(Luận văn tốt nghiệp) research of the instant tea production process from myxopyrum smilacifolium blume plant

THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY NGUYEN THI THU THAO RESEARCH OF THE INSTANT TEA PRODUCTION PROCESS FORM MYXOPYRUM SMILACIFOLIUM BLUME PLANT BACHELOR THE

THAI NGUYEN UNIVERSITY

UNIVERSITY OF AGRICULTURE AND FORESTRY

NGUYEN THI THU THAO

RESEARCH OF THE INSTANT TEA PRODUCTION PROCESS FORM

MYXOPYRUM SMILACIFOLIUM BLUME PLANT

BACHELOR THESIS

Study Mode : Full-time

Major : Food Technology

Faculty : Advanced education program office

Batch : 2016 - 2020

Thai Nguyen, 2020

Thai Nguyen University of Agriculture and Forestry Degree Program Bachelor of Food Technology

Student name NGUYEN THI THU THAO

Student ID DTN1654190015

Thesis Title Research of the instant tea production process from

Myxopyrum smilacifolium Blume plant

Supervisor MsC Dinh Thi Kim Hoa

Supervisor’s

signature

Abstract:

The research focused on the process of producing instant tea from

Myxopyrum smilacifolium plants with a laboratory scale, the research extraction method for extracting total polysaccarides from Myxopyrum smilacifolium root The instant tea product was made from solid Myxopyrum smilacifolium root

extract and other herbal extracts with the sensory quality was evaluated by the sensory panel and had the quite good quality

Keywords : Myxopyrum smilacifolium, instant tea, process, product

Number of pages:

Date of submission:

ACKNOWLEAGEMENT

I would like to express my sincere thanks to the teachers of Thai Nguyen University of Agriculture and Forestry and especially the and teachers in the AEP Faculty, the Faculty of Biotechnology and Food Technology for giving me the opportunity to interact and learn asked during the four years

I would like to thank Ms Dinh Thi Kim Hoa, a lecturer in the Department of Biotechnology and Food Technology, who always gave me a wholehearted guidance

in the process of studying and doing graduate research

I would like to thank teacher Luu Hong Son and teachers and friends in the Department of Food Technology Biotechnology for guiding and supplementing my knowledge of laboratory skills, always creating favorable conditions for you in the research process

Finally, I am extremely grateful to my family and friends for cheering me on during my graduation thesis

Thai Nguyen, December, 02, 2020

Sincerely

Nguyen Thi Thu Thao

TABLE OF CONTENTS

Acknowleagement iii

Table of contents iv

List of table vii

List of figuges ix

List of abbreviations x

Part I Introduction 1

1.1 Research rationale 1

1.2 Research objective 2

1.2.1 Overall objective 2

1.2.2 Detail objectives 2

1.3 Scientific and practical significance 2

1.3.1 The scientific meaning of the topic 2

1.3.2 The practical meaning of the topic 2

Part II Literature review 4

2.1 Overview about Myxopyrum smilacifolium (Wall.) Blume 4

2.1.1 Characteristics of Myxopyrum smilacifolium (Wall.) Blume 4

2.1.2 Overview of the chemical compositions of Myxopyrum smilacifolium (Wall.) Blume 6

2.1.3 Overview of biological effects (pharmacology) of Myxopyrum smilacifolium (Wall) Blume. 8

2.2 Overview of additional material for the production of instant tea 10

2.2.1 Lactose 10

2.2.2 Stevia rebaudiana Bertoni 11

2.2.3 Glycyrrhixa glabra L 13

2.3 Overview about extraction process 15

2.3.1 Overview of extraction solvents, extraction methods 15

2.3.2 Overview of extraction solvents 15

2.3.3 Extraction method 16

2.3.4 Extract 16

2.4 Overview about instant tea 17

2.4.1 Soluble tea products 18

2.4.3 The expected production process for soluble tea from Myxopyrum smilacifolium Blume 20

Part III Research contents and methods 22

3.1 Research object and research scope 22

3.1.1 Research object 22

3.1.2 Research scope 22

3.2 Place and time to proceed 22

3.3 Chemical, equipment 22

3.4 Research contents 23

3.5 Research methods 23

3.5.1 Experimental design method 23

3.5.2 Analytical method 28

3.5.3 Data statistical analysis methods 31

Part IV Results and discussion 32

4.1 The results of research results on the effect of ultrasonic treatment time on the efficiency of total polysaccharide from Myxopyrum smilacifolium Blume 32

4.2 The results of study on selecting solvent concentration to extract polysaccharides from Myxopyrum smilacifolium Blume 32

4.3 The results of selecting the ratio of raw materials Myxopyrum smilacifolium Blume/solvent 33

4.4 Research results of selecting the time to extract total polysaccharides from Myxopyrum smilacifolium Blume 34

4.5 Research results of selecting temperature for extraction of total polysaccharides from Myxopyrum smilacifolium Blume 35

4.6 Research on choosing the suitable ratios of Myxopyrum smilacifolium Blume extract and additive ingredients 35

4.7 The results of the research on the drying temperature of mixture of Myxopyrum smilacifolium Blume extract and additive ingridients to produce instant tea 37

4.8 The complete process for producing instant tea from Myxopyrum smilacifolium

Blume 38

Part V Conclusions and recommendation 42

5.1 Conclusions 42

5.2 Recommendation 42

REFERENCES 43

LIST OF TABLE

Table 3.1 Experiment chemicals 22 Table 3.2 Experimental equipment 22 Table 3.3 Laboratory instruments 23 Table 3.4 Experimental design to study the effect of ultrasonic treatment time on the

extraction efficiency of polysaccharides in Myxopyrum smilacifolium

Blume 24 Table 3.5 Experimental design to select solvents concentration for extracting

polysaccharides from Myxopyrum smilacifolium Blume 24 Table 3.6 Research on selecting the ratio of Myxopyrum smilacifolium Blume material

with solvent 25 Table 3.7 Experimental study on selecting the time of extraction 26 Table 3.8 The effect of temperature of extraction on efficiency of polysaccharides

extracted 26

Table 3.9 The mixing ratio of Myxopyrum smilacifolium Blume extract and herbal

additive extracts 29 Table 3.10: Experimental arrangement to investigate the effect of drying temperature

on instant tea sensory quality 28 Table 3.11 Instant tea weight coefficient for sensory evaluation 31 Table 3.12 Scores of quality levels 31 Table 4.1 Results about the effect of the ultrasonic treatment time on the efficiency of

extraction total polysaccharide from Myxopyrum smilacifolium Blume 32

Table 4.2 The effect of the solvent concentration on the efficiency of extracting total

polysaccharides from Myxopyrum smilacifolium Blume 33

Table 4.3 Effects of material/solvent ratio on efficiency of extracting total

polysaccharides in Myxopyrum smilacifolium Blume 33

Table 4.4 Effect of time on the efficiency of extracting total polysaccharides in

Myxopyrum smilacifolium Blume 34

Table 4.5 Effect of temperature on extraction efficiency of polysaccharides in

Myxopyrum smilacifolium Blume 35

Table 4.6 The effects of ratios of Myxopyrum smilacifolium Blume extract and

additive ingredients on the sensory quality of the product 38 Table 4.7 The effects of drying temperature on sensory quality of the product 37 Table 4.8 Ash and moisture contents of complete instant tea product 43

LIST OF FIGUGES

Figure 1 Myxopyrum smilacifolium Blume 4

Figure 2 Structural formula of lactose 11

Figure 3 Lactose powder 11

Figure 4 Stevia rebaudiana Bertoni 12

Figure 5 Glycyrrhixa glabra L 14

Figure 6 Instant tea products 18

Figure 7 Diagram of instant tea production process 39

LIST OF ABBREVIATIONS

TCVN Vietnam standard

PART I INTRODUCTION 1.1 Research rationale

Modern society leads to the increasing demand of people for quality of life, so many products are created to meet the needs of buyers

Tea is a cultural feature of Asia and has a very long tradition associated with history There are some studies that have found that tea originated from the Southwest China It is a drink bearing the essence of Vietnam in particular and Asia in general, and is a cultural feature of each region Tea used worldwide is also the second most popular beverage in the world after drinking water Currently, in Vietnam as well as in the world, the trend of using medicinal plants of natural origin is increasing Drink tea properly to help quench your thirst as well as digestion, avoid drowsiness, stimulate kidney function, enhance eyesight, help clear mind, dispel drowsiness [38]

Tea products are increasingly diversified and diversified from quality and design Besides the teas from the dried leaves, there are also tea bags and instant tea In an era

of development where time is not enough, the demand for beverages such as tea needs

to be convenient and time-saving, instant tea is really essential, and soluble tea proves

to be a lot of benefits It is beneficial for health and suitable for many customers and is extremely potential product for development

Since ancient times, plants belonging to ginseng group have been a precious herb, also known as a miracle herb, with a miraculous effect that ancient people recorded Indeed, researchers have revealed many valuable pharmacial features of these plants such as enhancing vitality, fostering health, preventing disease, prolonging life Currently there are many types of ginseng all over the world, based on the

natural conditions of Vietnam and Myxopyrum smilacifolium Blume has been found almostly in the Northwest region of Viet Nam Myxopyrum smilacifolium Blume

leaves have also been shown to contain compounds belonging to the group of alcaloid, carbohydrates, steroids, saponins, terpenoids, flavonoids, tannins and polyphenols [1], [2]

By blending some herbal extracts with Myxopyrum smilacifolium Blume extract,

the soluble tea is obtained with many health benefits and extremely useful benefits, promoting economic development to create jobs for workers, making the tea market diversified , meeting the demand for health and natural products Advantages of

Myxopyrum smilacifolium Blume are known to many people for its use and benefits

Based on discussion above, I conducted the project "Research on the process of

producing instant tea from the Myxopyrum smilacifolium Blume plant"

1.2 Research objective

1.2.1 Overall objective

The overall objective of the research is to build the process of producing instant

tea from Myxopyrum smilacifolium Blume plant, test production on a laboratory scale

and supply to the market to create high quality tea products, bring economic efficiency, contribute to the diversification of tea products in the market and increase

the application of Myxopyrum smilacifolium Blume plant

1.2.2 Detail objectives

- Determining the influence of the suitable conditions for the following factors: time, temperature, solvent extracted, material/solvent ratio and concentration solvent

on the amount of total polysaccharides of extraction;

- Identifying the mixing ratio of Myxopyrum smilacifolium Blume and addtive

ingredients to create instant tea;

- Completing the process of producing soluble tea from Myxopyrum smilacifolium Blume extract

1.3 Scientific and practical significance

1.3.1 The scientific meaning of the topic

- To provide more scientific information about the tree Myxopyrum smilacifolium

Blume

- To find out suitable parameters for the production process of instant tea from

Myxopyrum smilacifolium Blume plant at laboratory scale

1.3.2 The practical meaning of the topic

- To diversify products from Myxopyrum smilacifolium Blume to increase the

value of this kind of herbs

s;

- To create new products with good quality, good for the health of consumers and reasonable price;

- To create a new research direction on Myxopyrum smilacifolium Blume, and

improve the output for this herbs

PART II LITERATURE REVIEW

2.1 Overview about Myxopyrum smilacifolium (Wall.) Blume

2.1.1 Characteristics of Myxopyrum smilacifolium (Wall.) Blume

Myxopyrum smilacifolium Blume is a large woody twining shrub which is a

plant of the genus Myxopyrum, the Jasmine family (Oleaceae) In addition,

Myxopyrum smilacifolium Blume is also known as Nhuong Le Kim Cang which is a

plant capable of growing through rocks and precious herb only found in old forests in the Northwest region of Viet Nam such as Ha Giang, Lao Cai, Lai Chau where there is appropriate soil This plant lives mainly on the rock crevices of limestone mountains The main stem and roots develop under the black humus soil caused by the

decaying leaves of the Myxopyrum smilacifolium Blume tubers formed by the main

trunk, each main stem section will form 2 - 4 roots with a length of 3 - 8cm

The genus Myxopyrum includes 4 species originally distributed in tropical and subtropical regions of East Asia Its roots, stems, and leaves have a ton of medicinal effects and are used in many traditional medicine systems, especially in India The roots are used to treat diseases like scabies, cough, rheumatism, fever, and to treat wound healing Leaves are astringent, sweet, thermogenic, used as an analgesic, allergy, cooling, and tonic They are used in treating coughs, asthma, rheumatism, headache, fever, ear disease, and neuralgia [20]

Figure 1 Myxopyrum smilacifolium Blume

Classification of the genus Myxopyrum [39]

Domain: Eukaryota

Genus: Myxopyrum Blume

Species: Myxopyrum smilacifolium Blume

The stem of Myxopyrum smilacifolium Blume is Shrub, four-sided low stem,

rarely rounded Leaves are simple, opposite, oval - broad oval, the entire margin of one

or a half leaf, lateral veins 2-3 pairs stem from the leaf base, hairless, or with short, thin hairs The panicle clusters on leaf axils, many flowers Small flowers, bisexual, shaped cup, 4 lobes The color is yellow or pink, the tube is longer than the lobe or bell-shaped with the tube shorter than the lobe, 4 lobes, ovoid or broad, hairless Secondly, flowers are attached near the root of the colon Fruits have from 1 to 4 seeds [3]

The genus Myxopyrum is distributed in tropical and subtropical regions of Asia

such as Bangladesh, Cambodia, India, Indonesia, Laos, Malaysia, Myanmar, New Guinea, Philippines, Thailand, China, Vietnam [21], [22], [23]

Distribution: There are 4 species in the world, distributed in Asian regions such

as Bangladesh, Cambodia, India, Indonesia, Laos, Malaysia, Myanmar, New Guinea,

Philippines, Thailand, China; There are 3 species in Vietnam: M pierrei, M nervorsum, M smilacifolium [4], [3]

Overview of Myxopyrum smilacifolium (Wall.) Blume

Synonyms: Myxopyrum serratulum A.W.Hill [23], [24], or Myxopyrum ellipticifolium H.T Chang Vietnamese names: Nhuong Le Kim Cang, Duong Le Kim

Cang [3]

Distribution of Myxopyrum smilacifolium Blume

In the world: Myxopyrum smilacifolium (Wall.) Blume is distributed in regions

such as Hainan, Bangladesh, Cambodia, India, Assam, Laos, Myanmar, Thailand, Vietnam, Andaman, and the Nicobar Islands

In Vietnam, Myxopyrum smilacifolium is distributed in some areas such as Yen

Bai, Quang Ninh, Da Nang, Binh Duong, Thua Thien Hue, Dong Nai, Ho Chi Minh City, Ha Giang [3], [6]

2.1.2 Overview of the chemical compositions of Myxopyrum smilacifolium (Wall.) Blume

In recent years, in the world, particularly in India, there have been many studies

on the chemical compositions of the species Myxopyrum smilacifolium (Wall.) Blume,

including many published on chemical compositions of leaves and a few publications related to the stem and roots of this species

A various of preliminary surveys of Samu J et al 2014 showed that in the leaves

of Myxopyrum smilacifolium (Wall.) Blume has the presence of alkaloid, carbohydrate,

saponins, terpenoid, flavonoids, steroids, tannin, and polyphenols As published by

Raveesha Peeriga et al (2016), the leaves of Myxopyrum smilacifolium (Wall.) Blume

contains alkaloids, glycoside, tannin, saponin, terpenoid, carbohydrates, and

non-volatile oils [25] According to Vijayalakshmi (2016), the leaves of Myxopyrum smilacifolium (Wall.) Blume contain terpenoids, flavonoids, saponins, tannins, glycosides

and iridoid; in aqueous extracts, 26 compounds were identified [26]

Preliminary survey studies of Rajalakshmi K and Mohan VR (2016) have published in the ethanol extract of the body with alkaloid, catechin, coumarin, flavonoid, tannin, saponin, steroid, phenol, glycoside, terpenoid, xanthoprotein and

identified Identified 32 different compounds in the extract of the stem Myxopyrum smilacifolium (Wall.) Blume

Research by Praveen R.P and Ashalatha S.N (2014) has preliminarily

determined the chemical composition of Myxopyrum smilacifolium (Wall.) Blume

root has alkaloid,phenolic, glycoside, tannin, flavonoid and some other compounds

In 2015, the authors also published results of FTIR spectroscopy to compare functional groups in methanol extract of roots, fruits, and callus showed that all three extracts contain compounds carrying the same functional groups each other is alcohol, alkanes, alkenes, aldehydes,ketones, amines, carboxyl groups, alkyl halides

Iridoid: According to author Siju E.N (2017), the main active chemical

ingredient in Myxopyrum smilacifolium (Wall.) Blume is the iridoid glycosides From

the ethanol extract of the leaves Myxopyrum smilacifolium (Wall.) Blume, Henrik

Franzyk et al (2001) isolated iridoid glycosides: Myxopyrosid (9) and 6-o-acetyl-7-o- (E / Z ) –p methoxycinnamate myxopyrosid [21]

Alkaloid: In 2012, Gopalakrishnan S and Rajameena R isolated compounds dimethoxy-13-carbomethoxy-15-(3,4,5-trimethoxybenzoxy)-13,14

7,8-didehydroalloberban from leaf ethanol extract [27], [28] From the extract of the body

Myxopyrum smilacifolium (Wall.) Blume, by chromatography with mass spectrometry,

Rajalakshmi K and Mohan V.R (2016) determined the presence of 3 alkaloids: phenyl-3,5-pyrazolidinedion, 1-allyl-6,7-dimethoxy-3,3-dimethyl-1,2,3,4-tetrahydro isoquinoline, 3-dodecyl-4 - [((4-methoxybenzyl) oxygen] -2-methyl quinoline[29]

4-Research results of Sudharmini D and Ashalatha S N (2008) show that Myxopyrum smilacifolium (Wall.) Blume contains triterpenoid ursolic acid with a concentration of

0.175 mg/ml Research by Rajalakshmi K et al (2016) showed the presence of

triterpene compounds from Myxopyrum smilacifolium (Wall.) Blume: α- amyrin,

β-amyrin, betulin

Organic acids and derivatives: In 2016, authors Maruthamuthu V and Kandasamy R discovered the presence of many organic acids and derivatives from

Myxopyrum smilacifolium (Wall.) Blume [26] Besides, the research also found many

compounds belonging to this group In addition to the above main groups, the study

also identified that in the stem of Myxopyrum smilacifolium (Wall.) Blume also

contains steroid compounds 9.19-cyclocross-24 (28) -en-3-ol-4,14-dimethyl acetate, phenolic 4 - ((1E) -3-Hydroxy-1-propenyl) -2-methoxyphenyl, carbohydrates D-mannopyranose, ethyl α-D-glucopyranose, phytol,galactitol, amine (3-nitrophenyl) methanol isopropyl ether, vitamins E DL-tocopherol,aldehydes 5-hydroxymethylfurfural, ketones 7-ethyl-4-de-6-on…

Myxopyrum smilacifolium (Wall.) Blume also contains amines 2 [p-

(dimethylamino) phenyl] -1- (p-nitro phenyl cyclopropan carbonitril, 1 - [((2-methoxy phenyl) methyl] - 1,6-hexanediamin, anthranoid 1-phenyl-9,10-anthracenedion, molyb derivatives molybdenumcarbonyl- (2,4cyclopentadien-1-yl) - [P, Pdiphenyl-N- (1-phenylethyl) phosphinous amid] nitrosyl stereoisomer [28], derivatives of siliconic acid silicon, octamethyl cyclotetrasiloxane, dodecamethyl cyclohexasiloxane, naphthalene

hydrocarbon compounds, 2- methylbicyclo [2,2,1] heptane, neophytadien, 2- alcohols (cyclohexen-1-yl) ethanol, 2-pentadecyn-1 ol [26]

Currently, there is only an initial study of author Nguyen Minh Luyen (2017)

published on the chemical composition of Myxopyrum smilacifolium root, in the root

contains saponins, flavonoids, polysaccharides, amino acids, free reducing sugars, and feces established myxopyrosid compound from n-butanol fraction of root extract Thus, up to now, there has not been any research published in-depth about the

chemical composition of Myxopyrum smilacifolium growing in Vietnam

2.1.3 Overview of biological effects (pharmacology) of Myxopyrum smilacifolium (Wall) Blume

Recent studies in the world demonstrate that the species Myxopyrum smilacifolium (Wall.) Blume has remarkable pharmacological effects For example

Anti-oxidant [20], antibacterial, antifungal [30], [31], lower fever, anti-bronchodilator [32], wound healing[19], trigger toxic to cancer cells

Antibacterial, antifungal effect: In 2008, Sudharmini D and Ashalatha S N demonstrated triterpenoids isolated from the leaves of Myxopyrum smilacifolium (Wall.) Blume was resistant to Gram-positive bacteria Staphylococcus aureus, Bacillus subtilis, and Candida albicans, C glabrata

Antifungal activity is similar to antifungal drugs of chemical origin The results

of testing the antimicrobial activity of petroleum extract, benzene, chloroform, ethanol,

and water of Myxopyrum smilacifolium (Wall.) Blume by Gopalakrishnan S et al

(2012) showed that ethanol extract exhibits the best resistance to the test microorganisms The minimum inhibitory concentration (MIC) of ethanol extract ranges from 200 - 300 µg/ml for different types of microorganisms This study helps to elucidate the use of traditional medicinal herbs to heal wounds [30]

A 2014 study by Praveen et al compared the antimicrobial activity of methanol

extracts from roots, callus, and Myxopyrum smilacifolium (Wall.) Blume showed that:

+Antibacterial effect: All extracts have similar inhibitory effects of

Enterococcus faecalis and Bacillus subtilis The fruit extract was more effective against the bacterial strains tested except Staphylococcus aureus and at a concentration

of 50 µg/ml had a stronger effect against E faecalis than gentamycin at 20 mg/ml The

callus is effective against S.aureus but at concentrations greater than 100 µg/ml Root extract and callus tissue have no effect on Escherichia coli

+ Antifungal effect: Root extract does not inhibit Candida albicans callus and

fruit extract at 100 µg/ml had significant inhibitory activity but was weaker than clotrimazole 20 mg/ml [31]

Anti-oxidant effect

In 2013, the testing results of Rani T.S et al illustrate that the methanol extract

of the upper part of Myxopyrum smilacifolium (Wall.) Blume had a significant

antioxidant effect, with a test dose of 250 µg/ml MeOH extract showing a DPPH free radical scavenging effect 81.75% and nitric oxide and 84.95%, nearly equivalent to vitamin C at the same dose level (respectively 83.45% and 84.20%) This indicates that this is a potential source of antioxidants to prevent the progression of oxidative stress

As published by Siju E.N et al (2015), ethanol extract and water of the leaves

of Myxopyrum smilacifolium (Wall.) Blume has antioxidant activity but is inferior to

vitamin C By Praveen R.P and Ashalatha S N (2015) compared the antioxidant

effects of methanol extracts from roots, callus, and fruits of Myxopyrum smilacifolium (Wall.) Blume The results showed that all extracts showed significant DPPH free

radical scavenging capabilities In particular, the root extract has the highest DPPH and NOS inhibitory effects with IC50 of 37.5 µg / ml and 525 µg/ml, respectively, while the fruitextract has the inhibitory effect of hydroxyl radical with IC50 of 950 µg/ml [20]

Anti-inflammatory and antipyretic effect: Some research has shown that Myxopyrum smilacifolium (Wall.) Blume has a good antipyretic and anti-inflammatory

effect In 2014, research by Jolly Samu and fellow showed that ethanol extract and

water extract from Myxopyrum smilacifolium (Wall.) Blume both inhibited protein

denaturation and stabilized red blood cell membranes in test concentrations

In particular, ethanol extract at a concentration of 200 µg/ml inhibits 91.54% protein denaturation, almost equivalent to diclofenac concentration of 100 µg/ml; a water extract at a concentration of 200 µg/ml inhibits 69.60% hemolysis, stronger than

an ethanol extract at a concentration of 100 µg/ml (46.58%) but weaker than diclofenac at a concentration of 100 g/ml (97,059 %) [18]

The anti-inflammatory effect of extract dose of 400 mg/kg is higher than that of

200 mg/kg but lower than that of diclofenac at 10 mg/kg Ethanol extract with a dose

of 400 mg/kg has a stronger antipyretic effect than the dose of 200 mg/kg but is weaker than paracetamol at a dose of 150 mg/kg

Uses of Myxopyrum smilacifolium Blume

In the world: Myxopyrum smilacifolium (Wall.) Blume is an important medicinal

plant, widely used in the indigenous medicine system in India Roots are used to treat conditions such as scabies in children, cough, rheumatism, fever, and wounds Leaves are astringent, sweet, thermogenic, analgesic, allergic, cooling, and tonic Leaves are used to treat coughs, asthma, arthritis, headaches, allergies, fever, ear diseases, neuralgia, and wounds [36], [25], [20] In addition, the leaves are used as an antiseptic

to treat skin diseases [35] In India, people also use Myxopyrum smilacifolium (Wall.)

Blume leaves to alleviate itching, skin rashes, scabies, and helminth infections In

Thailand, the root Myxopyrum smilacifolium (Wall.) Blume is used for reducing fever

In Bangladesh, Myxopyrum smilacifolium (Wall.) Blume root is used to treat bronchial

asthma [34]

In Vietnam, there are no official documents on the use of this species According

to author Vo Van Chi, the leaves of Myxopyrum smilacifolium (Wall.) are used to treat

neurological disorders, rheumatism, asthma, cough

2.2 Overview of additional material for the production of instant tea

2.2.1 Lactose

Definication: Lactose has scientific name as O- β –D-galactopyranosyl- (1 → 4)

-D-glucopyranose A disaccharide, made up of one β D- galactose molecule and one β D-glucose molecule; linked together by β- 1-4 glicozid bonds, abundant in mammalian milk

Properties: Lactose is easy to dissolve in water, mild sweet taste, pleasant,

neutral and less hygroscopic, easy to granulate, easy to dry

Role: During digestion, lactose is hydrolyzed to galactose and glucose In which

galactose is an important nutrient in creating myelin membranes, it encapsulates nerves and speeds up the transmission of nerve signals, so it plays an important role in brain development Lactose is also considered a smart sugar Lactose is also used as substrate for beneficial dairy bacteria, facilitating them to multiply and eliminate

potentially pathogenic bacteria, helping to maintain the stomach environment, and make the stomach and intestines healthy and strong The final product of lactose metabolism is lactic acid, which is significant in helping to acidify the gut and aid in protein digestion [7]

- In the DEF pharma report, lactose is safe for diabetics, because of the amount

of carbohydrates consumed in the diet of the diabetic, as the amount of carbohydrates

in the diet of the diabetic can be up to 255g while the amount of lactose added to the drug is very small In addition, lactose is absorbed and digested more slowly than saccharose and glucose Furthermore, lactose does not cause tooth decay by enzymes The end product of lactose metabolism is lactic acid, which is significant in helping to acidify the gut and aid in protein digestion [7]

In general, lactose is used as a popular pharmaceutical adjuvant because of its low cost, availability, low hygroscopicity, chemical stability, mild sweetness, complete water solubility and minimal incompatibility with pharmaceutical substances and other excipients α-lactose is an adjuvant used in the manufacture of solid drugs such as tablets which are prepared by wet granulation or straight pressing (pelleting without moisture or heat treatment), capsules or powders inhaler (α-lactose acts as a carrier) β-lactose is used as a filler and stick in the straight pelletizing process

Figure 2 Structural formula of lactose Figure 3 Lactose powder

2.2.2 Stevia rebaudiana Bertoni

Origin :

- Scientific name: Stevia rebaudiana Bertoni

- Stevia is native to South America, commonly known as molasses, sugar

grass It was brought to Europe in 1887 Naturally native to the Amambay and Iquacu regions of the Brazilian and Paraguay border, today is grown in abundance around the world and is used as a medicinal herb its The Asians took the herb to dry and used it in everyday life as a herbal remedy, the part used mostly on the ground

Figure 4 Stevia rebaudiana Bertoni

Botanical characteristics:

- Genus Stevia has more than 100 varieties, but only Stevia rebaudiana Bertoni has sweet taste and can be grown Stevia is a perennial herb, grown more than 6

months and start to turn to wood after 6 months

- Roots: The roots of perennial Stevia are very strong, with feathers from 0-30cm

at a depth of 20-30 cm from the ground, spreading cluster roots with a diameter of 40cm

- Stem: dusty stem, round branches, hairy young stems, green old body purple with average height of either 50-60 cm or 80-90 cm when well developed with a diameter of 5-8mm

- Leaves have three prominent veins, the annex is feather-shaped, the leaf edge is serrated Both stems and leaves taste very sweet The first flowers grow at the leaf edge and cluster in the tops [9]

Stevia rebaudiana Bertoni flowers are clustered in a sphere containing many small ivory-white tubular flowers Each flower cluster is usually about 2cm long and has a gentle scent Flowers bloom in October and last until February next year [8], [9]

Distribution: Nowadays, Stevia is grown in many countries such as Brazil,

China, Japan, Mexico, Taiwan, Thailand, Argentina, Paraguay, Israel, USA,

Vietnam… [9]

Nutritional and chemical composition

- The nutritional composition of Stevia leaves contains protein content from

9.8% to 20.4%, fat from 1.9% to 5.9%, carbohydrates from 35.5 to 61.9% and

reducing sugar from 3.3% to 6.9% [10]

- The main ingredients in Stevia are diterpenoid glycosides, including main

types: Stevioside (5-10%), rebaudioside A (2-4%), rebaudisoside C (1-2%) and

dulcoside A (0.5- 1%) and two subtypes rebaudisoside D and E, Some

macronutrients and trace minerals are also found in Stevia : Ca, Mg, Fe, Mn, Sr, Cu,

Cr, Cd [11], [12]

- Stevia is 300 times sweeter than cane sugar because the main active ingredient

is steviol (it's a glycoside) Therefore, clover is used in foods to treat diseases such as

diabetes, high blood pressure [13] Steviozit in Stevia after hydrolysis will give 3

molecules steviol and isosteviol Steviol is 300 times sweeter than sucrose (Do Huy

Bich et al 2003) [13], low in energy, not fermented, not degraded but has a delicious

taste, can be used to replace sugar in the diet The important property of these

glucosides is that they can sweeten foods and beverages without being harmful to

humans, making it easy and economical to harvest

Uses of Stevia :

- Stevia is used to help people with diabetes significantly improve their health,

stabilize sugar gliders blood glucose content In addition, Stevia also works to reduce

blood pressure, reduce inflammation (gingivitis, stomach pain, ) [11]

- On the other hand, Stevia is also used for food production such as

confectionery, soft drinks It is also used in food production for diabetics and for

weight loss is used to replace the sweetness of sugar

- In the field of Stevia cosmetics with anti-inflammatory and antifungal effects is

used for shampoos, anti-inflammatory, acne reduction, sebum reduction to improve

skin look brighter

2.2.3 Glycyrrhixa glabra L

Origin :

- Scientific name: Glycyrrhiza uralensis fish and Glycyrrhixa glabra L,

belonging to the butterfly family (Fabaceae)[13]

- Morphological characteristics: Glycyrrhixa glabra L is a perennial plant, the

stem can be up to 1-1.5m high; The whole plant has very small hairs, double leaves, ovoid, pointed tip, intact edge is 2 to 2.5 cm long Licorice flowers when blooming are light purple, butterfly petals, flowers bloom in summer and autumn, long flowers 14-22mm The fruit is curved in sickle shape, 3-4cm long, 6-8cm wide, dark brown, the fruit surface has many long leaves, in the fruit there are 2-8 small flat seeds, grayish brown or light black green, shiny [14]

Figure 5 Glycyrrhixa glabra L

Chemical composition

- Glycyrrhizin belongs to the olean group which is a saponin, naturally found in

licorice root in the salt form of potassium and calcium It accounts for 10-14% in dry medicinal herbs, it has a sweet taste, 60 times sweeter than saccarozo sugar

- The second most important group of active ingredients in licorice is flavonoids

with a content of 3 - 4% The main flavonoids of Glycyrrhixa glabra L are liquiritin,

isoliquiritin and liquiritigenin

- In addition, in the roots of Glycyrrhixa glabra L, the main ingredients are

polysaccharice and some monosaccharides [33]

Uses of Glycyrrhixa glabra L :

- Glycyrrhixa glabra L has a very strong detoxifying effect on leukocytes toxins,

snake toxins, dizziness, tetanus poison [13]

- Effects as octison: Increases the volume of water and salt in the body, causing water retention, and at the same time treating ulcers in the digestive system

Glycyrrhixa glabra L is also combined with other herbs to treat coughs, stomach pain,

stomach ulcers, and boils [14]

- Glycyrrhixa glabra L is also used in medicine as a flavour Glycyrrhixa glabra

L root contains Glycyrrhizin which is a saponin belonging to olean group, content

from 10-14% in dry ingredients, only in the subsoil, has a very sweet taste (60 times

saccharose) This is the most important saponin of the root Glycyrrhixa glabra L [13]

In traditional medicine: Glycyrrhixa glabra L has a sweet taste, is calculated on 12

meridians, has the effect of clearing heat, detoxifying, laxative, spleen tonic, and regulating medicinal herbs [14]

2.3 Overview about extraction process

2.3.1 Overview of extraction solvents, extraction methods

Extraction is the process of separating one or more dissolved substances in a mixture of liquids or solids with another liquid called a solvent If the process of separating dissolved substances in a liquid with another liquid is called liquid-liquid extraction If the extraction process is dissolved in a solid with a liquid, it is called solid-liquid extraction In the extraction process, there are many factors that affect the extraction efficiency and the amount of active substance obtained These concentrations may be size, material, solvent, time, temperature, extraction time, extraction method, etc

2.3.2 Overview of extraction solvents

The selection of an extraction solvent is based on several factors:

- Characteristics of solvents: polarity, viscosity and surface tension Less polar solvents dissolve non-polar compounds and difficult to dissolve compounds with multiple polar groups On the contrary, strong polar solvents dissolve well substances with many polar groups and difficult to dissolve polar substances well Besides, the lower the viscosity of the solvent or the smaller the surface tension, the easier it will be absorbed into the medicinal plant and facilitate the extraction process

- Conditions for solvent selection:

+ Do not corrode equipment

+ Easy to find, cheap

+ No adverse reaction between solvents and solvents

+ Easily separates the substance to be separated from the solvent

+ Safe, non-toxic, no fire and explosion

2.3.3 Extraction method

- Depending on the nature of the medicinal material and solvents, the finished product standard as well as the conditions of the equipment and the production scale can use the methods: immersion, soak, upstream extraction or other methods In addition, it also requires other modern equipment such as supercritical liquid extraction, ultrasonic extraction

to penetrate the plant cells and dissolve the natural compounds of the sample The extraction solution is filtered (filtered through the filter paper) to recover the extract solution, recover the solvent and obtain extracts The solvent is added continuously until to the soaked sample several more times until the plant sample is completely extracted Efficiency can be increased by stirring, mixing, stirring, or using a shaker to shake the extraction mixture [15]

2.3.4 Extract

Medicinal extract is a preparation prepared by concentrating or drying the extracts obtained from plant or animal medicinal herbs with solvents suitable to certain physicality

Extract have three types:

- Liquid extract: That is a liquid, viscous, has a characteristic taste of the medicinal herbs used in which alcohol and water act as the main solvent (or preservative or both) If there are no other instructions, the convention of 1 ml of

liquid extract corresponds to 1 g of medicinal herbs used to prepare medicinal extracts

- Solid extract: That is a solid, non-stick touch The remaining solvent content in use is not more than 20%

- Dry extract: That is a homogeneous foam or dry powder, very easy to absorb moisture, moisture is less than 5%

Advantages of herbal extract

- Medicine extract with less impurity content and the percentage of active

ingredients are usually higher than active ingredients in the original herbal ingredients (except for liquid extracts with the ratio of active ingredients equal to those in herbal

ingredients)

- Herbal extract is compact, easy to transport an can be used immediately or as an intermediate to prepare other dosage forms such as capsules, tablets, round tablets, syrups

Disadvantages of herbal extract: The finished product is easy to contaminate

microorganisms (bacteria, mold, yeast) from the starting material, especially when prepared at low temperatures to protect active ingredients The medicinal herb itself is also a favorable nutrient environment for microorganisms to thrive

2.4 Overview about instant tea

In the 1885s, when John William Brown of Huddersfield United Kingdom patented the tea by making the tea a thicker and more diluted form by boiling water In the United States tea rooms gradually became popular in major cities in the early 20th century, but more than 10 years later tea rooms became obsolete and largely disappeared Melt was invented and it was quickly added to the diets of British and Canadian armies during World War II, but it was not until 1946 that it became commercialized and more popular based on tea processing technology more developed, but it was not until the 1950s that instant tea was officially commercialized

in the United States Because the US production and consumption of instant tea is greater than that of the rest of the world, soluble tea with cold water was created In

1969, methods of producing instant tea were patented and published Currently, the United States becomes the third largest tea importer in the world after the UK and Russia [40] [41]

The raw materials for the production of instant tea from green or black tea, extract the solute from the raw materials, carry the resulting fluid mixed with the

auxiliary materials at a certain concentration and then dried to create tea powder Today, instant tea is not only made from black tea and green carts but can also be made from other medicinal herbs, they also have more health promoting properties

2.4.1 Soluble tea products

The market today for instant tea products is abundant and made from a variety

of ingredients, varying in design and form All teas are mainly in two forms powder and seeds Famous and popular instant teas with whole water in Vier Nam such as Lipton, Nestea, warm, Matcha, iced tea, artichoke, instant milk tea,….The main function of instant tea is to refresh and rejuvenate, but in addition to being flexible according to the tea production materials that have the function of healing into medicinal herbs, herbs when combined together must be not match Examples of instant teas are as follows:

- Artichoke tea: Artichoke tea has the main ingredient of artichoke, so it is used

to increase energy, support, prevent cancer, detoxify, good for heart machineries, support diabetes

- Meditation Tea: that is used to treat diseases such as colds, coughs, stomach aches and alcohol help

There are 2 types of instant tea: additive instant tea and instant tea without additives

- Instant tea with additives: During the production process, some additives such

as flavour, tea flavour, vitamin C, carbohydrates are added to the production process

- Additive-free solubility: During the manufacture, packaging and sale without the addition or distribution of any additives This is a tea containing solute extracted from the ingredients so it has a flavour and character for the product

-

Figure 6 Instant tea products

2.4.2 Production and consumption of instant tea in the country and in the world

2.4.2.1 In the word market

Tea is a typical beverage of Asia, but in recent years, tea has gained popularity all over the world Due to the popularity of tea, the demand for it increased so instant tea was born as an addition to the quick requirements of the job, while satisfying the needs of refreshment and cooling for user

In May 2019, a premium tea company in Gujarat, India announced its 100th anniversary product range, featuring four flavours of masala, ginger, cardamom and lemongrass A month earlier, this company launched a series of new flavours such as orange, lemon, aloe vera, peach; Vetiver, litchi and dill

An Indonesian tea brand Sariwangi launched its three-in-one complete tea product in June 2019 The new product is a variation of powdered milk tea, Caramel and Teh Tarik Nestea India announced the product called EVERYDAY Chai Life that

is a line of instant tea products with various flavours such as ginger, saffron and cardamom, desi masala and lemongrass as the premium product line of Nestea [42]

2.4.2.2 In Viet Nam market

Instant tea in Vietnam is extremely dynamic and develops with the needs of its users In addition to the function of heat, beverage is also a product to enhance health and reduce the risk of obesity, cancer,

The instant tea product from Myxopyrum smilacifolium Blume has other effects

on inflammation, anti-cancer, with natural ingredients without additives will be a product that is expected to be a popular product because of its convenience

- Myxopyrum smilacifolium Blume raw matericals have to meet the requirements

of quality, fresh, not crushed damage;

- To cut slice: This process make raw material become smaller ingredients to facilitate the soaking process, shorten the injection process

- To soak and extract to extract the active ingredient in Myxopyrum smilacifolium Blume as much as possible Myxopyrum smilacifolium Blume is soaked in solvent,

then find out the suitable material size, the solvent concentration, the time, the material/solvent ratio, the suitable temperature when extracting to get the highest amount of total polysaccharides

Concentrates

Mixing

Drying

Packaging

- To concentrate to remove solvent from the extract to facilitate mixing and shorten drying time The rotavap evaporator system is used to concentrate the extract

in a certain ratio

- Mixing is to find out the most suitable ratio between extract of Myxopyrum smilacifolium Blume and additive materials To take the volume of concentrated Myxopyrum smilacifolium Blume and the auxiliary materials according to a certain

ratio, then mix together to create a homogeneous mixture

- Drying is to reduce the moisture content of the product, facilitate product quality and storage time

- Packaging: This is the final stage of the product This step is to dissolve tea in a jar containing a certain volume

PART III RESEARCH CONTENTS AND METHODS

3.1 Research object and research scope

3.1.1 Research object

- Main material: Stems of Myxopyrum smilacifolium Blume

- Additional ingredients: Lactose and sweet herbs like Stevia, licorice

3.1.2 Research scope

Research was carried out in the laboratory scale

3.2 Place and time to proceed

- Location: Laboratory of Faculty of Biotechnology and Food Technology, Thai Nguyen University of Agriculture and Forestry

- Implementation time: February 2020 to October 2020

3.3 Chemical, equipment

Table 3.1 Experiment chemicals Number Chemicals Origin of production

Table 3.2 Experimental equipment STT Experimental equipment Origin of production

7 Ultrasound system Elmasonic S 100 H Germany