methods as HPLC or
spectrophotometry. On other hand if the analyte is a major component
of the sample the classical method may be preferable.

2.

Degree of accuracy and precision required

The accuracy and precision required are of vital importance in the
choice of an analytical method and its performance, especially in small
and vital substances as hormones.

3.

Different components which are present in the sample

The chemical structure of these substances should be known because
they may interfere with the method, since the analysis is based on
reactions or properties share by several compounds, in this case prior
separation or selective method should be used.

4-Physical and chemical properties of the substance

The analyst should know the state of the substance at
ordinary conditions and whether losses by volatility, and
whether or not the sample is hygroscopic or efflorescent and
what sort of treatment is sufficient to decompose or
dissolve the sample without loss of the analyte.
5-Number of the sample to be analyzed
If there are many considerable times can be expended in
calibrating instruments, preparing reagents, assembling
equipments, the cost of these operations can be spread
over the large number of analyses.
On other hand, if a few samples are to be analyzed, a
longer and more tedious procedure involving a minimum of
these preparatory operations may actually prove to be the
wiser choice from the economic standpoint.
6-Availability of the instruments and equipments.
plays an important role in the selection of the analytical
method.