Paper No.

: 11
Paper Title: Food Analysis and Quality Control
Module-10: Refractometry Food Analysis

10.1 Introduction

Refractometry is a method of testing the physicochemical properties of a substance (commonly

liquids) bymeasuring its refractive index. Refractive index of a substance is the ratio of speed of light
in vacuum to the speed of light in that substance.
𝑆𝑝𝑒𝑒𝑑 𝑜𝑓 𝑙𝑖𝑔 𝑕𝑡 𝑖𝑛 𝑣𝑎𝑐𝑢𝑢𝑚
Refractive index of a substance (𝑛) = …. Eqn 10.1
𝑆𝑝𝑒𝑒𝑑 𝑜𝑓 𝑙𝑖𝑔 𝑕𝑡 𝑖𝑛 𝑡𝑕𝑎𝑡 𝑠𝑢𝑏𝑠𝑡𝑎𝑛𝑐𝑒

Normally refractive index is used to characterize liquids. Refractive index can be used to a) assess
purity of a substance by comparing its value with the standard value b) find the concentration of a
known substance in a solution by comparing the refractive index value with a standard curve for that
substance c) help identify a substance by comparing its refractive index with known values.
Refractive index also gives an idea about the polarizability of the substance. More polarizable the
substance, higher is the refractive index of the material.

10.2 Principle

Light generally travels in straight lines and at different speeds in different media. The denser a
medium, slower is the speed of light in that medium. When light travels from one medium to another
at an angle (other than 90)it changes not only its speed but also its direction at the interface of the
two media. This phenomenon is known as refraction.Observe the figure 10.1. There are two media A
and B. Let light travel at a velocity vAin medium A. This
beam of light is called incident ray. When this incident light
touches the boundary between the two media and starts
entering into the medium B, it bends, changes its direction
and its velocity changes to vB. This beam is called refracted
ray. If we consider a normal to the plane and observe the
angles made by the incident and refracted rays, A and B
will be the angles of incidence and refraction respectively.
Let nAandnBbe the refractive indices of the media A and B
respectively. According to Snell’s law,
𝑠𝑖𝑛 A 𝑛B
= … Eqn 10.2
𝑠𝑖𝑛 B 𝑛A

𝑣A 𝑠𝑖𝑛 A 𝑛B
Also = = … Eqn 10.3
𝑣B 𝑠𝑖𝑛 B 𝑛A

Figure 10.1 Refraction (Source: http://www2.ups.edu/faculty/hanson/labtechniques/ refractometry/)

If we assume medium A to be vacuum, then the eqn 10.3 becomes

𝐶 𝑠𝑖𝑛 A
= = 𝑛B …. Eqn 10.4 (as C is the velocity of light in vacuum and according to Snell nA = 1)
𝑣B 𝑠𝑖𝑛 B

The velocity of light in vacuum always remains the same. But its velocity in any other medium is less
as it is absorbed and re emitted by the atoms in that medium. The above equation shows that refractive
index of a substance can be found out by measuring the angles of incidence and refraction rather than
measuring the speed of light.
10.2.1 Critical angle

In this figure, there are two media water and air.

Light waves travel from water to air. Observe the
refracted rays. As the angle of incidence increases,
the angle of refraction also increases. At one
particular angle of incidence, the angle of refraction
r=90, i.e, the refracted ray is parallel to the
surface. Under such circumstances, the angle of
incidence is known as critical angle. The critical
angle is the angle of incidence that gives an angle of
refraction of 90°. It is impossible to have an angle
of refraction greater than 90°. It follows that for all
angles of incidence greater than the critical angle,
the light will experience total internal reflection.

Figure 10.2 Critical angle (Source: http://micro.magnet.fsu.edu/)

𝑠𝑖𝑛 C 𝑛1
From Eqn 10.2, = (according to Fig 10.2)
𝑠𝑖𝑛 90 𝑛2

𝑛1
𝑠𝑖𝑛C = ….. Eqn 10.5, where C is the critical angle.
𝑛2

Figure 10.3 (a) Figure 10.3 (b)

Source: (https://www.nationalmicroscope.com/)

Now imagine a prism in contact with a liquid surface. When light falls at various angles on the
interface of the prism and the liquid, as told above the rays with angle of incidence greater than
critical angle are totally reflected while others are refracted. Observe the Fig 10.3 (a). The ray A gets
refracted and C gets reflected. The ray B enters at critical angle and leaves parallel to the prism face.
The critical ray i.e, the ray B acts as the transition point between total reflection and refraction. When
viewed properly, critical ray can be seen as the boundary between dark and bright regions (Fig 10.3
(b)) and defines the refractive index of the liquid. In Eqn 10.5, n2 can be easily calculated by
measuring the critical angle as the n1 value is known and is constant. This serves as the basic principle
for the operation of many refractometers.
10.2.2 Factors affecting refractive index

There are three main factors that affect refractive index measurement viz., temperature, wavelength of
light used and concentration of sample.

1. Temperature
Temperature influences the refractive index of a medium primarily because of the accompanying
change in density. At higher temperature, the liquid becomes less dense and less viscous. As a result
light travels faster in the medium accompanied by a smaller refractive index value. At lower
temperature, the same medium has a higher refractive index. This is due to the liquid becoming denser
and more viscous thereby causing the light to move slowly. Usually temperature is regulated in
refractometers to avoid errors. Refractive index is reported at 20C. In order to adjust the observed
value of refractive index to that at 20C, the following formula can be used
nD20 = nDobserved + (T-20)*0.00045…… Eqn 10.6

2. Wavelength of light used

The refractive index of a transparent medium gradually decreases with increasing wavelength.The
refractive index varies with wavelength linearly because different wavelengths interfere with the
atoms of the medium to different extents. It is important to use monochromatic light to prevent
dispersion of light into different colours. The chosen wavelength should not be absorbed by the
medium. The sodium D line at 589 nm is the most frequently used wavelength of light for a
refractometer. In certain refractometers, when white light is used, a compensator (Fig 10.4) is also
used alongside to overcome dispersion. The compensator is made of Amici prisms, in such a way that
the dispersed radiation is converged to give a beam of light that travels in the path of yellow sodium D
line.

Figure 10.4 Compensator

(Source: http://amrita.vlab.co.in/)
Hence refractive index is usually measured at a known temperature (t=20C)and wavelength of 589
nm and is denoted as ntD.

3. Concentration of the liquid

As the concentration of dissolved solids in the liquid changes, the refractive index changes. Increasing
concentration of dissolved solids increases the amount of refraction while the amount of refraction
decreases with decrease in dissolved solids. It is this property which is used in many food industries
for checking the quality of food products.

10.3 Refractometer

Refractometer is the device used to find the refractive index of a liquid sample by measuring the
extent to which light is refracted when it moves from a medium into the sample.Refractometers are
versatile in design and are available in three important forms which are discussed below.

10.3.1 Bench top refractometers

Bench top refractometers are of the earliest designs of refractometers. They are stationary and are
accurate, but have to be kept out of harsh conditions. The results are reliable as temperature can be
regulated. Improved designs of bench top refractometers are also available.
10.3.1.1 Abbe’s refractometer

Ernst Abbé designed the first refractometer in 1869. Though it has undergone several modifications
according to the need, it is still the most convenient and widely used refractometer. Fig 10.3 shows
the components of Abbe’s refractometer. Here, the sample is sandwiched as a thin layer (~0.1mm)
between an illuminating prism and a refracting prism. The refracting prism is firmly mounted on a
bearing that allows its rotation (shown in dotted lines) by means of the side arm. The refracting prism
is made of a glass with a high refractive
index (e.g., 1.75) and the refractometer is
designed to be used with samples having a
refractive index smaller than that of the
refracting prism. The illuminating prism is
hinged to the refracting prism to permit
separation for cleaning and for introduction
of the sample. The bottom surface of the
illuminating prism is rough-ground.When
light source is projected through the prism,
each point on this surface effectively
becomes the source for an infinite number
of rays that pass through the sample at all
angles. The radiation is refracted at the
interface of the sample and the smooth-
ground face of the refracting prism.After
this, it passes into the fixed telescope. Two
Amici prisms that can be rotated with
respect to each other serve to collect the
divergent critical angle rays of different
colors into a single white beam, which
corresponds in path to that of the sodium D
ray. The eyepiece of the telescope is
provided with crosshairs.

Fig 10.5 Abbe’s refractometer(Source: http://amrita.vlab.co.in/)

While making a measurement, the prism angle is changed until the light-dark interface just coincides
with the crosshairs. The position of the prism is then established from the fixed scale. Thermosetting
is accomplished by circulation of water through the jackets surrounding the prism.

The Abbe refractometer owes its popularity to its convenience, its wide refractive index range (nD =
1.3 to 1.7), and to the minimal amount of sample needed. The accuracy of the instrument is about
±0.0002. The most serious error in the Abbe instrument is caused by the fact that the nearly glazing
rays are cut off by the arrangement of the prisms; the boundary is thus less sharp than is
desirable.Improvement in accuracy is obtained by replacing the compensator with a monochromatic
source and by using larger and more precise prism mounts. The former provides a much sharper
critical boundary, and the latter allows a more accurate determination of the prism position.

10.3.1.2 Pulfrich refractometer

In 1888 Carl Pulfrich, designed a refractometer that became commercially available in 19th century. It
is another type of critical angle refractometer.
 Fig 10.6Pulfrich refractometer
(Source-http://www.refractometer.pl/)

A glass prism of high

refractive index with two plane polished faces, which are perpendicular to one another is so placed
that one of these is vertical and the other horizontal. The substance (liquid) whose refractive index is
required is contained in a glass cell cemented to the horizontal surface. A beam of monochromatic
light is directed almost horizontally through the substance so that it meets the prism face at grazing
incidence (90). The emergent beam is bounded sharply by the grazing ray and the sharp boundary is
observed with a telescope attached to a divided circle. On this circle, whose axis of rotation is
horizontal, the angle of emergence of the beam from the vertical prism face can be read to one minute,
with the aid of a vernier. The hollow metal water jacket surrounding the prism is provided with a top
cover which is itself of hollow metal. These two are connected in train with the thermometer jacket
which dips into the cell for liquids. The prism and the substance experimented upon are thus
completely jacketed with the exception of the vertical prism face from which the light emerges, and a
small rectangular aperture by which the light enters the prism. The temperature can thus be
maintained very constant by pumping a stream of water at constant temperature through the system
described.

The automatic digitalbenchtoprefractometers use a light sensitive integrated circuit to detect the exact
position of the borderline, thereby minimising human error in reading.

10.3.2 Hand held refractometers

Hand held refractometers are one of the most popular designs of critical angle refractometers. They
are quick to use units and have the advantage of scales calibrated to read percentage of solids, sugars,
alcohol etc. depending on the products for which they are intended to be used. This kind of
refractometer is portable and can be taken to the site for measurement.

Fig 10.7 Hand held refractometer (Source: http://www.refractometer.pl/)

The main difference between bench top and hand held refratometers is that instead of having an
illuminating prism, hand held refractometers have an illuminator flap which allowslight at a grazing
angle. The flap also helps to keep the sample in place. Light passes through the sample, enters the
measuring prism and other lenses, and finally falls on the measuring scale (calibrated in required
units) from where it can be read. To take care of temperature differences, simple hand held
refractometers have to be either calibrated before taking measurements (using calibration screw and
distilled water), or corrections have to be made after measurement using temperature correction table.
In the latter case, separate temperature measurement is required. However, many refractometers have
built in temperature compensation, an additional optical wedge mounted on the bimetallic strip, which
bends when the temperature changes, compensating for changes of refractive index.

The measurement is also very simple as it requires a few drops of the sample to be placed on the
prism surface after opening the illuminator flap. When the illuminator flap is closed, the result can be
directly read from the scale.

A high concentration of
dissolved solids increases
the amount of refraction
while low dissolved solids
gives lower refraction.

Fig 10.8 Hand held refractometer calibrated in %Brix

10.3.3 In line refractometers

In-line refractometers are used for continuous monitoring of liquids like sugar syrups, jams,
marmalades, juice, juice concentrate, beverages, soft drinks,
wine, beer, coffee, milk products, jellies, tomato pastes,
liquid glucose, fructose etc.during processing. They can be
installed in pipings of equipments. The light from a LED
source is transmitted by a fibre optic onto one side of a
measuring prism and internally reflected to the interface of
the prism and the sample liquid. A portion of this light beam
is refracted while the rest is reflected back onto the opposite
side of the prism at an angle which depends on the refractive
index of the liquid at the interface. The position of this
reflected beam, as it exits the prism, changes with the critical
angle (which depends on the refractive index of the sample).
A linear diode array is used to accurately detect the varying
position of the reflected beam and this position is calibrated
with respect to refractive index.

Fig 10.9In line refractometer (www.topac.com)

They provide real-time data for quality control and Good Manufacturing Practice (GMP). Since
continuous monitoring is possible, the quality of the product is also high. The data from these can be
directly sent to a data logger (computer).
10.4 Applications in food industries

1. Refractometers are used to assess the ripeness of fruits, vegetables, sugar cane and beet before
harvesting - i.e., it serves to find the maturity index. This is mainly used for grapes in vineyards. It
measures the sugar content of the fruit in B. A liquid has one degree Brix (=1% Brix) if it has the
same refractive index as a solution of 1g sucrose in 100g of sucrose water solution.

2. They are used to check the alcohol content of wine and beer during fermentation in breweries. For
this purpose, special refractometers with scales calibrated to give the alcohol content directly are
available.

3. Quality of honey can be checked using special kind of refractometers which directly give the
amount of water in honey. Low water content in honey is preferable as it ensures better shelf life to
the product. If the % of water is high (>20%) then it leads to fermentation by wild yeasts. This kind of
refractometer is preferred by bee keepers.

4. The salinity meter is used in the food industry to measure the percentage of salt in condiments such
as soy sauce, ketchup, mayonnaise and in canning industries for brine solution.

5. Refractometers are used to monitor and control the quality of yoghurt, jam, fruit extract, syrup,
coffee extract, chocolate, milk, baby food etc. by measuring the total solids in them during processing.

6. The refractive index is an indicator of purity in products such as vegetable oils and animal fats.By
comparing the values with that of the standard the purity of the product can be assessed.

10.5 Advantages of refractometers

1. Refractometers give instant results.

2. Refractometers are versatile – available as bench top, hand held and in line models.
3. Hand held refractometers are portable and hence can be taken to the spot.
4. They are relatively inexpensive with low initial cost and low level of maintenance.
5. Refractometers, especially the hand held models are easy to operate. Hence they do not require
much skill to operate.
6. Also refractometerscan give direct readings such as B or salinity.

References
1. Robert L Bradley Jr, 2010, ‘Moisture and total solid analysis’, Food Analysis, Fourth
edition, Ed. Suzanne Nielson S, Springer Science + Bussiness Media, USA, ISBN 978-1-
4419-1477-4.