IAR Journal of Agriculture Research and Life Sciences ISSN Print : 2708-5090 | ISSN Online : 2708-5104 Frequency : Monthly Language : English Origin : Kenya Website : https://www.iarconsortium.org/journal-info/iarjals

36

Comparative studies of endogenous tea processed in NTHRI, Shinkiari

Pakistan

Abstract: Green, black and fresh tea leaves are all produced from leaves and shoots

of Camellia sinensis, the only difference is how they are processed. The aim of this study

was to compare the endogenous tea either produced or processed at National tea and High

Value Crops Research Institute Shinkiari. Fresh tea leaves, processed black tea and

processed green tea were selected for comparative analysis i.e. Biological activities and

secondary metabolites composition. Antimicrobial activity was determined by well diffusion

method against bacterial strains Pseudomonas aeruginosa and E.coli. Green tea showed

highest antimicrobial activity against both strains followed by fresh tea leaves and black tea.

Phytochemical analysis revealed the presence of secondary metabolites having medicinal

importance.

Keywords: Comparative Green, black and fresh tea leaves.

INTRODUCTION

Camellia sinensis (L.) is a very branched tree belonging to the

Theaceae family (Duarte & Menarim, 2006). Although it is originated in

China, Tibet and Northern India, today is widely cultivated throughout

the world. In Brazil, the tea plant has been cultivated in the Ribeira Valley, State of São Paulo (Lima et al., 2009).There are lot of chemical

constituents in the Camellia sinensis tea, such as polyphenols,

methylxanthines (caffeine, theophylline and theobromine), vitamins,

amino acids, carbohydrates, proteins, chlorophyll, volatile compounds,

fluoride, minerals, trace elements, and other undefined compounds

(Cabrera et al., 2003). Polyphenols are compounds of great interest

because they present potent antioxidant activity both in vitro and in vivo

due to its reducing properties (Wu & Wei, 2002).

Phenolics compounds are characterized by having one or more aromatic nuclei containing hydroxylated substituents

and/or its functional derivates; a group of phenolic compounds are flavonoids called catechins (Lima et al., 2009; Ashihara et al., 2010). Monomeric primary catechins that are present in high quantity in the teas of Camellia sinensis are:

(-) epigallocatechin gallate, (-) epigallocatechin, (-) epicatechin gallate, epicatechin and catechin (Ho et al., 1992; Nagle

et al., 2006).

The fermentation method used for obtaining the black tea results in the oxidation or condensation of primary

catechins, giving origin to dimers or polymers, called as theaflavins, theasinensins and thearubigins, which provides

peculiar and unique organoleptic characteristics, besides of aroma and color to the teas (Chan et al., 2007; Barcirova,

2010).The present study was undertaken to identify some physicochemical, phytochemical and biological potentialities in

comparison.

MATERIAL AND METHODS

Moisture Content

Moisture content of the green tea powders was determined by drying in an oven maintained at 105°C until

insignificant consecutive weight changes.

Ash Content

The ash content of the samples was determined by standard procedures (AOAC, 1995).

Crude Fiber

The crude fiber content of the samples was determined by standard procedures (AOAC, 1995).

Antioxidant Activity Antioxidant was determined by using dpph method.

Antibacterial Activity

Article History

Received: 11.12. 2020

Revision: 02. 12. 2020

Accepted: 19. 01 .2021

Published: 30. 01. 2021

Author Details Abdul Waheed, Madiha Bashir, Seemab Ali, Naveed Ahmed, Abbas Khan, Hussain Shah

Authors Affiliations Plant Science Division PARC, Islamabad

Corresponding Author* Abdul Waheed

How to Cite the Article: Abdul Waheed, Madiha Bashir, Seemab Ali, Naveed Ahmed, Abbas Khan, Hussain Shah (2021); Comparative studies of endogenous tea processed in NTHRI, Shinkiari Pakistan .

IAR J Agri Res Life Sci, 2(1) 36-40.

Copyright @ 2021: This is an open-access article distributed under the terms of the Creative Commons Attribution license which permits unrestricted use, distribution, and reproduction in any medium for non commercial use (NonCommercial, or CC-BY-NC) provided the original author and source are credited.

Research Article

Abdul Waheed, et al., IAR J Agri Res Life Sci; Vol-2, Iss-1 (Jan-Feb -2021): 36-40.

37

Antimicrobial activity was carried out against

Pseudomonas aeruginosa and E.coli. All microorganisms were obtained from the Microbiology

Laboratory, Hazara University Mansehra. The

antimicrobial assay was carried out by using agar well

diffusion method (Zain et al., 2012).Nutrient agar

media was prepared. The autoclave was carried out at

121°C for 15 minutes. After media plates preparation

they were placed under UV light for more than 20

minutes for sterilization. Inoculum was spread over

plates using wire loop. A 6mm standard cork barrier

was used for well preparation. Sample extracts were

introduced into wells by dropper. The filter paper discs

of specific antibiotics were placed on nutrient agar surface and use as a standard. The inoculated plates

were incubated at 37°C for 24 hours and inhibition zone

was measured in mm by mean of measuring scale after

24 hours.

Phytochemical Screening

Qualitative identification of phytochemicals i.e.

Tannins, Saponins, Flavonoids, Steriods, carotenoids,

Terpenoids, phenol, Alkaloids, phlobatannins and

glycosides were performed by using standard

techniques described by scientists. Extracts were made in two solvents Distilled water and Ethanol.

Test of Tannins

KOH test was performed for determination of

tannins by Evans et al., 2002 method.

Test of Saponins

Forth test was done for detection of saponins by

(Evan et al., 2012).

Test of Flavonoids

Method described by Trease and Evans., 1989.

Test of Steroids

H2SO4 test was performed described by Idu and Igeleke, 2012; Trease and Evan, 1983).

Test of Terpenoids

Method described by Harbone, 1973).

Test of Carotenoids

Detection of carotenoids was performed by method

described by Kudva et al., 1998.

Test of Phenol

Identification of phenol was performed by ferric

test defined by Sofowora, (1993).

Test of Alkoloids

Mayer’s reagent test was done described by (Idu and

Igeleke, 2012).

Test of phlobatannins

Determination of phlobatannins was performed by

test described by Sofowora, (1993).

Test of glycosides

Identification of glycosides was performed by method defined by Sofowora, (1993).

RESULTS AND DISCUSSION

Moisture Content

Moisture content of tea based on nature of tea and

drying time. Polyphenols of the tea eradicated during

process of fermentation as result moisture content is

retained. It was found that fresh tea leaves have high

content of moisture i.e. 40% as it is unprocessed. While processed black tea and green tea have 7.2 and 6%

moisture content.

Table 2. Moisture content of Green tea, Black tea and Fresh tea leaves

Samples Weight (gm) Dry Weight Percentage of moisture content (%

Green tea 5 0.3 6 Black tea 5 0.36 7.2 Fresh leaves 5 3 40

Ash Content

Ash content represents the total mineral content of the tea varied from 52, 50 annd 33 % in green, black and fresh tea

leaves.

Table 3. Ash Content of Green tea, Black tea and Fresh tea leaves

Samples Weight (gm) Dry Weight Percentage of Ash content (%) Green tea 3 1.56 52 Black tea 3 1.5 50 Fresh tea leaves 3 0.99 33

Crude Fiber

Abdul Waheed, et al., IAR J Agri Res Life Sci; Vol-2, Iss-1 (Jan-Feb -2021): 36-40.

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Table 4.Crude fiber of Green tea, Black tea and Fresh tea leaves

Samples Weight (gm) Dry Weight Percentage of Crude fiber (%)

Green tea 1 0.47 47 Black tea 1 0.48 48 Fresh tea leaves 1 0.25 25

Antioxidant activity of tea samples

The antioxidant of tea sample was determined by

using DPPH, free radical scavenging activity. Greater

dpph percentage was depicted in Green tea i.e.

94.4±1.10 follwed by Black tea and Fresh tea leaves.

Table 5. Antioxidant of Green tea, Black tea and Fresh tea leaves

Sample DPPH (%) Total phenol (%)

Green tea 94.4±1.10 33.65±0.07 Black tea 63±1.10 22.2±0.09 Fresh tea leaves 55.10±0.95 38.2±0.02

Zone of inhibition of Black tea against Pathogenic

Strains

Antimicrobial activity of tea varieties produced and

processed at NTHRI was determined by using Agar

well diffusion method, Tea extracts were obtained in

two solvents n-hexane and Aqueous. There

antimicrobial activity was determined against

Pseudomonas aeruginosa and E.coli in 50ppm and 100

ppm concentration.

Black tea also showed antimicrobial activity against

both strains i.e. 10±0.408mm in 50ppm, 13.5±0.408mm

in 100ppm in n-hexane and 15±0.408mm,

15.5±1.080mm in 50 and 100ppm against Pseudomonas

aeruginosa in Aqueous. , while it showed

10±0.408mm, 10.16±0.84mm in 50 and 100ppm in n-

hexane and 14.5±0.408mm, 13.8±0.62mm in 50 and

100ppm in aqueous against E.coli

Table 6. Antibacterial activity of Black tea

Zone of Inhibition(mm) Tea Varieties Pseudomonas aeruginosa E.coli 50ppm 100ppm 50ppm 100ppm n-hexane 10±0.408mm 13.5±0.408mm 10±0.408mm 10.16±0.84mm Aqueous 15±0.408mm 15.5±1.080mm 14.5±0.408mm 13.8±0.62mm

Zone of inhibition of Green tea against Pathogenic

Strains

Green tea showed highest antimicrobial activity against both strains i.e. 20±0.50mm in 50ppm,

18.5±0.708mm in 100ppm in n-hexane and

18±0.708mm, 17.5±1.88mm in 50 and 100ppm against

Pseudomonas aeruginosa, while it showed

12.05±0.70mm, 13.17±0.84mm in 50 and 100ppm in n-hexane and 15.5±0.50mm, 15.8±0.62mm in 50 and

100ppm in aqueous against E.coli,

Table 7. Antibacterial activity of Green tea

Zone of Inhibition(mm) Tea Varieties Pseudomonas aeruginosa E.coli

50ppm 100ppm 50ppm 100ppm n-hexane 20±0.50mm 18.5±0.708mm 12.05±0.70mm 13.17±0.84mm Aqueous 18±0.708mm 17.5±1.88mm 15.5±0.50mm 15.8±0.62mm

Zone of inhibition of Fresh tea leaves against Pathogenic Strains

Fresh tea leaves showed maximum antimicrobial

activity against both strains i.e. 16.50±2.12 in 50ppm,

17.50±3.12 in 100ppm in n-hexane and 10.08±1.08,

11.08±0.38 in 50 and 100ppm against Pseudomonas

aeruginosa in Aqueous. , while it showed 14.50±2.12,

16.50±1.00 in 50 and 100ppm in n-hexane and

7.32±1.13, 8.41±1.12 in 50 and 100ppm in aqueous

against E.coli.

Table 8. Antibacterial activity of Fresh tea leaves

Zone of Inhibition(mm)

Tea Varieties Pseudomonas aeruginosa E.coli 50ppm 100ppm 50ppm 100ppm n-hexane 16.50±2.12 17.50±3.12 14.50±2.12 16.50±1.00 Aqueous 10.08±1.08 11.08±0.38 7.32±1.13 8.41±1.12

Abdul Waheed, et al., IAR J Agri Res Life Sci; Vol-2, Iss-1 (Jan-Feb -2021): 36-40.

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Fig 1. Zone of inhibition of against Pathogenic Strains

Secondary metabolites of different processed tea

Green tea, Black tea and Fresh leaves were analyzed for phytochemical analysis. Two solvents were used

i.e. Ethanol and aqueous. Tannins, steroids, Reducing

sugar, phenols and phytosterols were found positive in

all sample of tea in both solvents. Carotenoids and

phlobatannins were found absent in all extracts of

black, green and fresh tea leaves. Saponins, alkaloids

and flavonoids were absent in ethanolic extracts of all tea samples while found positive in aqueous solvent.

Glycosides and terpenoids were found positive in

aqueous and ethanol extracts of almost all samples

except seen negative in black tea (ethanolic extracts and

aqueous extract).

Table.1 Phytochemical screening of Green tea, Black tea and Fresh tea leaves

Secondary Metabolites Extract Green tea Black tea Fresh leaves

Tannins Aqueous + + +

Ethanolic + + +

Saponins Aqueous + + +

Ethanolic - - -

Alkaloids Aqueous + + +

Ethanolic - - -

Flavonoids Aqueous + + +

Ethanolic - - -

Phenol Aqueous + + +

Ethanolic + + +

Glycosides

Aqueous + - +

Ethanolic + + +

Phlobatannins Aqueous - - -

Ethanolic - - -

Steroids Aqueous + + + Ethanolic + + +

Terpenoids Aqueous + + +

Ethanolic + - +

Reducing Sugars Aqueous + + +

Ethanolic + + +

Gums and Mucilages Aqueous + - +

Ethanolic + + +

Carotenoids Aqueous - - -

Ethanolic - - -

Phytosterols Aqueous + + +

Ethanolic + + +

+ = positive/ present - = negative/ absent

CONCLUSION

In present study selected tea varieties were

investigated, these tea varieties processes maximum

antimicrobial and antioxidant activities and are good

source of phytochemicals.

Presence of essential phytochemicals like alkaloids,

flavonoids, phlobatannins, glycosides, carotenoids

showed that it served as valuable material for

medicines.

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