IMPORTANT OF CHROMOTOGRAPHY IN FOOD ANALYSIS

Definition
• Chromatography is an important biophysical technique that enables the separation, identification,
and purification of the components of a mixture of qualitative and quantitative analysis.
• The Russian botanist Mikhali Tswett coined the term chromatography in 1906.
• The first analytical use of chromatography was described by James and Martin in 1952, for the use of
gas chromatography for the analysis of fatty acid mixtures.
• A wide range of chromatographic procedures makes use of difference in size, binding affinities,
charge, and other properties to separate materials.
• It is powerful separation tool that is used in all branches of science and is often the only means of
separating components from complex mixtures.

Principle of chromatography
• Chromatography is based on the principle where molecules in
mixture applied onto the surface or into the solid, and fluid
stationary phase (stable phase) is separating from each other
while moving with the aid of a mobile phase.
• The factors effective on this separation process include
molecular characteristics related to adsorption (liquid – solid),
partition (liquid – solid), and affinity or differences among their
molecular weight.
• Because of theses differences, some components of the mixture
stay longer in the stationary phase, and they move slowly in the
chromatography system, while others pass rapidly into the
mobile phase, and leave the system faster.

Three components thus from the basis of the chromatography technique.

I. Stationary phase :
This phase is always composed of a “solid” phase or “a layer of a liquid adsorbed on
the surface solid support.”

II. Mobile phase :

This phase is always composed of “liquid” or a “gaseous component.”

III. Separated molecules :

The type of interaction between the stationary phase, mobile phase, and substances contained in the mixture
is the basic component effective on the separation of molecules from each other.
• Substances can be separated on the basis of a variety of methods and the presence of characteristics
such as size and shape, total charge, hydrophobic groups present on the surface, and binding capacity
with the stationary phase.
• This leads to different types of chromatography techniques, each with their own instrumentation and
working principle.
1
 • For instance, four separation
techniques based on molecular
characteristics and interaction
type use mechanisms of ion
exchange, surface adsorption,
partition, and size exclusion.
• Other chromatography
techniques are based on the
stationary bed, including
column, thin layer, and paper
chromatography.

Commonly employed chromatography techniques include :

1. Column chromatography
2. Ion – exchange chromatography
3. Gel – permeation (molecular sieve) chromatography
4. Affinity chromatography
5. Paper chromatography
6. Thin – layer chromatography
7. Gas chromatography (GS)
8. Dye – ligand chromatography
9. Hydrophobic interaction chromatography
10. Pseudoaffinity chromatography
11. High – pressure liquid chromatography (HPLC)

Chromatography in food science technology

• With modern technology and facilities, our food supply is more diverse and more highly processed
than ever before.
• To ensure the safety and nutritional quality of our food many countries and organization have
promulgated regulations that stipulate acceptable levels for individual chemical additives, residues
and contaminants in food products.
• Other regulations require food packaging to list ingredients relating to nutritional content, such as
preservatives, artificial chemicals, unsaturated and saturated fat.
• Food manufactures and processors themselves must be able to assess product quality.
• Meeting all of these requirements is the functions of food analysis.

• Increasingly, food analysis methods are built around high – performance liquid chromatography
(HPLC), which has proven to be an optimal technology for detecting and/or quantifying the vast
majority of food analytes.
• These methods employ a stepwise approach while separating and analysis the sample, it first
removes the sample matrix, then isolates the analytes of interest and individually resolves them on a
chromatographic column.
• The efficiency of the separation depends on, among other things, the differential interaction of
analytes of interest with both mobile and column stationary phases during the separation.
• It is important to classify food analytes according to their relative volatility and polarity are factors
that must be considered when selecting an appropriate analytical method for their determination.

2
 • Gas chromatography (GC) is widely used in applications involving food analysis.
• High – performance liquid chromatography (HPLC) and to mention state-of-the-art GC techniques
used in the major applications pertaining to the quantitative and/or qualitative analysis of food
composition, natural products, food additives, flavor and aroma components, a variety of
transformation products, and contaminants, such as environmental pollutants, pesticides, fumigants,
natural toxins, veterinary drugs, and packaging materials.
• Among the several new qualitative and quantitative techniques being developed in food analysis
applications, fast-GC/mass spectrometry (MS) will have the most impact in the next decade.
• Three approaches to fast-GC/MS include low – pressure GC/MS, GC/time-of-flight)TOF)-MS and
GC/supersonic molecular beams (SMB)-MS.

 Spoilage detection and process control of foods

 Detection of food additives
 Applications in wine industry
 Determination of vitamin content in food
 Determination of vitamin C content in food
 Determination of nutritional quality of foods
 Application in dairy industry

Here bellow some applications of chromatography in food industry have been mentioned.

Spoilage detection

Chromatography can be used in flavor studies and to detect spoilage in foods. Determine the amount of
organic acids in foods provides key information about the quality of foods. Column chromatography is used
to detect and quantity spoilage indicator such as pyruvic acid in milk. pyruvic acid content is a measure of
psychrotrophic bacteria present in milk.

The same separation method is used to assess total organic acid

profile of milk and to measure lactose, which indicates the level
of sweetness. Chromatography enables rapid analysis when
compared with techniques such as bacterial plating, which may
take several days to yield results. Rapid analysis is crucial in the
industry to prevent outback of spoilage and to minimize possible
health risks.

Additive detection

Additives are added to foods to enhance their flavors or to

give them a visual appeal. For example, the presence of
added malic acid in apple juice is more difficult to detect
because apple juice naturally contains malic acid. However,
synthetic malic acid contains fumaric acid as a contaminant
and hence its level in an apple juice sample is an indicator
of the commercial malic acid. Chromatography has been
successfully used to detect and quantity fumaric acid in
apple juice.

3
Determining nutritional quality

Vitamin C depletion in foods can be an indicator of depletion of other nutrients and so the vitamin C content
of foods and beverages is closely monitored during all stages of food processing using column
chromatography. This analysis can be carried out rapidly using modern acid analysis columns coupled with
electrochemical detection even in complex samples. This technique is used to quantitative vitamin C in
juices, powdered drinks, and both fresh and frozen vegetables and fruits

Column chromatography

Subject : Food analysis and quality assurance

Name : M.D.M.De Silva
Registration number : GAM/FT/F/18/0039