Unit 6: Food Quality and Analysis

Unit 6
Food Quality and Analysis

UV-Vis spectroscopy
• UV-Vis spectroscopy is an analytical technique that
measures the amount of discrete wavelengths of UV or
visible light that are absorbed by or transmitted through
a sample in comparison to a reference or blank sample.
• This property is influenced by the sample composition,
potentially providing information on w
• Light has a certain amount of energy which is
inversely proportional to its wavelength. Thus,
shorter wavelengths of light carry more energy
and longer wavelengths carry less energy. hat is in
the sample and at what concentration.
UV-Vis spectrophotometer
work
A simplified schematic of the main components in a UV-Vis spectrophotometer

Light source
• A steady source able to emit light across a wide
range of wavelengths is essential.
• A single xenon lamp is commonly used as a high
intensity light source for both UV and visible ranges.
• Xenon lamps are, however, associated with higher
costs and are less stable in comparison to tungsten
and halogen lamps.
• A tungsten or halogen lamp is commonly used for
visible light.
• A deuterium lamp is the common source of UV
light.
Wavelength selection
• Monochromators - A monochromator separates light into a
narrow band of wavelengths. It is most often based on
diffraction gratings that can be rotated to choose incoming
and reflected angles to select the desired wavelength of light.
• Absorption filters - Absorption filters are commonly made
of colored glass or plastic designed to absorb particular
wavelengths of light.
• Interference filters - Also called dichroic filters, these
commonly used filters are made of many layers of dielectric
material where interference occurs between the thin layers of
materials. These filters can be used to eliminate undesirable
wavelengths by destructive interference, thus acting as a
wavelength selector
• Cutoff filters - Cutoff filters allow light either below
(shortpass) or above (longpass) a certain
wavelength to pass through. These are commonly
implemented using interference filters.
• Bandpass filters -Bandpass filters allow a range of
wavelengths to pass through that can be
implemented by combining shortpass and longpass
filters together.
Detection
• After the light has passed through the sample, a detector is used to
convert the light into a readable electronic signal. Generally,
detectors are based on photoelectric coatings or semiconductors.
• A photoelectric coating ejects negatively charged electrons when
exposed to light. When electrons are ejected, an electric current
proportional to the light intensity is generated. A photomultiplier
tube (PMT) is one of the more common detectors used in UV‑Vis
spectroscopy. A PMT is based on the photoelectric effect to initially
eject electrons upon exposure to light, followed by sequential
multiplication of the ejected electrons to generate a larger electric
current. PMT detectors are especially useful for detecting very low
levels of light.
• When semiconductors are exposed to light, an electric current
proportional to the light iantensity can pass through. More
specifically, photodiodes and charge‑coupled devices (CCDs) are two
of the most common detectors based on semiconductor technology
Principle
Beer–Lambert's law is especially useful for obtaining the concentration of a
substance if a linear relationship exists using a measured set of standard
solutions containing the same substance.
• The absorbance (A) is equal to the logarithm of a fraction involving the

intensity of light before passing through the sample (Io) divided by the
intensity of light after passing through the sample (I). The fraction I
divided by Io is also called transmittance (T), which expresses how much
light has passed through a sample. However, Beer–Lambert's law is
often applied to obtain the concentration of the sample (c) after
measuring the absorbance (A) when the molar absorptivity (ε) and the
path length (L) are known. Typically, ε is expressed with units of L mol‑1
cm‑1, L has units of cm, and c is expressed with units of mol L‑1. As a
consequence, A has no units.
DATA Analysis
Strengths and limitations
of UV-Vis spectroscopy
• The technique is non‑destructive, allowing the sample
to be reused or proceed to further processing or
analyses.
• Measurements can be made quickly, allowing easy
integration into experimental protocols.
• Instruments are easy to use, requiring little user
training prior to use.
• Data analysis generally requires minimal processing,
again meaning little user training is required.
• The instrument is generally inexpensive to acquire and
operate, making it accessible for many laboratories.
Limitations
• Stray light - In a real instrument, wavelength selectors are not perfect and a small
amount of light from a wide wavelength range may still be transmitted from the light
source,1 possibly causing serious measurement errors.9 Stray light may also come from
the environment or a loosely fitted compartment in the instrument.1
• Light scattering - Light scattering is often caused by suspended solids in liquid

samples, which may cause serious measurement errors. The presence of bubbles in
the cuvette or sample will scatter light, resulting in irreproducible results.
• Interference from multiple absorbing species - A sample may, for example, have
multiple types of the green pigment chlorophyll. The different chlorophylls will have
overlapping spectra when examined together in the same sample. For a proper
quantitative analysis, each chemical species should be separated from the sample and
examined individually.
• Geometrical considerations - Misaligned positioning of any one of the instrument's

components, especially the cuvette holding the sample, may yield irreproducible and
inaccurate results. Therefore, it is important that every component in the instrument is
aligned in the same orientation and is placed in the same position for every
measurement. Some basic user training is therefore generally recommended to avoid
misuse.

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Unit 6

Food Quality and Analysis

A simplified schematic of the main components in a UV-Vis spectrophotometer

• The absorbance (A) is equal to the logarithm of a fraction involving the

• Light scattering - Light scattering is often caused by suspended solids in liquid

• Geometrical considerations - Misaligned positioning of any one of the instrument's

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