UNIVERSITY OF CAPE COAST

THE FORMULATION AND EVALUATION OF POLYHERBAL TEA BAGS

AFUA ANYIMEDUA OHEMENG

2025
 UNIVERSITY OF CAPE COAST

THE FORMULATION AND EVALUATION OF POLYHERBAL TEA BAGS

BY

AFUA ANYIMEDUA OHEMENG.

Dissertation submitted to the Department of Pharmacognosy and Herbal

Medicine of the School of Pharmacy and Pharmaceutical Sciences, College of

Health and Allied Sciences, University of Cape Coast in partial fulfillment of

the requirements for the award of Doctor of Pharmacy (Pharm D) degree.

MONTH AND YEAR OF AWARD

 DECLARATION

The work described herein was carried out in the Department of

Pharmacognosy and Herbal Medicine, School of Pharmacy and

Pharmaceutical Sciences, University of Cape Coast. I declare that this project

work is my independent work, and all works of others have been cited

appropriately.

……………………….. …………………

Student's full name Date

(Student)

…………………………… …………………

Supervisor's full name Date

(Supervisor)

iii
 ABSTRACT

Traditional herbal medicines have played a critical role in human

healing practices for centuries, with diverse regions and cultures

around the globe relying on plant-based remedies for their healing

properties (Gupta et al., 2023). This study aimed to formulate a herbal

preparation combining three plant materials, which are Loeseneriella

Africana, Hibiscus sabdariffa, and Pyrus communis. The formulation

was prepared by combining these three plant materials in the ratio

1:2:1, respectively. A general formulation was prepared, which

consisted of all plant materials except pear seed. Qualitative analysis,

such as physicochemical, phytochemical, and microbial tests, was

carried out on the teabag.

iv
 ACKNOWLEDGEMENTS

I thank God Almighty for guidance and strength throughout the work.

I would like to express my sincere gratitude to all the students in my group for

their hard work, dedication, and cooperation throughout the project. Each

member brought their skills to the table, which ultimately led to the success of

our endeavor.

I also want to extend my thanks to our supervisor for guiding us and providing

valuable feedback along the way. Your support and encouragement helped us

stay on track. Your expertise and encouragement were instrumental in helping

us reach our full potential.

v
 DEDICATION

I dedicate this write-up to myself.

vi
TABLE OF CONTENTS

DECLARATION..............................................................................................................iii

ABSTRACT....................................................................................................................iv

ACKNOWLEDGEMENTS................................................................................................v

DEDICATION................................................................................................................vi

CHAPTER ONE..............................................................................................................1

INTRODUCTION............................................................................................................1

1.1 Background of Study....................................................................1

1.2 Statement of Problem..................................................................1

1.3 Justification..................................................................................1

1.4 Aims and Objectives...........................................................................................1

1.4.1 Aim..............................................................................................................1

1.4.2 Specific objectives.......................................................................................2

CHAPTER TWO.............................................................................................................3

LITERATURE REVIEW....................................................................................................3

2.1 Overview............................................................................................................3

2.2 Polyherbal Formulations....................................................................................3

MATERIALS AND METHODS.........................................................................................9

3.1 Materials............................................................................................................9

3.1.1 Instruments and equipment........................................................................9

3.1.2 Chemicals and reagents..............................................................................9

vii
 3.1.3 Test organisms/animals...............................................................................9

3.2 Methods...........................................................................................................10

3.2.1 Collection of plant samples.......................................................................10

3.2.2 Formulation of polyherbal teabag.............................................................10

3.2.3 pH determination......................................................................................10

3.2.5 Bulk and tapped Density...........................................................................11

3.2.6 Microscopy................................................................................................11

3.2.7 Thin Layer Chromatography......................................................................11

3.3 Phytochemical screening..................................................................................12

CHAPTER FOUR..........................................................................................................14

4.0 Results..............................................................................................................14

4.1 Master Formula................................................................................................14

Table 4.1.................................................................................................................14

4.2 Weight of teabags after production.................................................................14

Table 4.2.................................................................................................................14

4.3 Bulk and Tapped Density..................................................................................15

4.4 pH determination.............................................................................................16

Table 4.4 Ph in alcohol...........................................................................................16

Ph in water.............................................................................................................16

4.5 Microscopy.......................................................................................................17

4.6 Refractometry..................................................................................................18

CHAPTER FIVE............................................................................................................19

viii
DISCUSSION, CONCLUSION AND RECOMMENDATIONS.............................................19

5.1 Discussion........................................................................................................19

5.2 Conclusion........................................................................................................19

5.3 Recommendations...........................................................................................20

REFERENCES...............................................................................................................21

ix
TABLE OF TABLES

Table 1: List of instruments and equipment used.----------------------------------4

Table 2: Population of various genders in a popular Ghanaian tertiary

institution---------------------------------------------------------------------------7

x
TABLE OF FIGURES

Figure 1: Distribution of the male population in the various tertiary institutions

---------------------------------------------------------------------------------------8

Figure 2: Gender distribution amongst selected tertiary institutions in Ghana. 8

Figure 3: A diagrammatic comparison of efflux pump families, with examples

of medications and substances that act as substrates (Munita and Arias,

2016).-------------------------------------------------------------------------------9

xi
 CHAPTER ONE

INTRODUCTION

1.1 Background of Study

Herbal teas have garnered widespread acceptance for years due to the
widespread availability of plant materials and the health benefits recognized
by our predecessors. They are consumed as breakfast, refreshments, and also
for their medicinal properties. Polyherbal formulations

1.2 Statement of Problem

Over the years, despite the growing demand for herbal tea formulations, they

have often been designed to contain single constituents. This practice reduces

the therapeutic benefits the formulations could offer if they included a variety

of constituents.

1.3 Justification

The surge of interest in natural products over the years, instead of the orthodox

products, due to costs and other reasons has created a market for these natural

formulations. Introduction and usage of a polyherbal approach allows a

broader range of therapeutic benefits as compared to the limited range a single

constituent formulation would provide.

1.4 Aims and Objectives

1.4.1 Aim

This experiment aims to formulate a safe polyherbal tea using Loeseneriella

Africana, Hibiscus sabdariffa and Pyrus communis as constituents. This blend

would offer broad therapeutic benefits.

1
1.4.2 Specific objectives

i. To formulate a poly-herbal tea bag

ii. To determine the phytoconstituents present in the formulation
iii. To perform natural products evaluations
iv. To ensure microbiological quality and safety of the formulation
v.

2
 CHAPTER TWO

LITERATURE REVIEW

2.1 Overview

Herbal teas are indeed a presentation of herbal medicines. They consist of

mono or poly-herbal materials that are brewed as decoction or infusion and

drank for their therapeutic benefits (Poswal et al. 2019). In recent times, there

has been an apparent upsurge in the popularity and use of herbal teas (McKay

&Blumberg 2002; Khan & Mukhtar 2013). This may be related to the

acclaimed benefits in managing many chronic diseases (Builders2019). Thus,

herbal teas belong to a rapidly expanding market of wellness beverages

(Byeon & Han 2004). The herbal materials that make up the recipe are often

characterized by the presence of various secondary metabolites which are

responsible for their pharmacological activities and health benefits. Indeed,

many folk recipes that are used for managing especially chronic diseases are

currently presented as herbal teas (Cohen & Ernst 2010; Park et al. 2014).

Polyherbal tea bags have gained popularity due to their potential health

benefits and unique flavor profiles. Research suggests that polyherbal teabags

may offer synergistic effects.

2.2 Polyherbal Formulations

Polyherbal formulation (PHF) involves using more than one herb in a

medicinal preparation. Multiple herbs are combined in specific ratios to

formulate potent mixtures for the treatment of illnesses. This concept was

developed in Ayurvedic and other traditional medicinal systems as far back as

AD 1300 (Subramani et al., 2014). However, a major hindrance to the

3
integration of herbal formulations in modern medical practice is the lack of

scientific and clinical data proving their efficacy and safety (I.O. Lawal et

al,2024). The high efficacy of PHF in the treatment of many diseases has

ensured its popularity. They have a wide variety of treatment choices (effective

at low doses and safe at high doses), fewer side effects, and are more

environmentally friendly, cheaper, and readily available. Polyherbal

formulations have a reputation for communicating high viability in a wide

range of ailments. Due to the presence of numerous phytoconstituents,

homegrown medicine has a healing effect. The effects are further amplified

when viable herbals are combined in PHFs (Dwivedi and Daspaul, 2013,

Pushpendra et al., 2022).

2.2.1 Advantages of such formulations

The therapeutic capability of a polyherbal greatly depends on the individual

constituents incorporated to make a blend. For this study, three plant materials

were used in making the formulation. Loeseneriella Africana, Hibiscus

sabdariffa and Pyrus communis were combined to make the formulation,

which we call Hibis Tea. The name Hibis tea was used because hibiscus

formed a greater percentage in the formulation.

2.3 Review of Plant materials

Hibiscus sabdariffa, also known as roselle, is a nutritive and medicinal herb

from the Malvaceae family. It is an annual summer shrub with a deep
penetrating taproot that grows upright and generally branches. Its leaves range
in colour from green to red, and its enormous short-peduncled blooms have a

4
dark centre (Abou-Sreea et al., 2022). The presence of anthocyanins, a
coloured byproduct of the flavonoid pathway, is thought to be responsible for
H. sabdariffa's purported medical benefits. Antioxidant activity has been
discovered in the anthocyanins present in H. sabdariffa, which provide defense
against cancer and atherosclerosis. Additionally, they are related to liver
protection and improved cholesterol action. It is an annual crop utilised in
pharmaceuticals, nutraceuticals, cosmeceuticals, food, and animal feed.
Acidic flavour can be found in the calyces, stems, and leaves. Due to the
calyces' high concentration of vitamin C, anthocyanins, and other
antioxidants, the juice is advertised as a beverage that improves health
(Mohamed et al., 2012). The dried calyx is used to make a tasty and nutritious
beverage. It formed 50% of the formulation and was used in its powdered
form.

Loeseneriella africana is distributed in continental tropical Africa and South

Africa, including countries such as Ghana, Senegal, Ethiopia, Burkina Faso,
Nigeria, Ivory Coast, Togo, Benin, Kenya, Tanzania, Malawi, and Zimbabwe
(Burkill, 1985). It is also distributed in India, Sri Lanka, Myanmar, and Laos.
In Ghana, Loeseneriella africana is distributed in Cape Coast, Obuase, Kade,
and Kwahu (Jiofack-Tofokou and Bosch, 2011). In Ghana, the stem decoction
is used for treating malaria, wounds, oedema, menstrual pains, and in
combination with other plants for infectious diseases and hypertension.
Previous reports show that Loeseneriella africana and related species have
antidiabetic and hypolipidaemic, analgesic, anti-inflammatory, antipyretic,
antioxidant, antidiarrheal, and antinuclear activities. Its elongated stems and
glossy, heart-shaped leaves allow it to weave through the understory, seeking
sunlight. The plant produces small, bell-shaped flowers that may attract
specific pollinators, contributing to its role in the ecosystem. Loeseneriella
africana's ability to thrive in humid, shaded conditions demonstrates its
adaptation to dense forest habitats. The presence of alkaloids, triterpenoids,
phytosterols, flavonoids, tannins, and coumarins, which could be responsible
for the medicinal uses of the plants.

Pyrus communis (for European pear) or Pyrus pyrifolia (for Asian pear) of
thefamily Rosaceae. The grounded seeds were used in formulation. Pear seeds

5
are small, brown, and oval-shaped, located within the core of the pear fruit.
Like apple seeds, they contain amygdalin, a cyanogenic glycoside that can
release small amounts of hydrogen cyanide when metabolized.

However, the quantity in normal consumption is not harmful (Kritchevsky,

2000). Pear seeds are rich in polyphenols, antioxidants, and essential fatty
acids, contributing to their potential anti-inflammatory and antimicrobial
properties (Zhang et al., 2021).

2.4 Phytochemical screening

Phytochemicals(from the Greek word phyto, meaning plant) are biologically

active, naturally occurring chemical compounds found in plants, which
provide health benefits for humans further than those attributed to
macronutrients and micronutrients[1]. They protect plants from disease and
damage and contribute to the plant’s color, aroma, and flavor. In general, the
plant chemicals that protect plant cells from environmental hazards such as
pollution, stress, drought UV exposure, and pathogenic at UV exposure and
pathogenic attack are called phytochemicals. Recently, it has been known that
they have roles in the protection of human health when their dietary intake is
significant. Plants consist of various kinds of chemical constituents that
include tannins, phenols, flavonoids, alkaloids, terpenoids, and saponins,
among many others. They are known as secondary metabolites. (Journal of
Pharmacognosy and Phytochemistry,2013.)

2.4.2 physicochemical parameters (Ph determination)

The pH of consumable products is essential in production evaluation. This

determines the alkalinity or acidity of a formulation, which can affect its

therapeutic effects and shelf life. pH affects the stability of the products, which

6
in turn determines shelf life. A highly acidic product may affect the stomach’s

gastric lining since the stomach already contains acid. A balanced pH is

advised. Hence, it is a crucial parameter for evaluation.

2.4.3 Microbial analysis

The safety of herbal medicines is of great importance from a regulatory

perspective. Plant-derived crude drugs are subject to contamination by
microbial species, which include bacterial and fungal spores as well as viable
forms. If the quality of tea is not managed correctly, it could pose a health
risk.

2.4.4 Thin Layer Chromatography

TLC analysis is also used for the classification of materials on the basis of the
chemicals present in them. Widely used in the analysis and quality control of
crude drugs. Both qualitative and quantitative evaluations can be performed
with this technique for the presence or absence of any particular secondary
metabolite. Generally, TLC helps maintain the safety, efficacy, and
consistency of tea bag formulations.

2.4.5 Microscopy

Microscopy is essential for examining the internal structure, composition, and

inclusions of plant and animal cells in detail. It plays a crucial role in detecting

adulterants and contaminants in herbal preparations, ensuring their

authenticity, purity, and quality. This technique helps in the standardization

and quality control of herbal drugs, making it a valuable tool in

pharmacognosy and drug evaluation.

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2.4.6 Refractometry

The food and beverage industry uses refractometers for measuring the soluble

solids content (SSC) in a solution, including sugar, salt, protein, and acids.

Understanding the content of a food or beverage can help improve its quality

and consistency, ensuring correct composition in each batch.

2.4.7 Bulk and Tapped densities

These parameters affect flowability and storage of the teabag. They ensure
uniform filling of teabags as well. It also ensures products stay stable during
their shelf- life period.

8
 CHAPTER THREE

MATERIALS AND METHODS

3.1 Materials

3.1.1 Instruments and equipment

Table 1: List of instrument and equipment used.

Item Manufacturer/Supplier/Place
Autoclave Arnold and Sons Limited, London, UK
Eppendorf tubes Eppendorf, Hamburg, Germany
Forceps Labkem Ltd, Dublin, UK
Hot air oven Gallenkamp Ltd, London, UK
Incubator Gallenkamp Ltd, London, UK
Laminar air flow cabinet Labconco Corporation, Kansas City, USA
Microscope Holmaec Opto-Mechatronics,Dermark
Hotplate Philips Electronics, China.
Refractometer Atago Co., Ltd, Tokyo, Japan
Chromatank Provided by SOPPs UCC
Beaker Provided by SOPPs UCC
pH meter Provided by SOPPs UCC
Water bath Provided by SOPPs UCC
Petri dishes Provided by SOPPs UCC
TLC plates Provided by SOPPs UCC
UV machine Provided by SOPPs UCC
Weighing Balance Provided by SOPPs UCC

3.1.2 Chemicals and reagents

Vanillin, Phloroglucinol,1% Lead acetate, Conc HCl, Conc H2S04, Alcohol,

Iodine Solution,

3.1.3 Test organisms/animals

No test organisms were used

9
3.2 Methods

Experimental procedures

This section describes step by step the various models and experimental

procedures utilized in conducting the study

3.2.1 Collection of plant samples

Plant samples were readily available in powdered form.

3.2.2 Formulation of polyherbal teabag

The plant materials were weighed in the ratio 2:1:1, with Hibiscus forming

50% of the formulation. The tea bag weighed 2.5g, but room of excess of 0.5g

was made to prevent losses. The plant samples were triturated in a glass

mortar to ensure uniform formulations. The porcelain mortar was used

because of the colours of the plant materials to prevent staining. 2.5g of the

mixture was transferred to the tea bag and sealed. This was done for ten tea

bags and all ten bags were weighed.

3.2.3 pH determination

The herbal tea was extracted with warm water and 100 mL of ethanol using
the infusion method, and was allowed to cool before the pH readings were
taken using a digital pH meter

3.2.4 Refractometry

The refractometer was calibrated with distilled water, an infusion was made

with hot water and allowed to cool to room temperature. A few drops of the

extract were placed onto the refractometer’s prism using a dropper, the prism

10
cover was closed, and the readings were taken thrice, and the average

compared to the refractive index of distilled water.

3.2.5 Bulk and tapped Density

Bulk density

10.002 grams of the powder was weighed and transferred into a measuring
cylinder, the bulk volume was then noted. The bulk density was then
calculated using the formula

Mass/Bulk volume = Bulk density,

Tapped density

The measuring cylinder was tapped on a clean napkin 100 times to get the
consolidated volume. The value obtained after tapping was noted. Tapped
density was calculated by the given formula.

Tapped density = Mass of powder (g) / Tapped volume

3.2.6 Microscopy

Samples of the powders from the dried leaf and stem were mounted with water
and subsequently stained with either phloroglucinol in HCl or N/50 iodine.
Characteristic cell structures and contents, including cork cells, stone cells,
fibres, starch grains, and calcium oxalate crystals observed under the
microscope, were captured and characterized as such (Evans, 2009; Anokwah
et al., 2021)

3.2.7 Thin Layer Chromatography

About 5g of each powdered sample was cold macerated in 50 ml of ethyl

acetate for 24 h. The extract was spotted on TLC silica gel plate. The plates
were developed in pre-saturated chromatank containing Petroleum ether: ethyl

11
acetate (7: 3) as mobile phase. The resolved spots on the developed TLC
plates were dried and observed for characteristic fluorescence and quenching
under UV lamp at 365 nm and 254 nm, and pictures were taken. The
components of the extracts were further determined by spraying the plate with
vanillin in conc. H2SO4 and pictures were taken (Anokwah et al., 2023).

Vanillin is commonly used as a TLC visualization reagent to detect a wide

range of organic compounds. It reacts with functional groups in many
phytochemicals, producing colored spots that help identify and differentiate
compounds.

3.3 Phytochemical screening

Alkaloid Test

Add 1 ml of Dragendorff’s reagent to 2 ml of leaf extract, an orange red

precipitate indicates the presence of alkaloids.

Flavonoids Test

a) Prepare aqueous extracts of the samples provided by maceration.

b) Filter and dip a strip of filter paper in the liquid extracts.

c) Dry and expose to ammonia solution and observe the appearance of an

intense yellow colour which indicates Flavonoids may be present.

Confirmatory Test

a) Dissolve about 0.2g of extract in 1 ml of ethanol.

b) Add few drops of concentrated HCL and magnesium turnings. The
present of flavonoids is detected with the development of a pink or
magenta-red colour

Tannin Test

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 a) Boil 0.4g of powdered drug with 20 ml of water for 5 min. Cool, filter

and adjust the volume of the solution to 20 ml.

b) To 1 ml of the extract, add 10 ml of water and 1 to 5 drops of 1% lead

acetate solution. A white, yellow or amber precipitate indicates that

tannins may be present.

Confirmatory Test

a) To 1 ml of the extract, dilute with 10 ml of water.

b) To 3 ml of 1% gelatine solution, add 2 ml of 10% sodium chloride

solution and then add 3 ml of diluted extract. White precipitate

indicates the presence of Tannins.

Test for saponin

Froth flotation test: 2 ml of sample was added in a test tube with few ml of
water, a froth observed and persisted on constant shaking confirms the
presence of saponins.

Test for coumarins

Few drops of FeCl3 were added in 2 ml of sample; yellow coloration confirms

the presence of coumarins.(Ashish Phuyal et al,2019)

13
 CHAPTER FOUR

4.0 Results

These results were obtained after all experiments were performed.

4.1 Master Formula

To make room for excess, 0.5g was added to the total weight (2.5g) to make it
3g

Scale factor = 3/100 = 0.03 Therefore, new amount to be measured = initial

amount * scale factor

Table 4.1

Plant Initial Scaled Percentage X10 for

material amount Quantity(x0.03 bulk
) production

L.africana 0.625g 0.01875g 25% 0.1875g

Hibiscus 1.25g 0.0375g 50% 0.375g

Pear seed 0.625g 0.01875g 25% 0.1875g

4.2 Weight of teabags after production

Table 4.2

TEABAG WEIGHT

NUMBER

1 2.709g

2 2.704g

3 2.717g

4 2.701g

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 5 2.696g

6 2.687g

7 2.715g

8 2.728g

9 2.722g

10 2.695g

Average weight=2.7074g

4.3 Bulk and Tapped Density

Table 4.3

BULK DENSITY TAPPED DENSITY

22Ml 17mL

21mL 16mL

22mL 15.5mL

Averge 21.6mL Average 16.16mL

Density 0.46g/mL Density 0.618g/mL

15
4.4 pH determination

Table 4.4 Ph in alcohol

EXTRACT 1 2 3

L.africana 5.36 5.29 5.41

Pear seed 4.78 4.50 4.62

Hibiscus 3.37 3.31 3.23

Mixture 3.39 3.35 3.36

Ph in water

EXTRACT 1 2 3

L.africana 5.54 5.61 5.74

Pear seed 5.00 5.29 5.36

Hibiscus 2.21 2.17 2.17

Mixture 2.35 2.34 2.43

16
 4.5 Microscopy

Fig 4.5.1 Pear seed with water Fig 4.5.2 Pear seed with iodine

Fig 4.5.3 Hibiscus with iodine Fig 4.5.4 Hibiscus with water

Fig 4.5.5 L.africana with water Fig 4.5.6 L.africana with iodine

17
 Fig 4.5.7 L.africana with phloroglucinol

4.6 Refractometry

VOLUME 1 2 3

350mL 1.3330 1.3330 1.3330

300mL 1.3330 1.3330 1.3330

250mL 1.3335 1.3335 1.3335

200mL 1.3335 1.3335 1.3335

150mL 1.3335 1.3335 1.3335

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 CHAPTER FIVE

DISCUSSION, CONCLUSION AND RECOMMENDATIONS

5.1 Discussion

This experiment was

5.2 Conclusion

The results showed that…

19
5.3 Recommendations

Provide recommendation on the implementation of the results.

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 REFERENCES
1. Jiofack Tafokou, R.B. and Bosch, C.H., 2011. Loeseneriella africana
(Willd.) N.Halle. In: Brink, M. and Achigan-Dako, E.G. (Eds.)
PROTA (Plant Resources of Tropical Africa/Ressources Vegetables
De L’afrique Tropicale), Wageningen, Netherlands. Accessed 4
December 2019.
2. Abou-Sreea, A. I. B., Roby, M. H., Mahdy, H. A., Abdou, N. M., El-
Tahan, A. M., El-Saadony, M. T., El-Tarabily, K. A., & El-Saadony,
F. M. (2022). Improvement of selected morphological, physiological,
and biochemical parameters of roselle (Hibiscus sabdariffa L.) grown
under different salinity levels using potassium silicate and Aloe
saponaria extract. Plants, 11(4), 497.
3. Mohamed, B. B., Abdelatif, A. S., & Abdelhafiz, A. D. (2012).
Roselle (Hibiscus sabdariffa L.) in Sudan, cultivation and their uses.
Bulletin of Environmental Pharmacology and Life their uses.
Bulletin of Environmental Pharmacology and Life Science, 1(6), 48-5
4.

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