ANALYTICAL TECHNIQUES IN

BIOTECHNOLOGY LAB (BM3708)

EXPERIMENT NO: 6

TITLE: Measuring refractive index using Refractometer

DEGREE:B.Tech

SUBMITTED BY

GROUP- 1

SUBGROUP-5
SI NO. NAME REGISTRATION NO. SIGNATURE

1 Karthik Peddi 120BT0571

2 Satabdi Beuria 120BTO569
3 Ashna Reyaz 120BT0032

DATE OF EXPERIMENT DATE OF SUBMISSION

DEPARTMENT OF BIOTECHNOLOGY AND MEDICAL ENGINEERING

NATIONAL INSTITUTE OF TECHNOLOGY
ROURKELA-769008
ODISHA

MARKS
SIGNATURE REMARK
AWARDED
 DEPARTMENT OF BIOTECHNOLOGY & MEDICAL ENGINEERING
NATIONAL INSTITUTE OF TECHNOLOGY, ROURKELA
COURSE: BM3708 Analytical Techniques in Biotechnology Laboratory

ANALYTICAL TECHNIQUES IN
BIOTECHNOLOGY LAB (BM3708)
EXPERIMENT NO: 6

TITLE: Measuring refractive index using Refractometer

DEGREE: B.Tech

SUBMITTED BY

GROUP- 1

SUBGROUP-5

SI NO. NAME REGISTRATION NO.

1 Karthik Peddi 120BT0571
2 Satabdi Beuria 120BT0569
3 Ashna Reyaz 120BT0032

DATE OF EXPERIMENT DATE OF SUBMISSION

 DEPARTMENT OF BIOTECHNOLOGY & MEDICAL ENGINEERING
NATIONAL INSTITUTE OF TECHNOLOGY, ROURKELA
COURSE: BM3708 Analytical Techniques in Biotechnology Laboratory

DEPARTMENT OF BIOTECHNOLOGY AND MEDICAL ENGINEERING

NATIONAL INSTITUTE OF TECHNOLOGY
ROURKELA-769008
ODISHA

EXPERIMENT-3
AIM: Measuring refractive index of sample, to calculate its concentration
using Refractometer.

THEORY:
A refractometer is an optical device used for measuring the extent to which light is bent,
or refracted when it moves through a substance. It works because light travels at different
velocities in different mediums. The principle of Refractometer is:
When a beam of visible light passes through the interface between two different
substances, the phenomenon of refraction occurs. In an extreme refractive condition
(total reflection), the light will not enter the interface of the second substance, but will be
reflected as if it encountered a mirror. The incident angle of light when this phenomenon
occurs is called the "critical angle".
The size of the critical angle is strictly determined by the refractive index of their two
substances. If the refractive index of one substance is known, plus the measurement of
the critical angle, then the refractive index of the other substance can be determined.
Refractive index is the inherent physical property of each substance. The analysis of
liquids is based on the analysis of solute concentration.

There are mainly 2 types of refractometers:

1)optical refractometers
2)Digital refractometers.
In our lab we basically use digital refractometer, which gives 3 kinds of reading namely
ND, ND20 ,BRIX. These terms commonly used in the field of refractometry to describe
the properties of liquids.
ND (Refractive Index): The ND (refractive index) value of a liquid is a measure of the
amount by which light is bent when it passes from air into the liquid. The ND value is a
 DEPARTMENT OF BIOTECHNOLOGY & MEDICAL ENGINEERING
NATIONAL INSTITUTE OF TECHNOLOGY, ROURKELA
COURSE: BM3708 Analytical Techniques in Biotechnology Laboratory

dimensionless number that is used to describe the refractive index of a liquid and is
typically expressed as a decimal value. The higher the ND value, the more the light is
bent when it enters the liquid.
ND20: ND20 is a unit of measurement used to describe the refractive index of a liquid at a
specific temperature (usually 20°C). This value is commonly used in the food and
beverage industry to measure the sugar content of liquids, such as fruit juices and syrups.
Brix: Brix is a unit of measurement that is commonly used in the food and beverage
industry to describe the concentration of sugar in a liquid. The Brix value is a measure
of the amount of sugar in a solution and is typically expressed as a percentage. Brix values
are used to determine the sweetness of a liquid and can be useful in a variety of food and
beverage applications, including the production of soft drinks, fruit juices, and syrups.
DIFFERENT PARTS OF DIGITAL REFRACTOMETER:
1. Light source: A light source, such as an LED, is used to illuminate the liquid sample.
2. Photodetector: A photodetector, such as a photodiode, is used to measure the angle of
refraction of the light as it passes through the liquid sample.
3. Microprocessor: An onboard microprocessor is used to calculate the refractive index of
the liquid based on the measurement from the photodetector.
4. Display: An electronic display is used to show the refractive index value of the liquid.
5. User interface: A user interface, with a screen that display readings, an ON/OFF
button, READ button to read the sample and switch between ND, ND20 ,BRIX and
ZERO button to turn the reading to zero 0.

METHODOLOGY:
 DEPARTMENT OF BIOTECHNOLOGY & MEDICAL ENGINEERING
NATIONAL INSTITUTE OF TECHNOLOGY, ROURKELA
COURSE: BM3708 Analytical Techniques in Biotechnology Laboratory

1. Prepare the liquid sample: To prepare the liquid sample we first make the stock solution
of 15% by weight sugar solution total volume 100 ml, To prepare it we mix 15 grams
of sugar in 100 ml of water. Now we prepare 5 different concentrations of sample (20 ml
each) 5%, 7%, 10%, 12.5%, 14% from our stock solution by adding 6.66, 9.33, 13.33,
16.66, 18.66 ml of stock solution respectively and rest water.
2. Load the sample: Load the liquid sample into the measurement cell of the refractometer,
taking care not to spill or contaminate the sample we also cover the measurement cell to
minimise possibility of light interaction. After this we press the ON/OFF button.
3. Set the standard value of the refractive index: Here we select the standard as air and give
its refractive index a value of 1.00.
4. Start the measurement: Press the READ button to start the measurement. The refractive
index(ND) of the liquid will be calculated and displayed on the electronic display. If we
press the READ button again we get a value of ND20 and if we press once again we get a
Brix value.
5. Read the value: Read the refractive index value from the electronic display. The value
will be displayed in the selected unit (e.g., ND, Brix, and ND20.).
6. Clean the cell: After the measurement, clean the measurement cell thoroughly to avoid
contamination of future samples.
RESULT:
We can get the result by Measuring the samples at different Concentartions using Digital
Refractometer .
Concentration Stock (in ml) Water(in ml) Sugar% (Brix Refractive
value) Index
5% 6.66 13.33 4.5% 1.3392
7% 9.33 10.66 6.5% 1.3420
10% 13.33 6.66 9.3% 1.3463
12.5% 16.66 3.33 11.5% 1.3498
14% 18.66 1.33 12.9% 1.3519
 DEPARTMENT OF BIOTECHNOLOGY & MEDICAL ENGINEERING
NATIONAL INSTITUTE OF TECHNOLOGY, ROURKELA
COURSE: BM3708 Analytical Techniques in Biotechnology Laboratory

CONCLUSION:
By plotting the graph of Known concentration vs Refractive Index we got the
line Equation y = 0.0014x + 1.3321
Using this line Equation we can calculate the unkown Sample Concetration .
The unknown Sample Refractive Index = 1.3521==y
X = 14.285 %
So the unkown sample Concentartion is 14.285%.