ANALYSIS OF DRUG AND

EXCIPIENTS IN THE SOLID STATE

Chromatographic Technique
Gas Chromatophy (GC)

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Chromatographic Technique

• Chromatography is a technique of separating chemical species based on their polarities

or size.
• Chromatography can be performed to chemicals in either liquid or gas form.
• The technique involve two phases;
• Mobile phase (contains analyte)
• Stationary phase (contains adsorbent)
• The common chromatographic techniques;
• Thin layer chromatography (TLC)
• Column chromatography (CC)
• Gas-Liquid chromatography (GLC)
• High performance liquid chromatography (HPLC)
• Size exclusion chromatography (SEC)
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Gas Chromatography

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 Terminologies
• Analyte  mobile phase
• Analytical chromatography  Preparative chromatography
• Bonded phase  Retention time
• Chromatogram  Sample
• Chromatograph or aerograph  Solute
 Solvent
• Eluent
 Stationary phase
• Eluate
 Detector
• Effluent

https://en.wikipedia.org/wiki/Chromatography 4
 Definition of terms

• Chromatography is a laboratory technique for

the separation of a mixture.
• The mixture is dissolved in a fluid (gas or solvent) called
the mobile phase, which carries it through a system (a column, a
capillary tube, a plate, or a sheet) on which a material called
the stationary phase is fixed.
• The different constituents of the mixture have different
affinities for the stationary phase.

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 Definition of terms

• Chromatogram – the visual output of the

chromatograph. In the case of an optimal separation,
different peaks or patterns on the chromatogram
correspond to different components of the separated
mixture.
• Chromatograph or aerograph – an instrument that
enables a sophisticated separation, e.g. gas
chromatographic or liquid chromatographic separation.

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 Gas chromatography (GC)

GLC = Gas Liquid chromatography

GSC = Gas solid chromatography
Schematic Representation of GLC

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 Gas chromatography (GC)

• GC can be used for both qualitative separation

and quantitative analyses.
• The stationary phase for GC is usually an organic
polymer coated on the inside of the very long tube,
and a mobile phase is an inert gas , such as helium.

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 Question

What is the difference between

chromatogram and chromatograph?

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chromatogram

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example

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 Application of GC in
Pharmaceutical Analysis
• Accurate and precise analysis of official pharmaceutical
substances
i. Assay of Drugs,
ii. Determination of specific organic compounds as
impurities in official pharmaceutical substance,
iii. Determination of related substances in official drugs,
iv. Determination of water in drug, and
v. Determination of chloroform with head-space
chromatography.
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 The Key Advantages of GC

• It has high frequency of separation and even complex

mixtures may be adequately resolved into constituents.
• It has a very high degree of sensitivity in detection of
components i.e., only a few mg of sample is enough for
complete analysis.
• Speed of analysis is quite rapid.
• Gives reasonably good accuracy and precision.
• The technique is fairly suitable for routine analysis
because its operation and related calculations do not require
highly skilled personnel. 14
 GLC Samples

Liquid Samples :
• They are usually injected by hypodermic syringes
through a self-sealing silicon-rubber septum into
a preheated-metal-block flash evaporator.
• The sample is vapourized as a ‘plug’ and carried
right into the column by the respective carrier
gas.
• Amount of sample ranges between 1–10 μL.
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 GLC Samples

Liquid Samples : …
• The column size may range: long (60-100 m), medium
(25 – 30 m) and short (5 – 10 m)

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 GLC Samples

Solid Samples :
• These are either dissolved in volatile liquids (solvents) or
temporarily liquefied by exposure to infra-red heat.
• Solid phase samples can prove to be a difficult analysis
because of the sample preparation involved (extraction).
• The solid sample (or slurry type, viscous samples that do
not inject using a standard syringe) is loaded into a quartz
tube which is then inserted into the pyroprobe.
• The filament of the probe is heated up to 1400 oC at
injection. (e.g. Agilent 7890 GC)
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 Types of Column

• The stationary phase for GC is usually an organic

polymer coated on the inside of the very long tube

Packed Column Capillary column

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Fundamentals of GC Columns | Agilent

Video clip
 Types of Column
• Packed Column: Are Stainless steel or glass tube filled
with particulate packing material (an adsorbent material,
or a support material coated or impregnated with a solid
phase).
• Internal Diameter: 2 to 4 mm
• Length: 0.5 to 5 m (most commonly 2 m)
• Packing: Support material with 0.5 to 25 % liquid phase
(partition material) or no liquid phase (adsorbent material)
• Liquid Phase: Multiple types available

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Fundamentals of GC Columns | Agilent

Video clip
Type of Column

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 Type of Column…
• Capillary Column: A typical capillary column is a thin,
fused silica glass tube, lined with a liquid phase or
adsorbent material or having a chemical bonding layer.
Thin metal tubes are also sometimes used as capillary
columns.
• Internal Diameter: 0.1, 0.25, 0.32, 0.53 mm
• Length: 5 to 100 m (most commonly 30 m)
• Material: Fused silica glass
• Liquid Phase: Good separation but less variety than
packed columns
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Type of Column…

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 Type of Column

• Packed column
• Capillary column

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Fundamentals of GC Columns | Agilent

Video clip
Columns
WCOT = wall-coated open tubular
SCOT = Supported- coated open tubular
PLOT = Porous –layer open tubular 24
Packed Column Capillary column
Stationary phase is coated Stationary phase is coated
directly in the column with the inner wall of the
column

Applied for both GSC and GLC Applied only for GLC

Liquid phase is adsorbed onto Liquid stationary phase is

the surface of the beads in a thin immobilized on capillary
layer or onto the solid inner tubing walls
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packing
 Fused Silica, SiO2 Glass Properties

• Fused silica is a noncrystalline (glass) form of silicon

dioxide (quartz, sand).
• Typical of glasses, it lacks long range order in its
atomic structure.
• It’s highly cross linked three dimensional structure
gives rise to it’s high use temperature and low
thermal expansion coefficient.

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 Fused Silica, SiO2 Glass Properties

• Key Fused Silica Properties includes; Near zero

thermal expansion, Exceptionally good thermal
shock resistance, Very good chemical inertness

• Typical Fused Silica Uses; High temperature lamp

envelopes, Temperature insensitive optical
component supports, and other.

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• Type and thickness of solid phase
• Type and linear velocity of carrier gas
• Column internal diameter
• Column length
• Oven temperature

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Column internal diameter and length

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Packed Column Capillary column
✓Produce broad peaks ✓Produce sharp and taller peaks
✓It has low detection sensitivities ✓It has high detection of lower
concentrations

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 Assessment of the peak area

✓ The time (retention time) until the injected sample

reaches the detector is a characteristic value of each
component.
=> determine type of component (qualitative analysis).

✓ The size of the component peak : Area and height

=> determine how much of the component (quantitative
analysis).

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Features of peaks in a
chromatogram

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 Assessment of the peak area
quantitative analysis

qualitative analysis
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 Column
Polarity Length Internal Coating
diameter thickness
Column selection
General guide to column selection

Non/weak
Strong

Thick
Long

short

thick
Thin

thin
Analysis of non-polar compounds 
Analysis of polar compounds 
Compounds with large difference in boiling 
points (b.p)

Isomers /cpds with little difference in b.p 

High resolution separation  
Shorter analysis time   
Analysis of low b.p compounds  
Analysis of high b.p compounds   36
 WORKING TECHNIQUE FOR
GLC
The three working techniques that provides optimum
accuracy and precision for the quantitative
analysis of pharmaceutical substances:
(i) Area Normalization,
(ii) Internal Standard Method, and
(iii) Comparison Method.
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Feature of peaks in a chromatogram

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 Area normalization

Where, A = Peak area; f = Response factor, defined as the ratio between the
concentration of a compound being analysed and the response of the detector to that
compound.
A chromatogram will show a response from a detector as a peak.
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f = Peak area (A) /conc.
 How to calculate peak area, A
Consider the following:
• The area of a peak is proportional to amount of the
compound that is present.

• The area can be approximated by treating the peak as a

triangle.

• The area of a triangle is calculated by multiplying the height

of the peak times its width at half height.

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How to calculate peak area, A
Rel. amount in %

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 Determine the % ge
composition of mixture

Total Area = Area A + Area B

𝐴𝑟𝑒𝑎 𝑜𝑓 𝐴
% 𝐶𝑜𝑚𝑝𝑛. 𝑜𝑓 𝐴 = × 100
𝐴𝑟𝑒𝑎 𝐴 + 𝐴𝑟𝑒𝑎 𝐵
Component A = 22.7%
Component B = 77.3%
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Determine the percentage of component y from the
following GC chromatogram: Show calculations.

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 Salient Features of Area
Normalization Method
(i) Very suitable for routine type of samples where
the variations in composition are only marginal i.e.,
in such cases where the response factors need to be
checked periodically only when necessary, and
(ii) An obligatory condition of this method being that
all the components of the sample should elute and
be recorded.

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 Detectors used in GC

(i) Thermal conductivity detector (TCD)

(ii) Flame ionization detector (FID)
(iii) Electron capture detector (ECD)
(iv) Thermionic detector (NP–FID)
(v) Flame photometric detector (FPD), and
(vi) Photoionization detector (PID).

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DOI: 10.13140/RG.2.2.35663.07844
 Problem

1. Discuss the strength and limitations of GC

techniques.
2. Write short notes on the common problems with
GC and suggest the ways to overcome
a) Leaks

b) Activity

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