STUDY ON BIOLOGICAL CHARACTERISTICS, DISTRIBUTION AND …gust.edu.vn/media/25/uftai-ve-tai-day25971.pdf · 2017-10-30 · 1 INTRODUCTION 1. Rationales of study Amongst the flora of

MINISTRY OF EDUCATION VIETNAM ACADEMY OF SCIENCE

AND TRAINING AND TECHNOLOGY

GRADUATE UNIVERSITY SCIENCE AND TECHNOLOGY

LE DONG HIEU

STUDY ON BIOLOGICAL CHARACTERISTICS, DISTRIBUTION

AND CHEMICAL COMPOSITION OF ESSENTIAL OF

PIPERACEAE SPECIES IN THE NORTH CENTRAL VIETNAM

ABSTRACT OF A THESIS DOCTOR OF PHILOSOPHY IN BIOLOGY

Major: Botany

Code: 62.42.01.11

HA NOI, 2017

This thesis was fulfilled at Graduate University of Science and Technology

Supervisors: 1. Assoc. Prof. Dr. Tran Minh Hoi

2. Prof. Dr. Tran Dinh Thang

Reviewer 1:

Reviewer 2:

Reviewer 3:

The thesis will be defended at the University of Science and Technology,

18 Hoang Quoc Viet, Cau Giay, Ha Noi City, 2017

The thesis can be found at: The National Library of Vietnam

Library University of Science and Technolog

1

INTRODUCTION

1. Rationales of study

Amongst the flora of Vietnam, the group of oily plants is very rich and varied.

To date, around 657 species belonging to 357 genera and 114 families (accounting

for 6.3% of total species, 15.8% of total and 37.8% of families) have been recorded.

Piperaceae have 4 genera, 50 species (Lepianthes - Lepianthes umbellatum,

Peperomia - 6 species, Piper - 42 species, Zippelia has 01 species); They are

commonly vines with single leaves (mainly spiny or sprouting, rarely growing) and

most of them have archers. Flowers form thick dense rings (sometimes shaped like

squirrels); no petals; antherogenic outward direction and with tiny stipule. These

plants mainly grow in the tropics, especially in Southeast Asia and tropical America.

The current trend among scientists around the world is to focus not only on the

morphology but especially on the chemical compounds presenting in this family for

medical application purposes.

Folk experience shows that for many species of Piperaceae, different parts are

used by ethnic minorities to make medicines, vegetables, etc. Therefore, the study of

Piperaceae, as a scientific basis for the sustainable exploitation and utilization of

plant resources, has been one of the great concerns of mankind. Amongst the groups

of plant resources, the group of plants containing essentialstands ranks a crutial

position. This is an essentialresource for a large number of industries such as

cosmetics, food and pharmaceuticals ... Currently, most species in the family Pepper

(Piperaceae) are capable of biosynthesing and accumulating natural substances,

especially essential oil.

The North Central Vietnam is one of the centers of biodiversity not only in

Vietnam but also worldwide. Here, there may be various new plants and animals.

Among them, the Piperaceae family can imbibe a large number of new species and

which are extremely rich and diverse resources.

The results of investigations, studies, data collection on morphological,

biological, ecological and chemical characteristics to fully evaluate the potential of

their diverse resources (Piperaceae) is the reason for which the author of the thesis

2

has chosen the topic: Study a number of biological and distributional

characteristics, chemical composition of essential oils of species in Piperaceae

family in the North Central Vietnam”

2. Objective of study

To effectively evaluate the diversity of species composition, biological and

distributional characteristics, chemical composition of essential oils of a number of

species in Piperaceae family in the North Central Vietnam.

CHAPTER 1

GENERAL OVERVIEW

1.1. Research on the plant family Piperaceae

1.1.1. On the worrld

Piperaceae is a large family of the Magnoliophyta; distributing mainly in the

tropics around the world, with about 10 genera and 2,000 species. Of which 2 genera

including Piper and Peperomia account for about 90% of total species. A conclusion

on the grouping of the species and the genera has not yet reached unity. Miquel

(1843-1844) divided the family into two groups, Piperneae, consisting of 15 genera

with 304 species and Peperomeae with 5 genera with 209 species. De Candolle

(1869) identified this family consisting of two genera Piper and Peperomia with over

1,000 species. According to Rendle (1956), the Pepper family consists of two large

genera, Piper with over 700 species and Peperomia with over 600 species, plus seven

more. In the meanwhile, according to Lawrence (1957), there are 10-12 genera with

two large genera, Piper and Peperomia. According to the classification of Takhtajan

(2009), the Piperaceae family consists of 7 genera and over 2,000 species.

Research on the classification of the Piperaceae began with the publication of

"Plants" by Linnaeus (1753). He described 17 species, all of them are included in the

Piper genus. Later a number of works published by Hooker (1885), Quisumbing E.

(1930), Ridley (1967), Long (1984), Huber (1987), ...

3

1.1.2. In Vietnam

Piperaceae in Vietnam was mainly studied by the French with the first study

conducted by J. Loureio (1793), Gagnepain (1908), Le Kha Ke et al. (1975), Pham

Hoang Ho (1993, 2000), Le Tran Chan et al. (1999), Nguyen Kim Dao (2003), ...

1.2. Uses of species in the family Piperaceae

A large number of species of Piperaceae, which contain essential, have been

grown as spices, as stimulants and as medicines in folk medicine. Many parts of the

species belonging to the genus Piper L. can be used as medicine. However, in

different countries, different species or different parts of the same spicie are used.

The most common use of this genus, according to folk experience, is to treat

gastrointestinal diseases (gastritis, flatulence, diarrhea, constipation), pain relief

(rheumatism, back pain, tooth pain) and antibacterial, anti-inflammatory (bronchitis,

skin lesions, inflammation of the gastrointestinal tract).

Ingredients in a number of herbal processed products contain extracts from some

species of the genus Piper L. which are used to treat diseases such as Eupolins

(contains concentrates of Piper betle and Chromolaena odorataleaves which is

beneficial for gingivitis, inflammation teeth); Sakantin contains fruit extracts from

two species of Piper nigrum L., Piper longum L. and ginger to stimulate digestion;

Armorex T (contains pepper, garlic Essential, sesame Essential..., antimicrobial,

antioxidant, reduce spasm and cure diarrhea). In particular, some products contain the

extract from Piper methysticum G. Forst (also known as kava), which has a sedative

effect and reduces anxiety. However, there have been recent reports of liver toxicities

when using these products, so caution should be exercised when using them.

1.3. Essential oils

This section raises the concept of plant Essential, natural state and distribution,

characteristics and chemical composition of essential, the use value, the importance

of Essentialand Essential-containing material.

1.4. Research on the chemical composition of Piperaceae

This section summarizes the studies on the essentialof the Piperaceae family on

the world and in Vietnam.

4

1.5. Natural, economic and social conditions of study site

This section has described the geographical, topographical, climatic, social and

floristic characteristics of study site.

CHAPTER 2

MATERIALS, CONTENTS AND METHODOLOGY OF STUDY

2.1. Materials and scope of study

Objects of the study are species in the Piperaceae family in the North Central

Vietnam.

Scope of the study: Mainly National Parks (Ben En, Pu Mat, Vu Quang, Phong

Nha - Ke Bang, Bach Ma) and nature reserves (Xuan Lien, Pu Luong, Pu Hoat, Pu

Huong , Ke Go, Dak Krong) in the North Central. There is also sampling in some

locations of the districts with the distribution of the plants belonging to this family.

2.2. Content of study

- To study biological characteristics (morphology and ecology), distributional

characteristics and diversity of species in the Piperaceae family in the North Central

Vietnam.

- To identify the use value of the above species in the Piperaceae family in the

North Central Vietnam.

- To identify the content and chemical composition of essentialof some species

in the Piperaceae family in the North Central Vietnam.

2.3. Study methodology

2.3.1. Botany methods

2.3.1.1. Inheriting data method

This section has described the geographical, topographical, climatic, social and

floristic characteristics of study site, sample in Museum national and international

and previously published works.

2.3.1.2. Field survey methods

To conduct the main survey routes for research in each study site:

5

+ Ben En National Park: Song Trang - Bai Tranh route; Dong Muoi - Song Muc

Lake - Dien Ngoc route; Dong Muoi - Xuan Thai route.

+ Pu Mat National Park: the Mon Son route; Khe Bu route; Khe Kem route;

Tam Dinh - Tam Hop route; Khe Choang route.

+ Vu Quang National Park: Huong Quang - Doc Do route; Huong Dai route;

Huong Son route.

+ Phong Nha - Ke Bang National Park: East and West intersection Truong Son -

U Bo route.

+ Bach Ma National Park: the center of the Park - the hydroelectric dam – Da

Dung waterfall route; Tri Sao - Bach Ma peak - Do Quyen route; South East route

(Huong Phu, Thuong Nhat, Hung Loc).

Xuan Lien Nature Reserve: Van Xuan route, Yen Nhan - Bat Mot route.

+ Pu Luong Nature Reserve: Phu Le route; Co Lung - Lung Cao route.

+ Pu Huong Nature Reserve: Chau Thai - Nam Son - Bac Son - Binh Chuan

route; Chau Hoan - Dien Lam - Quang Phong route.

+ Pu Hoat Nature Reserve: Tri Le - Nam Dam route; Hanh Dịch - Thông Thu -

Đồng Văn route.

+ Ke Go Nature Reserve: tourist center to the areas running along South – North

direction.

2.3.1.3. Sample collection and species identification methods

At least two or three specimen per individual per species were collected from

the same study site. All traits such as color, resin, characteristics of flower, fruit and

leaf, which may change due to subsequent procedures of sample processing (e.g.

drying), were documented in details. The plants were also pictured by a digital camera

(Canon). Then, these samples were dried and stored in botany lab of Vinh University.

More than 500 samples were collected in this study, of which about 300

samples were species identified. Samples are keeping in Institute of Ecology and

Biological Resources, Vietnam Academy of Science and Technology, Botanical

laboratory, Department of Medical Materials, Vinh Medicine University.

The species identification based on following documents:

6

- Pham Hoang Ho (1999), Vietnamese Herbs, Book II; H. W. Li (1982), Flora of

China; Cheng Y., N. Xia & M.G. Gilbert (1999), Flora of China, Vol. 4, Piperaceae.

2.3.1.4. Identification of the use value of the species in the Piperaceae

Diversity of useful value of flora: Statistical calculating of useful species from

botanical monograph documents such as: Vo Van Chi (2012), Do Tat Loi (1995), Le

Tran Duc (1997), Trieu Van Hung et al. (2007), Do Huy Bich et al. (2004),

Chaveerach A. et al. (2006) and PRA (Parcitipartory Rual Appraisal).

2.3.2. Methods to identify chemical composition of essential oil

2.3.2.1. Sampling and essential oil distilling

Samples for essential oil distillation were collected separately from different

parts of trees (leaf, stems, roots, flower, fruit). Each sample was about 0.5 – 3.0 kg of

fresh materials. Samples were chopped into small parts and distilled by

hydrodistillation for 2-4h at normal pressure, according to the Vietnamese

Pharmacopoeia IV (2010).

2.3.2.2. Methods quantitative of essential oils

Essential oils of different parts to be quantified by the method I of Vietnamese

Pharmacopoeia IV (2010).

X(%) = (d<1)

Or

X(%) = (d>1)

a the volume of essential oils in ml

b the volume of the sample in grams.

2.3.2.3. Methol of essential oil

Gas chromatography (GC) analysis was performed on an Agilent Technologies

HP 6890 Plus Gas chromatograph equipped with a FID and fitted with HP-5MS

column (30 m x 0.25 mm, film thickness 0.25 m, Agilent Technology). The

a x 0.9

b x 100%

a

b x 100%

7

analytical conditions were: carrier gas H2 (1 mL/min), injector temperature (PTV)

250oC, detector temperature 260

oC, column temperature programmed from 60

oC (2

min hold) to 220 oC (10 min hold) at 4

oC/min. Samples were injected by splitting and

the split ratio was 10:1. The volume injected was 1.0 L. Inlet pressure was 6.1 kPa.

An Agilent Technologies HP 6890N Plus Chromatograph fitted with a fused

silica capillary HP-5 MS column (30 m x 0.25 mm, film thickness 0.25 m) and

interfaced with a mass spectrometer HP 5973 MSD was used for the GC/MS

analysis, under the same conditions as those used for GC analysis. The conditions

were the same as described above with He (1 mL/min) as carrier gas. The MS

conditions were as follows: ionization voltage 70eV; emission current 40 mA;

acquisitions scan mass range of 35-350 amu at a sampling rate of 1.0 scan/s. The MS

fragmentation patterns were checked with those of other essential oils of known

composition with Wiley (Wiley 9th

Version), NIST 08 Libraries (on ChemStation

HP), with those in the literature, and also with standard substances.

2.4. Statistical analysis method

Data were analyzed statistically using Microsoft Office Excel 2010

CHAPTER 3

RESULTS AND DISCUSSION

3.1. Biological characteristics of species in the Piperaceae

3.1.1. Morphological characteristics of species in the Piperaceae

+ Plant stem: Stand straight or branched, standing with climbing roots on the stem,

clinging to the other plants; Occasionally there are cobia on the ground or parasitoid

and are usually fragrant.

+ Leaves: simple, raw edges, opposite or alternate, may or may not have stipule

(premature fall leaving scar tissue). Leaves vary in shape and size among species.

The leaves are in the form of: sword shaped, ovate, heart shaped, elliptical ... On the

same plant can have many different leaf shapes with the characteristics of older

leaves are usually larger and more prominent than the remaining leaves . The lamina

8

might be thin or thick, smooth, wrinkled or rough and hairy. The leave hair is either

brown or white, may be single or multicellular, covering the leaf surface and or the

whole stem. The sheat is symmetrical or deflected, in the form of circles or hearts.

The petiole is often short or several centimeters long. The leaf ribs usually form pairs;

Some foliage have a secondary vein system that runs along the midrib, which is curved

from the root to the top of the leaf, has a propeller shape or has a feathered portion.

+ Flower: Originating from some locations such as the sheat, next to the sheat,

middle or upper point of midrib. opposite to leaf or axillary; Seldom grows on the top

of the branch and rarely forms a cluster of many flowers growing in the axillary.

Inflorescences are straight or slightly curved and may be drooping when ripe. Bare

flowers, often simple (most of the original, rarely of the same) or bisexual, stemless.

Flowers may be pink, chestnut, blue-gray, yellow-green, opaque white. The stipule is

opposite the flower, sometimes stick with the flower axis; either shield, triangle or

circle shaped; either smooth or hairy. Stamen 2-6, short filament; capel 2, 2-4 lobes.

The stigmatic ovary is loose or sometimes embraces the axis, 1 cell, 1 ovary; Stigma 2-5.

+ Fruit: nuts, with or without stem; oval, spherical, ovate, or triangular section,

rarely oval; ripe fruits are usually red or yellow. Fruit is usually smooth or sometimes

a little hairy ; each fruit has one seed. Seeds are nearly spherical with thin seed pods;

small embryos, extruded and hardened emulsions.

3.1.2. Ecological characteristics, distribution of species in the Piperaceae

+ Locality: The species mainly crawl on other plants such as: Piper acre Blume,

Piper arboricola C. DC., Piper bavinum C. DC., Piper betle L., Piper cambodianum

C. DC., Piper cf. caninum Blume, Piper chaudocanum C. DC, Piper griffithii C. DC.,

Piper cf. maclurei Merr., Piper nigrum L.,… Some species grow on the ground or in

karst limestone hollows such as: Peperomia pellucida (L.) Kunth., Peperomia

parcicilia C. DC., Piper boehmeriifolium Wall. ex Miq., Piper boehmeriifolium var.

tonkinensis C. DC., Piper lolot L., Piper sarmentosum Roxb., Zippelia begoniifolia

Blume ex Schult. & Schult. f.,…

+ Flowering season and fruit season: Depending on the species, flowering season

varies from one to another, but the common reason is either from February (March)

9

to May (June), with fruits from April to August, typically Peperomia pellucida (L.)

Kunth., Peperomia parcicilia C.DC, Piper cambodianum C.DC, Piper cf. caninum

Blume, Piper hainanense Hemsl., Piper hymenophyllum Miq., Piper harmandii C.

DC., Piper cf. maclurei Merr., Piper laosanum C. DC… Some of these species

flowering in July-December, with fruits from October to April of the following year:

Piper cubeba L.f., Piper gymnostachyum C. DC., Piper sarmentosum Roxb.,…

3.1.3. The diversity of the Piperaceae in the North Central Vietnam

The results of the investigation and collection of specimens of species in the

Piperaceae family in North Central have identified three genera of 36 species and

species; and recorded the distribution for the North Central Vietnam for 28 species.

Results are presented in Table 3.1.

Table 3.1. List of species in the Piperaceae family in the North Central Vietnam

No Scienctific name

Name in

Vietnamese

Tree of

type Use value

1 Peperomia pellucida (L.)

Kunth.*

Rau càng cua Th THU, ĂNĐ

2 Peperomia parcicilia C.DC. Càng cua ba lá Th ĂNĐ

3 Piper acre Blume*

Tiêu gắt L CTD

4 Piper albispicum C. DC.*

Tiêu gié trắng Th CTD

5 Piper arboricola C.DC.*

Tiêu thượng

mộc

L THU, CTD

6 Piper baccatum Blume* Tiêu phì quả L CTD

7 Piper bavinum C. DC.*

Tiêu ba vì L CTD

8 Piper betle L. Trầu không L THU, CTD

9 Piper boehmeriifoliumWall. ex

Miq

Tiêu lá gai Th THU, CTD

10 Piper boehmeriifolium var.

tonkinensis C. DC.

Tiêu lá gai bắc

bộ

Th THU, CTD

11 Piper bonii C.DC.*

Hàm ếch rừng L CTD

12 Piper brevicaule C. DC.* Tiêu thân ngắn L CTD

13 Pipercambodianum C.DC.*

Tiêu cam bốt L CTD

14 Piper cf. caninum Blume*

Tiêu chó L CTD

15 Piper carnibracteum C.DC.*

Tiêu lá hoa

mập

L CTD

16 Piper chaudocanum C.DC.

Tiêu châu đốc L THU, CTD

17 Piper cubeba L.f.*

Tiêu thất L THU, CTD

18 Piper griffithii C. DC.*

Tiêu griffith L CTD

10

19 Piper gymnostachyum C. DC.* Tiêu gié trần L CTD

20 Piper hainanense Hemsl.*

Tiêu hải nam L CTD

21 Piper harmandii C. DC.*

Tiêu harmand L CTD

22 Piper hymenophyllum Miq.*

Tiêu lá mỏng L CTD

23 Piper laosanum C. DC.*

Tiêu lào L CTD

24 Piper lolot L. Lá lốt Th THU, CTD,

ĂND

25 Piper longum L. Tiêu lá tím L THU, CTD

26 Piper cf. maclurei Merr.*

Tiêu maclure L CTD

27 Piper majusculumBlume*

Tiêu to L CTD

28 Piper mutabile C.DC.*

Tiêu biến thể L THU, CTD

29 Piper nigrum L. Hồ tiêu L THU, CTD,

ĂNĐ, GV

30 Piper pendulispicum C. DC.*

Tiêu gié thòng L CTD

31 Piper pierrei C. DC.*

Tiêu pierre Th CTD

32 Piper pubicatulum C.DC.* Tiêu sóng có

lông

L CTD

33 Piper retrofractumVahl*

Tiêu dội L THU, CTD

34 Piper sarmentosum Roxb.*

Lốt Th THU, CTD

35 Piper saxicola C. DC.*

Tiêu trên đá Th CTD

36 Zipppelia begoniifolia Blume

ex Schult.& Schult. f.*

Tiêu rận Th THU, CTD

Notes: * recorded species distributed in the North Central Vietnam; L:

climbing; Th: herbaceous; THU: medicinal plants; CTD: Essentialy plants; GV:

Spicy plants; ĂNĐ: edible plants.

In order to see the diversity of Piperaceae in the North Central Vietnam, the

results are compared with the total number of species currently known in Vietnam

(Nguyen Kim Dao, 2003) (Table 3.2).

Table 3.2. Composition of the number of species in the Piperaceae in North Central

Vietnam with the total number of species known in Vietnam

Taxon North Central

Vietnam (1) Vietnam (2)

Percentrage %

between (1) and (2)

Genus 3 4 75,00

Species and varieta 36 50 72,00

(2) Nguyen Kim Dao (2003)

11

The results presented in Table 3.2 show that the species composition of

Piperaceae in the North Central Vietnam is quite diverse. Of these, there are 3 genera

than to 4 genera, accounting for 75% of total genera and 36 species and varietas, than

with 50 species and varietas, accounting for 72.00% of total species and varietas

presenting in Vietnam. This shows that, although only distributed in a region with a

relatively modest area compared to other regions in the country, species in this family

are quite diverse and rich.

3.1.4. Species in the Piperaceae family acknoleged additional distribution zones

for the Northern Central flora and added to the flora of Vietnam

Compared to the list of species belonging to the Piperraceae family in the

North Central of Vietnam by Nguyen Kim Dao (2003), the study results recorded the

distribution of 28 species and varieties to the flora of the North Central.

28 species and varieties are newly recorded species in the North Central

Vietnam which presented in the table are distributed as follows: 7 species that are

distributed from Gia Lai Back to Kien Giang is: Peperomia parcicilia C. DC., Piper

arboricola C. DC., Piper cf. caninum Blume, Piper cubeba L.f., Piper harmandii C.

DC., Piper hymenophyllum Miq., Piper pierrei C. DC.; The 7 species that are

distributed from Ninh Binh Province are: Piper bavinum C. DC., Piper bonii C. DC.,

Piper brevicaule C. DC., Piper carnibracteum C. DC., Piper mutabile C. DC., Piper

pendulispicum C. DC., Piper pubicatulum C. DC.; the 6 species and varieties that are

distributed in both North and South are: Piper gymnostachyum C. DC., Piper

hainanense Hemsl., Piper cf. maclurei Merr., Piper retrofractum Vahl., Piper

saxicola C. DC., Zippelia begoniifolia Blume ex Schult. & Schult. f.; 8 species that

are found in the North Central are: Piper acer Blume, Piper albispicum C. DC., Piper

baccatum Blume, Piper cambodianum C. DC., Piper griffithii C. DC., Piper

laosanum C. DC., Piper majusculum Blume, Piper sarmentosum Roxb..

3.1.5. Diversity of use values

The use value of the species in the Piperaceae is identified and determined by

both domestic and international publications. 36 species and varieties that are

12

recorded in the North Central are all valuable, accounting for 100% of the total

number of species and varieties, belonging to three groups of different uses. Groups

of essentialwith 34 species, the largest share which accounts for 94.4%; followed by

medicinal plants with 14 species and 38.89% and edible plants and spices with 4

species, accounting for 11.11%.

- Groups of essential oils (CTD)

Almost all species in the Piperaceae contain essential. However, the

accumulation of Essentialcontent varies from one to another depending on the species

and genus. Essentialfrom species of the genus Piper L. and Zippelia Blume ex Schult.

& Schult.f. are found of high value so they are used in the production of cosmetics,

pharmaceuticals, medicine,.... We studied 18 essential of Piper, distilled 48

specimens and analyzed 36 samples.

- Group of medicinal plants (THU)

With 14 species and varieties, medicinal plants are mainly used to treat common

diseases such as digestive health, digestive diseases, etc., such as: Peperomia

pellucida (L.) Kunth., Piper arboricola C. DC., Piper betle L., Piper

boehmeriifolium (Miquel) Wallich ex C. de Candolle in A. de Candolle, Piper

boehmeriifolium var. tonkinensis C. DC., Piper chaudocanum C. DC., Piper cubeba

L. f., Piper lolot L., Piper longum L., Piper mutabile C. DC., Piper nigrum L., Piper

retrofractum Yahl, Piper sarmentosum Roxb., Zippelia begoniifolia Blume ex Schult.

& Schult. f..

- Group of Edible Plants and Spice Plants: (edible and spice)

The four species used by people today are Piper lolot, Piper nigrum, Peperomia

pellucida and Peperomia parcicilia. These species are widely planted in nationwide.

Especually, Piper nigrum is now a staple crop planted in many mountainous and

Central Highlands regions of Vietnam and is seen a highly profitable export crop,

helping to alleviate poverty.

13

3.1.6. Characteristic discription of species in the Piperaceae

This section mentioned about characteristics of identification, ecological,

distribution, samples and use values.

3.2. Chemical composition of essentialof some species of Piperaceae in the North

Central Vietnam

1. Chemical composition of essential oils of Piper boehmeriifolium

Leaf and stem specimens were collected in Bach Ma National Park in August

2012 (LĐH 335). Essentialcontent is 0.20% fresh weight, light yellow, lighter than

water. 49 compounds accounts for 91.2% of total Essentialcontent. Essentialare

characterized by sesquitecpen accounts for 54.1%; monotecpen accounts for 32.8%;

other compounds account for negligible proportions. Α-copaen (28.3%), α-pinene

(7.4%) and 1.8-cineol (5.7%) are the main components of the essential. It is the first

species to be studied for essential.

2. Chemical composition of essential oils of Piper brevicaule

Sample of Piper brevicaule (LĐH 351) was collected in Pu Mat National Park

in March 2014, reaching 0.15% and 0.12% of fresh weight. Sample no. 453 was

collected in Pu Mat National Park in June 2014; Essential content was 0.22% and

0.17% of fresh weight.

In sample LĐH 351 (leaves and stems), monotecpen hydrocarbons (29.7% and

21.0% respectively), sesquitpenes containing oxygen (25.3% and 31.7%) and

aromatic compounds (22.2% % and 33.8%). The main components of the two

essentialwere sabinene (17.9% and 13.5%), β-eudesmol (13.8% and 8.4%) and

benzyl benzoate (20.6 and 32.5%).

In the leaf samples, 47 compounds were identified and they account for 91.7%

of total essential. The main components of the essentialare benzyl benzoate (20.5%)

are sabinene (17.9%), β-eudesmol (13.8%) and β-phellandrene (3.6%).

In the samples from the stem, 46 compounds were identified accounting for

93.6% of total essential. Benzyl benzoate (32.5%), sabinene (13.5%), β-eudesmol

(8.4%) and farnesol (5.9%) are the main components of the essential. This is the first

species to be studied for essential.

14

3. Chemical composition of essential oils of Piper cambodianum

Specimens of this species were collected in Pu Mat National Park, Nghe An in

May 2013 (LĐH 393). Essential content was 0.23% and 0.18% of fresh weight.

Essential was light yellow and lighter than water. Leaf essential was characterized by

sesquitecpenium compounds accounting for more than 70%; sesquitecpenes

containing oxygen accounts for 64.1%, monotecpenic compounds accounts for

negligible proportions. The main components of the essential were δ-cadinen

(10.3%), bicyclogermacrene (9.7%), bicycloelemen (8.4%), α-humulene (8.1%), α-

pinene %). Essential was also characterized by sesquitrate containing 51.6% oxygen;

Other compounds accounted for negligible proportions. Β-pinene (11.7%), α-pinene

(8.8%), bicyclogermacrene (7.8%), β-caryophyllene (6.4%), α-humulene are the main

part of the essential. This was the first species to be studied for essential.

4. Chemical composition of essential oil of Piper cf. caninum

Samples of leaves and stems were collected at Ke Go Nature Reserve in July

2013. Sample number (LĐH 364). Essential content was 0.2: 0.18: 0.15 and 0.30% of

fresh weight; the Essential was light yellow, lighter than water. Essentialin 4 parts

(leaf, stem, root and fruit) were characterized by sesquitecpen hydrocarbons which

account for 45.1% to 65.6% of total essential; monotecpen accounts for 10.1% to

17.3%; Other components made up negligible proportions. The main components of

the four obtained essentialwere 4-allyl-1,2-diacetoxybenzene (6.4% -24.6%),

bicyclogermacrene (9.4% -17.1%), bicycloelemene (2.5% Β-elemen (3.7% -7.3%), β-

caryophyllene (2.8% -7.2%), α-humulene (2.5-6.7%).

When compared to the Salleh plant in Malaysia, leaf essential was

characterized by safrole (17.1%) but was not detected in the sample; and β-pinene

(8.9%) but in the study sample was very low (0.6%) and linalool (7.0%) in the

sample was 1.6%. Stem’s essential was also characterized by safrole (25.5%) and β-

caryophyllene (9.8%), but in the sample there was none or very low. Thus, climatic

and essential conditions have affected the accumulation of essential.

5. Chemical composition of essential oil of Piper carnibracteum

Leaf and stem specimens were collected in Vu Quang National Park in August

15

2014 with sample number - LĐH 384. Essential content of 0.22 and 0.18% of fresh

weight; The Essential was light yellow. The essential of this species was

characterized by monotecpen hydrocarbons and sesquitecpen hydrocarbons. In

particular, the specific components of the two essentialwere α-pinene (18.3% -

28.1%), β-pinene (15.5% -17.1%), germacrene D (1.5% - 12.2%), β-caryophyllene

(9.0% -10.1%). This was the first species to be studied for essential.

6. Chemical composition of essential oils of Piper chaudocanum

Samples of leaves and stems were collected at Vu Quang National Park in

August 2014 with sample number is LĐH 382. The Essentialcontent was 0.22 and

0.18% of fresh weight; The Essentialis light yellow, lighter than water. In this

essestial Essential, sesquitecpen hydrocarbons accounts for 50.1%; Monotecpen

hyrocacbon accounts for 35.1%; Other compounds account for negligible

proportions. Bicycloelemen (17.2%), bicyclogermacrene (14.3%), β-myrcene

(16.3%), (E) -β-ocimene (10.8%) are the major components of this essential.

Sesquitecpen contains oxygen (54.2%), which is the main component of the

essentialfrom the stem. Other components make up negligible proportions. The main

constituents of bastellon are bicyclogermacren (11.8%), cuparen (15.8%),

bicycloelemen (8.3%), β-myrcen (8.2%), β-caryophyllene (7.5% , (E) -β-ocimene

(7.3%). This is the first species to be studied for essential.

7. Chemical composition of essential oils of Piper gymnostachyum

Essentialand leaf essentialare collected in Pu Mat National Park (LĐH 332)

reached 0.15% and 0.10% of fresh weight. 60 compounds identified in leaf essential

accounting for 96.4% of the total essential content. The main components of the

essential are bicyclogermacren (10.7%), bicycloelemene (9.9%) and -muurolol

(6.8%). From Essentialin the stem has identified 33 compounds accounting for 99.0%

of total Essentialcontent. Limonen (33.6%), α-phellandrene (27.8%) and α-pinene

(18.6%) are key components of essential. This was the first species to be studied for

essential.

16

8. Chemical composition of essential oils of Piper hainanense

Samples of leaves and stems were collected in Ke Go Nature Reserve in July

2013. Sample number is LĐH 359. Essentialcontent of 0.30% and 0.25% of fresh

weight; The essential was light yellow. Essential was characterized by sesquitecpen

hydrocarbons (31.0%), monotecpen hydrocarbons (12.0%), sesquitecpen containing

oxygen (10.0%) and other compounds (17.2%). The main components of the

Essentialare β-caryophyllene (16.7%), 2-tridecanone (4.6%), sabinene (4.4%).

55 compounds found in the stem’s essential accounting for 86.6% . Essential

are sesquitecpen hydrocarbons (37.6%), other compounds are from 3.6% to 13.5%.

Β-caryophyllene (10.5%), 2-decenal (7.4%), germacrene D (4.6%), limonene (4.5%)

are the major components of the essential. This was the first species to be studied for

essential.

9. Chemical composition of essential oils of Piper harmandii

Sample Piper harmandii leaves and stalks are collected in Pu Mat National

Park, Nghe An Province in August 2013 (LĐH 343). The corresponding essential

content was 0.20% and 0.17% of fresh weight. It’s essential was yellow, lighter than water.

The main components identified from the leaves and stems were monotecpen

hydrocarbons (21.6% and 25.2%), sesquitpenones containing oxygen (20.2% and

11.2%) and aromatic compounds (34.1 % and 53.7%). The main components of the

two essentialwere sabinene (14.5% and 16.2%), benzyl benzoate (20.0% and 29.4%)

and benzyl salicylate (14.1% and 24.3%) This was the first species to be studied for

essential.

10. Chemical composition of essential oils of Piper longum

Sample of Piper longum leaves was collected in Pu Mat National Park, Nghe

An in August 2013 (LĐH 342). The content of essential was 0.20% and 0.15% of

fresh weight, the essential was yellow, lighter than water. The results showed that in

essential sesquitecpen accounted for 89.5% and 90.3% respectively for leaves and

stems, the sesquitecpen contained oxygen, for 48.6 and 67.2% respectively; other

components made up negligible proportions.

17

Compared to the published works in Malaysia, the leaves are characterized by

β-caryophyllene (10.2%). Samples studied were characterized by fonenol (40.5%),

elemol (8.2%); Β-caryophyllene accounts for a very low 1.6%.

11. Chemical composition of essential oils of Piper cf. maclurei

Leaves and stems were collected in Bach Ma National Park, Thua Thien Hue

Province in August 2012 (LĐH 337). Essential content of 0.25% and 0.20% of fresh

weight, yellow Essential, lighter than water. (E) -cinnamic acid (37.4%) and (E) -

nerolol (19.4%) are key components of leaf essential. (E) -cinnamyl acetate (17.2%)

and (E) -cinnamaldehyde (8.8%) and phytol (12.2%) were the main component of the

essential. This was the first species to be studied for essential.

12. Chemical composition of essential oils of Piper majusculum

Leaf specimen of Piper majusculum was collected in Pu Mat National Park,

Nghe An Province in August 2013 (LĐH 322). Essential content was 0.15% of fresh

weight, yellow Essential was slightly lighter than water. 36 compounds were

identified and account for 92.9% of total essential. The sesquitecpen hydrocarbons

(76.5%) and the sesquitpenes containing oxygen (10.7%) with the main components

of the Essentialwere β-caryophyllene (20.7%), germacrene D (18.6% %) And β-

elemen (11.3%).

13. Chemical composition of essential oils of Piper mutabile

Leaf specimens and Piper mutabiles were collected in Phong Nha-Ke Bang

National Park in May 2013 (LĐH 319). The essential content was 0.25 and 0.20%

respectively, of fresh weight. Essentialare light yellow, lighter than water.

Essentialare characterized by sesquitecpen 88.5%; Therein, sesquitecpen containing

oxygen accounts for 62.2%; other compounds accounted for negligible proportions.

Spathoulenol (28.5%), α-guaiol (9.8%), β-caryophyllene (9.1%), aptendren epoxide

(6.5%) were the main constituents. Stemo Essentialis characterized by sesquitecpenes

containing oxygen (28.9%), sesquitecpen hydrocarbons (28.6%), monotecpen

hdrocacbon (24.6%) and other components account for negligible proportions. The

main components of the Essentialare sabinene (12.8%), β-caryophyllene (6.4%), -

18

selenene (5.8%), (E) -nerolide (4.6%). This was the first species to be studied for

essential.

14. Chemical composition of essential oils of Piper pendulispicum

Leaf and stem specimens were collected at Vu Quang National Park in August

2013 (LĐH 381). Essential content was of 0.28 and 0.21% of fresh weight; The

essential was light yellow. The Essentialis characterized by sesquitecpen which

accounts for (75.2%); Oxygenated sesquitecpen accounts for 71.9%; Sesquitecpen

hydrocarbons accounts for 3.3%; Other compounds accounted for negligible

proportions. The main component in the essential sesquitecpen (59.8%);

sesquitecpenes containing oxygen accounted for 32.4% and sesquitecpen

hydrocarbons accounted for 27.4%. This was the first species to be studied for

essential.

15. Chemical composition of essential oils of Piper pierrei

Leaf and stem specimens were collected in Pu Mat National Park in May 2014

(LDH 305). The Essentialcontent of leaf and stem of Pierre (Piper pierrei C. DC.)

were 0.15% and 0.12%, respectively. Research results show that, even on the same

plant, the accumulation of essentialvaries from one part to another. The ratio of major

components also varied considerably in the two components. Specifically, in the

leaves α-gurjunen accounts for 18.5% while in the stems it was 10.0%. By contrast,

β-caryophyllene in the stems was 14.0%, while in the leaves it was 10.1%. In

addition, other compounds weew similar to α-copa- and cis-calamenes. The common

components of the two essentialwere α-gurjunen (18.5% and 10.0%), β-

caryophyllene (10.1% and 14.0%), α-copaen (8.0 and 10, 2%) and calamenen (6.7%

and 6.8%).

16. Chemical composition of essential oils of Piper pubicatulum

Samples of leaf and stem were collected at Ke Go Nature Reserve in July 2013

(LĐH 358). essential content of 0.25 and 0.2% of fresh weight; the Essential was

light yellow. The characteristic components in leaf and stem samples were

bicyclogermacren (11.5% -11.9%), β-caryophyllene (6.5% -11.8%), germacren D

19

(6.6% Α-humulen (5.3% -10.7%), β-elemen (9.8% -11.0%). This was the first species

to be analyzed for essential.

17. Chemical composition of essential oils of Piper retrofractum

Piper retrofractum Vahl. leaves were collected from Ke Go Nature Reserve,

Ha Tinh Province in August 2012 (LDH 334). Essential content was 0.2% of fresh

weight, yellow essential, lighter than water. Main components of this Essentialare

were mainly monotecpen and sesquitecpen, accounting for over 30%. In particular,

the aromatic compounds accounted for 18.2% of the composition is benzyl benzoate.

This was the first species to be studied for essential.

18. Chemical composition of essential oils of Piper sarmentosum

Piper sarmentosum leaves were collected from Ke Go Nature Reserve, Ha

Tinh in August 2012 (LDH 336). Essential content was 0.2% of fresh weight, yellow

essential, lighter than water. Essentialare aromatic compounds with benzyl benzoate

(49.1%), benzyl alcohol (17.9%), 2-hydroxybenzoic acid phenylmethyl ester (10.0%)

and 2-butenylbenzene , 9%) was the main component.

Results of the study of 36 samples of essential from various parts such as

leaves, stems, roots, and fruits from the 18 species in the family of Pipeaceae are

summarized in Table 3.22.

Table 3.3. Key components in essential of various parts in a number of species of

Piperaceae in the North Central

No Species

Part of

plant

Content

(%)

Identified

components

Percentage of essential

components

1 Piper

boehmeriaefolium Leaves 0.20 49

α-copaen (28,3%), α-pinen

(7,4%) and 1,8-cineol (5,7%)

2 Piper brevicaule

Leaves 0.15 49

α-gurjunen (18,5%), β-

caryophyllen (10,1%), α-

copaen (8,0%) and cis

calamenen (6,7%)

Stems 0.12 40

β-caryophyllen (14,0%), α-

copaen (10,2%), α-gurjunen

(10,0%), bicyclogermacren

(9,2%)

Leaves 0.22 47 benzyl benzoat (20,5%),

sabinen (17,9%), β-eudesmol

20

(13,8%), β-phellandren (3,6%)

Stems 0.17 46

benzyl benzoat (32,5%),

sabinen (13,5%), β-eudesmol

(8,4%), farnesol (5,9%)

3

Pipercambodianum

Leaves 0.23 45

δ-cadinen (10,3%),

bicyclogermacren (9,7%),

bicycloelemen (8,4%), α-

humulen (8,1%), α-pinen

(7,4%)

Stems 0.18 50

β-pinene (11,7%), α-pinen


(7,8%), β-caryophyllen (6,4%),

α-humulen (6,3%)

4 Piper cf. caninum

Leaves 0.20 50


bicycloelemen (8,4%), β-

elemen (7,3%), 4-allyl-1,2-

diacetoxybenzen (6,4%)

Stems 0.18 50

4-allyl-1,2-diacetoxybenzen


(10,8%), β-caryophyllen

(7,2%), -curcumen (6,3%)

Fruit 0.30 36



(17,1%), limonen (7,1%), β-

elemen (4,3%)

Roots 0.15 46




α-humulen (6,7%)

5 Piper carnibracteum

Leaves 0.22 42

α-pinen (28,1%), β-pinen


(9,0%)

Stems 0.18 37

α-pinen (18,3%), β-pinen

(15,5%), germacren D


(10,1%)

6 Piper chaudocanum

Leaves 0.22 40

bicycloelemen (17,2%),

bicyclogermacren (14,3%), β-

myrcen (16,3%), (E)-β-ocimen

(10,8%)

Stems 0.18 29


cuparen (15,8%),

bicycloelemen (8,3%), β-

21

myrcen (8,2%), β-caryophyllen

(7,5%), (E)-β-ocimen (7,3%)

7 Pipergymnostachyum

Leaves 0.15 60


bicycloelemen (9,9%) và -

muurolol (6,8%)

Stems 0.10 33

limonen (33,6%), α-

phellandren (27,8%) và α-

pinen (18,6%)

8 Piper hainanense

Leaves 0.30 26

β-caryophyllen (16,7%), 2-

tridecanon (4,6%), sabinen

(4,4%)

Stems 0.25 55

β-caryophyllen (10,5%), 2-

decenal (7,4%), germacren D

(4,6%), limonen (4,5%)

9 Piper harmandii

Leaves 0.22 34

benzyl benzoat (20,0%), α-

cadinol (17,0%), sabinen

(14,5%), benzyl salicylat

(14,1%)

Stems 0.17 27


benzyl salicylat (24,3%),

sabinen (16,2%), farnesol

(5,3%)

10 Piper longum

Leaves 0.20 43

fonenol (40,5%), elemol

(8,2%),calamenen (4,1%), δ-

cadinen(3,9%) và α-cadinol

(3,9%)

Stems 0.15 35

fonenol (42,3%), α-cadinol

(9,5%), β-eudesmol (6,8%), -

panasinsen (5,4%), elemol

(5,2%)

11 Piper cf. maclurei

Leaves 0.25 40 E)-cinnamic acid (37,4%) và

(E)-nerolidol (19,4%)

Stems 0.20 21

(Z)-9-octadecanoic acid methyl

ester (28,0%), (E)-cinnamyl

acetat (17,2%),(E)-

cinnamaldehyt (8,8%), phytol

(12,2%)

12 Piper majusculum Leaves 0.15 36

β-caryophyllen (20,7%),

germacren D (18,6%) và β-

elemen (11,3%)

13 Piper mutabile Leaves 0.25 36

spathoulenol (28,5%), α-guaiol


aromadendren epoxit (6,5%)

22

Stems 0.20 58

sabinen (12,8%), β-

caryophyllen (6,4%), -selinen

(5,8%), (E)-nerolidol (4,6%)

14 Piper pendulispicum

Leaves 0.28 36

germacren D (24,3%), β-

caryophyllen (15,0%), β-

elemen (6,1%),

bicyclogermacren (5,3%)

Stems 0.21 45

Acorenon (20,6%), limonen


(7,1%), germacren D (6,9%),

sabinen (6,4%), α-terpinolen

(6,1%)

15 Piper pierrei

Leaves 0.15 49

α-gurjunen (18,5%), β-

caryophyllen (10,1%), α-

copaen (8,0%), cis calamenen

(6,7%)

Stems 0.12 40

β-caryophyllen (14,0%), α-

copaen (10,2%), α-gurjunen


(9,2%)

16 Piper pubicatulum

Leaves 0.25 39



elemen (9,8%)

Stems 0.20 14


caryophyllen (11,8%), β-

elemen (11,0%), α-humulen

(10,7%), bicycloelemen

(10,3%)

17 Piper retrofractum Leaves 0.20 65


myrcen (14,4%),


bicyclogermacren, (7,0%) và

β-caryophyllen (5,3%)

18 Piper sarmentosum Leaves 0.20 19


benzyl alcohol (17,9%), 2-

hydroxy-benzoic acid

phenylmethyl ester (10,0%), 2-

butenyl-benzen (7,9%)

From the results of the analysis of 36 sample in essential extracted from parts

such as leaves, stems, roots, fruits of 18 species in the Piperaceae, it can be seen that

the essential content varies from 0.10% - 0.30% of the fresh weight. The extracted

23

essential were from yellow to light in color, and lighter than water. The number of

compounds identified are between 14 and 65, accounting for 73.9% - 99.4% of total

essential content. It also indicates that the main components in the essential of the

species are also rich and varied. However, the essential are characterized by

monotecpenes and sesquitecpenes.

CONCLUSIONS AND SUGGESTIONS

1. Conclusions

Research on some biological and distributional characteristics and chemical

composition of essential oils of the Piperaceae in North Central Vietnam has drawn

the following conclusions:

1. Identify 36 species and varieties of 3 genera, of which Piper genus is the

most diverse with 33 species. A new distribution area of 28 species for the Northern

Central Vietnam.

2. The general biological characteristics of the species studied is the flowering

season, mainly from February to June; and the fruiting season is from May to

September.

3. Most of the species studied in the Piperaceae family are valuable; 34 species

of oily plants, 14 species of medicinal plants, 4 species of spice plants and edible

plants.

4. Compositions and contents of essential of 36 samples belonging to 18 species

in the Piper genus are identified and analised. For the first time, content and chemical

composition of essential oil of 13 species have been identified: Piper

boehmeriifolium, Piper brevicaule, Piper cambodianum, Piper carnibracteum, Piper

chaudocanum, Piper gymnostachyum, Piper hainanense, Piper harmandii, Piper cf.

maclurei, Piper mutabile, Piper pendulispicum, Piper pubicatulum, Piper

retrofractum.

5. The main chemical compositions in the essentialof a number of Piperaceae are

characterized by monotecpenes and sesquitecpen. Especially monotecpenes

24

containing oxygen and sesquitecpenes containing oxygen such as Piper

boehmeriifolium, Piper brevicaule, Piper cambodianum, Piper harmandii. Essential

from these species has the potential for practical application.

2. Suggestions

- There should be further complete studies on the content and composition of

essential at different stages of plant growth of the same plant in the same place and

same part of plant in different locations to learn about the Essential's accumulation

dynamics of each species. From that, a basis for assessment of plant resources should

be provided. Study the bioavailability of essential of high-essential species to assess

their value is also recommended.

- Further research and evaluation of essential content, quality of essential,

economic value of some species with practical potential should be conducted. This

will be significantly helpul to local authorities and policy makers in term of

developing essentialplant planting areas in the region for improving the local

economy, creating new jobs and improving the local living standards purposes.

3. New contributions of the thesis

- Full examination of specimens and composition of Piperaceae species in the

North Central Vietnam.

- New distribution area of 28 species and the North Central Vietnam.

- The provision of data of the content and chemical composition of essential oils

in parts namely leaves, stems, roots and fruits of 36 samples belonging to 18 species

with the major monotecpenoid and sesquitecpenoid compounds.

- The provision of data on essential oils of 13 species for the first time: Piper

boehmeriifolium, Piper brevicaule, Piper cambodianum, Piper carnibracteum, Piper

chaudocanum, Piper gymnostachyum, Piper hainanense, Piper harmandii, Piper cf.

maclurei, Piper mutabile, Piper pendulispicum, Piper pubicatulum, Piper

retrofractum.

25

PUBLISHED WORKS

1. Le Dong Hieu, Tran Dinh Thang, Tran Minh Hoi (2013), Chemical Composition

of essential in Piper gymnostachy C. DC. from Pu Mat National Park, 5th National

Conference on Science, Agriculture Publisher, Hanoi, 22/10/2013, 1031-1036.

2. Le D. Hieu, Tran M. Hoi, Tran D. Thang, Isiaka A. Ogunwande (2014), Chemical

composition of essential from four species of Piper (Piperaceae), Journal of Oleo

Science, 63 (3): 211-217 (SCIE).

3. Le Dong Hieu, Tran Dinh Thang, Tran Minh Hoi (2014), Chemical composition

of essential of Piper brevicaule C. DC. from Pu Mat National Park, Nghe An,

Journal of Science, Hanoi National University, 30 (6S-A); 184-188.

4. Le D. Hieu, Tran M. Hoi, Tran D. Thang, Isiaka A. Ogunwande (2015), Volatile

constituents of three Piper species from Vietnam, Natural Product Communications,

10(11): 1997-1998 (SCIE).

5. Le Dong Hieu, Tran Minh Hoi, Tran Dinh Thang (2015), Chemical composition

of Pierre pepper (Piper pierrei C.DC) in Nghe An, 6th National Conference on

Science, Nature and Technology, Hanoi, 22 October 2015, 125-129.

6. Le Dong Hieu, Tran Minh Hoi, Do Ngoc Dai (2016), Use values of species in the

Piperaceae family in Vietnam, Scientific Report on Biological Research and

Teaching in Vietnam , 2nd National Science Conference, Da Nang, May 20, 2016,

971-975.

7. Le Dong Hieu, Tran Minh Hoi, Do Ngoc Dai (2016), Diversity of Species in the

Piperaceae in the North Central Coast, Journal of Agriculture and Rural

Development, 2nd March 2017 109-115.

Documents

STUDY ON BIOLOGICAL CHARACTERISTICS, DISTRIBUTION AND …gust.edu.vn/media/25/uftai-ve-tai-day25971.pdf · 2017-10-30 · 1 INTRODUCTION 1. Rationales of study Amongst the flora of