Extraction of Catechin from Different Plants (Palm Oil shoots, Pereskia Bleo,

Centella, Coriander and Salak)

Syahiriah Mustafar Bakri, Norsalliana Shaari, Norkamruzita Saadon
Faculty of Chemical Engineering, Mara University of Technology, Sura Hujung, 23000 Dungun
Terengganu Darul Iman
Abstracts
Herbal tea, tisane, or ptisan is a herbal or plant infusion and usually not comes from the leaves of the tea bush
(Camellia sinensis). Generally, herbal tea is simply the combination of hot or boiling water and dried fruits, flowers,
leaves or herbs. There is still lack of research of plants or herbs that contain catechin and also not many of them
realized the benefits of catechin. The state when the plants were extracted was manipulated by extracting the plants
in fresh conditions and dried conditions. In this research, the Palm oil shoot (PS), Coriander leaves (CL), Centella
leaves (CEL), Pereskia leaves (PL) and Salak peelings (SP) were processed according to green tea processing
procedures. The polyphenol component which is catechin was extracted and analysed using high performance liquid
chromatography. The analysis recorded that the dried PS contain higher catechin followed by SP, PL, CEL and CL.
The fresh samples analysis gives out that PS still have the higher catechin followed by SP, CEL, CL and PL. As a
conclusion, the samples have gone through the analysis to determine the concentration of the catechin in it. Research
shows that, from all the samples, pereskia bleo contain the highest concentration from others both in fresh and dried
states. The different condition also gives effect to the catechin in the plant as the dried plants give more
concentration than the fresh plants; depend on the types of the plants itself.Keywords: Catechin, herbal tea, palm oil,
centella, pereskia, salak, coriander, different states.
Keywords: Herbal tea, different states.

1. Introduction
Polyphenolic is one of the components in a
tea herbs or ptisan that contain catechin, more or less
depend on the types of the herbs or plants itselves.
Catechin is one of the components of polyphenolic
antioxidant plant metabolite. It commonly used to
refer the related family of flavonoids or simply
flavanols. Catechin compounds have been of focus
for the strong sulfated effect and anti-cancer function
(Row and Jin, 2005).

Herbal tea, tisane, or ptisan is a herbal or

plant infusion and usually not made from the leaves
of the tea bush (Camellia sinensis). Generally, herbal
tea is simply the combination of hot or boiling water
and dried fruits, flowers, leaves or herbs. Herbs such
as chives, coriander, hyssop, chamomile and so much
more are belong specifically to certain botanical
families such as Alliaceae, Apiaceace and Lamiaceae.
Botanical evidence shows that India and China were
among the first countries to cultivate tea. Today, tea
is the most widely consumed beverage in the world,
second only to water. Hundreds of millions of people
drink tea around the world, and studies suggest that
green tea (Camellia sinesis) in particular has many
health benefits. For centuries, herbs have been used
as food and medicinal purpose. Many researches
focuses on various herbs that contain hypolipidemic,
antiplatelet, antitumor, or immune-stimulating
properties that may be useful adjuncts in helping
reduce the risk of cardiovascular disease and cancer.
In different herbs, a variety of active phytochemicals,
including the flavonoids, terpenoids, lignans, sulfides,
polyphenolics, carotenoids, coumarins, saponins,
plant sterols, curcumins, and phthalides have been
identified (Craig, 1999).

There are two states for the extraction of

catechin which are in dry state or fresh condition. As
it is known for herbs, the highest levels are found
expectedly in dried herbs, especially from the
Lamiaceae. The highest levels for the other botanical
families are found in dried Lauraceae, Asteraceae and
dried Fabaceae. The contents in fresh herbs are lower
such as in fresh common types of Lamiacae, but Chan
et al. (2006) state that fresh tea leaves are very rich in
catechins, which may constitute up to 30% of dry
weight (Graham, 1992). Principal catechins of young
tea leaves are epigallocatechin gallate (EGCG),
epigallocatechin (EGC), epicatechin gallate (ECG),
gallocatechin (GC), epicatechin (EC) and catechin.
Content of catechins varies with climate, season,
horticultural practices, leaf age and variety. Chen et

al. (2003) reported that young tea leaves were richer

in caffeine, EGCG and ECG than were mature leaves.
Old leaves had higher levels of theanine, EGC and
EC.

be constant which are 80C and 40 minutes. The

solutions start heated and stirred at 300 r.p.m.
After 40 minutes, the solutions are left to
cool at room temperature. Then, the solutions are
filtered using 110mm filter paper so that there is no
residue left on the solutions. Lastly, 10mL of the
solutions is kept in the sample bottles and stored in
4C fridge for further analysis.

2. Materials and Methods

2.1. Raw Material
There are five types of plants and herbs used
as raw materials. They are coriander, centella,
pereskia bleo, salak peelings and palm oil shoots.
The coriander, salak and centella can be found at
Pasar Besar Dungun at Kuala Dungun whereas the
pereskia bleo is getting from one of the dealer at the
morning market. The palm oil leaves is obtained from
the Rasau kertih palm oils plantation, Dungun. For
dried process, the process were done using
microwave at college.

2.5. Samples Analysis for Catechin (HPLC)

Firstly, about one gram catechin is weighed and
added into 10mL distilled water. Then, 1/10 from the
solution is taken and diluted with 10mL distilled
water. The process repeated until the solution is less
concentrated. The samples will be analyzes by using
High Performance Liquid Chromatogrphy. This
equipment has a mobile phase, a stationary phase and
detector. The mobile phase is pumped continuously at
a fixed flow rate through the system. Then, the
injector is used to introduce a plug of sample into the
mobile phase without needing to stop the mobile
phase flow and introducing air into the system. The
mixture then carried in a narrow band to the top of the
column. The detector is used to respond to a psychochemical property of analyte digitally amplified and
sent to a data system which will be recognized as
chromatogram. Here are the requirements to run the
analysis: Each of the samples was run for 8 minutes
with the flow rate 1mL/min. The column used was
C18 with the temperature 35C and the mobile phase
of 80% distilled water and 20% acetonitrile. The
graph will gives out the peak areas where it indicates
the concentration of catechin in tisane and in the
standard solution.

2.2. Chemical
During the process, there is no chemical
used. For the sample analysis, the catechin standard is
used. About one gram catechin will be diluted with
100mL distilled water and 1/10 from the solution will
be taken and diluted with 100mL distilled water until
get the solution needed.
2.3 Apparatus and Equipment
The apparatus used are weighing scale which
used to weight the samples, the heating mantle used
to heat the samples with distilled water. For the
equipment, high performance liquid chromatography
(HPLC) is used to determine the catechin in the
samples.

3. Results and Discussion

2.4. Extraction of Catechin

3.1 Peak Areas

For this experiment, five gram of five raw

materials is used. They are centella, coriander,
pereskia bleo, salak peelings and palm oil shoots. The
raw materials are prepared for fresh and dried. The
leaves and peelings are cut into small pieces and
mashed using pestle and mortar so that more the
leaves and peelings can release more juice. For dried
samples, before it mashed, the leaves and peelings are
dried using microwave for two minutes. The time is
constant for all samples.
As the first step is finished, the mashed
leaves then is added to 100mL of distilled water and
put into round bottom flasks. The ratio of samples to
distilled water 1:20 where one gram of sample
equivalent to 20mL of distilled water. Then, the
round bottom flask is put into the heating mantle for
heating process. The temperature and time are left to

Table 1 below shows the concentration of

the standard in part per billion and the peak area for
each of them. The standard one have the no peak area
as it is the most dilute solution, compare to the second
standard and so on. This is because the standard was
prepare from the less dilute which is standard five
which contain the highest catechin concentration.
During analysis process, standard one will be injected
first because it is the most diluted solution. This is
because, if the standard five is injected first, the
column needs to be clean in order to proceed to the
standard four. This is because if the column is not
clean first, it would contaminate the other standard
resulting the inaccurate reading. As it should be, as
the peak area increase, the concentrations of the
standard also increase.

Standards

Concentration
(ppb)

Peak Area

6.83

0.00

75.13

0.19

samples also affected the result because it appeared

that the stability of catechins during long-term storage
was optimum at low temperature (4 C) and acidic pH
(pH 4.0) (Bazinet et al., 2010). Concerning the
temperature during storages, researchs showed that a
lower storage temperature extended appreciably
catechin half-life.

826.45

18.99

Table 2: Peak area for fresh leaves

9090.90

40.82

100000.00

70.78

Table 1: Peak area for the standards catechin

Fresh plants

Table 2 and 3 shows the peak area for each

dried and fresh leaves of the palm oil shoots, pereskia
bleo, coriander, centella and salak. The concentration
of the catechin in the samples can be calculated from
the peak area from the analysis. The temperature and
the time that is kept constant during the extraction are
80 C and 40 minutes respectively. It can be seen that
when the operating temperature was 50 C, the
recovery of the catechin compounds was lower, while
at higher temperatures of 80 C and 100 C, resulted
in higher recovery. However, at 100 C, another peak
labeled as GCG, which is a product of EGCG after
epimerisation (the right peak of EGCG). Therefore an
operating temperature of 80 C was chosen. When the
temperature was 80 C, better recovery at the
extraction time of 40 min could be obtained than that
of 15 min (Row and Jin, 2005). That is why the
temperature is set to 80 C during the extraction
process because it is proven to be the optimum
temperature so do the time for extraction.

Peak area

Palm Oil shoots

0.56

Pereskia Bleo

1.87

Coriander

0.01

Centella

0.24

Salak

0.04

Table 3: Peak area for dried leaves

Dried plants

The results showed the all the dried leaves

samples have more high peak area values than the
fresh leaves samples. According to Liang et al.
(2006), the catechin is easily dissolved in the hot
water treatment. The leaf cell membranes were
further damaged in the subsequent drying process,
which would increase the extractability of catechins.
The leaf might be partially bruised by the stirring
action during pan fixing process and so catechins in
the pan fixed leaf became more easily extracted than
in the fresh leaf. This is the same when the samples
leaves are cut smaller before the drying process. As
Liang et al. (2006) stated that the leaf might be
partially bruised during the drying process and so
catechins in the pan fixed leaf became more easily
extracted than in the fresh leaf. Thats become one of
the reasons when comparing these two results.
Bazinet et al. (2010) stated, when the fresh leaf was
extracted in hot water, compare to the catechin, the
caffeine could diffuse and dissolve into water more
easily than catechins because of its higher solubility
in water and smaller molecule size. The storage of the

Peak area

Palm Oil shoots

8.08

Pereskia Bleo

10.97

Coriander

1.26

Centella

8.52

Salak

0.57

3.2 Calibrations Curve of the Standards

Five standard solutions were prepared with
different concentration in order to determine the
concentration of the catechin in the samples. The
calibration curve for the standard was plotted by area
versus the standard concentration (ppb). As shown in
the graph, the standard linear line is drawn and the
equation of the graph is y = 18.219x 28.501. The
equation then is used to determine the concentration
of the samples.

times. The fresh palm oil shoots takes about 6.301

minutes. This means that the catechin is detected
during that time. Pereskia bleo takes 5.527 minutes in
dried states while for fresh states it takes 5.559
minutes followed by the coriander and centella which
in dried condition it takes about 4.659 and 4.673
minutes respectively while for their retention time in
fresh sates are 5.646 and 5.915 minutes respectively.
The retention time in dried salak is 6.040 and in fresh
states it takes about 5.996 minutes.

80
70.78

70

y = 18.219x - 28.501

60

Peak Area

50
Calibration
Curve for
Standard
Solutions

40.82

40

Table 5: Retention time of samples

30
20

Linear
(Calibration
Curve for
Standard
Solutions)

18.99

10
0

0.19
Std 1 Std 2 Std 3 Std 4 Std 5

Retention Time
(min) for Dried
Plants

Retention Time
(min) for Fresh
Plants

Palm Oil shoots

6.206

6.301

Pereskia Bleo

5.527

5.559

Coriander

4.659

5.646

Centella

4.673

5.915

Salak

6.040

5.996

-10
-20

Types of Plants

Standard Concentration (ppb)

Fig 1: Calibration
concentration

curve

for

the

standard
3.4 Concentration of Catechin in the Samples

3.3 Retention Time

The calibration curve of peak area versus

standard concentration determines the concentration
of the standard in the samples corresponding to their
peak areas by using this standard equation line for the
calibration curve, y = 18.219x 28.501 where y is the
peak areas and x is the concentration in the samples.
These experiments have gives out the value of the
concentration of catechin in the samples but as unit
part per billion is used, the numbers of concentration
of the samples detected are smaller.

The catcehin standard shows the retention

time for the first standard, which is 4.63 minutes. The
second standard shows 5.51 minutes while the third
standard shows 5.46 minutes followed by the next
standard, 5.48 minutes and lastly 6.47 minutes. The
retention time increase proportionally to the
concentration because as the solution becomes more
concentrated, the time needed for the HPLC detector
seeing the component in the standard is longer.

Different graphs have been plotted based on

the equation from the standard line of the
concentration of the standard which can be referring
to the figure 1 above for dry and fresh samples, palm
oil shoot (PS), coriander leaves (CL), centella leaves
(CEL), pereskia leaves (PL) and salak peelings (SP).

Table 4: Retention time of standards catechin

Standards

Concentration
(ppb)

Retention time
(min)

6.83

4.63

75.13

5.51

826.45

5.46

9090.90

5.48

100000.00

6.47

The graph below show the concentration of

the dried samples versus types of plants. The salak
peelings show the lowest concentration which is
1.5956 ppb while the highest shows the pereskia
leaves which 2.1165. The table below shows the peak
area of the sample and its concentration (ppb), where
there is a conclusion can be made which the greater
the area, the higher the concentrations.

Refer to the to the table 5 below, the dry

palm oil shoots takes about 6.206 minutes retention

Concentration of Ctechin in Fresh Samples (ppb)

Concentration of Catechin in Dried Samples (ppb)

3.00

2.50
2.0078

2.032

2.1165

2.00
1.5956

1.6335

1.50

1.00

0.50

3.00
2.50
2.00
1.5649 1.5666 1.5775 1.5954

1.667

1.50
1.00
0.50
0.00

0.00
Salak

Coriander Palm Oil Centella Pereskia

Types of plants

Types of Plants

Fig. 3: Graph of concentration of the fresh samples

versus types of plants

Fig. 2: Graph of concentration of the dried samples

versus types of plants

Table 7: Peak area and concentrations of fresh plants

Fresh plants

Peak area

Concentrations
of Catechin in
dried samples
(ppb)

Concentrations of
Cetchin in fresh
samples (ppb)

Palm Oil shoots

0.56

1.5954

Pereskia Bleo

1.87

1.667

Coriander

0.01

1.5649

Centella

0.24

1.5775

Salak

0.04

1.5666

Table 6: Peak area and concentrations of dried plants

Dried plants

Peak area

Palm Oil
shoots

8.08

2.0078

Pereskia Bleo

10.97

2.1165

Coriander

1.26

1.6335

Centella

8.52

2.032

Salak

0.57

1.5956

4. Conclusion
The objectives of this experiment are to
extract the catechin from different plants (palm oil,
pereskia bleo, coriander, centella & salak) and to
determine the effects of different state of plants on
catechin extraction which is dry and fresh. As results,
research shown that, from all the samples, pereskia
bleo contain the highest concentration from others
both in fresh and dried states. Traditionally, this plant
is eaten raw after the leaf is plucked from the plants.
Pereskia bleo (Kunth) DC. (Cactaceae), commonly
known as Jarum Tujuh Bilah in Malaysia has been
used as natural remedy in cancer-related diseases,
either eaten raw or taken as a concoction brewed from
fresh plant. P. bleo is a spiny shrub which can reach
up 2 to 8 m (Sim et al, 2010).

The graph below shows the fresh samples

where the concentration is sorts by the types of
plants. The coriander leaves have the lowest
concentration of catechin while pereskia bleo, again,
have the highest value. The value can be seen in the
graph below. The details of the fresh samples are
tabulated in the table 7.

The different condition gives effect to the

concentration of the catechin in the plants as the dried
plants give more concentration of the catechin than
the fresh plants. The dried state gives more catechin
than the fresh state mainly because of the processes
involves damaged the leaves cells and produce more
catechin than the fresh leaves. The processes such as
drying, cutting and stirring help produce more
catechin.
Acknowledgements
Sincere gratitude to supervisor, Miss
Norsalliana Binti Shaari and for the one who also
contributes lots of ideas, Miss Norkamruzita Saadon.
Special thanks to UiTM Terengganu for providing
fund to do the research.
References
Bazinet. L., Araya-Farias. M., Doyen. A, Trudel. D.
& Tetu. B., 2010. Effect of Process Unit
Operations and Long-term Storage on
Catechin Contents in EGCG-enriched Tea
Drink. Food Research International, 43:
1692-1701.
Chan. E. W. C., Lim. Y. Y. & Chew. Y. L., 2006.
Antioxidant Activity of Camellia Sinensis
Leaves and Tea from a Lowland Plantation
in Malaysia. Food Chemistry, 102: 1214
-1222.
Chen. Z. Y., Su.Y. L. & Leung. L. K., 2003. Stability
of Theaflavins and Catechins. Food
Chemistry, 83: 189-195.
Craig. W. J., 1999. Health-promoting Properties of
Common Herbs. American Society for
Clinical Nutrition,70: 491s-9s
Graham, H. N., 1992. Green tea composition,
consumption, and polyphenol chemistry.
Preventive Medicine, 21: 334350.
Liang. H., Liang. Y., Dong. J., Lu. J., Xu. H. &
Wang. H., 2006. Decaffeination of Fresh
Green Tea Leaf (Camellia Sinensis) by Hot
Water Treatment. Food Chemistry, 101:
1451-1456.
Row. K. H. & Jin. Y., 2005. Recovery of Catechin
Compounds From Korean Tea By Solvent
Extraction. Bioresource Technology, 97
(2006): 790793.
Sim. K. S., Sri Nurestri. A. M. & Norhanom. A. W.,
2010. Phenolic Content and Antioxidant
Activity of Crude and Fractionated Extracts
of Pereskia Bleo (Kunth) DC. (Cactaceae).
African Journal of Pharmacy and
Pharmacology, 4(5): 193-201.