Industrial Training

at

Tea Research Institute of Sri Lanka

Thalawakelle
by
A. A. D. M. Adhikari(S/08/905)

External Supervisors: Dr. K. M. Mewan

Mr. G. A. A. R. Perera

Internal Supervisor : Dr. C.V. Hettiarachchi

Content
1) About TRI.
2) About Biochemistry Division.
4) Analysis of tea samples.
5) Development of a method for manufacturing
paper from refuse tea.

About TRI...........................

The Tea Research Institute (TRI) was started in

1925.
TRI has been a driving force behind the numerous
successes in developing the Sri Lankan tea industry
over the years.
The two estates of TRI, St. Coombs in Talawakelle
and St. Joachim in Ratnapura, are primarily meant to
provide facilities to undertake research in cultivation
and processing, while attempts are also made to run
them as viable units on commercial lines.
Reference:http://www.tri.lk/tri/tri/7

The areas of research performed by TRI

Breeding for crop improvement

Alternative energy sources and energy saving techniques

Soil fertility improvement

Mechanization

Factory development and quality improvement

Integrated Pest Management (IPM)

Value addition and by-products

Productivity improvement

Technology Transfer

Societal - Economics

Research divisions of the TRI

a) Agriculture Economics
b) Agronomy
c) Biochemistry
d) Entomology and Nematology
c) Plant Breeding
d) Plant Pathology
e) Soil and Plant Nutrition
f) Process Technology

About the Biochemistry Division

The biochemistry division conducts studies
on chemical and biochemical aspects of tea.
The biochemistry Division contains,
Two biochemistry laboratories
A pesticide laboratory
A biotechnology unit
A pilot plant

Reference: http://www.tri.lk/research-and-development/biochemistry

The areas of research performed by the

Biochemistry division

Product development and value addition

Development of standards of tea

Biological effects of tea consumption

Application of molecular technology to

support germplasm conservation and crop
improvement
Chemical ecology of tea pests and diseases
Estimation of pesticide residue levels of
made tea.

Manufacturing black tea in

St. Coombs tea Factory
Plucking
Plucking tea
tea

-Pluck
-Pluck tea
tea as
as bud
bud with
with two
two leaves
leaves

Rolling
Rolling and
and CTC
CTC
-Brake
-Brake the
the tea
tea leaves
leaves into
into small
small particles
particles

Fermentation
Fermentation

-- Keep
Keep the
the tea
tea for
for oxidation
oxidation of
of polyphenols
polyphenols

Firing
Firing (Drying)
(Drying)

-Reduce
-Reduce the
the moisture
moisture of
of fermented
fermented tea
tea to
to 2%-3%
2%-3%

Grading
Grading

-Grade
-Grade the
the tea
tea based
based on
on particle
particle size
size and
and amount
amount of
of
fiber
fiber

Grades of tea in St. Coombs

tea factory

Dust 1

BOPF

BOP

Peko

Dust 2

BM

Refuse tea
Refference:http://ceylonteainfo.com/recent-developments

Manufacturing instant tea

from refuse tea
Preparation
Preparation of
of tea
tea extraction
extraction

--Extract
Extracttea
teainto
intohot
hotwater
waterfrom
fromrefuse
refusetea
tea
Filtering
Filtering and
and centrifuging
centrifuging

--Removal
Removalof
ofparticle
particleof
ofspent
spenttea
tea

Concentrating
Concentrating

-Concentrate
-Concentratethe
theextract
extractuntil
untilsolid
solidparticle
particlelevel
levelreaches
reaches24%
24%

Spray
Spray drying
drying

-Powder
-Powderof
ofinstant
instanttea
teais
isprepared
prepared

Analysis of tea samples

a)

Determination content of total

polyphenols in tea

b)

Determination of antioxidant activity of a

tea sample

c)

Determination of TF, TR in a tea sample

(Roberts and Smith method)

Determination content of total

polyphenols in tea
Introduction:
Tea contains polyphenolic compounds such catechine,
theoflavin ...etc. .
OH

OH
OH

OH

OH

HO

OH

H
O

HO

HO

OH

OH
OH

OH

(-)-catechin

OH

theaf lavin

OH
OH

OH

H
OH
HO

O
H
OH

(-)-epigallocatechin

Analysis of tea polyphenols used as the quality

parameters or indicators of tea .

Theory

In this experiment total polyphenol content is given as

gallic acid equivalent.
The reagent used to oxidize polyphenols is FolinCiocalteu.
This reagent is formed from a mixture of
phosphotungstic acid (12WO3.H3PO4) and
phosphomolybdic acid, (12MoO3..H3PO4).
After oxidation of the phenols, the reagent is reduced
to a mixture of blue oxides of tungsten, W 8O23, and
molybdenum, Mo8O23.
Maximum absorption is in the region of 765 nm.
Reference:
http://www.springerimages.com/Images/LifeSciences/1-10.1007_s11103-011-9739-3-5
http://www.sigmaaldrich.com/catalog/product/sigma/ht153?lang=en&region=LK

OH
HO

HO

OH

2(12WO 3.H3PO 4)(aq)

3(W 8O 23)(aq)

phosphotungstic acid

(aq)

(aq)

3H 3PO 4(aq)

COOH

COOH

gallic acid

OH

HO

HO

OH

2(12MoO 3.H3PO4) (aq)

(aq)

3(Mo 8O23) (aq)

(aq)

phosphomolibtictic acid

3H3PO4(aq)
COOH

gallic acid

COOH

All the measurements are based on colorimetric

methods and Beer-Lambert Law.
Where,
A=
lC
where ,
A- Absorbance

olar extinction coefficient

C- Concentration
l- Path length
fig 2: Graph of A vs Concentration

The total polyphenol content, WT, expressed as a

percentage of sample dry matter mass, is given by
the formula:
WT= (D

sample

- Dintercept)Vsample

S m
std

sample

d100

10000 w

DM, sample

where,
Dsample - the optical density obtained for the sample
test solution;
Dintercept- the optical density at the point the best-fit
linear calibration line intercepts the y-axis;

Sstd

- the slope obtained from the best-fit linear

calibration;

msample - the mass, in grams, of the sample test

portion;
Vsample - the sample extraction volume, in milliliters
(50 ml for instant tea and 10 ml for leaf tea);
d

- the dilution factor used prior to the

colorimetric determination (typically 1,0 ml to
100 ml, thus a dilution factor of 100);

wDM, sample-the dry matter content, expressed as a mass

fraction in percent, of the test sample.

Procedure :
Preparation of samples
The
The tea
tea leaves
leaves were
were ground
ground

Moisture
Moisture of
of the
the tea
tea sample
sample was
was determined
determined

0.200g
0.200g of
of the
the sample
sample was
was measured
measured

10.00
10.00 ml
ml extraction
extraction of
of 70%
70% methanol
methanol was
was prepared
prepared
(Solution
(Solution A)
A)
1.00
1.00 ml
ml of
of AA was
was diluted
diluted up
up to
to 100.00
100.00 ml
ml with
with distilled
distilled
water
water(Solution
(SolutionB)
B)

Standard preparation.
Series of gallic acid solutions were prepared
according to the below table.
Table 1: Concentration of gallic acid solution
Gallic acid standard
solution

Concentration of
solution/(g/ml)

10.01

20.02

30.03

40.04

50.04

Taking measurements:
1.00
1.00 ml
ml from
from solution
solution was
was poured
poured into
into aa graduated
graduated test
test
tube
tube
5.00
5.00 ml
ml of
of 10%
10% Folin-Ciocalteu
Folin-Ciocalteu phenol
phenol reagent
reagent was
was
added
added

After
was
After 44 minutes
minutes 4.00
4.00 ml
ml of
of 7.5%
7.5% Na
Na22CO
CO3(aq)
was
3(aq)
added
added

The
The solution
solution was
was mixed
mixed and
and kept
kept for
for 60
60 minutes
minutes

Absorbance
Absorbance was
was measured
measured at
at
765
765 nm
nm

Results :
Table 2: Average absorbance of standard solutions

Gallic acid
standard
solution

Absorbance
Concentration
(0.0001)/
of solution/
Arbitrary
(g/ml)
units

10.01

20.02

30.03

40.04

50.04

0.1667
0.2709
0.3943
0.5283
0.6461

Graph 1:Graph of absorbance vs concentration of gallic acid y = 0.0122x + 0.0364

R = 0.9984

0.7

Absorbance/
arbitori unit

0.6
0.5
0.4
0.3
0.2
0.1
0
0

10

20

30

40

50

Concentration of gallic acid solutions/(micro gram/ml)

60

Table 3:calculated total polyphenol content of

each sample

Sample no

Absorbance
Mass of tea
total
(0.0001)/
(0.0001)/
polyphenol
(arbitori content (%)
g
units)

0.2004

0.5051

20.32

0.2008

0.5540

22.39

0.2003

0.4785

19.17

Average % of total polyphenol content = 20.63%

Conclusion :
Average % of total polyphenol content based on dry
mater of sample is 20.63%

http://www.google.lk/url?sa=t&rct=j&q=&esrc=s&frm=1&source=web&cd=2&sqi=2&ved=0CDUQFjAB&url=http%3A%2F
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Development of a method
for
manufacturing paper
from
refuse tea

Development of a method for

manufacturing paper from
refuse tea
Introduction:
Paper is manufactured from plant material
containing cellulose fiber such as straw, timber of
some plants...etc.
At present manufacturing paper has become a
environmental problem.

Refuse tea also contains a higher amount of fiber,

therefore it is possible to manufacture paper from
it.
In this test an attempt was taken to develop a
manufacturing method by finding conditions at
which the best quality paper is obtained.

Procedure :
Pulp preparation.
Refuse
Refuse tea
tea was
was mixed
mixed with
with boiling
boiling water
water for
for 10
10
minutes
minutes
Extract
Extract was
was filtered
filtered and
and washed
washed with
with 200
200 ml
ml of
of
boiling
boiling water
water
the
the residue
residue was
was added
added to
to aa beaker
beaker containing
containing NaOH
NaOH
and
and boiling
boiling water
water

The
The mixture
mixture was
was mixed
mixed
The
The solution
solution was
was filtered
filtered
The
The residue
residue was
was ground
ground step
step wise
wise with
with
water
water
The
The mixture
mixture was
was filtered
filtered and
and washed
washed with
with water
water
Pulp
Pulp
http://homepage.cs.uiowa.edu/~jones/prairiepaper.htm

Table 3: Condition of each procedure

Procedure

Used
refuse tea
Amount

Used
NaOH
amount

/g

/g

5.00
8.00
25.00

3.500
2.000
1.250

(0.0001) (0.001)
A
B
C

Mixing
time/
minutes

Grinding
Time/
minutes

5
30
30

1
3
3

Producing paper:
In procedure A:
The
The pulp
pulp was
was put
put on
on to
to aa net.
net.

Another
Another net
net was
was laird
laird on
on it.
it.
It
It was
was pressed
pressed and
and water
water was
was removed
removed

Paste
Paste was
was kept
kept to
to dry.
dry.

Paper
Paper

In A and B procedure:
The
The pulp
pulp was
was put
put on
on to
to aa sieve
sieve dipped
dipped in
in water
water

Particles
Particles were
were allowed
allowed to
to suspended
suspended

It
It was
was kept
kept it
it to
to drain
drain water
water for
for over
over night
night

It
It was
was pressed
pressed

It
It was
was kept
kept to
to dry
dry

Paper
Paper

Results :
Table 1: NaOH used per 100g of refuse tea for
each procedure
Procedure

Used refuse Used NaOH NaOH used

tea mass
tea mass
per 100g of
(0.0001)/ (0.0001)/ refuse tea/g
g
g

5.000

3.500

70.000

8.000

2.000

25.000

25.000

1.250

5.000

Table 2: Quality of the papers

Paper of
Paper of
Paper of
Procedure A Procedure B Procedure C
Smoothness Low
of surface

High

High

Strength

High

High

Low

Colour

Golden
yellow

Brown wish Brown

yellow

Table 2: Ranking of paper base on each quality

Quality

Paper of
Procedure
A

Paper of
Paper of
Procedure Procedure C
B

Smoothness
of surface

Strength
Colour

1
1

1
2

2
3

Conclusion:
NaOH amount added for pulp preparation and the
time of mixing and grinding the pulp should be
high to get a best quality paper.
Grind refuse tea is not suitable for getting paper
with higher strength

Suggestion:
In this process the tea extraction ,extracted
previously can be used to manufacture instant tea.
The NaOH extraction can be used to extract
protein. So that the paper is manufacture from
waste product of above processes.

The paper obtained in second process is the best

one from the others and it is better to use as
paper which is used in greeting cards, invitation
cards ..etc.
As this papers contains some amount of
polyphenols it is resistance to fungus. Therefore it
is not necessary to add anti fungal reagent to it.

In next stage it is suggested to find

the minimum amount of NaOH which gives the
best paper
a way to bleach the paper
the area of the paper manufactured from unit
mass of refuse tea
optimum pressure used to press the pulp
optimum temperature and time used in pulp
preparation

u
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