Research on the production process of tea bag from myxopyrum smilacifolium leaves

The experimental formula for surveying the content of supplementary materials: Stevia, Herba passiflorae, Licorice.. Overall objectives Research to build the production process of tea b

THAI NGUYEN UNIVERSITY

UNIVERSITY OF AGRICULTURAL AND FORESTRY

LE THI THUY

RESEARCH ON THE PRODUCTION PROCESS OF TEA BAG FROM

MYXOPYRUM SMILACIFOLIUM LEAVES

BACHELOR THESIS

Study Mode : Full time

Thai Nguyen, 5/12 /2020

Thai Nguyen University of Agriculture and Forestry

Degree Program Bachelor of Food Technology

Student name Le Thi Thuy

Student ID DTN1654190009

Thesis Title Research on the production process of tea bag from

Myxopyrum smilacifolium leaves

Supervisor Ms Dinh Thi Kim Hoa

Abstract:

In Vietnam and around the world, the trend of using medicinal plants of natural origin is increasing Including many kind of herbal tea, an indispensable staple in the tea culture of Vietnamese people The use of herbal tea has been shown to have many medicinal uses Drinking herbal tea properly can help quench thirst, aid digestion, avoid sleepiness, stimulate kidney function, help clear the mind,

dispel sleepiness Myxopyrum Smilacifolium leaves are a precious specialty of the

Northwestern mountains and forests It also has many valuable effects on health The leaves are also shown to contain compounds belonging to the group of alkaloids, glycosides, tannins, phenols, saponins, terpenoids, carbohydrates, vegetable oils, steroids, flavonoids, anthropoids, and amino acids The use of

Myxopyrum Smilacifolium leaves with some other herbs to produce tea bags will

bring convenience and many good medicinal properties to consumers Moreover,

it helps consumers to have more choices in buying, improve efficiency, and

contribute to popularizing products from the leaves of the Myxopyrum smilacifolium tree

Number of Pages 70 pages

Date of Submission December 05 , 2020

Supervisor’s

signature

I would like to express my gratitude to Ms Dinh Thi Kim Hoa - Lecturer of Faculty of Biotechnology and Food Technology, Thai Nguyen University of Agriculture and Forestry, dedicated and guided throughout the project

I would like to express my thanks to the teachers and civil servants in the School of Biotechnology and Food Technology and all the teachers in Thai Nguyen University of Agriculture and Forestry, who taught and led, guided me

in my time at school

So I would like to express my sincere thanks to my parent and friends for encouraging and helping in my study

Thank you sincerely!

Thai Nguyen, date month year 2020

Student

Le Thi Thuy

TABLE OF CONTENT

List of figure 1

List of table 2

PART I INTRODUCTION 3

1.1 Research rationale 3

1.2 Research’s objectives 4

1.2.1 Overall objectives 4

1.2.2 Detail goals 4

1.3 Scientific and practical significance 4

1.3.1 The scientific significance of the topic 4

1.3.2 Practical significance of the topic 4

PART II LITERATURE REVIEW 5

2.1 Overview of Myxopyrum smilacifolium 5

2.1.1 Reconstructive origin and morphological characteristics 5

2.1.2 Chemical compositions of Myxopyrum smilacifolium 6

2.1.3 Pharmacological effects and effects of Myxopyrum smilacifolium: 8

2.2 Overview of additional ingredients for teabag production 11

2.2.1 Stevia 11

2.2.2 Herba passiflorae: 13

2.2.3 Licorice 14

2.3 Overview about tea bags 15

2.3.1 Introduction to tea bags 15

2.3.2 Tea bag filter products 16

2.3.3 Production and consumption of tea bags in the country and in the world 16

2.3.4 Expected production process of Myxopyrum smilacifolium leaf tea bag: 18

PART III CONTENTS AND METHOD 20

3.1 Materials 20

3.1.1 Research scope 20

3.2 Place and Timeline 20

3.3 Chemical, equipment and laboratory equipment 20

3.4 Research contents 21

3.5 Research Methods 21

3.5.1 Experimental design method 21

3.5.2 Biochemistry methods 28

3.6 Data processing methods 37

PART IV.RESULTS AND DISCUSSION 38

4.1 Results of determining the chemical composition of the raw materials 38

4.2 The results of studying on the affect of drying temperature on product quality 39

4.3 The results of investigation the influence of raw material size on product quality 41

4.4 The results of research and select the mixing ratio of supplementary ingredients to the product quality 42

4.5 The results of the study on the effect of temperature on the solubility of tea bags 44

4.6 The results of research on the influence of tea brewing time on the sensory quality of tea bag 46

4.7 The complete production process of tea bag from Myxopyrum smilacifolium leaves 47

4.8 Some quality indexes of final Myxopyrum smilacifolium tea bag 49

4.8.1 Moisture, ash and tannin components 49

4.8.2 Microbiological evaluation of complete tea bag product 50

4.9 Results of preliminary assessment of product costs: 51

PART V CONCLUSION AND RECOMMENDATION 52

5.1 Conclusion 52

5.2 Recommendation 52

REFERENCES 53

LIST OF FIGURE

Figure 2.1 Pictures of Myxopyrum smilacifolium 5

Figure 2.2 Pictures of Stevia 11

Figure 2.3 Pictures of Herba passiflorae 13

Figure 2.4 Pictures of Licorice 14

Figure 2.4 Some pictures of tea bags in the Viet Nam 16

Figure 2.5 Some pictures of foreign tea bags 17

Figure 2.6 Planned production process diagram of Myxopyrum smilacifolium tea bags 18

Figure 3.1 Experimental arrangement diagram of the effect of drying temperature 22

Figure 3.2 Experimental layout to determine the size of raw materials 24

Figure 3.3 The experimental diagram of surveying mixing ratio of Stevia, Herba passiflorae, Licorice 25

Figure 3.4 Experimental arrangement diagram to survey the temperature of tea-making water 27

Figure 4.1 Pictures of dried Myxopyrum smilacifolium leaves at different temperatures 39

Figure 4.2 The graph shows the effect of the drying temperature on the sensory quality of the product 40

Figure 4.3 The chart shows the total organoleptic scores of tea bag products with different size of raw materials 42

Figure 4.4 The graph of the total sensory scores of product with different ratios of added ingredients 43

Figure 4.5 The graph shows the effect of temperature on the solubility of tea bags 45

Figure 4.6 The graph shows the influence of tea brewing time on the sensory quality of tea bags 47

Figure 4.7 Diagram of production process of Myxopyrum smilacifolium filtered tea bag 48

LIST OF TABLE

Table 3.1 Chemical experiment 20

Table 3.2 Experimental equipment 20

Table 3.3 Laboratory instruments 21

Table 3.4 Experimental formula to investigate the drying temperature of raw materials 23

Table 3.5 Raw material size experiment formula: 24

Table 3.6 The experimental formula for surveying the content of supplementary materials: Stevia, Herba passiflorae, Licorice 26

Table 3.7 Experimental formula to investigate the temperature of the tea-making water 27

Table 3.8 Experimental formula to investigate tea brewing time 28

Table 3.9 Weight coefficient of tea bag products 36

Table 3.10 Scores of quality levels 36

Table 4.1 Results of determination of chemical composition in raw materials 38

Table 4.2 Table of effects of drying temperature on sensory quality of the tea bag 40

Table 4.3 Result of sensory evaluation of material size 41

Table 4.4 Effect of mixing ratio of Stevia, Herba passiflorae, Licorice 43

on sensory quality of the product 43

Table 4.5 The effect of temperature on solubility of tea bags 45

Table 4.6 Effect of tea brewing time on tea solubility 46

Table 4.7 Moisture, ash and tannin components of Myxopyrum smilacifolium leaves tea bags 49

Table 4.9 Results of preliminary assessment of product costs 51

PART I INTRODUCTION

1.1 Research rationale

In the period of industrialization and modernization of the country, continuous economic and scientific development has contributed to improving the quality of life.Life is getting higher and higher and the need to choose healthy, clean food is being concerned with society when food safety is an urgent issue Therefore, the use of natural products that are good for health is favored by consumers

Currently, in Vietnam as well as in the world, the trend of using medicinal plants of natural origin is increasing In which tea is a familiar drink of Vietnamese people in particular as well as East Asians in general, it has cultural characteristics of each region.Tea is used all over the world, and is considered a global beverage Tea originates from the Asian region, mainly the southwestern region of China The use of tea has been shown by the medical world to have a lot of healing power Drinking tea properly helps to quench your thirst as well as digestion, avoid drowsiness, stimulate kidney function, enhance eyesight, help clear mind, and dispel drowsiness

Along with the development of society, tea products also have many changes to better suit the increasing consumer demand of people Ingredients for tea processing in addition to the fresh tea leaves also have teabags Instant tea is more popular because of its convenience such as instant tea, filter bag tea, bottled tea,…Filter bag tea is a favorite item in the market but has great growth potential

Through the process of learning, it can be seen that Myxopyrum smilacifolium leaves are a precious herbal of the Northwestern Mountains The leaves of Myxopyrum smilacifolium have also been shown to contain compounds belonging to the group of alkaloids, glycosides, tannins, phenols, saponins, terpenoids, carbohydrates, vegetable oils, steroids, flavonoids, anthracoid and amino acids [2], [8]

It can be seen that the use of Myxopyrum smilacifolium leaves with some other herbs to produce tea bags can bring convenience and many good medicinal properties to consumers At the same time, it helps consumers to have more choices in buying, improve efficiency, and contribute to popularizing products

from the leaves of Myxopyrum smilacifolium tree Stemming from the above problems, I conducted the project "Research on tea bag production process made from Myxopyrum smilacifolium leaves"

1.2 Research’s objectives

1.2.1 Overall objectives

Research to build the production process of tea bags made from

Myxopyrum smilacifolium leaves to create high-quality tea products, bring economic efficiency, contribute to diversifying tea products in the market, and improving income for growers

1.2.2 Detail goals

- Develop a technological process to produce tea bags from Myxopyrum Smilacifolium , Stevia, Herba passiflorae and Liquorice in laboratory conditions

- Study the effect of the raw material drying temperature on product quality

- Study the effect of material size after grinding on product quality

- Study to select the mixing ratio of supplementary materials to product quality

- Study on the effect of temperature on the solubility of teabags

- Study the tea brewing time to the solubility of teabags

1.3 Scientific and practical significance

1.3.1 The scientific significance of the topic

- Provide more scientific information about Myxopyrum smilacifolium leaves

- The data in the research topic can be a reference for the manufacturers of similar products

- Find out the appropriate parameters for the production process of tea bags from Myxopyrum smilacifolium leaves on a laboratory scale

1.3.2 Practical significance of the topic

- Diversify products from Myxopyrum smilacifolium leaves to increase the

value of raw materials

- Create new products with good quality, good for the health of consumers, and a reasonable price

- Support and prevention for consumers

- Create new research directions on Myxopyrum smilacifolium trees; create outputs for the crop industry

PART II LITERATURE REVIEW

2.1 Overview of Myxopyrum smilacifolium

2.1.1 Reconstructive origin and morphological characteristics

2.1.1.1 Botanical origin and characteristics

- Scientific name: Myxopyrum smilacifolium (Wall.) Blume, is a climbing shrub, woody of genus Myxopyrum, family Jasmine (Oleaceae)

- Synonyms: Myxopyrum serratulum A.W.Hill or Myxopyrum

is 0.5 - 1.5cm long [13]

The inflorescence is lobe-shaped, grows in leaf axils, 10 - 12cm long, with thin hair; filamentous bracts, 2 to 3 mm long hairs Radio smooth cup, 0.5mm-1mm long, 4 oval lobes, 0.5mm long The corolla is white, the flower tube is 1 - 1.5mm long, 4 lobes are oval shaped, 1- 1.5mm long, hairless Stamen 2, attached near the throat of the corolla; the oval pollen envelope is wide and 0.5

mm long The ovary is ovoid, hairless, and 1mm long; the pistil split in half, 0.2 -

0.4mm long The fruit has a spherical nodule shape, size 3 - 5 × 8 - 10mm Seed

2, and hemispherical, size 4-5mm [13]

2.1.1.2 Distribution situation, cultivation season and collection

 Distribution:

Trees usually grow in thin forests along the stream, at an altitude of 500 -

1000 m [13] Myxopyrum smilacifolium tree usually grows in deep forests, cliffs with special climatic and soil conditions Trees are found in many areas in the

provinces of Ha Giang, Lao Cai, Thai Nguyen, Yen Bai, of our country [13]

Also distributed in: Thua Thien-Hue, Ho Chi Minh City, Quang Ninh, Dong Nai Also in India, Cambodia, Thailand, Malaysia, China, Bangladesh, Laos, Myanmar [9] Due to the growing conditions the tree can only grow in certain places; it increases the rarity of this species of Myxopyrum smilacifolium

 Seasons of cultivation and collection:

Myxopyrum smilacifolium are harvested all year round However, the standard harvest season is mainly autumn, from September to November

2.1.2 Chemical compositions of Myxopyrum smilacifolium

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

studies on the chemical composition of the species Myxopyrum smilacifolium

(Wall.) Blume; There are many publications on chemical composition of leaves and some claims regarding stems and roots of this species

Some preliminary surveys of Samu J et al 2014 showed that in

Myxopyrum smilacifolium leaves (Wall.) Blume had the presence of alkaloids,

carbohydrates, steroids, saponins, terpenoids, flavonoids, tannins and polyphenols [18] According to the publication of Raveesha Peeriga et al (2016),

the leaves of Myxopyrum smilacifolium (Wall.) Blum e contain alkaloids,

glycosides, tannins, saponins, terpenoids, carbohydrates and non-volatile oils [24] According to research by S Gopalakrishnan et al (2012), the leaves of

Myxopyrum smilacifolium (Wall.) Blume contain terpenoids, flavonoids,

saponins, tannins, glycosides and iridoid [25]

Praveen R.P and Ashalatha S Nair in 2014 have preliminarily determined

the chemical composition of the root extract of Myxopyrum smilacfolium showed

that there are alkaloids, phenolic, glycoside, cardiac glycoside, tannin, flavonoid,

carbohydrates, protein, phytosteroid, phytosterol, saponin [22] In 2015, the above authors continued to study to analyze and compare functional groups in methanol extracts of roots, fruits, and callus FTIR spectroscopic analysis results showed that all 3 extracts contain compounds with similar functional groups: alcohol, alkanes, alken, aldehydes, ketones, amines, carboxyl groups, alkyl halogen [34]

The preliminary survey study by Rajalakshmi K and Mohan VR (2016) announced that in the ethanol extracts of stalks there are alkaloids, catechin, coumarin, flavonoid, tannin, saponin, steroid, phenol, glycoside, terpenoid, xanthoprotein and confirmed There are 32 different compounds that were

identified in the extracts of Myxopyrum smilacifolium (Wall.) Blume [30]

 Alkaloid:

In 2013, Gopalakrishnan S and Rajameena R isolated compounds dimethoxy-13-carbomethoxy-15- (3,4,5-trimethoxybenzoxy) -13,14-didehydroalloberban from ethanol extracts of leaves [32 ] From the extract of the

7,8-body of Myxopyrum smilacifolium (Wall.) Blume, by mass spectrometric

chromatography, Rajalakshmi K and Mohan V.R (2016) determined the presence of 3 alkaloids: 4-phenyl-3,5-pyrazolidinedion,1-allyl-6,7-dimethoxy-3,3-dimethyl-1,2,3,4-tetrahydro isoquinolin, 3-dodecyl-4 - [(4-methoxybenzyl) oxygen] -2-methyl quinoline [31]

 Iridoid:

From the ethanol extraction of the leaves of Myxopyrum smilacifolium

(Wall.) Blume, Henrik Franzyk et al (2001) isolated iridoid glycoside: Myxopyrosid (9) and 6-o-acetyl-7-o- (E / Z) -p methoxycinnamoyl myxopyrosid [30]

 Organic acids and derivatives:

In 2016, by the chromatographic method combined with the format and HPTLC method, the authors Maruthamuthu V and Kandasamy R discovered the

presence of many organic acids and derivatives from Myxopyrum smilacifolium

(Wall.) Blume leaves [33]

Besides, research [31] also discovered many compounds of this group Besides the main groups of substances above, research [31] also identified in the

body of Myxopyrum smilacifolium (Wall.) Blume also has steroid compound

9,19-cycloergost-24 (28) -enol-4, 14 -dimethyl acetate, phenol 4 - ((1E) Hydroxy-1-propenyl) -2-methylphenol, D-mannopyranose carbohydrates, ethyl α-D-glucopyranoside, phytol, galactiol, amine (3-nitrophenyl) isopropyl methanol ether, vitamin E DL-tocopherol, aldehyd 5-hydroxymethylfurfural, ketone 7- ethyl-4-Decen-6-on… [31]

-3-Currently, in Vietnam, there is only the original study of Nguyen Minh Luyen (2017) published on the chemical composition of Myxopyrum smilacifolium, the roots contain saponins, flavonoids, polysarcharids, amino acids, free sugar, and feces A myxopyroside compound was established from the n-butanol fraction of the root extract Thus, up to now, there has not been any research that has an in-depth publication on the chemical composition of

Myxopyrum smilacifolium growing in Vietnam [17]

2.1.3 Pharmacological effects and effects of Myxopyrum smilacifolium

 Pharmacological effects

The studies on Myxopyrum smilacifolium showed that this species has

pharmacological effects such as: anti-oxidation of ethanol [19], [24]; antibacterial, antifungal [21], [27]; antipyretic, anti-inflammatory [19], [20]; bronchiectasis [29]; anti-cancer [23]; wound healing of ethanol extracts [26], [28]

 Antioxidant effect

In 2013, the test results of Rani T.S et al showed that the methanol

extract of the aboveground portion of Myxopyrum smilacifolium (Wall.) Blume

had a significant antioxidant effect by studying the DPPH free radical removal capacity of nitric oxide versus vitamin C Test dose Test 250 µg / ml of MeOH extract showed a DPPH free radical removal effect of 81.75% and nitric oxide and 84.95%, nearly equivalent to vitamin C at the same dose (83.45% and 84 respectively, 20%) [24]

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

Myxopyrum smilacifolium leaves (Wall.) Blume has antioxidant activity but less

than vitamin C [19]

 Antibacterial and antifungal effect

According to the study [27], the results of testing the antimicrobial activity

of kerosene, benzene, chloroform, ethanol, and water ether extracts of the leaves

Myxopyrum smilacifolium (Wall.) Blume by Gopalakrishnan S et al ( 2012)

showed that the ethanol extract exhibited the best resistance against microorganisms tested The minimum inhibitory concentration (MIC) of ethanol extracts ranged from 200-300 µg / ml with different microorganisms [27]

A 2014 study by Praveen et al Comparing the antimicrobial activity of methanol extracts from roots, callus, and Myxopyrum smilacifolium (Wall.) Blume fruit showed:

Antimicrobial effect: All extracts have similar inhibitory effects on

Enterococcus faecalis and Bacillus subtilis The extract was more effective against strains of bacteria tested except for Staphylococcus aureus and at a concentration of 50 µg / ml was more effective against E faecalis than

gentamycin at 20 mg/ml Callus has anti-S aureus effect but at concentrations higher than 100µg / ml Root and callus extracts are ineffective against

Escherichia coli

Antifungal effects: The root extract has no inhibitory effect on Candida albicans The extract of callus and fruit at a concentration of 100 g / ml had a

significant but weaker inhibitory activity than that of clotrimazole 20 mg/ml [21]

 Anti-inflammatory and antipyretic effects

Study show that Myxopyrum smilacifolium (Wall.) Blume has quite good

antipyretic and anti-inflammatory effects In 2014, research by Jolly Samu et al

Showed that ethanol extract and water extract from leaves Myxopyrum smilacifolium (Wall.) Blume both had the effect of inhibiting protein

denaturation and stabilizing red blood cell membranes in Test concentrations The results showed that ethanol extract at the concentration of 200 µg/ml inhibited 91.54% of protein denaturation, nearly equivalent to diclofenac at a concentration of 100 µg/ml; High extraction of water at 200 µg/ml inhibited hemolysis 69.60%, stronger than ethanol extract at 100 µg/ml (46.58%) but weaker than diclofenac at 100 µg/ml (97,059 %) [19]

In another study by Manu Skiaria Varughese et al (2015), ethanol extracts

from leaves Myxopyrum smilacifolium (Wall.) Blume exhibited

anti-inflammatory and antipyretic effects in experimental animals [20]

In 2015, Praveen et al Evaluated the cytotoxic effects of methanol extract

from roots and callus of Myxopyrum smilacifolium (Wall.) Blume on liver cancer

cell line HepG2 showed that: On Trypan blue staining model, the dose-dependent cytotoxic response and cytotoxic activity were strongest at a concentration of 200 μg/ml When using the MTT method, methanol extract of roots and callus inhibits the growth of HepG2 cells with IC50 of 63.75 μg / ml and 98.75 μg / ml respectively Thus, in both trials, the extracts exhibited significant cytotoxic effects [23]

 Effects on wound healing:

According to the study of Gopalakrishnan S et al (2013), ethanol extract

from leaves of Myxopyrum smilacifolium (Wall.) Blume has wound-healing

effect In the mouse wound model, the experimental results showed that the epidermal time and wound healing time when treated with ethanol extract were

17 and 19 days, respectively, equivalent to celandine ointment treatment At 400 mg/kg the weight of granulation tissue was 88.7 ± 13.0 mg while with celandine

50 mg the weight of granulation tissue was 146.4 ± 4.43 mg [26]

According to traditional medicine, Myxopyrum smilacifolium has a sweet

taste, mild aroma, is equal to the heart, can be equal to the Korean ginseng thousand years According to Pham Hoang Ho in Vietnamese herb, it has been recorded to use leaves to treat nervous disorders, rheumatism, and cough [9]

According to an Indian document, the leaf is used as medicine for asthma, cough, cold, fever, headache, rheumatism, neuralgia, bites, and wounds The roots are used to treat pruritus and scabies in children, fever cough, rheumatic diseases, bites, and wounds [3], [9], [19]

But this plant has been rarely studied both in the world and in Vietnam In some ancient documents, there are also records of the use of this plant, but it is

only used to treat rheumatism, cough, and asthma, but it is unclear how much that effect In addition, some ethnic communities in the mountainous areas have also used this plant in the following cases: orange treatment in children and osteoarthritis pain in adults

2.2 Overview of additional ingredients for teabag production

2.2.1 Stevia

2.2.1.1 Origin, characteristics and distribution

 Source:

Stevia (Stevia rebaudiana (Bert.) Hemsl Eupatorium Bert) is sugar grass,

nectar, or sweet daisies Naturally originating in the Amambay and Iguacu regions of the Brazilian and Paraguay border, today many countries around the world have been developing using fresh grass in their daily life The part used is

the above-ground part [12]

Figure 2.2 Pictures of Stevia

 Characteristics:

Stevia is a perennial herb, 20 - 60 cm high; the stem can branch more or grow

straight The stem section is round, small, with longitudinal grooves with many fine hairs, the leaf axils have protrusion The leaves have three prominent veins, the feathery appendages, 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 [12]

The flowers of Stevia are in the form of a cluster of 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 [1], [12]

 Distribution:

Nowadays, Stevia is grown in many countries such as Brazil, China,

Japan, Mexico, Taiwan, Thailand, Argentina, Paraguay, Israel, USA, and Vietnam… [12]

2.2.1.2 Chemical composition

The main ingredients in the Stevia are diterpenoid glycosides, including 4

main types: Stevioside (5-10%), rebaudioside A (2-4%), rebaudisoside C (1-2%), and dulcoside A (0.5-1) %) and two subtypes are rebaudioside D and E The herb also contains a very low amount of substances: Stigmasterol, sitosterol, campesterol, 8 flavonoids, besides cosmosin and 2 volatile substances caryophyllen, spathuienol Several macronutrients and trace minerals are found

in sweet grass: Ca, Mg, Fe, Mn, Sr, Cu, Cr, Cd [3], [8]

The main active ingredient in the Stevia is stevioside (glucoside) 250-300

times sweeter than saccharose, but stevioside does not produce energy The

Stevia side content in Stevia varies depending on the climate, the time of harvest,

and ranges from 3-20% This sugar source is characterized by no caloric intake and is very stable at a high temperature of 198 ° C (388 ° F), free from darkening and caramelization [8], [12]

In the food industry, it is used relatively widely in Japan as a concoction to increase the sweetness of various foods

In the beauty industry and cosmetics, Stevia is used to smooth hair, soften

the skin, help the body to regenerate new skin on the entire skin surface, while fighting infection and eliminating fungus [3]

2.2.2 Herba passiflorae:

2.2.2.1 Origin, characteristics and distribution

 Source:

Scientific name is Passiflora foetida L In addition, with many different

regions and regions, it is often called with many names such as longan, persimmon, gourd wire, net wire, pistil [12 ]

Figure 2.3 Pictures of Herba passiflorae

 Characteristics

Herba passiflorae is a type of vine, soft, hollow with many thin hairs The

leaves are staggered and distributed about 7cm long, about 10 cm wide, divided

into 3 pointed lobes, fluffy edges and two sides, curled tassel Herba passiflorae

has white flowers, light purple in the middle, with filament-shaped appendages Ripe fruit is red or yellow, is a favorite dish of rural people in Vietnam [12]

 Distribution:

Herba passiflorae is a fairly easy-to-find tree, usually growing in

wasteland, hills and forests

2.2.2.2 Chemical composition

Herba passiflorae has chemical components such as alkaloids, flavonoids, saponins

2.2.2.3 Uses of Herba Passiflorae

Prescriptions containing Herba Passiflorae:

Curing depression, insomnia, anxiety: 50g peanut, 30g lotus, 2.2g lotus, 10g strawberry leaves, 90g sugar, 100ml water, benzoic acid to preserve and just enough alcohol to dissolve benzoic acid [12]

2.2.3 Licorice

2.2.3.1 Origin, characteristics and distribution

 Source:

Licorice is a common name for Glycyrrhiza glabra, a species of flowering

plant in the Fabaceae family Licorice is a legume, native to Western Asia and

Southern Europe It is also used as a flavoring in confectionery and tobacco, especially in some European and Western Asian countries [13]

Figure 2.4 Pictures of Licorice

 Characteristics:

Licorice is a perennial herbaceous plant, 0.3-1m high The long roots are

pale yellow in color The body has short, soft hair The leaves are double, alternate, consisting of 9-17 egg-shaped leaflets, the edges are whole The structure is sturdy, very difficult to break, the crack is light yellow with a lot of longitudinal fibers, and has starch Light yellow to brown-yellow powder The inflorescence grows in the middle of the leaves to form flowers, pale purple flowers in a butterfly shape.The fruit is like a bean, curled in a crescent shape, 3-

4 cm long, 6-8mm wide, dark brown, with thick hairs, containing 2-8 small, flattened, pink-brown seeds [13]

 Distribution:

The northern licorice tree was not previously available in our country, and the trees grew wild in the Soviet Union, China In many countries, the plant grows as a wild grass that is difficult to kill By 1958, the trees were tested in some places in Vietnam, the trees grew well in the spring, summer, and autumn,

and in the winter they died and less developed, the next year the tree would grow well again

Planted with seeds or rhizomes, after 4-5 years can be harvested [13]

2.2.3.3 Chemical composition

In Licorice has a sugar content of 4-6% (glucose, sucrose), starch content

is 25-30%, asparagine content is 2-4%, Vitamin C, resin 5%, and a little essential oil, flavonoids 1% But the main active ingredient in licorice is glyxyridine (glycyrrhizin) at the rate of 6-14%, sometimes up to 23% [13]

Some of the effects of Licorice: detoxifying effect; effects on the liver, kidneys;

effects on respiration; effects on the digestive tract… [13]

2.3 Overview about tea bags

2.3.1 Introduction to tea bags

This product line sounds new, but it has a history of development for more than a century In the early nineteenth century, a business named Thomas Sullivan in New York sought to improve the marketing capabilities of its tea-making business He sent his customers small tea bags for them to taste A lot of customers dipped this tea bag directly in water to make it and then sent letters to thank Sullivan for this new discovery However, they complained that the silk bag was so thick that a lot of the tea scent did not penetrate the material while making the tea Sullivan continued to refine her findings, but it was not until Joseph Krieger perfected them that the tea bags were widely accepted by the public, especially after World War II Today, tea bags with different flavors are enjoyed and enjoyed all over the world and compete significantly with warm brewed tea in the ordinary way

On the market today, tea products are becoming more and more diversified and diversified in terms of types, especially with the development trend of functional foods Most herbal teas in addition to their refreshing effects

also have a tonic effect and improve the health of the user Compared with many types of traditional tea products, tea bags are considered as a new product, more convenient, saving tea making time to meet the needs of life [14]

2.3.2 Tea bag filter products

Currently, tea bag products are sold a lot, not only diverse in packaging, designs, but also raw materials for making tea bag, which helps consumers to have more choices Tea bag filter products are sold in a variety of types from popular to high-end for the purpose of more convenience to meet the needs of life

Figure 2.4 Some pictures of tea bags in the Viet Nam 2.3.3 Production and consumption of tea bags in the country and in the world

2.3.3.1 Production and consumption of tea in the world

Over the past decades, the global tea industry has seen rapid growth, with

a growing number of consumers globally as well as in many national markets For 2016, the FAO pegged global tea production at 5.73 million tons [35]

From 2007 to 2016, tea production grew at an average annual rate of 4.4% Global tea consumption stood at 5.53 million tons in 2016, up from 5.08 and 5.29 million tons in 2014 and 2015 From 2007 to 2016, consumption

increased rate Annual growth 4.55%, three-quarters of global output is consumed locally, demonstrating the important role played by major producing countries in expanding current demand Consumption is driven by population growth, urbanization and rising incomes, especially in developing countries [35]

Tea bags are increasingly strongly developed because of its fastness, convenience and health benefits; there are also a variety of flavors available for consumers to choose from According to statistics, in today's teas, tea bags make

up 12% of the tea consumed in the world Tea bags are available in most countries around the world and are very popular in North America and Europe , Asia-Pacific, South America, Middle East and Africa [35]

Figure 2.5 Some pictures of foreign tea bags

2.3.3.2 Domestic tea production and consumption

In the early 2000s, tea bags appeared on the Vietnamese market After more than ten years of appearing on the market, tea bag products are becoming more and more diversified in types, especially with the trend of developing functional foods Most of the herbal teas in addition to beverage functions are advertised for their uses such as liver tonic, lung cooling, fat loss,…However, in order for these products to be developed for international markets, Vietnamese enterprises need to invest in more in-depth research on tea bag products

Tea bags processed from Myxopyrum smilacifolium leaves with many pharmacological effects such as antipyretic, anti-inflammatory [19], [20]; anti-cancer [23], create an explosion of new trends in Vietnam That is an opportunity to develop and expand the product business market At the same time, bringing convenience to modern life is not only a beverage with health and

spiritual value, but also a functional food that is convenient and full of convenience, quality assurance

2.3.4 Expected production process of Myxopyrum smilacifolium leaf tea bag

Through articles and references on the tea bag production process, we can give the production process of tea bag filtered from Ginseng Stone leaves as follows [6], [7], [11]:

Figure 2.6 Planned production process diagram of Myxopyrum smilacifolium

tea bags

Packaging products

Myxopyrum smilacifolium

Clean

Drying

Crushed

Mixing Herba passiflorae

Stevia

Tea bag

Product Licorice

Process description:

- Raw materials: Raw materials must meet the quality requirements, not

be crushed or damaged

- Cleaning: raw materials are then washed away to remove impurities

- Drying: Raw materials are dried to 10 - 11% humidity, ensuring quality and storage time

- Crushing: crushing the ingredients to the appropriate size to facilitate the extraction of solutes in tea bags

- Mixing: materials after grinding are mixed with additional ingredients to create more harmonious colors, flavors and flavors for the product

- Packing: after mixing tea, tea will be packed into a dedicated filter bag that has been prepared with the volume of 2g/bag, sealing the bag, pairing wire and packing into a paper box

PART III CONTENTS AND METHOD

3.1 Materials

Main raw materials are Myxopyrum smilacifolium leaves that were purchased in Ha Giang province

Additional materials were purchased at the traditional pharmacy store in

Thai Nguyen province Includes: Stevia, Herba passiflorae and Licorice

3.1.1 Research scope

The research was implemented in the laboratory

3.2 Place and Timeline

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

Implementation time: July 2020 to November 2020

3.3 Chemical, equipment and laboratory equipment

Table 3.1 Chemical experiment

Table 3.2 Experimental equipment

Table 3.3 Laboratory instruments

3.5.1 Experimental design method

Experiment 1: Determination of chemical composition of Myxopyrum smilacifolium leaves

Proceed: Selected fresh leaf ingredients are washed, drain, and then slice thinly to 0.5cm size The leaves of Myxopyrum smilacifolium were crushed, put

to determine the chemical composition of the raw materials: tannin, moisture, ash Based on the analysis results, we can determine the chemical composition of

Myxopyrum smilacifolium leaves

The experiment was repeated 3 times, taking the average results

Experiment 2: Studying the effect of drying temperature on product quality

Carry out drying of Myxopyrum smilacifolium leaves at different

temperatures to reach the moisture of 9-10% Each experiment was repeated 3 times under the same conditions

The experimental design formulas are as follows:

Accurately weigh 200g of fresh leaf material into drying with the formula

of drying temperature 50˚C, 60˚C, 70˚C, 80˚C, 90˚C, drying until the humidity reaches 10-11% Analyze the content of nutrients and evaluate sensory quality to choose the appropriate drying temperature

The experiment is arranged according to the following diagram:

Table 3.4 Experimental formula to investigate the drying temperature

Experiment 3: Studying the effect of material size on product quality

After determining the appropriate drying time, use the ingredients treated in experiment 2 and then grind them with a blender with different sizes: d ≤ 1 mm, 1

<d ≤ 3mm, x ≥ 3mm.Use the sieve to classify Packaging into a dedicated filter bag 2g/bag and mix in hot water 100˚C with time 5 minutes and then conduct the sensory assessment The recipes were made at the same time and water temperature Each experiment was repeated 3 times under the same conditions

Monitor criteria: the sensory value of color, odor, taste, and state of tea The experiment is arranged according to the following diagram:

Figure 3.2 Experimental layout to determine the size of raw materials

Table 3.5 Raw material size experiment formula:

Experiment 4: Researching and choosing the mixing ratio of supplementary materials to ensure product quality

Purpose: Add ingredients Stevia, Herba passiflorae, Licorice appropriate

ratio to increase the sensory value of the product

Implementation: Mix the ingredients in the right ratios to choose the best recipe to increase the value of the product The ratio of mixing ingredients by total weight is 2g

With the main ingredients of Myxopyrum smilacifolium leaves accounting

for 80%, adding Stevia, Herba passiflorae, Licorice is formula 9 (1.6g / 0.14g /

0g / 0.26g); Formula 10 (1.6g/0g / 0.26g / 0.14g); formula 11 (1.6g / 0.1g / 0.1g / 0.2g); Formula 12 (1.6g / 0.1g / 0.2g /0.1g) was performed with 4 replicates

The experiment is arranged according to the following diagram:

Figure 3.3 The experimental diagram of surveying mixing ratio of Stevia,

Herba passiflorae, Licorice

Table 3.6 The experimental formula for surveying the content of supplementary materials: Stevia, Herba passiflorae, Licorice

Formula

Myxopyrum smilacifolium leaves (%)

Stevia (%)

Herba passoflorae

(%)

Licorice (%)

Material handling: washed, dried, and crushed Myxopyrum smilacifolium,

Stevia, Licorice, and Herba passiflorae leaves

Weigh 2g of raw materials including Myxopyrum smilacifolium , Stevia, Licorice, and Herba passiflorae leaves with the additional content as the above

experimental layout table Extract with 120ml boiling water for 5 minutes, conduct sensory evaluation for tea just obtained.The experiment was repeated 3 times under the same conditions and recorded the results with the evaluation criteria

Sensory evaluation criteria: The score for sensory evaluation of the color, smell, taste and state of tea

Based on the analytical results, choosing the appropriate material mixing ratio, the results of experiment 4 are used for the next experiment

Experiment 5: Effect of temperature on the solubility of tea bags

Procedure: Experiment with the mixing formula found in experiment 4 with the temperature of the mixed water changing according to the formulas, corresponding to Formula 13: 70˚C, Formula 14: 80˚C, Formula 15: 90˚C, Formula 16: 100˚C and repeat 3 times

The experiment is arranged according to the following diagram:

Figure 3.4 Experimental arrangement diagram to survey the temperature

of tea-making water

Weigh 2g of ingredients including Myxopyrum smilacifolium , Stevia, Licorice, and Herba passiflorae leaves with the ratio of additional ingredients as

in experiment 4 Make tea at the above temperature

Table 3.7 Experimental formula to investigate the temperature

of the tea-making water

Formula The temperature of the tea-making water (ºC)

Experiment 6: Studying the effect of brewing time on the solubility of tea bags

Purpose: To find the most suitable tea-making time for products to have good color and taste, as a basis for recommendations to consumers

Implementation: Experimentation of making tea bags with filter bags weighing 2g/pack, with known parameters in experiments 4, 5; With the time of

4, 5, 6 minutes, respectively, formula 17, formula 18, formula 19 and repeat 3 times under the same conditions

Extract tea packages with about 120ml / pack, water mixed according to temperature for the best sense in experiment 5

Table 3.8 Experimental formula to investigate tea brewing time

Formula Tea brewing time (minutes)

3.5.2.1 Method of determination of moisture: According to the drying method to

constant weight according to TCVN 5613: 1991

 Principle: Use the heat to separate the moisture in the material, while retaining all the substances in the material Therefore the drying temperature should not be too high or too low If the temperature is too high, the moisture will evaporate quickly, but it is easy to change the chemical properties of the substances, especially volatile substances, which will affect the analysis results

If the temperature is too low, the steam is released slowly, causing time and substances to change Moisture was determined by drying to constant mass [37]

- Determinations of moisture follow TCVN 5613 – 1991 standard

- Principle of the method: The method is based on drying the tea samples

to constant mass under defined conditions

To conduct moisture determination must use:

- Analytical balance with permissible deviation not exceeding 0.001g

- The oven can be controlled to adjust the temperature (105˚C) or (120˚C)

- The weighing bowl is made of glass, porcelain or aluminum with a diameter of 50mm and a lid

- Desiccator

 Procedure:

Take samples and crush them, use a analytical balance to weigh 5g of

Myxopyrum smilacifolium leaves, with an error not exceeding 0.001g, put them

in a bowl of known weight Place the bowl containing the sample in the oven and raise the temperature to 105ºC Dry for about 6 hours, take the bowl containing the sample and place it in the desiccator, make it cool Then weigh and record the results After the first weighing, re-dry the sample at the above temperature for 1 hour until the mass is constant

When necessary, repeat these operations until the difference in results between two successive weighing does not exceed 0.005g

 Calculate results:

Moisture is determined by drying to constant mass

Moisture (W) is calculated as a percentage of mass by the formula:

W = 100

Inside:

- W: Moisture of food (%)

- m: Weight of sample before drying (g)

- m1: Weight of sample after drying (g)

The test result is the mean of two parallel determinations, the difference between them not exceeding 0.2%

3.5.2.2 Determination method of ash

 Principle: Fire ingredients or food products at a temperature of 500 -

600 °C of the burning and volatile organic substances, the remaining inorganic matter is the ash component [14], [16]

Instruments and equipment used to conduct the test should be used:

- Analytical balance with error not exceeding ± 0.01g

- The crucible is made by porcelain or metal

- The furnace can be adjusted to 500 - 600ºC

- Desiccator, below to desiccant matter

- Electric stove or alcohol lamp

- 10 volumes H2O2 or HNO3 concentrated

 Proceed: Fire a porcelain crucible or metal bowl washed, in the oven with temperature from 500 to 600ºC to constant weight Cool in a desiccator and weigh on the analytical balance to the nearest 0.0005g

Place in the crucible about 5 g of the test sample Weigh them all on the analytical balance with the above accuracy Put everything in the furnace and increase the temperature slowly from 500 to 600ºC Ignite until the ash is white, which means that common organic matter has been removed for 6-7 hours

In the case of black ash left, make it cool, add a few drops of concentrated

H2O2 or HNO3, and fire it to white ash Make it cool in the desiccator, and weigh

on the analytical balance Continue firing at the above temperature for 30 minutes, and then make it cool in the desiccator After that, weigh and repeating until the weight remains constant The results between the two firing times and the next weighing should not be more than 0.0005g apart

- G1: Weight of crucible and sample (g)

- G2: Weight of crucible and white ash (g)

3.5.2.3 Determination method of tannin content: According to Leventhal titration method

Tannin is a reducing compound, oxidized by KMnO4 in an acidic environment, with the Indigocarmin indicator to form carbon dioxide and

water[10] At the same time loss of the blue color of Indigocarmin according to the following reaction:

Tannin + KMnO4 CO2 + H2O

- Prepare aqueous extract:

Use boiling water to extract solutes of tea to obtain a concentration of 10g tea/liter

8-Step 1: Weigh accurately on 10g tea analytical balance into clean paper, lightly dry at 500C for one hour

Step 2: Put the dried tea in a porcelain mortar, crush it finely and transfer

it to a 250ml conical flask, add 100ml of distilled water, soak for 15 minutes,shake gently every 5 minutes, then filter through a filter funnel into a 1-liter volumetric flask Continue to add to the conical flask 100ml of boiling water, soak for 15 minutes Repeat 5-7 times to drain all solutes from the tea, all the filtered tea water is transferred to the original volumetric flask, allowed to cool, then add distilled water to the point of the flask, shake well

Use the solution (solution A) to determine the chemical components of tea

 Prepare:

- Raw materials: tea extraction

- Chemistry:

+ Indigocacmin 0.1%: Dissolve 0.1g Indigocarmin in 5ml of concentrated

H2SO4, transfer to a 100ml volumetric flask, then add distilled water to the 100ml point, shake well, filter for a clear solution

+ H2SO4 ¼ (25%) (Concentrated acid solution): 25ml H2SO4 + 75ml water + KMnO4 0,1N:

Dilute the solution KMnO4 0,1N:

Concentration mol/l of KMnO4: CM=CN/5=0,1/5=0.02M

=) n KMnO4= 0.02*0.25= 0.005 (mol)

=) n KMnO4= 0.005*158= 0.79 (g) Weigh 0.79 (g) of KMnO4 dissolved in distilled water, place in a 250 ml volumetric flask, rinse the cup several times, add water to the point 250 ml, cover, and shake well

- Tools and equipment:

+ Pipette + White ceramic bowl of 1-liter + Glass rod

+ Burette

 Conduct experiments:

Step 1: Use a pipette to suck exactly 10ml, put solution A into a white ceramic bowl of 1-liter capacity, add 750ml of distilled water, and 10ml of 0.1% Indigocacmin indicator solution and 10ml of H2SO4 ¼

Step 2: Fill the burette with 0.1N KMnO4 to the mark

Step 3: Titrate the solution with 0.1N KMnO4 During the titration stir vigorously with a glass rod Titrate at a rate of 1 drop/s

Step 4: Read the result: When the solution appears greenish-yellow (mixed color in a ceramic bowl, gradually change from green to grayish-green, light blue, yellow-green) stop titration and read the result of KMnO4 0.1N consumption

Note: - It is necessary to titrate 2 - 3 samples under the same conditions

- Titrate a blank sample in parallel (replace 10 ml of solution A with 10 ml

- a: Volume of KMnO4 0.1N used to titrate tea samples,

ml

- b: Volume of 0.1N KMnO4 used for blank titration, ml

- V1: Volume of solution A: V2 = 1000ml

- W: The amount of tea dry matter in the original sample

3.5.2.4 Method of evaluating microbiological criteria

Limits of bacteria and fungi (3113/1999/QD-BYT dated October 11,

1999, of the Minister of Health)