2-2. 2 国間ワークショップ 「日本・モンゴル、日本・インドネシア：生物遺伝資源とその取扱い」(1) 日本・モンゴル 2 国間ワークショップ「モンゴルにおける生物遺伝資源とその取扱い」
2011 年 9 月 30 日、JBA は、モンゴルよりモンゴル国自然・環境・ツーリズム省/持続可能な開発及び戦略計画部の Banzragch Tsesed 局長と、植物の専門家であるモンゴル国立大学のJavzan Batkhuu 教授を招聘し、野村コンファレンスプラザ日本橋（東京）において 2 国間ワークショップを開催した。Banzragch 局長にはモンゴルの生物遺伝資源に関する現状と将来について（講演 1）、Batkhuu 教授にはモンゴルの薬用植物の利用について（講演 2）お話いた
だいた。
講演 1：
「Convention on Biodiversity in Mongolia: Implementation of Nagoya Protocol andfuture approaches」（発表資料 1 参照）
モンゴル国内には多様なエコシステムがある。モンゴルは 1993 年に CBD を批准し、1996
年には国の生物多様性戦略、行動計画を決めた。絶滅危機にある動植物を自然環境で保護し、
現在約 14％が保護地域（保護地域を 4 つのレベルに設定し、70 カ所を指定）となっている。
2030 年にはこれを 30％にしたい。現地住民を保護活動に参加させることが重要で、130 万 ha
を現地住民の組合に委譲し保護活動を委任している。モンゴルは社会主義から自由主義の経済
成長の時代に移行し、地下資源の開発が盛んになってきた。これを自然環境の保護とどのように対処させるかが重要である。2009 年、河川の源流での地下資源開発を禁止した。これに対し開発会社が反対活動を続けているので、法の実現は非常に難しい。モンゴルの生物多様性は
年々減少している。この要因は、野生生物を狩猟し海外へ輸出する、また、遊牧民による森林
資源の燃料化によるものである。
遺伝資源は非常に豊かであるが、国の最大関心事は地下資源・家畜であり、遺伝資源の経済的な評価はされていない。種の多様性も豊かであるが、あまり科学的研究もされていない。限
られた研究の中でも、ほとんどの動物は登録されている。海外との共同研究により、新種発見も増えている。
モンゴルにおける遺伝資源に関する法律

憲法（1992）：条項 6 では、資源は国家の財産であるとされた。



環境保護法：条項 15 では、保護機関は自然環境省とされた。



動物相に関する法、植物相に関する法、狩猟に関する法：資源の状況により用途を決め、制限する。

－ 57 －

遺伝資源に関する法律（Future goal）
 「名古屋議定書」は内閣の承認を得て、国会に提出中。本年中に署名したい。
 その後、モンゴルの遺伝資源法（案）を国会に提出する予定。
 遺伝資源法（案）に含めること：中央政府の権限、地方政府の権限、利用許可システムの在り方、支払いシステムの在り方、伝統的知識の扱い方（モンゴル自然環境省が担当）
遺伝資源法（案）に関し、Banzragch 局長は、日本から意見提案をして欲しい旨述べた。また、彼は「利益配分など、モンゴルにとってどのようなシステムが良いのか一緒に考えて欲しい。教育、人材育成の分野でも協力してほしい。モンゴルの遺伝資源データ・ベース構築を大きなプロジェクトとしたい」と日本への期待を述べた。

発表資料 1
Geography of Mongolia

Convention on Biodiversity in Mongolia
Implementation of Nagoya Protocol and future
approaches●

BANZRAGCH Tsesed/PhD/
Director-General of Sustainable Development andStrategic Planning Department,
The Ministry of Nature, Environment and Tourism
September 30, 2011

1

2

Main natural ecosystem types

Implementation of Biodiversity Convention in
Mongolia► Mongolia joined the Convention on Biodiversity in

1993► Mongolian Government is approved National
Biodiversity Strategy and Action Plan /NBSAP/ in 1996
► Major activities:► Enhancing the survey on rarity of animals and plants► Reducing the adverse impact on rare animals and plants► Limiting usage of rare animals and plant species

3

4－ 58 －

► To protect biodiversity promoting in-situ conservation.

Currently 14.4 % of total territory or 22.94 million ha of land
is under state protection. There are 70 specific areas ofprotected areas which administrated by 28 Protected Areas
Management Administration.
► To enhance rehabilitation of endangered species.
► Reintroduction of wild horses /Prijivalskii horse/► Implementation of reforestation projects nationwide .

56• To introduce community based natural
resources management.
- 1.3 million ha forest protected and theirplants, animals and pastureland preserved- 541 forest local communities are existing7• In 2009, Mongolian Parliament approved the Law on
restricting mining in the river basin and forest areas.
•The law provides legal base to protect biodiversity inriver basin and forested areas.

8

Biodiversity loss

Mongolian economic basis• Mineral resources /gold, copper, coal,• Pouching and illegal export of animaland plant species and their products

etc,./• Animal husbandry /42 millions
livestock/• Genetic resources /rich inbiodiversity species/ however
economic value not determined• Illegal collection of medical plants andmass usage of plants and trees for energy
source and construction materials.
• Increasing demand for animals andtheir organs for commercial andhousehold purposes. For example: deerhorn, some kind of birds, wild boarorgans910－ 59 －

Mongolian biodiversity species compared to others

Mongolian biological resources

Biological Class► Mongolian land is about 1% of world total

land territory► 19000 species of biodiversity is recorded in
Mongolia.

11

Russia

World

1574

9500

40.000

Fungi

900

?

100.000

Lichen980

3000

25.000

Moss

445

1370

12.000High Plants

2946

11.400270.000

Insects

615

3000

32.000

Worm

4

1000

10.000

Snake

36

2000

70.000

Crab

16

2000

40.000Spider451

10.00075.000Insects

13.000100.000

950.000

Fish

76

669

30.000Amphibians

8

27

5000

Reptiles

22

75

7300

Birds

468

732

9500

Mammals

132

320

4500

12

However limited studies undertaken, large animals mostly
registered :► For example:► parasite of insects:

Percentage of export by main natural andgenetic resources
Total export:

592 species registered in 1990
615 species registered in 2008
► Butterfly:
153 species registered in 1990
257 species registered in 2008
► Ant:
40 species registered in 1990
72 species registered in 2008
► soil acarus:
43 species registered in 1990
335 species registered in 2008

► copper 27.9%,► coal 27.7%,► Iron 8.2,
► gold 7.1%,► crude oil 6.1%,► raw cashmere 7.1%,► other

export 6.4 % but genetic resource is notregistered.

13

14

Use of biological resources

Legal basis for Genetic resources
National Constitution of 1992:
Clause 6:► Use for domestic household consumption► Sportand special purpose hunting byforeign hunters► Use plant species for medicine► Use of plants and animals for scientificresearch purposes

15

Number of species
Mongolia

Algae

1. Mongolian land, its undergroundresources, forest, water, animal, plants andother natural resources are national property ofpeople and subject to protection by National
Government.
2. Land except owned by Mongolian citizen,
its underground resources and all natural
resources, forest, water resources, animals arecommon property of Mongolian State.

16－ 60 －

Law on Environmental protection:

Law on Fauna, Law on Flora, Law on Hunting:Clause 15:
Authority of Central Government:

The main regulations are as follow:► Animals and plants are classified in 3 classes:► Very rare
► Rare► Rich► The usage of animal and plant species are classified in 3 prurposes:► Commercial► Household► Special purposes and medicine production► Those law also legalized usage, protection, rehabilitation andselection process of animal and plants.

Article 6.
To provide environmental information for citizen,
companies and organizations, and to support
process for sustainable usage of biological andgenetic resources and its practice of knowledge
and new idea.

17

18

Goals for Future

Future goal

► Convention on Biodiversity:► Nagoya Protocol:

1/ To protect biological and genetic resources
2/ To provide sustainable use of biological and genetic
resources
3/ To distribute economic and future efficiency of
biological and genetic resources as a equal.

1/ To prepare process to join Nagoya protocol
2/ To complete the process of joining the Nagoya
protocol this year
3/ To develop a new law on Genetic resources

19

20

Future goal
► Basic issue of the Law on Genetic resources:

1/ To determine management power of Government
and local administrations for genetic resources
2/ To establish provisional and usage permission
system3/ To establish usage payment system of genetic
resources
4/ To establish legal condition of traditional knowledgeof genetic resources21

Proposed cooperation:- Enhance the cooperation on capacity building for distribution of theeconomic and future efficiency of biological and genetic resources as aequal.
- Establish database for genetic resources and its traditional knowledge- Participation in “SLEEPING MICROBIAL BEAUTIES “ project22－ 61 －

講演 2：「Access to Mongolian plants: In vitro bioassay screening」（発表資料 2 参照）
モンゴルの薬用植物は古くから様々な病気の治療や予防に幅広く利用されている。今日では
近代分析技術やインビトロ薬理試験により、植物の特性や有効性を明確にする必要がある。その活性評価のためのインビトロ試験では少量の抽出物、フラクション、あるいは化合物が必要となる。モンゴル植物相はまだ体系的に生物学的活性を選別されていない。そこで、演者等は2006 年よりモンゴル植物の生理活性スクリーニング（抗菌・抗酸化・抗変異活性試験、アセ
チルコリンエステラーゼやリパーゼ阻害活性）を実施している。以下それらの試験結果を発表した。
モンゴル植物 130 種から 350 以上のメタノール抽出物を得、生理活性を調べた。
抗菌スクリーニング（ S. aureus 、 M. luteus 、 E.coli 、 E. faecalis 、 Ps. aeruginosa 、 S.

epidermidis）では、Larix sibirica Ldb. (stem)、Juniperus sabina (aerial parts)、Comarum
Salesovianum (aerial parts)、Caryopteris mongolica (root)及び Potentilla viscosa (leaves)が強い活性を示した。その内、Larix sibirica Ldb からは活性物質としてイソピメリン酸を得た。
さらに、抗菌物質として、 Comarum Salesovianum(aerial parts)からは 2 つの化合物、2-hydroxy-6-nonil benzoic acid, 2-hydroxy-6-non-8-enil benzoic acid（新規物質）を得た。
モンゴルでは家畜の炭疽菌感染（Bacillus anthracis）がしばしば起こり問題となっているので、炭疽菌に対する活性も調べている。Larix sibirica Ldb.は、抗炭疽菌活性を有している。
遊牧民は昔、この木の樹脂を家畜に与えていた。

Chamaerhodos erecta (L.) Bge (aerial parts)、 Dasiphora parvifolia (aerial parts)、
Nymphaea candida (aerial parts)及び Geranium pseudosibiricum (aerial parts)は、高い抗酸化活性を有する植物であることがわかった。Chamaerhodos erecta (L.) (aerial parts)からは、1 新規化合物と 11 の既知化合物が単離された。

Leptopyrum fumaroides (whole plant)、Juniperus sibirica Burgsd (aerial parts)、及び
Pulsatilla flavescens (leaves)は、高い抗変異活性を有していた。Pulsatilla flavescens (leaves)からは、12 の化合物（2 つの新規物質を含む）を得た。Juniperus sibirica と Leptopyrum

fumaroides の抗酸化活性の活性成分はクロロフォルム画分の cetylic alcohol であった。
Carduus crispus L. (aerial parts)、Bergenia crassifolia (L.) (leaves and root)、Juniperus
communis L. var saxatilis Pall. (aerial parts)及び Patrinia rupestris (aerial part)は、アセチ
ルコリンエステラーゼ阻害活性を有していた。Carduus crispus からはいくつかの phenolicacids を単離同定した。

Agrimonia pilosa (aerial parts) 、 Cotoneaster melanocarpa (stem) 及 び Pteridium
aquilinum (stem)は、強い抗リパーゼ活性を示した。
スクリーニングの結果に基づき、ハーブティーの商品化の可能性も検討した。

－ 62 －

発表資料 2

Our research interest• Field survey on distribution and natural resources ofMongolian medicinal plants (collection and identification of plants, also educate young

Access to Mongolian plants: In vitro bioassay screening

people in the field…)• Preparation of extracts (Bank of Mongolian plant extracts)

J.Batkhuu• Primary screening for evaluation of biologicalactivities by using in vitro assays ( We are required to introduce

Ts.Dash Memorial Laboratory of Bioorganic Chemistry and Pharmacognosy
Department of Biochemistry and Bioorganic Chemistry
School of Biology and Biotechnology, National University of Mongolia (NUM)

assays demanding little money and big skill of young sensitive students, for example antimicrobial,
antioxidant, enzyme inhibition and mutagenic)• Selection of active plants (may be patent application for dietary supplement,
cosmetics etc. with domestic and foreign companies , Monos, Monchemo, Kao, GIST, VTT…)

1

2

Screening methods employed in the Lab

Our research interest (continued)• Activity-guided isolation of active compounds byusing simplest methods as CC and TLC• Structure determination – NMR, MS (if we obtained pure active compounds,
structure determination is possible with foreign colleagues , then may be patent application for functional food, cosmetics etc.
with domestic and foreign companies , Monos, Monchemo, Kao, GIST, VTT…)• In vitro and clinical studies → patent application for drug
→ production and marketing with pharmaceuticalcompany• Cultivation of active plants by classic method and tissue
culture (cell suspension culture)• To search ways to use plants on the basis of traditional
medicine and experiment results

Antibacterial
Activity

Disc Diffusion
MethodS.aureus, M.luteus
E.coli, E.faecalis

LipaseInhibition Assay

BALB-DTNB
methodPancreatic lipase,
BALB, DTNB,
Spectrometry-412nm

Anti-mutagenic
Activity

Ames test

S.typhimurium TA1537 (his¯)Standart mutagen:9-aminoacridine

DPPH radical
scavenging method

DPPH, rutin
Spectrometry-517nm

β-carotene
bleachingmethodLinoleic acid, trolox,
Twin40, β-carotene
Spectrometry-470nm

Ellman‘s method

AChE, ACh, DTNB,
Spectrometry-412nm

Antioxidant
Activity

AChE Inhibition
assay

3

4

Comarum Salesovianum (Steph.) Aschers. et Gr.

Screening on antibacterial activity of Mongolian plants

Diagram 1 Extraction and isolation procedure of C. Salesovianum
C. Salesovianum, 1615 g

•
•
•

•
•

Since 2004
Method: Disc diffusion assay
Test microorganisms:
1st screening: S. aureus, M. luteus, E. coli, Ps. aeruginosa, E, faecalis
2nd screening: S. epidermidis, B. anthracis, H. pilory
Positive control: Kanamycin
594 extracts of 226 plant species were tested. Briefly, 125 samples of 76
plants showed antibacterial active at least against one bacterial strain.

Maceration

80% EtOH extract of C. Salesovianum (418 g)
Solvent-solvent partition

DCM fraction, 120 g

n-BuOH fraction, 68 g

Columnchromatography

Water residue, 230 g

Molecular mass = 264
C16H24O3

Compound Cs-1
HOHigh active plants:
Comarum Salesovianum (aerial part), Caryopteris mongolica (root),
Agrimonia pilosa (root), Larix sibirca (stem), Potentilla viscosa (stem)

HPLC

O

2-hydroxy-6-nonil benzoic
acid (Cs-1E)

OH

Molecular mass = 262
C16 H22O3

Cs-1E and Cs-1D
HO

Spectroscopic analyses

5

6－ 63 －OOH

2-hydroxy-6-non-8-enil benzoic
acid (Cs-1D)

Antibacterial activity of Comarum Salesovianum(Steph.) Aschers. et Gr.

Anti-Bacillus anthracis activity of some Mongolian plants
PlantsBacillus anthracis strains
Parts 90 107 120 126 132 147
leaf
― 10
10
8
10
―
stem
― 15
8
14 10
―
leaf
12 ―
12
― ―
13
Abies sibirica
stem
14 ―
13
― ―
12
Betula platyphylla
leaf
10 ―
13
― ―
13
Caryopteris mongolica
root
20 ―
18
― ―
18
Chelidonium majus
stem
12 ―
13
― ―
13
Ephedra sinica
leaf
12 ―
11
― ―
11
Ferula Bungeania
stem
13 ―
14
― ―
10
Ferula feruloeoides
root
10 ―
14
― ―
13
leaf
― 15
18
13 12
―
Juniperus sabina
stem
― 16
13
15 15
―
leaf
― 15
16
16 13
―
Juniperus sibirica
stem
― 16
0
20 15
―
leaf
― 16
0
21 16
―
Pinus sylvestris
stem
― 10
14
18 17
―
Larix sibirica
stem 11 0
10
11
12
11
stem
10 ―
10
― ―
0
Pulsatilla flavescens
leaf
11 ―
12
― ―
0
Polygonum divaricatum
areal part 10 ―
10
― ―
10
Rhododendron dahuricum
leaf
14
15
―
0
0
Sanguisorba officinalis
root
― 13
11
12 12
―
Spiraea salicifolia
leaf
10 ―
10
― ―
10
Trollius asiaticus
areal part 14 ―
12
― ―
13
Vaccinum vitis-idaea
leaf
― 0
11
0
0
―
Name

Artemisia frigida

Table 1.
Name ofsample80% EtOH

Antibacterial activity of fractions and isolated compounds from
C. Salesovianum
Inhibition zone (mm)C(µg/disc)500

S. aureus

M. luteus

E. coliE. faecalis

Ps. auruginsa

S. epidermidis

20,4±0,14

21,1±0,01NA

12,2±0,18NA17,4±0,3620.2±1DCM

500

24.8±0.08

24.8±0.03NA16.5±1.2

NA

n-BuOH500

10.9±0.01

NA

NA

NA

NA

NA

WR

500

NA

NA

NA

NA

NA

NA

Cs-1

500

33

35.2

11

28.3

9.7

37

Cs-1

50

22.1

22.6

NA

17.3

NA

12.2

10

15

12,7

16,6

9

15,2

КМ

•

We tested totally 67 samplesof 49 plant species on
Bacillus anthracis strains by
disc diffusion method.
25 samples of 19 plant
species inhibited growth of
Bacillus anthracis.
Further we studied Larixsibirica Ldb.

••

NA=no activity. Inhibition zone (in mm) including the diameter of disc (the diameter of disc = 8 mm);
C=concentration; DCM=dichlorometane; n-BuOH=n-Butanol; WR=water residue; KM=Kanamycin

7

178
―
―
12
12
14
19
13
11
11
14
―
―
―
―
―
―
11
13
13
10
―
―
10
14
―

8

Inhibition activity of Las 1 compound of the Bacillus anthracis (clear zone, mm)

Methanolic extractfrom stem of Larix sibirica ldb. (70g)Part I
soluble in water (20g)

SamplePart II
insoluble in water (70g)

Inhibited growth of
B.anthracis

Las1

Conc

500µg/ disc
Control (amp)

Fractionation by Silica gel
Column Chromatography

Bacillus anthracis strains
90
120
147
178
17
15
15
18
17
15
17
17
17
17
15
17
36

Mass spectra of Las 1 compound
Fr. 1

Fr. 2

Fr. 3 (2.85g)

Fr. 3-1

Fr. 4

Fr. 5

Fr. 6

Fr. 7

Fr. 8

Fr. 16

Fractionation by Silica gel
Column Chromatography

Highly inhibited growth of
B.anthracis from other fractions

Fr. 3-10

Fr. 3-4 (0.83g)

Fractionation by Silica gel
Column Chromatography

Fr. 3-4-1

Fr. 3-4-2 (0.67g)

Fr. 3-4-3Isopimaric acid
C20H30O2

Fr. 3-4-7

Preparation TLC

Las 1 (0.022g)

910

13С NMR of Las 1 compound

1Н NMR of Las 1 compoundСН3 (С17)

С7

С15

С8
С18

11

С16

С9

С14

С5

С17 С20С2СН3 (С19)

С13
С4 С1 С12 С10
С3

С19
С11

12

－ 64 －

СН3 (С20)

Screening on anti-mutagenic activity of Mongolian Plant

Isolation of active compounds Pulsatilla flavescens leaves

1st screening for “Antimutagenic activity of Mongolian Medicinal Plants”

Dry leaves 680g of P.flavescens

Since 2007
Method: Modified Ames test
Test strain: Salmonella typhimurium TA1537（His¯）Standart mutagen: 9-aminoacridine (20µg/plate)
Inhibition ratio: (IR)=A-B/C×100

Hexane, (5.7g)

EtOAc, (17.8g)

Water , (175g)

EA (8ｇ)Water (20ｇ)

MPLC,
Hi-Flash silicagel column,
60x180mm, CHCI3 :MeOH

EA1.. EA5.. EA8.. EA10..

Chamaneron angustifolium (green leaves),
Rubus Sachalensis (stem)
Pulsatilla flavescens (leaves)
Arabis pendula L. (stem)
Leptoperum fumaroides L. Reichb (A.part)
Juniperus sibirica (leaves)
Rhinanthus songaricus (leaves, stem)

MPLC condition:
Column: silicagel column,
Sol: Hex-EtOAc or
CHCI₃-MeOH
F.rate: 20ml/min
Det: UV 254nm

Antimutagenic
activities were
IR>40%

Ulcer pus, Heals infected wounds,
Neutralize poisoning, Gland tuberculosis(scrofula), Hemorrhoid

Methanol extract 215g

N:48º05.159´, E:106º18.433´
H:1374m (2nd June 2010)

Screened samples: 141 different samples of 82 plant species.
(100µg/plate, 10%DMSO)

Use in The Mongolian Traditional Medicine:

FR1

EA14

OCC ,
Diaion HP-20,
Sol : H2O→EtOH

FR2

FR3

FR4

HPLC condition:
Column: L-column HB, ODS
10x250mm, 5µm
Sol: A - 0.1% HCOOH,
B - CH3 CN
Isocratic 20%B
F.rate: 1.2ml/min (×3)
Det: UV 254nm and IR-500
Inject: 10mg/400µl ×8Comp 1 (73mg)
Comp 2 (20.2mg)
Comp 3 (18mg)
Comp 4 (78mg)
Comp 5 (17mg)
Comp 6 (4.5mg)

FR5Comp 7 (176mg)
Comp 8 (53.7mg)
Comp 9 (41.6mg)
Comp 10 (5.4mg)
Comp 11 (12mg)
Comp 12 (29mg)

Antimutagenic activity of fraction were tested on Salmonella typhimurium TA98, B[a]P with S9, AF-2 with buffer, Average±SD1314

Molecular structure of isolated compounds

Molecular structure and anti-mutagenic activity
of isolated compounds from P.flavescens

OH

O
OH

NH
HO

O

HO
O

O

O
O

O

OH

O

Dihydro-5-(hydroxymethyl)2(3H)-furanone
Comp[2]IR=9,3%

H

H
HO H

COOH
H

O

O

H
OH

OH

H
OH

O
H
OH

β-sitosterol glucoside
Comp[3]
IR=37,1%OHO

N-trans-Caffeoylphenylamide
Comp[7]
47,8%

H
OH

H

OH

AnemoninComp[1]
IR=61,8%

OH

O

OH

O

H

Quercetin 3-O-β-D-glucoronide
Comp[11]
IR=89,6%

O
O
CH2OH

CH2OH
O
H
H
H OH
OH
H
OH H

HO

HO

Caffeic acid
Comp[4]
IR=55,6%

OH

R1OOC

O

OCH3
OH

OH

O

Methyl caffeateComp[5]
IR=65,8%

OH

O

OH
OH

L-Chicoric
COOR2 acid

Our study confirmed that major components ofleaves of Pulsatilla flavescens were anemonin,caffeic acid related compounds and triterpene
saponins which could be responsible forvarious biological activities of the plant.

OH

15

Comp[12]
20,1%

O

H3C
HO

O
O

OH

O

O

HO

Caffeic acid β-glucopyranoseComp[6]
IR=11,8%

OH

OH OH

HO

OH

HO

O
O

O

OH

OH

HO

16

1H and 13C NMR spectrum of triterpene saponin

1H and 13C NMR spectrum of N-trans-caffeoylphenylamiderhamnoseС6 - 3Н (d)Solvent
Pyridine-d5

1 H NMR(DMSO, 600MHz)
Solvent

Hederagenin:С13 Н

(Glucone part)Glu-glu-rham С2-С6
H(Aglucone part)hederagenin С1-С30
H

27.7

NH
Rham С1
Glu С1 Н (d)Н (d)С2 С5 С6 and С3' С4‘ Hd

O

water

5

C1'-2H
m, m

HO 4

1
2
3

1'
8

9

Glu‘ С1
Н (d)

N
H

2'

3'

121.9
18.9

4''

5'

1H NMR(Pyridine-d5, 600MHz)OH(aglucone part)
Hederagenin С1-С30
Solvent
Pyridine-d5

N-trans- caffeoylphenylamide(glucone part)
glu-glu'-rham
С1 atoms

HO 74.4

C3' C3''
C9

C5' C

4

C3
C7

Hederagenin
С28

C1 C6
C 2'

C8

O

C1'

18

－ 65 －

102.8OH

74.3

OH

OH

76.8

68.5
74.1 O

72.5

OH

103.6

OH

HO

76.1
79.7

OH

Hederagenin

Hederagenin
С13

13C NMR(Pyridine-d5, 150MHz)

17

93.9

OH

76.7

78.5

DEPT

O
61.2 74.7

O73.5

59.676.1

HO
O

DEPT

29.1
21.0
35.3

HO

(glucone part)
glu-glu'-rham
С2-С6

19.9

C4' C4''

O

O

20.5
65.4

13.3

30.0

177.2
27.4

39.9

47.6

40.7

47.3
42.2

47.8 42.2

38.2
45.7

3''

27.7

31.0

27.7

4'

OH

13C NMR(DMSO, 150MHz)

144.2

25.0
21.7
33.5

7

6
C7-H dC 8-H d46.4

H28-O-α-L-rhamnopyranosyl(1→4)β-D-glucopyranosyl(1→4)β-D-glucopyranosyl ester
C48H78O18

Fractionation and Isolation of Leptoferum fumaroides (aerial part)

Extraction, fractionation and column chromatography
separation of Juniperus sibirica aerial parts

Dry leaves 160g of L.fumaroides

Antimutagenic
activity=%

Powdered aerial parts ofJuniperus sibirica (280g)

Methanol extract 44.5g

Extracted with methanol

Methanol extract (77g)

Suspended in water

44%

CHCI₃ (5g)48%

Undissolved in water (54g)

Column chromatography, Silicagel Wakogel C-200

65%

Fr.1

1
HO

3
2

FR1…

Fr.2

49%

Fr.3

56%

Fr.4

Cetyl
alcohol55%

β-Sitosterol

40%

5

7
6

4

11

9
10

8

13
12

Fr.5…

FR4

FR5

FR6..

FR10..

Lef-1 (150mg)
[65%]

1
CH3

HO

16

Cetyl alcohol

Water (33.2g)

FR15

FR16..

FR30..

FR32…Fr.2915
14

n-BuOH (5.4g)

Column chromatography
SiOH column

23%

Dissolved in water (23g)32β-Sitosterol

5
4

7
6

11

9
8

Lef-2 (11mg)
[26%]

10

13
12

Lef-3 (5mg)
[34%]

15
14

CH3

16

Cetyl alcohol[IR,%]=antimutagenic activity

19

20

Some high antioxidant active plants

Screening on antioxidant effect of Mongolian plant extracts
DPPH radical scavenging method

β-carotene bleaching method

•
•

•
•

Since 2007
We studied 324 Mongolian
plant methanol extracts
belonging to 186 species andwe evaluated their inhibitory
effect against free radicals.

••

Chamaerhodos erecta•Upper part IC50 (μg/ml) = 59.24±0.04
Ephedra sinica•Leaf IC50 (μg/ml) = 52.38±1.91

Since 2010
By Miller (1971) β-carotene
bleaching method
Auto-oxidation of Linoleic
acid
60 plant extracts belonging
37 species of 18 families.

Geranium pseudosibiricum J. Mayer•Leaf IC50 (μg/ml) = 51.18±0.2
Geranium pratense L.
•Upper part IC50 (μg/ml) = 52.12±0.69

High antioxidant active plants:
Chamaerhodos erecta, Agrimonia pilosa, Ephedra sinica, Cotoneastermelanocarpa Lodd, Nympaea candida, Dasiphora parvifolia (Fisch.) Juz.

Vaccinium vitis-idaea L.
•Stem IC50 (μg/ml) = 52.52±0.222122Isolation and chracterization of antioxidant compounds fromaerial parts of Chamaerhodos erecta

Comp.1 4,5-dihydroxybenzaldehyde-3-O-β-D-glucopyranoside

HMBC

IC50(μg/mL)

Water (23g)

27.8

H-1’
d,
J=8Hz

n-ButOH (19g)165.6

CHCl3 (18g)H-7
singlet59.2

H-2 d,
H-6 d,
J=2Hz
J=2Hz

Methanol extract (67g)

167.8

H
t

Powdered aerial parts of
Ch.erecta (450g)

H-6’ dd, J=12Hz and
H-2’
J=5.5Hz,dd,H-J=12Hz and
H-4’
J=2Hz 3’ H5’
multiple

1H-NMR

Extracted with methanol
Suspended in water

IC50(μg/mL)

CO

Column chromatography, Sephadex LH-20

Fr.1-Fr.3

Fr.4

Fr.5

Fr.6

Fr.7

Fr.8-923

12
3 (50mg)4
5 (21mg)
6
1013C-NMRFinal
fraction

ODS column,
HPLC

CC

1 (267mg)
2 (90mg)
4 (7mg)
6 (7mg)
10 (282mg)
11 (3mg)
12 (3ｍｇ)Fr.10-15

13
4
8 (0.8mg)
9 (0.3mg)
10

1
8
9

7 (1.8mg)

HO

24－ 66 －

OH

O
OH

HO
HO

13 (6.7mg)
14 (2.6mg)
15 (14.1mg)
16 (3.3mg)
17 (10.6mg)
18 (7.5mg)
19 (20mg)

O126.55OHMS

IC50(μg/mL)
R1

O
HO

O
HO

OH

HO

OH

OH
O

2
3
4
5
6
7
8
9

R1
Н
ОН
ОН
Н
H
H
OH
H

O

OH
O

HO

OH

O

O

OH
OH

HO

O

13. Strictinin

7.44
10
Ellagic acidR2
β-D-(6”-O-trans-p-coumaroyl) glucopyranoside
β-D-glucuronide
β-D-glucopyranoside
β-D-glucopyranoside
β-D-glucuronide
β-D-glucuronide methyl ether
H
H

(S)

(S)

OH

(R)

O

(S)

O

(S)

(S)

OH

(S)

HO

O

(R)

HO

O

HO

O

OH
O
O

O

HO
HO
O

OH

HO
HO

O

O

O

OR2

O

O

O

OH
HO

HO

HO

O

OH

OH HO

HO

OH

OH

>200

14. Eugeniin

HO

13.9
6.19

HO

O
HO

O
HO

O

O

103

HO
HO

ND
ND
19.7
24.8

HO
HO

O

O

O

(R)

HO

OH
O

(S)

(S)

OH

O

O

O

(R)

(R)

O

O

O

OH
O

OH

HO

(S)

O
HO

O

O

O

(S)

OH

(R)

OH
O

HO
O

O

HO

HO

OH

OH HO

HO

OH

OH

OH

OH

OH

OH HO

HO

HO

HO

O

HO

HO

12
Euscaphic acid>200
11
Tormentic acid

OH

OH

15.
1,2,3,4,6-O-penta-O-galloyl-β-O-glucose

O

16. Casuarictin

>200

25

26

HO

Extraction and fractionation of Dasiphora parvifoliaO

17. Potentillin

HO

•

HO

O
HO
HO
O

OCH3

O

O

O

(R)

(R)

HO

(S)

O

HO

OH
O

(R)

O

O

HO

O

Dasiphora parvifolia Fisch. Juz (Leaf)

OH O

OH

with MeOH at room temperature (W/V, 1:10)

18. Methyl gallate

OH

Extraction for 5 days

OH

OH HO

HO

We selected leaf extract of Dasiphora parvifolia (Fisch.) Juz. based on results.
Fig 1. Leaf of Dasiphora parvifolia (Fisch.) Juz. fractionated by liquid-liquid partition.

OH

OH

Filtration

OH

OH

OH

OH
HO

HO
HO
O

Concentrated with a rotary evaporator

OH

HO

HO

O

O

O
O

HO
HO
O

(R)

O

HO

O
(S)

O

(R)

O

HO

O

HO

(R)

O

Dasiphora parvifolia (Fisch.)
Juz.

O

(S)

(R)

O

OH

O
(R)

OH
O

(R)

Methanol extract(37 g)

O

O

OH
OH

O
O

O

O

OH

O
H

O

OH

Dichloromethane(7 g)n-butanol (17 g)

Water residue (13 g)

IC50=67.3±0.3

IC50= 33.4±0.5

IC50=200<

OH
OH
HO

HO
OH

OH
OH

19. Agrimoniin

Isolate active compounds

27

28

Screening on acetylcholinesterase inhibition activity

Investigation on acetylcholinesterase inhibitor
effect of Carduus crispus

Since 2009
Method: Modified version of the colorimetric method of Ellman.
Screened samples: 134 different samples prepared from 82 plant species.

Active plants:
Carduus crispus (leaves) activity=81.6%
Patrina rupestris (aerial part) activity=49.6%

Methanol extract 44.5g

Hexane

CHCI₃

EtOAc

n-BuOH

Water[34.5%][40.1%][48.0%][81.6%][45.7%]

MeOH extract

HO

Isolation
RP-HPLC

COOH

Artemisia annua (flowers) activity=52.1%
Filipendula palmata (seeds) activity=63.8%
CHCI₃

n-BuOH

R1O

water

OR
OH

compoundR

neochlorogenicacid

H

IC50(μg/ml)

R₁
O

Isolation
Column chromatography

p-coumarylquinicacid
β-sitosterolsterol

chlorogenic acid

HO

400

HO

O

H

HO

205

Pure compounds:
1 neochlorogenic acid
2 p-coumarylquinic acid
3 chlorogenic acid

O
HO

HO

H

˃1000
[AchE inhibitor effect,% at 1000μ/ml]

2930－ 67 －

Evaluation of lipase inhibitor activityof Mongolian plant extracts

Evaluation of lipase inhibitor activityof Mongolian plant extract

Method: BALB-DTNB method• Since 2006

Reaction mechanism:• We screened124 plant extracts from prepared 81 plant species+ 3.H 2 O

Lipase

CH 2

OH

CH

SH

+

3 HO C CH 2 CH 2 CH 3

CH 2 SH• Active plants:
Cotoneaster mongolica (stems 55%), Pteridium aquilinum(stems 36.8%), Ephedra equistina (leaves 68.3%), Abies
sibirica (leaves 16%) showed higher anti-lipase activity thanother extracts.

O

2,3-Dimercapto-1propanol (BAL)
2,3-Dimercapto-1-propanol
tributyrate (BALB)., 334.5
HOO
C

CH2 OHCH SH
CH2 SH

HO

O
C

+

O2N

2,3-Dimercapto-1-

propanol (BAL)S S

5,5’-Dithiobis(2nitrobenzoic acid).,(DTNB)396.35

NO2
C

O
OH

-H2

O2N

OH
CH2
CH S SNO2S S CH2C

Active-guided isolation ofactive compounds from
Cotoneaster mongolica (stems)
is going on.

O
OH

31

32

Possibilities of making product based on our screening results

Herbal tea
•
•

•

•

Herbal teas have been used fordaily needs since ancient times.
All types of teas, including
green, black, red and herbal
teas, have antioxidant
properties because of thepolyphenols.
These antioxidants in herbal
tea are known to provide thebody with protection against
free radicals.
Therefore, the aim of this study
was to investigate antioxidant
activity of herbal tea infusions
prepared from some
Mongolian medicinal plants
using DPPH free radical
scavenging method.

Preparation against
smell of armpit sweat

Composition of herbal tea infusion
We chose following plants based on their high antioxidant activity.

Staphyllacoccus epidermidis

Fig. 1. Chamaenerion
angustifolium (L.) Scop.

Comarum Salesovianum (aerial part) showed
high activity against S. epidermidis.

33

Fig. 2. Dasiphoraparvifolia (Fisch.) Juz.

Fig. 3. Geranium
pseudosibiricum J. C.
Mayer.

Herbal tea preparation:
We mixed these plants with various ratios
1.200 ml of hot water (95ºC) was added to 2 g of dried herbs mixture and extracted it for
15 min at room temperature.
2.200ml water was added to 2g of dried herbs mixture and boiled it for 15 min.
Measure antioxidant activity

34

Fig 1. Antioxidant activity of herbal tea infusion

Tab. 1. Antioxidant activity of 2A, 5A herbal tea infusion andother commercial herbal tea№

Furthermore, we
selected 2A and 5A
extracts based results.

35

Fig. 4. Vaccinium
vitis-idaea L.

36－ 68 －

5A

IC50 values(µg/ml)54.06±0.4

2A67.29±0.13

Seabuckthorn herbal tea

100.54±1.03

Chinggis Khaan herbal tea

200<

Huvsgul herbal tea

164.47±0.54

Karkade herbal tea

195.6±3.08

Introduction of Ts.Dash Memorial Laboratory of Bioorganic Chemistry
and Pharmacognosy, NUM

Domestic collaboration:

1991

Tne laboratory was established with keen effort by prof .
Ts.Dash as Laboratory of Protein Chemistry under the Instituteof Chemistry, Mongolian Academy of Science

1997

moved to Faculty of Biology, NUM

 Laboratory of Natural Product1997

Investigation on biological active metabolites of Mongolian
plants is started

 National Center for Infectious Diseases with Natural Foci, Ministry of Health2002

Screening of Mongolian plants for biological activities

2010

The lab named Ts.Dash Memorial Laboratory of Bioorganic
Chemistry and Pharmacognosy

2011

Renewal open of Laboratory of Plant Biotechnology at this lab(Obtaining of secondary active metabolites by cell suspension culture) Department of Plant Taxonomy, Institute of Botany, Mongolian Academy of Science

Chemistry, Institute of Chemistry and Chemical
Technology, Mongolian Academy of Science Monos Co., Ltd Otoch Manaramba University

37

38International collaboration:
Japan: Tohoku Pharmaceutical University
Toyama University
Toho University
Kao Co., Ltd
Regional Promotion Support Center, Japan

Thank you for your attention

Austria: Graz University
China: Inner Mongolia University (Institute of Macromolecular Chemistry andMongolian Medicine)
Peking University (Lab of Pharmacognosy, School of Pharmaceutical Sciences)E-mail:bathuu@mobinet.mn
jbatkhuu@hotmail.com

Republic of Korea: Gwangju Institute of Science and Technology
Finland: Sector of Plant Biotechnology, Technical Research Centre
Belarus: Institute of Biology, Academy of Sciences

39

40－ 69 －