€¦ · Conservation and Sustainable Use of Medical Plants in Kayin State-GAP and GCAP-KENJI IRIE...

Department of Food and Drug

Administration

Hpa_an , Kayin State

Conservation and Sustainable Use of Medical

Plants in Kayin State-GAP and GCAP-

KENJI IRIE

(Tokyo Nodai)

The role of Myanmar Seed Bank

Myanmar Seed Bank

➢Seed storages with additional experimental facilities were constructed in 1990.

➢Project-style technical cooperation has been conducted from 1997 to 2002 .

Plant Genetic Resources Management System

Exploration & collection

Temporary conservation

Multiplication & characterization

Active seeds in short storage

Base seeds in base storage

Data management

Detailed Implementation Plan (DIP) of Myanmar Seed Bank Project.

1.Exploration and Collection

1)Exploration planning

2)Field survey and collection

3)Distribution map making

4)Isolation and purification

2.Classofication and evaluation

1)Classification and identification of collected materials

2)Establishment of methodologies and standards for evaluation

3)Evaluation of conserved accessions

4)Evaluation of collected accessions

3.Conservation and Multiplication

1)Multiplication of collected accessions

2)Establishment of methodologies for desiccation and conservation

3)Germination test of seeds

4)Rejuvenation

4.Data Management

1)Standarization of data

2)Design of information system

3)Construction of database

4)Cataloging

5.Exchange of genetic resources and information

1)Training

2)Preparation of exchange system for genetic resources and information

Plant Total Accessions

Rice 7,350

Wild rice(5 species) 184

Black gram 126

Chickpea 547

Pigeon pea 143

Mung bean 189

Cowpea 180

Soybean 80

Lima bean 68

Lablab bean 5

Kidney bean 69

Lentil 8

Wild vigna 101

Maize 98

Wheat 1,807

Sorghum 327

Millet 142

Sesame 42

Groundnut 665

Sunflower 16

Niger 3

Safflower 1

Mustard 7

Jute 42

Vegetable 175

Total 12,375

No. of accessions conserved in MyanmarSeed Bank 2016.

Topix

Ⅰ. GAPs：Good Agricultural Practices

Ⅱ. Good Agricultural and Collection Practices (GACPs) for Medicinal Plants

Ⅲ. Suggestion of Sustainable Use for Medicinal Plants

GAPs：Good Agricultural Practices


◼Sustainable Agriculture

Sustainable agriculture is a farming system that meets the need for safe,

nutritious, and affordable food for the world population in a way that

progressively conserves the environment and natural resources. It involves

seeking to optimize skills and technologies to achieve long term

productivity and profitability for stakeholders of the agricultural industry in

order to ensure that future generations can also experience the same

prosperity that we enjoy today.

Ecological concerns

•Soil productivity (Erosion, depletion of top soil)

•Water (Depletion, groundwater usage, contamination)

•Pest and Disease resistance to pesticides

•Greenhouse effect and Climate Change

Economical and social concerns

•Price of food

•Income of the small and rural farmers

Impacts on human health

•Food safety and food hygiene

•Farm workers health and welfare

◼Three major concerns


◼What are GAPs?

GAP is a production pathway that identifies critical control points

and establish compliance standareds at varying degrees to

elimenate hazareds and prevent accidents in order to progressively

promote safe and hygenic fresh produce at farm with minimum

negative impacts to the environment.

GAPs are a set of principles, regulations and technical

recommendations applicable to production, processing and food

transport, addressing human health care, environment protection

and improvement of worker conditions and their families.


◼What are GAPs?

The basic concepts of GAPs are to guide the

production systems towards a sustainable agriculture

and ecologically safe, obtain harmless products of

higher quality, contribute to food security generating

income through the access to markets and improve

working conditions of producers and their families.

Security for

People

•Improve worker and

consumer conditions

•Enhance the

Agricultural Family

welfare

•Improve food security

Environment

•No contamination of

water and soils

•Rational handling of

agro-chemicals

•Concern about

Biodiversity

Food Safety

•Healthy food, not

contaminated and of

higher quality to

improve nutrition and

food consumption

Animal Wellfare

•Animal care

•Adequate feeding


◼ What do the GAPs foster?

◼ The Benefits for Farmers from GAPs Certification


◼ Enhanced food safety through the improvement of coordination

with suppliers

◼ Reduction of risks and liabilities in production, leading to fewer

recalls and withdrawals of products during distribution

◼ Improved cost management and good practices for IPM

◼ Improved productivity through the introduction of efficient

operations management

◼ Competitive advantages related to costs, the market, credibility,

and price

◼ Market access and recognition by buyers

◼ Improvement of farm ecology through good management of input

resources


GLOBALG.A.P.

• The GLOBALG.A.P standard is based on a monitoring system of risk assessment.

JAPAN GAP (JGAP) / ASIAGAP(former JGAP Advance)

• Inspired by growers supplying to supermarkets / Concern for export markets

Malaysia GAP (MyGAP:former SALM)

• Initiated by authorities to advande competitiveness

ASEAN GAP

• Harmonizes ASEAN countries GAP systems

ChinaGAP

• Concern for food safety / Concern for export markets

ThaiGAP

• Initiated by regulators / Adopted from research cluster

◼ GAPs in ASIA

Good Agricultural and Collection Practices (GACP) for Medicinal Plants

◼ Promotion of sustainable use and cultivation of medicinal plants

◼ protection natural resources of medicinal plants


Objects

1. Contribute to the quality assurance of medicinal plant materials

used as the source for herbal medicines to improve the quality,

safety and efficacy of finished herbal products;

2. Guide the formulation of national and/or regional GACP

guidelines and GACP monographs for medicinal plants and

related standard operating procedures; and

3. Encourage and support the sustainable cultivation and collection

of medicinal plants of good quality in ways that respect and

support the conservation of medicinal plants and the

environment in general.



Good agricultural practices for medicinal plants

Identification/authentication of cultivated medicinal

plants• Selection of medicinal plants

• Botanical identity

• Specimens

Seeds and other propagation materials

Cultivation

•Site selection

•Ecological environment and social impact

•Climate

•Soil

•Irrigation and drainage

•Plant maintenance and protectionHarvest

Personnel


Good collection practices for medicinal plants

Permission to collect

Technical planning

Selection of medicinal plants for collection

Collection

Personnel


Common technical aspects

Post-harvest processing

•Inspection and sorting / Primary processing / Drying / Specific processing / Processing facilities

Bulk packaging and labelling

Storage and transportation

Equipment

•Materials / Design, construction and installation / Identification

Quality assurance

Documentation

Personnel (growers, collectors, producers, handlers, processors)

•General/ Health, hygiene and sanitation

Inventory

• Diversity of medicinal plants / Traditional knouledge

Domestication

• Establishment of cultivation techniques

ABS (Access and benefit sharing)

• Intellectual property rights and benefits-sharing

Value Chain

• Production/Collection→Processing/Preparation→Distribution→Man

ufacture

Suggestion of Sustainable Use for Medicinal Plants

http://oshiete.goo.ne.jp/qa/5899562.html

Tome-asu

Management of Plant Genetic Resources in the Amazon region of Brazil-A case study of a farmer of Agroforestry practice in Tomé-Açu, Pará-

Collaborative research with UFRA, Brazil

Acerola Soursop

Field（agroforestry）

Forest

Home

No. of useful plants in land

エリシール・パレゴリコPortuguese：Elixir-paregóricoFamily：Piperaceae(コショウ科)Species：Piper callosum Ruiz et Pavon

ブラジルナッツ、パラグリEnglish：Brazil nut, Para nutPortuguese：Castanheiro-do-paráFamily：Lecythidaceae(サガリバナ科)Species：Bertholletia excelsa Humb. Et Bonpl.

Food：72

Medicinal：

41

Industrial

：19

Lumber：

35

Oil:5

Ornamen…

Spices:6Other:7

168 useful plants

Collecting medicinal plants from the wild.

Loss biodiversity (Genetic erosion)

Domestification

Establishment of cultivation techniques in Chin state (JICA-Makino Botanical Garden)

Collecting from the wild

A case of Wa-U (Konnyak)


The Nagoya Protocol on Access to Genetic Resources and the Fair and Equitable Sharing of Benefits Arising from their Utilization (ABS) to the Convention on Biological Diversity is a supplementary agreement to the Convention on Biological Diversity. It provides a transparent legal framework for the effective implementation of one of the three objectives of the CBD: the fair and equitable sharing of benefits arising out of the utilization of genetic resources.

The Nagoya Protocol on ABS was adopted on 29 October 2010 in Nagoya, Japan and entered into force on 12 October 2014, 90 days after the deposit of the fiftieth instrument of ratification. Its objective is the fair and equitable sharing of benefits arising from the utilization of genetic resources, thereby contributing to the conservation and sustainable use of biodiversity.


What is PIC?• Access permission mechanisms based on national

legislation• Permit：Access Permit, Use Permit, Export Permit etc.

• License：Hunting License, Fishing License, Picking License etc.

• Based on Environment Protection Act, Wildlife Protection Act etc.

• Conditions for Permit or License be attached• No transfer of property right for genetic resources

• Only permit or license to access, export and use

• Need another permission for new use or third party transfer

• Violation has various punishment such as fines, imprisonment, seizure etc.

What is MAT?

• Mutual agreement between provider and acquirer (user) such as:• Memorandum of understanding (MOU)

• Material transfer agreement (MTA)

• Research collaboration agreement

• Must include:• Access conditions such as no transfer of property right

• Benefit-sharing conditions such as non-monetary and monetary benefits

PGRAsia” is an international joint research project between the Japanese Research Group and genebanks of Asian countries. PGRAsia project aimed to promote conservation and sustainable use of plant genetic resources.（2014－2017, 2018-2022)

PGRAsia-Exploration and Collection for Vegetable Genetic Resources

in the Family Brassicaceae-

Characterization Allyl isothiocyanate

PGRAsiaEvaluation of Mustard genetic resources

Diversity of Cultivated Hibiscus Species（CHIN-BAO) in Myanmar Home Garden

I. Plant genetic resources and related traditional knowledge in semi-autonomous ethnic minority areas with political and geographic isolation (Dr. K. Watanabe)

II. Sustainability of seed procurement for small sclae farmers in Asia (Dr. Y. Nishikawa)

SURVEY AND COLLECTION (Materials & Methods)

Market Field & Home garden Seed shop & Seed bank

・SURVEY ① 22,Aug.~17,Sep.2015② 15,Dec.2015 ~ 21,Feb.2016Using a car to move between towns

・COLLECTION Seed and Specimen

・INTERVIEWING Farmers and Market traders（Characteristics of cultivation, usage, history, Price, Local name, Habitat, traditional knowledge）

36

Myintha(134~136)

Kyaukse(132.133)

Htauk kyant(9)

Dedaye(179.180)

Hmawbi(1~8)

Hlegu(12~14)

Kungyangon(181.182)

Bago(10.11.15)

Yangon

Mandalay(128~131)

Hsipaw(150~154)

Kyaukme(155~157.159.160)

Pyinoolwin(161~164)Nawnghkio(137.158)

Lashio(138~149)

Shwebo (121~125.184.185)

Singu (126.127)

Sagain(165.166)

Taungdwingyi(89~91.108.109)

Padein(64~66)

Pwintbyu(69)

Minbu(61~63)

Yenangyaung(76~79)

Salin(70~75)

Chauk(80.81)

Monywa(112~120)ChaungU(110.111)

Labutta(173~178)

MyaungmyaPantanawThegon(93~97)

Pyay(92.99~104)

Shwedaung(98)

Padaung(105~107)

Pathein(167~172)

Magway(46~60.67.68.188)

Yezin(18~37.183.186.187.189~194)

Pinmana(16.17.195~204)

Naypyitaw(205)

Muti(82~84)

Takon(38~45)

Myothit(85~88)

ミャンマー連邦共和国

1/2,469,500

0 400km

Fig.1 Collection sites in Myanmar.

Field survey ⇒ 6 Divisions and state 49 Townships

Accessions ⇒ Total 232

Dry area Mountainous area

・H.cannabinus and H.acetosella have high wide area adaptability

・Many species and hybrids had been maintained by the selection of the land to be cultivated by natural environmentby artificial selections

SURVEY AND COLLECTIN (Result)

TAXSONOMIC ANALYSIS

Fig.1 Characteristics of Hibiscus genus plant in Myanmar

H.sabdariffa, H.cannabinus,H.radiates, H.acetosella4 species are collectively called“CHIN-BAO” in Myanmar.

Species Hibiscus radiatus Hibiscus acetosella Hibiscus surattensis

Loc.

name

CHINBAO-ni / -pyu

kathe-CHINBAOCHINBAO-ni / -pyu recha-CHINBAO

birra-CHINBAO

CHINBAO-hin

CHINBAO-ka

recha-CHINBAO

CHINBAO-ka

CHINBAO-wainrecha-CHINBAO CHINBAO-ni

CHINBAO-ka

CHINBAO-pyu

Calyx

Stem

colorred / pink / green red pink / green deep red red / green red /green green deep red green

Leaf

shape3-5Palmatilobed 3-5Palmetely parted 3-5Palmetely parted 3-5Palmatilobed

3-5Palmetely parted /

Serrate palmatifidSerrate palmatifid 3-5Palmetely parted 3-5Palmetely parted 3-5Palmatilobed

Young

leaf

Old leaf

Hibiscus sabdariffa Hibiscus cannabinus

Species Cultivated Market Wild Others Total

H.sabdariffa 126 36 5 167

H.cannabinus 21 12 14 47

H.radiatus 13 3 16

H.acetosella 1 1

H.surattensis 1 1

Total 161 51 1 19 232

Table2. Summary of collected samples in Myanmar.

SURVEY AND COLLECTIN (Result)

Flowercolor

Calyxshape

Leafshape


Production/

Collection

Processing/

PreparationDistribution Manufacture

Establishment of value chain for medicinalPlants

Ex.

Tadasu Furusho/PhD

Depar tment o f N u t r i t i o n Sc ience &

Depar tment o f I n t e r n a t i o n a l Food and

Agr i cu l t u r e Sc ience

Tokyo Univer s i t y o f Agr i cu l t u r e2018.8

The shokuiku is one of the education about the maintenance or improvement of human health from the side of the dining habits.

For t h i s

1.Study to acquire correct dining habits2.Various studies concerning food and health 3.Succession of gastronomic culture of home country

An in crease o f l i f e s t y l e disease

p a ti e n t by l i f e s t y l e m o d i f i c a t i o n

High b lood pre ssu re, diabe t i c , and cancer

e t c . ・・・May o b e s i ty by an energy excessi ve

i n t a k e (west e r n i z a t i o n o f food) be a cause?

Review of “Dietary habits"

Succession ofGastrono m ic c u l t u r e

I m p roveme n t ofd ie ta r y h a b i ts

Secur ing o f saf e t y Diversification of "Food“!?

Shokuiku

Shoku i ku '?

The Ministry of Education, Culture, Sports, Science and

Technology (JPN): Children are brought up to a mentally

and physically rich, healthy adult.

Knowledge concerning "Food"

Custom of thinking about "Food"

Judgment that selects "Food"

･Achievement of healthy eating habits

･Succession of gastronomic culture in Japan

･Securing of health etc.

Shokuiku

New mode l

Loca t i on o f Shoku i ku i n School Educat ion

Old mode l

Then how should I develop food education at school?・Interest in eating, tell the importance of meals.

・Cultivate vegetables and make them cookthemselves.

・Check living habits including eating habits to clarifythe problem.

・Devote food and make efforts to eliminate 'likes anddislikes'.

・Understand national traditional meals.

・Acquire knowledge on processed foods and ability to select foods.

・Think about health. etc.

Practical education is required.

・Creating a curriculum with clear purpose

· Improve teacher's knowledge about food

· Development of teaching materials

according to grade level

・ Improve awareness of parents' diet and

nutrition

・ Collaboration with local communities

Poi nt s to make Shokui ku successf ul i n school educat i on

The M in i s t r y o f Agr i cu l t u r e , F o r e s t r y and Fish e r i es (JPN):

Promot ion o f p r i m a r y i n d u s t r y (agr i cu l t u r e , f o r e s t r y and

f i s h er ies )

･Succession of traditional gastronomic culture･Establishment of environmental symbiosis type agriculture･Activation of rural village, forest village and fishingvillage･Securing of safety of "Food"

Improvement of self-sufficiency in food

One of the extremely

important plan in the

farm policy.

1965 1975 1985 1995 2005 2015

Year

Transition of rate of food self-sufficiency

(JPN)

73

54 53

4340 39

100

90

80

70

60

50

40

30

20

%

Enagy base

1960 1970 1980 1990 2000 2015Year

Transition of population of agricultural work force (JPN)

30.1

17.9

9.8

6.44.5

1.7

35

30

25

20

15

10

5

0

%

Problems o f Agr i cu l t u r e in JPN

1. Decrease i n popu l a t i o n o f

agr i c u l t u ra l work f o r c e

2. Aging o f a g ri c u l t u r a l3. Lack o f Agr i c u l t u ra l

worke rs

successors

4. Decrease i n r a t e o f foo d

s e l f - s u f f i c i e ncy

5. The r i s k i s h igh i n a new

agr i c u l t u ra l e s tabl i s h m e n t

app l i ca n t .

How should we so lve t h e prob lem o f agr i cu l t u r e ?

1.The f a r mer 's f r e e marke t expans ion

The Government and t h e Loca l Pub l i c Oganizat ion

s u p p o r t

t he f a r mers.

2. Sending o f i n f o r m a t i o n on occup a t i o n o f

a g r i c u l t u r e (Food Educa t i o n )

The charm o f a g r i c u l t u r e and th e impor tance o f t he

product i on o f f ood are educated.

3 .P romo t ion o f l o c a l p r o d u c tio n f o r consumpt ion o f l o c a l

This i s an approach t o connec t t he consumers

and the f a r mers.

Sixt h i n d u s t r i a l i z a t i o n i s coo p e r a ti on and in teg ra t i o n o f p r ima r y

i n d u s tr y , secondary i n d u s tr y , t e r t i a r y i n d u s tr y .

Six t h in dust r i al i zat i on pol i cy of agr i cul t ur e, fo r est r y and

f i sher y in dust r y (pr i mar y in dust r y ) i n JPN

The purpose o f t he s i x t h

i n d u s tr i a l i z a t i o n i s t o connect w i t h

v a r iou s i n d u s tr i e s such as r eg iona l

resources, fo od i n d u s t r y, t o u r i s m

i n d u s tr y , energy i n d u s tr y and so on.

And by deve lop ing reg ion a l business

and p r o m o t i ng expans ion o f sales

channels, i t i s t o improve inco me and

crea t e a p lace o f employment i n t he

r u r a l a r ea.

Primaryindustry

Secondary(Processing)

Tertiary(Distribution)

Sixth industriali-zation

A group of farmers operate fromproduction to processing and sales.

To r e a liz e t h i s ...

Humanresources・ Education (Agliculture Science/Food Science/ Economics/ Business administration etc.)

Funds・Gaverment

・ Localpublic entity・ Bank

Expert of Production

Expert of Food

Processing

Expert of Distribution

Efficient Production

Place andEqipment

Channel of Distribution

6th

Industriali-zation

Young power i s

necessa r y

An in crease i n

p op u l a t i o n o f

sen io r c i t i z e n

・ A decrease in producingpopulation・ A decrease in taxrevenu

Lowered tandard of living

M in i s t r y o f Healt h , Labor and Wel fare (JPN): Maintenance and

i m p rovement o f pu b li c h e a lt h

Decrease in finance

・ D e lay of pension payment age・ Price hike of medical expense

The public dissatisfaction

is enhanced.

New law f o r t he l i f e s t y l e

d is ease p a ti en t ' s

decrease and med ica l

expense co n t r o l

Bas i c Act o f

Food and

N u t r i t io n

Educati on2005.6.17

Basic Actof shokuiku

A c tu a l l y

This i s n o t a law f o r t he hea lt h o n ly o f

c h i l d r e n . Th is i s p r o v ided as a law f o r

a l l Japanese c i t i zen 's h e a lt h .

Then… ・ E x t e n s ion of healthy lifelongevity・ An increase in producingpopulation・ Securing of revenue

Maintenance and development of state of country

Guidance t o t h e a d u l t who comple ted t h e d in i ng

hab i t and the l i f e s t y le i s n o t easy.

The education concerning food and health is executed tochildren.

Acc o r d ing t o t he My anmar Diabet es Ass o c i a t io n, i t i s

sai d tha t t he m or t a l i t y r a t e o f non- p a ti e n t w i t h an

i n f e c t iou s d isease such as cancer, th e diabet es, and h e a r t

disease e t c . w i l l account f o r 59% i n t h e deceased p e rson a y ear

i n Myanmar .

M i n i s tr y o f Hea l th o f Myanmar has rep o r t e d th a t abou t 2.7

m i l l i o n o f Myanmarese ( f r om 20 to 72 y e a rs o ld ) are dia b e tes.

Moreov e r, i t is warned t h a t t he prediabe t es are 5 mi l l i o n peopl e

o r more.

And th e h igh b lood

press u re synd rome p a t i e n ts are 6.3 m i l l i o n

peop le accord ing t o s t a t i s t i c s , and th e o b e s i ty synd ro me

p a ti e n t s are 5.3 m i l l i o n people .

10.3 m il l i o n people are us ing a k ind

o f t he chewing t ob acco tha t i s ca l l ed kimma

reg u l a r l y .

(I n My anmar , t he smoking r a t e under t he age o f 15 is 7.7%, wh i ch

Do you need Shokuiku i n Myanmar?

What i s fea tu r e o f Myanma r (Burmese) c u is ine ?Mochinger Khauk swe kyawShan Khao swe

Amedahin

Taminjo Oho no Khao swe

Chin'ieWetahin Chetahin

Various types of sweets

I t i s a mea l st y l e o f bas ic My anma r t hat e a ts 1- 2 k in ds o f

s ide d ish and a la r ge amount o f r i c e 3 t im e s o f JPN). I n the

mor n i n g , Nood les (Mochinge r, Oho no Khao swe e t c .), Naan, F r i ed

dough s t i c k are o fte n eaten as break f as t a t t he st a l l

and t h e d in in g. And, a l a r ge amount o f o i l i s o f t e n used i n t he

Myanmar d is h.

I n My anm ar, peop le are w i l l i n g l y eat i n g the cake tha t

uses

a l o t o f suga r and o i l .And more, Tea and t he s o f t d r i n k w it h a st r ong sweetness are l i k e d .

Energy i s h igh thou gh meal o f Myanmar i s r i c h .

Obesity+

Diabetes

I t is we l l known t h a t obe s i ty, h igh b loo d pressure, t h e dys l i p e mia,

and t he diabet es are t he r i s k f ac t o r s o f th e card i o v asc u l a r d is ease.

The r i s k r i s es sy n e rg is t i c i n t he me t abo l i c syndrome

t ha t comp l i ca tes w i t h two o r more these cond i t ion s.

Concomitant disease

High blood pressure

Diabetes is stressed on the liver.

A decrease in the liver function

depraves the diabetes.

Hepatoma is a diabetic phenotype in the liver

Normal Simple fatty live NASH Cirrhosis Hepatoma

+

I mprovement o f d i e t and exerc ise

are e f fe c t ive f o r t he me tabo li c

syndrome

The use o f f u n c t i o n a l foods

increases e f fec t s

2nd Diet control

3rd Smokingcessation

Appropriate food andnutrition education

Shokuiku1st Exercise

The last is medicine

Metabo l i c syndrome ~ i t s c o p ing method ~

Conclus ion

1. Shokuiku i s necessary e d u c a t ion f o r c h i l d r en t o

become h e a lt h y a d u l t s bo th m e n t a ll y and phy s i c a l l y.

2.Shoku iku i s necessary educa t ion t o succeed the

food cul t u r e o f t he home coun t r y .

3.Shoku iku i s necessary educa t ion f o r t he p r o mo t i o n

o f a g r i c u l t u r e i n the home coun t r y .

Shokuiku i s necessary educa t ion f o r t he hea lt h o f

t he p e o p le and th e prosp e r i t y o f t he coun t r y .

TOKYO UNIVERSITY OF AGRICULTURE(Tokyo NODAI)

Radish “Daikon”

Functional nutrients and chemicals from foods resources

Tokyo University of Agriculture

Yuji YAMAMOTO Ph.D.

1891

Foundation of the Department of Agriculture,

Ikueiko School, based on Tokugawa Ikuei Kai

Foundation emphasizing "Practical Science"

The founder was Mr. ENOMOTO, Takeaki, who

served as Minister of Foreign Affairs, Post and

Telecommunications, Agriculture, Education, and

the Chairman of the Tokugawa Ikuei Kai

Foundation.

Since its establishment in 1891, Tokyo University of Agriculture, with its education and research based on the principle of practical science, has fostered students’ individual talents and skills.

Background of Tokyo NODAI

Return Man to the Farm

Tokyo University of Agriculture and Tokyo NODAI Graduate School

☞ 3 Campuses☞ 2 Graduate Schools (20 majors)☞ 6 Faculties☞ 23 Departments☞ 150 Laboratories

Location of 3 campuses

Hokkaido Okhotsk Campus

Setagaya Campus

Atsugi Campus

「Why do we HAVE TO eat ？」

Question

１） resource of ENERGY

２） Body composition

ANSWER

nutrient

Carbohydrate ; Energy

Lipid; Energy

Protein; Body component

Mineral; Body component

Vitamin; Balance the metabolism

Functional nutrients: ex. derived from amino acids

Tree types of Rice grain

Vitamin B2

Vitamin B1

Iron Magnesium

Niacin

Zinc Protein

Lipid

Dietary fiber

Folic acid

Calcium

Refined Grain

Whole GrainEnergy

Nutritional composition comparison

Of Whole Grain with Refined Grain

Brown rice includes many functional components.

・inositol hexaphosphate（IP6）:from sugar

・γ-oryzanol

・γ‐aminobutyric acid(GABA):from amino acids

Background -Pregelatinized brown rice-

Functional nutrients

any of various organic substances that are essential in minute quantities to the nutrition of most animals and some plants, act especially as coenzymes and precursors of coenzymes in the regulation of metabolic processes but do not provide energy or serve as building units, and are present in natural foodstuffs or sometimes produced within the body

Definition of Vitamins

Fat-soluble

Vitamin A、 Vitamin D 、 Vitamin K、 Vitamin E

These vitamins are not soluble in water but are readily soluble in fat dissolving organic solvents. Theyneed presence of fats for their transport and metabolism. They are found in foodstuffs in associationwith lipids. The fat soluble vitamins are absorbed along with dietary fats. Adequate bile flow andgood micelle formation favor absorption of fat-soluble vitamins. These vitamins are excreted primarilyin the faeces via bile. The following are the characteristics of fat soluble vitamins,1)Adsorbed with dietary fat in micelles.2)Excreted much more slowly.3)Stored in adipose tissue & liver and hence they pose a greater risk of toxicity when consumed inexcess4)Chylomicrons containing fat-soluble vitamins are transported via the lymph to the bloodstream andeventually to the liver.

Classification of vitamins

13

Vitamin A

Chemical structures of important functional forms of vitamin A

・Retinal is essential in vision

・Retinoic acid is involved in growth and cellular differentiation.

Retinoids activate gene expression through nuclear receptors

Water-soluble

1. Vitamin B complex (includes B1, B2, B3, B5, B6, B7, B9, B12)2. Vitamin C (Ascorbic acid)

Water soluble vitamins are are the ones which dissolve in water. These vitamins are sent out of the

body through urine. The ups and downs in the fat absorption do not affect the absorption of water soluble vitamins. The following are the main characteristics of water soluble vitamins,1)Dissolve in water2)Readily excreted by kidney3)Function as a coenzyme & in energy metabolism4)Hydrophilic compounds and water leach them from vegetables5)Marginal deficiency more common

Classification of vitamins

Vitamin B2

Vitamin B1

Iron Magnesium

Niacin

Zinc Protein

Lipid

Dietary fiber

Folic acid

Calcium

Refined Grain

Whole GrainEnergy

Nutritional composition comparison

Of Whole Grain with Refined Grain

Brown rice includes many functional components.

・inositol hexaphosphate（IP6）:from sugar

・γ-oryzanol

・γ‐aminobutyric acid(GABA):from amino acids

Background -Pregelatinized brown rice-

Functional nutrients

Vegetable

http://www.google.co.jp/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&cad=rja&uact=8&ved=0ahUKEwj8qOvG5L_KAhUBMJQKHQS7CDgQjRwIBw&url=http://tst.japan-topics.com/archives/26&psig=AFQjCNH3WrSA130fTVzoWeo2yB030_YzWw&ust=1453633390011957

S-methylmethionine[(CH3)3S(CH2)2CH(NH2)CO2H]+

is also called vitamin U

S-Methylmethionine sulfonium (SMMS) is a derivative of the amino acid methionine, and is synthesized in a variety of plants. SMMS is widely referred to as vitamin U because of its potent therapeutic effect on gastrointestinal ulceration. 。

https://www.google.co.jp/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&uact=8&ved=0CAcQjRw&url=https://ja.wikipedia.org/wiki/S-%E3%83%A1%E3%83%81%E3%83%AB%E3%83%A1%E3%83%81%E3%82%AA%E3%83%8B%E3%83%B3&ei=dc-RVcGFMsa7mwXhvqaIDQ&bvm=bv.96783405,d.dGY&psig=AFQjCNGSzCXEPdf7whjCIrqDuJGb7TRT-Q&ust=1435705582511596

トマトトマト（学名：Solanum

lycopersicum）は、南アメリカのアン

デス山脈高原地帯（ペルー、エクアドル）原産のナス科ナス属の植物。また、その果実のこと。多年生植物で、

果実は食用として利用される。緑黄色野菜の一種である。日本語では唐柿（とうし）、

赤茄子（あかなす）、蕃茄（ばんか）、小金瓜（こがねうり）

などの異称もある。トマトという呼び名は「膨らむ果実」を意味する「トマトゥル」からきていますウィキペディア

エノキタケ

ナメコ

シイタケ

Type 1 diabetesOccurs when the immune system mistakenly attacks and kills the beta cells of the pancreas

Type 2 diabetesOccurs when the body can’t properly use the insulin (insulin resistance)

What is Type 2 diabetes and life style related disease

24

obesity is a main concern in also Myanmar

Metabolic syndrome

Insulin resistance

diabetes Hyperlipidemia

Arteriosclerosis

Obesity

Eating habit Lack of exercise

hypertension

平成28年4月307件平成29年4月927件現在は1300件超

P

培養細胞に薬草抽出物を添加

AMPKの基質

数値化、薬草の選抜

生活習慣病に改善効果を示す薬草の探索（担当：鈴木）

AMPKの活性化を基質のリン酸化状態で評価

AMPKタンパク質

リン酸化

2型糖尿病モデルマウスに薬草を投与

経口糖負荷試験などで血糖値の低下作用を評価

96well plate

Six-week-old male Zucker rats and their lean fed each diet for 10 week.

*Cornstarch of AIN-93G was replaced with each pregelatizined rice.

Group Leptin receptor gene Diet

Lean （＋/？） AIN-93G

Control （－/－） AIN-93G

WR （－/－） AIN-93G＋pregelatinized white rice 53％*

BR （－/－） AIN-93G＋pregelatinized brown rice 53％*

Experimental Design

×10

×

40

Brown Rice

White Rice

ControlLeanL1 W1 B1C1

血清肝臓炎症マーカー

TG NFEA

TC HDL GLU Ad 脂肪面積

T-Lipid

TG TC ALT LDH T-Bil AST ALP AMY

WhiteVs control

－－－－－－－－－

BrownVs control

－－－

BrownVs white

－－－－－－－－－－－

32

p<0.05

p<0.01SUMMARY

Daw Tha Zin Nwe Oo

30.8.2018

1

Screening on some Bioactivities, Elemental Analysis and

Isolation of some Organic Constituents

in Pterocarpus macrocarpus Kurz ( Padauk) Barks

2

This research work aimed to investigate of some chemical constituents,

antioxidant and antimicrobial activities in barks of Pterocarpus macrocarpus Kurz

(Padauk). Some phytochemical constituents such as -amino acids, carbohydrates,

flavonoids, glycoside, organic acids, phenolic compounds, reducing sugars, saponins,

starch and tannins were observed in the sample by using the standard methods. The

EDXRF spectral results revealed the presence of Ca, K, P, Fe, S, Sr, Mn, Rb, Zn, Cu

and Y. Some metal concentrations determined by AAS method were found to be

Ca (125.3 ppm), K (22.04 ppm), Fe (11.29 ppm), Mg (9.904 ppm), Zn (2.388 ppm),

Cr (0.422 ppm) and Pb (0.229 ppm), respectively. The various crude extracts from the

sample like PE (1.64 %), EtOH (4.12 %), EtOAc (1.07 %) and H2O (2.54 %) were

prepared respectively by using successive solvent extraction method. By column

chromatographic separation technique, compound I (1.4 mg, 0.0006%), compound II

(1.1 mg, 0.00044 %) and compound III (1.0 mg, 0.0004 %) were isolated from EtOAc

Abstract

3

crude extract of the sample. Based on FT IR spectral data, the functional group of

compounds I, II and III may be aromatic ,–unsaturated ketone with hydroxyl

groups, aliphatic polyhydroxy ,–unsaturated ketone and aliphatic unsaturated

alcohol. In the investigation of the antioxidant activity screening, the ethanol and

watery extracts were found to possess the mild antioxidant activity by DPPH assay

method. According to the results of antimicrobial activity, the ethanol extract from

the sample was found to inhibit only Pseudomonas aeruginosa (inhibition zone

diameter = 12 mm) while the watery extract did not exhibit the antimicrobial activity

against six microorganisms such as Bacillus subtilis (N.C.T.C-8236),

Staphylococcus aureus (N.C.P.C-6371), Pseudomonas aeruginosa (6749), Bacillus

pumilus (N.C.I.B-8982), Candida albicans and Escherichia coli (N.C.I.B-8134) by

agar well diffusion method.

Key words : Pterocarpus macrocarpus Kurz (Padauk) barks, phytochemical

constituents, antioxidant activity, antimicrobial activity

Outline

-Aim and Objectives

-Introduction

-Materials and Methods

-Results and Discussion

-Conclusion

-References

4

Aim and objectives

Aim

To investigate of some chemical constituents, antioxidant

and antimicrobial activities in barks of Pterocarpus

macrocarpus Kurz (Padauk, PD)

5

Objectives

❖ To collect the sample

❖ To identify the collecting sample

❖ To perform the preliminary phytochemical tests

❖ To analyze the some elemental constituents by using EDXRF and AAS methods

❖ To prepare various crude extracts from collecting sample such as PE, EtOH,

EtOAc and watery extracts by using successive solvent extraction method

❖ To isolate some organic constituents from EtOAc crude extract by using thin layer

chromatography and column chromatographic separation techniques

❖ To determine the physicochemical characteristics of isolated compounds by using

appropriate methods6

7

❖ To detect the functional group nature of isolated compounds by using FT IR

spectroscopic method

❖ To screen the antioxidant activity of ethanol and watery extracts by using DPPH

assay method

❖ To screen the antimicrobial activity of ethanol and watery extracts by using Agar

Well Diffusion method

Introduction

Medicinal plant

❖ have a recognized medicinal use

plant’s seeds, berries, roots, medicinal properties Traditional medicine

leaves, bark or flowers

Traditional medicine one of the oldest forms of medical

treatment in human history

Benefits

1.They cost less - the rising cost of prescription drugs have led the people to

look for alternatives.

2.They may have fewer side effects - while the side effects of any herbal

medication depend on the drug in question, many have fewer side effects than

conventional medicine.

3.There is a choice on how to use them - medicinal herbs can be used in a

variety of ways, depending on the kind of herb that is to be used in a variety of

ways, depending on the kind of herb that is to be used. Some herbs can be

mixed with food. Some can be made into tea, and there are some that are

available in capsule or tablet form.

4.They are good for more than one condition - most prescriptive drugs are

designed for one specific health problem. By contrast, many herbal medicine act

on several parts of body at once.

10

Botanical Aspects of Pterocarpus macrocarpus Kurz

Botanical name - Pterocarpus macrocarpus Kurz

Myanmar name - Padauk

English name - Myanmar Padauk

Family - Fabaceae

Genus - Pterocarpus

Species - macrocarpus

Parts used - bark

Distribution - Native to Southeast Asia in northeasten India,

Myanmar, Laos, Thailand and Vietnam

11

Figure 1 Photographs of flowers, leaves and fruits of Pterocarpus macrocarpus

Kurz (Padauk)

Pterocarpus macrocarpus Kurz (Padauk)

Flowers FruitsLeaves

12

Figure 2 Photographs of barks and plant of Pterocarpus macrocarpus

Kurz (Padauk)

BarksPlant

13

Plant description

➢ medium-sized tree growing to 10-30 m tall, with a trunk up to 1.7 m diameter

➢ Barks flaky, grey brown

secretes a red gum (if cut)

➢ Leaves 20-30 cm long, pinnate, with 9-11 leaflets

➢ Flowers yellow, fragrant produced in racemes 5-9 cm long

➢ Flowering period March-May (in Myanmar)

➢ Fruits a pod surrounded by a round wing 4.5-7 cm

diameter containing two or three seeds

➢ Seeds flat, red-brown, 0.8-1 cm long with leathery seed coat

14

Chemical Composition

❖ pterocarpin,

❖ pterocarpol,

❖ homopterocarpin,

❖ liquiritigenin,

❖ isoliquiritigenin,

❖ pterostibelline,

❖macrocarposide,

❖ –sitosterol,

O

O

O

O

H3CO

OH

HO

H

OO

OO

H

H

O

OH

O

HO

OH

O

OH

HO

OH

OCH3

H3CO

O

O

O

OH

OHOHOH

OH

HO

HO

HO

HO

H H

H

H

15

Uses

✓ valued for its toughness,

✓ stability in use,

✓ decorativeness,

✓ used for furniture,

✓ construction timber,

✓ cart wheels,

✓ tool handles and posts (wood is durable and resistant to termites)

✓ dyes

• Sample collection

Pterocarpus macrocarpus

Kurz (Padauk) bark

• Sample Preparation

-dried samples

-ground into powder

-stored in air tight container

• Identification

Identified at the Department of Botany, Mawlamyine University

16

Mawlamyine University Campus,

Mawlamyine Township, Mon State

Materials & Methods and Results & Discussion

17

Sample extracted

with

1 % HCl 1%HCl extract Alkaloids

SteroidsPE

PE extractCHCl3 Terpenoids CHCl3 extractEtOH EtOH extract Flavonoids

H2O H2O

extractPhenolic compounds

Glycosides

Saponins

Tannins

-amino acids

CarbohydratesCyanogenic glycosides

Reducing sugars

Starch

• Preliminary Phytochemical Investigation of Sample

Organic acids

18(+) = presence, (−) = absence

Sr.No. Tests Extracts Test Reagents Observations Remark

1 Alkaloids 1% HCl

(i) Dragendorff's reagent No orange ppt −

(ii) Sodium picrate solution No yellow ppt −

(iii) Wagner's reagent No brown ppt −

(iv) Mayer's reagent No white ppt −

2 -amino acids H2O Ninhydrin reagent Purple colour +

3 Carbohydrates H2O10% -naphthol,

Red ring +conc:H2SO4

4 Cyanogenic glycosides H2O Sodium picrate No brick red colour −

5 Flavonoids EtOHMg turning,

Pink colour +conc : H2SO4

6 Glycosides H2O 10% lead acetate White ppt +

7 Organic acids H2O Bromocresol green Blue colour +

8 Phenolic compounds H2O 1% FeCl3 Deep blue +

9 Reducing sugars H2O Benedict solution Brick-red ppt +

10 Saponins H2O Distilled water Frothing +

11 Starch H2O I2 solution Deep blue +

12 Steroids PE Acetic anhydride, conc:H2SO4 No green colour −

13 Tannins H2O FeSO4 Deep blue +

14 Terpenoids CHCl3 Acetic anhydride, conc:H2SO4 No pink colour −

Table 1 Phytochemical Test Results of Barks of Pterocarpus macrocarpus Kurz (Padauk)

Determination of Ash Content

Dried powder sample

(5 g) in a preweighed

porcelain crucible

heated

at 6000C

grey

colour of

ash

cooled,

weighedConstant weight

of ash

Ash content (%) Weight of ash (g)

Weight of dried

powder (g)

x 100

Elemental Analysis from Barks of Pterocarpus macrocarpus Kurz (Padauk)

Sr.No. Sample weight

(g)

Ash weight

(g)

Ash

content (%)

Average

ash content

(%)

1.

2.

3.

5

5

5

0.2541

0.2685

0.2926

5.082

5.370

5.852

5.43

Table 2 Ash Content in Barks of Pterocarpus macrocarpus

Kurz (Padauk)

Elemental Analysis on Barks of Pterocarpus macrocarpus Kurz (Padauk)

20

21

- Elemental contents were measured by EDXRF method at URC

(Universities’ Research Centre)

Some Elemental Analysis from the sample by using EDXRF and

AAS methods

• Qualitative Determination of the Contents of Elements by EDXRF Method

• Quantitative Determination of the Contents of Elements by AAS Method

- Elemental contents were measured by AAS method at URC

(Universities’ Research Centre)

Table 3 Relative Abundance of some Elements in Barks of Pterocarpus macrocarpus

Kurz (Padauk)

Elements Relative abundance (%)

Ca 71.929

K 20.255

P 4.649

Fe 1.932

S 0.494

Sr 0.371

Mn 0.169

Rb 0.070

Zn 0.067

Cu 0.054

Y 0.01122

Qualitative Determination of some Elements by EDXRF Method

23

No. Elements Metal concentration (ppm)

1. Ca 125.3

2. K 22.04

3. Fe 11.29

4. Mg 9.904

5. Zn 2.388

6. Cr 0.422

7. Pb 0.229

Table 4 The Concentrations of some Elements in Barks of

Pterocarpus macrocarpus Kurz (Padauk)

Quantitative Determination of some Elements by AAS Method

24

residue

concentrated by evaporation

extracted with PE (800 ml)

Soxhlet extractor (7 h)

extracted with EtOH (800ml),

Soxhlet extractor (14 h)

partitioned between

EtOAc / H2O

concentrated by rotatory

evaporator

residue

PE layer

PE extract

(4.108 g)

95% EtOH soluble matter

defatted 95% EtOH

extract (10.31g)

EtOAc layer aqueous layer

EtOAc extract

(2.664 g) H2O extract

(6.337 g)

Powdered sample (250g)

Figure 4 Flow diagram for the preparation of various crude

extracts from barks of P.macrocarpus Kurz (Padauk)

Preparation of Crude Extracts

concentrated by rotatory evaporator

concentrated by rotatory

evaporator

25

Plant sample Solvent Extract weight (g) Yield (%)

PD Bark

PE 4.11 1.64

95%EtOH 10.31 4.12

EtOAc 2.66 1.07

H2O 6.34 2.54

Table 5 Yield Percent of Crude Extracts from Barks of Pterocarpus macrocarpus

Kurz (Padauk)

Preparation of Crude Extract

26

EtOAc extract (2.664g)

PE:EtOAc

(9:1,v/v, 800ml)

Compound I

White powder

1.4 mg, 0.00056%

Rf = 0.54 (PE:EtOAc = 2:1,v/v)

CC on silica gel

PE:EtOAc gradient elution

Total 366 fractions

PE:EtOAc

(2:1, v/v,600ml)

f71-f130

FIV

Removal

of solvent

Solid material

-Washed with PE

-Crystallized from

PE:EtOAc = 9:1, v/v

Compound II

white powder

1.1 mg, 0.00044%

Rf = 0.23 (EtOAc only)

f191-f256

FVI

Removal

of solvent

Solid material

-Washed with PE

-Crystallized from

PE:EtOAc = 2:1, v/v

PE:EtOAc

(1:1,v/v,500ml)

Compound III

white powder

1.0 mg, 0.0004%

Rf = 0.2 (EtOAc only)

f257-f322

FVII

Removal

of solvent

Solid material

-Washed with PE

-Crystallized from

PE:EtOAc = 1:1,v/v

Figure 5 Procedure for the separation of EtOAc crude

extract from barks of PD by column chromatography

Isolation of some Organic Constituents by Column Chromatography

27

Physical properties

Isolated compounds

I II III

Solubility in

PE − − −

EtOAc + + +

EtOH − − −

MeOH − − −

CHCl3 − − −

H2O − − −

Rf value 0.54

(PE : EtOAc =

2:1, v/v)

0.23

(EtOAc only)

0.2

(EtOAc only)

Physical state White powder White powder White powder

Yield (%) 0.0006 0.00044 0.0004

Table 6 Some Physical Properties of Isolated Compounds

Isolation of some Organic Compounds from EtOAc Crude Extract of Barks of PD

(+) = soluble,

(−) = insoluble

28

Table 7 Some Chemical Properties of Isolated Compounds

Isolated

compounds

Test reagents Observations Remark

I

1% FeCl3 No colouration Not a phenolic compound

Vanillin – H2SO4 No colouration Not terpenoids / phenol derivatives

Liebermann-Burchard No colouration Not steroids

II


Vanillin – H2SO4 Blue black Terpenoids / phenol / steroids derivatives

Liebermann-Burchard Pink Steroids

III


Vanillin – H2SO4 Blue black Terpenoids / phenol / steroids derivatives

Liebermann-Burchard No colouration Not Steroids

29Figure 6 FT IR spectrum of isolated compound I

Study on FT IR Spectra of Isolated Compounds

3070

30

Table 8 Assignment for FT IR Spectral Data of Isolated Compound I

Significant frequencies (cm-1) Assignment

3443 O-H (alcohol)

3070 =CH (aromatic)

2918 C-H in CH2 and CH3 (antisymmetric)

2850 C-H in CH2 and CH3 (symmetric)

1730 C=O (ketone)

1637 C=O and C=C in ,–unsaturated ketone

1620, 1599, 1570, 1512 C=C (ring) in aromatic

1456 C-H in CH2 and CH3 (antisymmetric)

1381 C-H in CH2 and CH3 (symmetric)

1315, 1276 O-H (in-plane) (alcohol)

1099, 1082, 1028 C-O (alcohol)

C-H (in-plane) (aromatic)

1028 C-O (in cyclic alcohol)

970 =CH (out-of-plane) in alkene

721 C-H (out-of-plane) (aromatic)

549 C=C (ring) (aromatic)

31Figure 7 FT IR spectrum of isolated compound II

32

Table 9 Assignment for FT IR Spectral Data of Isolated Compound II


3333 O-H (alcohol)

3223 O-H (alcohol)

2918 C-H (antisymmetric) of CH2 and CH3

2848 C-H (symmetric) of CH2 and CH3

1726 C=O (ketone)

1620 C=O and C=C in , –unsaturated ketone

1465 C-H of CH2 and CH3

1280 O-H (in-plane) (alcohol)

720 =CH (out-of-plane) in alkene

Figure 8 FT IR spectrum of isolated compound III33

Table 10 Assignment for FT IR spectral Data of Isolated Compound III


3394 O-H (alcohol)

2958, 2920 C-H (anti-symmetric) of CH2 and CH3


1641 C=C (alkene)

1465 C-H (anti-symmetric) of CH2 and CH3


1165, 1105, 1072 C-O (alcohol)

34

35

Antioxidant Activity –DPPH Assay Method

Materials required

- 2,2-diphenyl-1- picryl- hydrazyl ( DPPH ) ,95% EtOH, 50% EtOH

- Cell (5 ml), vortex mixer, Spectrophotometer,

electric balance, pipette ( 10 ml)

Standard

- BHT (Butylated Hydroxy Toluene)

Samples

- EtOH and H2O extracts

Procedure

Control

1500l (60 M DPPH in EtOH)

+

1500 l 95 % EtOH

Blank

1500 l (sample)

+

1500 l 50 % EtOH

Sample

1500 l (60 M DPPH in EtOH)

+

1500 l sample

mix thoroughly

with vortex mixer

30 min

R.T

mix thoroughly

with vortex mixer

30 min

R.T

mix thoroughly

with vortex mixer

30 min

R.T

measured the

absorbance at 517 nm

Figure 9 Procedure for DPPH assay 36

37

Where, % inhibition = percent inhibition of test sample

DPPH alone = absorbance of DPPH in EtOH solution

Sample = absorbance of sample + DPPH solution

Blank = absorbance of sample + EtOH solution

DPPH alone - ( Sample - Blank ) 100

DPPH alone% inhibition =

IC50

(50% inhibitory concentration)

38

Antioxidant Activity Test

DPPH only EtOH extract

only

H2O extract

only

(a) Before the reaction (b) After the reaction

DPPH only DPPH + EtOH

extract

DPPH + H2O

extract

Figure 10 Photographs of the changes in colour of DPPH free radical against

ethanol and watery extracts of barks of PD

39

Table 11 Results of Antioxidant Activity Screening of Ethanol and

Watery Extracts of Barks of PD by Using DPPH Assay Method

Samples EtOH

extract

H2O

extract

Remark

Activity +++ +++ Sample concentration

200 µg/ml

− - no change in colour of DPPH free radical

+ - slowly change in colour of DPPH free radical

+++ - immediately change in colour of DPPH free radical

40

Antimicrobial Activity Test- Agar Well Diffusion Method

- PE, 95% EtOH solvent

- Tryticase soy agar, Tryticase soy broth

Samples

- EtOH and H2O extracts

Test organisms

- Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus pumilus,

Candida albicans , Escherichia coli and Bacillus subtilis

Materials required

(measured at CRDC)

Sterile petridish (1) Extract (1 g) was introduced

(ii) Dissolved in 1 ml of its

respective solvent

Concentrated extract

Bacterial suspension

from trypticase soyagar plates

cultured on the agar

medium

agar well with 10 mm

sterile cork bore

inoculum

had been dried

(5 min)

the agar

disc were

removed

the well

were filled

with sample

after overnight

incubation at

37C

Inhibition zone

diameters

41

Procedure

Bacillus subtilis (N.C.T.C-8236) Staphylococcus aureus (N.C.P.C-6371) Pseudomonas aeruginosa (6749)

Bacillus pumilus (N.C.I.B-8982) Candida albicans

A – EtOH control B− H2O control Agar well – 10 mmC – EtOH extract D− H2O extract 10 mm ~ 14 mm (+), 15 mm ~ 19 mm (++), 20 mm above (+++)

Figure 11 Inhibition zone diameters for EtOH and watery extracts of barks of PDagainst six species of microorganisms

E.coli (N.C.I.B-8134)

A

C B

D A

C B

DA

C B

D

A

C B

DA

C B

DA

C B

D

43

Table 12 Inhibition Zone Diameters of Ethanol and Watery Extracts of

Barks of Pterocarpus macrocarpus Kurz (Padauk) against Six

Microorganisms by Agar Well Diffusion Method

Agar well - 10 mm Organisms

10 mm~14 mm (+) (1) Bacillus subtilis (N.C.T.C-8236)

15mm~19 mm (++) (2) Staphylococcus aureus (N.C.P.C-6371)

20mm above (+++) (3) Pseudomonas aeruginosa (6749)

(4) Bacillus pumilus (N.C.I.B-8982)

(5) Candida albicans

(6) Escherichia coli (N.C.I.B-8134)

Antimicrobial Activity Test

Sample Extracts

Organisms and inhibition zone diameters (mm)

B.subtilis S.aureus P.aeruginosa B.pumilus C.albicans E.coli

PD Bark

EtOH - - 12

(+)

- - -

H2O - - - - - -

Conclusion

From overall assessment of the research work concerning about the some chemicals and

biological investigation in the Pteroccarpus macrocarpus Kurz ( Padauk) barks, the following

inferences could be drawn.

1. Preliminary phytochemical tests → -amino acids, carbohydrates, flavonoids, glycosides,

organic acids, phenolic compounds, reducing sugars, saponins, starch and tannins

2. Relative abundances of some elements by EDXRF method → Ca (71.929 %), K (20.255 %),

P (4.649 %), Fe (1.932 %), S (0.494 %), Sr (0.371 %), Mn (0.169 %), Rb (0.070 %),

Zn (0.067 %), Cu (0.054 %) and Y (0.011 %)

3. The concentrations of some elements by AAS method → Ca (125.3 ppm), K (22.04 ppm),

Fe (11.29 ppm), Mg (9.904 ppm), Zn (2.388 ppm), Cr (0.422 ppm) and Pb (0.229 ppm)

44

45

4. The various crude extracts prepared by using successive solvent extraction method

→ PE (1.64 %), EtOH (4.12 %), EtOAc (1.07 %) and H2O (2.54%)

5. Three isolated compounds : compound I (1.4 mg, 0.0006 %, white powder form),

compound II (1.1 mg, 0.00044 %, white powder form) and compound III (1.0 mg,

0.0004 %, white powder form)

6. The functional groups of three isolated compounds → compound I (aromatic ,–

unsaturated ketone with hydroxyl groups), compound II (aliphatic polyhydroxy ,–

unsaturated ketone), compound III (aliphatic unsaturated alcohol)

7. Antioxidant activity → both of the ethanol and watery extracts had the

mild antioxidant activity (by DPPH assay method)

8. The poor antimicrobial activity of the ethanol extract of barks of PD → only

Pseudomonas aeruginosa (inhibition zone diameter = 12 mm)

Anderson, J. and Fitzgerald, C. (2010). “Iron : An Essential Nutrient”, Food and Nutrition

Series/ Health, Colorado State University Extension, Fact Sheet No. 9,356, 1

Chow, K. W. (1980). “ Quality Control, in Fish Feed Manufacturing”, In Fish Feed

`Technology, FAO/ UNDP Training Course, College of Fisheries, University of

Washington, ADCP / Rep / 80 / 11, 369-385

Cruickshank. (1960). Handbook of Bacteriology, Edinburgh: 10thEd, E&S. Living Stone

Ltd., 256

Francis, J.K. (1989). Pterocarpus macrocarpus Kurz. Burma padauk, pradu

Leguminosae, Legume family. Silvics Manual, No. SO-ITF-SM-19, New Orleans,

LA, USA: USDA Forest Service, Southern Forest Experiment Station, Institute of

Tropical Forestry, 673

Harborne, J.B.(1984). Phytochemical Methods A Guide to Modern Techniques of

Plant Analysis, London : 2nd Ed, 37, 101 46

47

Hatano, T., Edamatsu, R., Hiramatsu, A.,Fujata, Y.,Yasubara, T., Yashida, T. and

Okuda, T. (1989). “Effects of Tannins and Related Polyphenols on

Superoxide Anion Radical and on 1,1-Diphenyl-2- Picryl Hydroxyl Radical”,

Chem. Pharm.Bull, 37,2016-2021

Lavin, M., Pennington, R.T., Klitgarrd B.B., Sprent, II., de Lima, H.C. and Gasson,

P.E. (2001). “The Dalbergioid Legumes (Fabaceae) : Delimitation of a

Pantropical Monophyletic Clade”, Am J Bot, 88 (3), 503-33

M-Tin Wa. (1972). Phytochemical Screening, Methods and Procedures,

Phytochemical Bulletin of Botanical Society of America, Inc., 5(3),4-10

Marini- Bettolo, G.B., Nicole Hic, M. and Palamia, M. (1981).“Plant Screening by

Chemical and Chromatographic Procedure under Field Conditions”, J.

Chromate, 46(2), 359-363

Muyskens, M. and Vitz, E.D. (2006). “The Fluorescence of Lignum nephriticum : A

Flash Back to the Post and a Simple Demonstration of Natural Substance

Fluorescence”, Journal of Chemical Education, 83 (5), 765

Nay Mar Soe. (2005). Isolation and Characterization of Chemical Constituents

Possessing Radical Scavenging Activity of a Selected Myanmar Medicinal

Plant, PhD (Dissertation), Department of Chemistry, University of Yangon,

20-22

Robinson, T. (1983). The Organic Constituents of Higher Plants, North America: 5th

Ed, Cordus Press, 63-68

Shriner, R.L., Fuson, R.C., Curtin, D.Y. and Morrill, T.C.(1980). The Systematic

Identification of Organic Compounds, A Laboratory Manual, New York: John

Wiley and Sons, 15-20

Trease, G.E. and Evans, W.C. (1980). Pharmacognosy, London: Spottis Woode

Ballantyne, 62248

Vogel, A.I. (1966). A Text Book of Practical Organic Chemistry. London: 3rd Ed,

Longmans, Green & Co. Ltd., 112

Wikipedia. (2004). Occupational Allergic Contact Dermatits Caused by Padauk

Wood, http://www. en. wikipedia. org/ wiki/ Pterocarpus soyauxii Taub.

(Accessed 25 November 2014)

Wikipedia. (2005). Pterocarpus indicus (narra), http://www. en. wikipedia. org/ wiki/

Pterocarpus indicus (Accessed 20 October 2014)

Wikipedia. (2013). Purdue University New Crops : Pterocarpus indicus,

http://www.en.wikipedia.org/wiki/Pterocarpus indicus (Accessed 20 October

2014)

Wikipedia. (2014). Pterocarpus macrocarpus, http://www. en. wikipedia. org/ wiki/

Pterocarpus macrocarpus (Accessed 10 September 2014)

49

Exportability of Medicinal Plantsto Japan

The Institute of Pharma Food Co., Ltd.

President UNO, Tomoko

（Pharmacist）

①Commercially Producible Medicinal Plants

cultivated in Kayin State and

their Typical Medicinal Properties

Medicinal

PlantsTypical Medicinal Properties

Turmeric Curcuminoids（Curcumin）

Ginger Gingerol，Shogaol，Galanolactone

Capsicum(Red pepper)

Capsaicin

Yam Diosgenin

②Principal Pharmacological Effect

of Typical Medicinal PropertiesMedicinal Plants Typical Medicinal

PropertiesMedical Efficacy

Turmeric Curcuminoids

（Curcumin）

Promoting Bile Secretion

→ Choleretic Action

Ginger Gingerol Stomachic Action,

CholereticAction

Shogaol Elevating Body Temperature,

Analgesic Action, Antiallergy

Galanolactone Antiemesis

Capsicum(Red pepper)

Capsaicin Improve Blood Circulation, Aperitive

Yam Diosgenin Improving and/or

Preventing Dementia

③About an Effective Ingredient ”Diosgenin” contained in

the Yams under growing test in KMPRC

Botanical Names The Content of Diosgenin

(mg/100g)

D.alata N.D.D.alata

D. opposita

D. japonica

D. esculenta

D.bulbifera

D.opposita 0～2

D.japonica 0～2

D.esculenta 8～10

D.bulbifera N.D.

④The Imported Amount of

Medicinal Plants to Japan(comparison of 2012 and 2017)

（ton）

（Created by The Institute of Pharma Food Co., Ltd. using Trade statistics (Ministry of Finance) as a reference.）

Medicinal

Plants

The Total Amount Quantity and Ratio ofImports from China

2012 2017 2012 2017

Turmeric 4,338 3,824 629 14.50% 723 18.90%

Ginger 71,592 66,873 50,211 70.13% 47,826 71.52%

Capsicum 11,475 12,399 9,670 84.27% 10,617 85.63%

Yam 1,111 1,469 918 82.65% 1,241 84.48%

⑤ Conclusion

◎ To export the medicinal plants cultivated in Kayin State

⇒Cultivate the medicinal plants which contain

a fixed quantity of medicinally effective component.

⇒ Prevent the generation of defective

by improving processing technologies.

⇒Establish the agricultural methods by the entire region

without relying on chemical fertilizers and

agrochemicals and conserve environment.

◎ Increasing exports↓

Acquisition of foreign currencies↓

Farmers, State, and Nation will become better off.

A Distribution Channel of Crude Drugs

Farms Local collector and processorsin local a herbal medicine market

Local brokers

Wholesale dealers

（A example of imports from China）China

Local exporters or Joint ventures between a local exporter and a Japanese company

purchase

JapanHerbal medicine wholesalers and trading companies in Japan

In case of exclusively used as drugs, they have to retain import license

every crude drugs from the Ministry of Health, Labor and Welfare.

Processors make sterilized powder, slice and slice for essence, and extract essence.

Products are sold as manufacturing materials or are sold to pharmacies for preparation.

Consumers

Kayin State Medicinal Plant Resource Center(KSMPRC) 1

GHPP and QA/QCDr. T. Matsuo/Technical Advisor to KSMPRC


What kind of information do you need on the raw material?

What kind of information do you need onthe raw material?1. Where is it harvested?

2. How is it selected?

3. Foreign matters contamination?→stone, sand, insect, bacteria

heavy metals, pesticide

→how is the washing(method/time)

→analytical results(bacteria/heavy metal/pesticide)

4. How is the identification? →macro/microscopic observation

5. Content of active ingredient(eg.; curcumin in turmeric)?

Above are requirementsKaoyinfStatce MleideicinaltPlasnt/ResionurcteeCenrtenr(KSaMtPRiCo) nal community. 3


MedicinalPlant

from growto harvest

Herbal raw material

Cut/dried Powdered

Dosage form/Traditional

Med./Health Suppl.

WHO/GACP WHO/GHPP GMP

Selection/processing

produce dosage form/extract

Integrated International/ASEAN Requirements

Today’s seminar covers;

1. The safety and quality should be ensured at every step of herbal production processthrough GACP(medicinal plant), GHPP(herbal processing) and GMP(manufacturing finished product) in the quality assurance (QA) and control(QC) of herbalmedicines.

2. What are QA and QC?

QA and QC of KSMPRC are introduced.

3. What is GHPP?

GHPP is applied to produce herbal(raw) materials(annex 1) , the herbal

preparation(annex 2 )and herbal dosage form(annex 3, to which GMP is

also applied)

4. SOP

GXPs require SOP.

5. SOP to produce primary herbal material at KSMPRC, which should be complied withGHPP Kayin State Medicinal Plant Resource Center(KSMPRC) 6

Organization Chart of Processing Facility ofKSMPRC


MM

Manufacturing staffs

selection washing cutting drying milling packaging labelling cleaning

QM

QC staffs

physical microbial heavy metal pesticide

characteristics

Manufacturing (processing) Facility

QAU

What are QA and QC?• Quality Assurance:

"To assure whether the standards, processes, and procedures are

appropriate for the project and are correctly implemented“

key words; QAU, inspection/audit

• Quality Control:

"To check that the project follows its standards, processes, and

procedures, and that the project produces the required internal and

external (deliverable) products“

key words; QC check

• Thus, Quality Assurance focuses on the process of quality, whileQuality Control focuses on the quality of output.


What does GHPP require us?

Annex 1; Example of production of a herbal material(SOP)

1. Objective of the SOP protocol

2. Scope

3. Procedures

1) Sampling and quality testing; morphology, identification,

water content, total ash

2) Quality control assay

(1) marker compound(eg.; curcumin/turmeric)

(2) HPLC analysis Kayin State Medicinal Plant Resource Center(KSMPRC) 9

What does GHPP require us? -continued-

3) initial sorting

4) washing

recovery%=weight after washing/weight before washing

5) cutting/sectioning/comminuting

6) drying of processed herbal material

recovery%=weight after drying/weight before drying

7) final sorting

8) packaging, labelling, and storageKayin State Medicinal Plant Resource Center(KSMPRC) 10

Kay in S ta te M e dic in a l P lan t Re sou rc e Ce n te rQ u a lity Von tro l L abo ra to ry

U n iqu e Spe c if ic a tio n Sh e e t

M e dic in a l R aw M a te ria ls ; □ c u t, □ pow de re dP lan t n am e (sc ie n tif ic )

□ f ru it , □ ro o t, □ ba lk , □ tru n kN am e o fQ M

Lo t n o ./ dry w e igh t(Kg)

s ign a tu re P rin t D a te

Te st ite m s Spe c if ic a tio n c on fo rm e d to spe c if ic a tio n

□ ye s □ n oif n o , w rite ac tu a l da ta

1 . P h ys ic a l c h a rac te r is t ic1 ) M ac ro sc opic e xam in a tio n □ ye s □ n o

2 ) O rgan o le ptic c h a rac te r ist ic s □ ye s □ n o

3 ) M ic r isc opic e xam in a tio n □ ye s □ n o

4 ) Cu t s ize in m m

□ ye s □ n o

5 ) Fo re ign m a tte r n e ga tiv e □ ye s □ n o

6 ) W a te r c on te n t le ss th an ??% (to be de f in e d

□ ye s □ n o

2 . M ic ro b ia l lim its TAM C (CFU / g o r CFU / m l)

N M T 2 x1 0 4 □ ye s □ n o

TYM C (CFU / g o r CFU / m l)

N M T 2 x1 0 2 □ ye s □ n o

Spe c if ie d m ic ro o rgan ism s・Abse n c e o f Esc h e ric h ia c o li □ ye s □ n o

in 1 g o r 1 m l

・Staph y lo c oc c u s au re u s; n o t don e

・Sa lm on e lla ; n o t don e

3 . H e avy m e ta l n o t don e

4 . A rse n ic n o t don e

Summary1.In the context of Quality assurance and control of herbal medicine,GACP, GHPP and GMP are integrally linked each other.

2.GHPP provides technical guidance on good processing practices of preparing herbal materials, herbal preparations and herbal dosage forms for therapeutic applications.

3. GHPP concerns ensuring the quality of the herbal materials, herbalpreparations and herbal dosage forms produced.

4. GHPP is to promote safety, efficacy and sustainability of herbalmedicines.

5.For this purpose, KSMPRC created SOP complied with GHPP, where QA and QC at KSMPRC play the important roles not only to promote herbal farmers’ quality, but also to bridge the technicalissues between farmers aKnayidn Stathe MeedricibnalaPlalnt RiensoudrceuCesntetr(rKSiMePRsC)

in Myanmar.12


Medicinal plant Selection

Washing

Drying

Labelling

Packaging

CuttingThank you for your attendance!!!

Analysis of Curcumin level of Turmeric

Presented by

Zin Nge Nge Tun

Q.C Officer

Kayin State Medicinal Plant Resource Center

Different Uses of Turmeric

1. Used as Food

2. Used as Dye

3. Used as Traditional

Medicine

4. Used as Food Supplement

5. Used in Cosmetic

6. Used in Skin Care

Heal wounds

Alzheimer's

disease

Anti-inflammatory

Anti-cancer

Omega3 Booster

Anti-Oxidant

Prevent skin damage

Prevent cancer growth

cells

Some of the benefits of Curcumin

(Ref: health line)

(Ref: health line)

(Ref: health line)

(Ref: health line)

(Ref: health line)

(Ref: health line)

(Ref: health line)

Different townships that can produce of Turmeric

in Kayin State

1. Kyarinnseikgyi

2. Kawkareik

3. Taungkalay

4. Thandaungyi

5. Htee lone

Types of Q.C test of Turmeric in Center

1. Macroscopic characters

2. Microscopic characters

3. Moisture content

4. Foreign matters

5. Microbial limit

6. Thin Layer Chromatography

7. UV Vis Spectrophotometer

8. HPLC

1. Identification of the species by

microscopes

2. Checking the moisture content of

the materials

Stages of Turmeric Test in Center

3. Checking the Microbial

contamination

5. Qualitatively identification of

the active components by UV Vis

Spectrophotometer

4. Qualitatively identification of the

active components by Thin Layer

Chromatography

6. Quantitatively identification of

the active components by High

Performance Liquid

Chromatography

Depends on the different township, Turmeric are different

in color, size and appearance.

1. Kyar Inn Seik

Kyi

4. Kaw Ka Reik 5. Htee Lone

2. Than

Daung Gyi3. Shan State

Result of Curcumin level according to the different

townships

No. Townships Quantity of

purchased(kg)

Colour Curcumi

n Level

1. Kyar In Seik Kyi 976.05 Orange 2.6%

2. Kaw Ka Reik 10,310.75 Orange 4.9%

3. Taung Ka Lay 244.8 Orange 4.4%

4. Than Daung Gyi (1) 12,601.96 Light

Yellow

0.4%

5. Than Daung Gyi (2) 16,184 Orange 2.9%

6. Than Daung Gyi (3) 25,036.64 Orange 2.7%

7. Than Daung Gyi(4) 6,601.05 Orange 3.8%

8. Than Daung Gyi (5) 6,570.85 Orange 2.7%

9. Htee Lone 834.75 Orange 3.3%

10. Shan 4,206.45 Orange 2.2%

(2) Steeping

Flow Chart for Turmeric Processing

(1) Selection and Primary washing

(3) Foreign matter detection

before packaging and labeling

(4) Package of dried turmeric

Photos of Sliced and dried Turmeric

1. Botanical Name

Curcuma longa L.

2. Moisture content

< 7%

3. E.coli

Nil

4. Foreign matter

Nil

Chemical

free

Contamina

tion free

Analysis of

active

component

s

Safety of

food

Conclusion

Analysis of active components is one of the food and drug

safety process. Therefore, it may lead to value added

product and higher market potential.

GACP GHPP Q.CValue added

product

Thank You

For Your

Attention !

May I have Any

Questions?

The Quantitative Analysis of

Medicinal Properties

in Medicinal Plants

Authorized NPO R&D Agency for Curative Natural Products

WATANABE, Yukari

（Pharmacist）

In Kayin State Medicinal Plant Resource

Center （KMPRC), a staff performs growing

tests, primary processing, and shipping of

medicinal plants.

We aim to supply market with excellent raw

materials. For that purpose, we determine

medical components and ensure quality of

medicinal plants as a part of consistent quality

control, “cultivating”→”processing”→”shipping”.

We introduce to you the important method for

quantitative analysis of medicinal component

that KMPRC is concentrating on.

There are several types of method for quantitative analyzing

medicinal properties.

1.Thin-layer

Chromatography 2.Paper

Chromatography 3.Gas

Chromatography 4.Liquid

Chromatography 5.Mass

Spectrometry

（Combined use of 3 and 5, or 4 and 5）

✓1 and 2: They are formerly used.

✓3: In order to analyze a sample, we have to vaporize it at

a high temperature. But this method is employed to

analyze

evaporable essential oil and agrochemicals.✓4: We can analyze a sample in its liquid state. In the case

of a solid body, if we liquefy it, we can analyze. An analysis is

possible at ordinary temperature.

High Performance LiquidChromatography; HPLC

This quantitative analyzing method can

carry out highest analytical accuracy and is

the most versatile in the world.

Liquid Chromatography: When components

pass through a column(stationary phase) with

the solution (mobile phase), components are

separated in a column based on each

component’s affinity and molecular size.sample

Component separation

Sample

introducing

Mobile

phase

Signal from detector

Retention time

Detection

HPLC Method

FEEDING

LIQUID

SAMPLE

INJECTIONSEPARATION DETECTION ANALYSIS

Eluent

(mobile phase)

Pump Injector

Data processor Column

Conventional particle packing column

Detector

Waste liquid

Chart

(Chromatogram)

APPARATUS

High Performance Liquid Chromatographs

The Quantitative Analysis of

Medicinal Components at KMPRC

We consider determining Curcumin in

Turmeric, and 6-gingerol and 6-shogaol

in Ginger.

Ms. Zin Nge Nge Tun(KMPRC staff,

Pharmacist) is going to present a flow

and practice of the quantitative analysis.

€¦ · Conservation and Sustainable Use of Medical Plants in Kayin State-GAP and GCAP-KENJI IRIE...

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