study on biological characteristics, distribution and...
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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
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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
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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), ...
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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.
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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:
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+ 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:
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- 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%
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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
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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)
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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
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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)
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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
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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.
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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.
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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
(8,8%), bicyclogermacren
(7,8%), β-caryophyllen (6,4%),
α-humulen (6,3%)
4 Piper cf. caninum
Leaves 0.20 50
bicyclogermacren (12,3%),
bicycloelemen (8,4%), β-
elemen (7,3%), 4-allyl-1,2-
diacetoxybenzen (6,4%)
Stems 0.18 50
4-allyl-1,2-diacetoxybenzen
(12,9%), bicyclogermacren
(10,8%), β-caryophyllen
(7,2%), -curcumen (6,3%)
Fruit 0.30 36
4-allyl-1,2-diacetoxybenzen
(24,6%), bicyclogermacren
(17,1%), limonen (7,1%), β-
elemen (4,3%)
Roots 0.15 46
4-allyl-1,2-diacetoxybenzen
(14,6%), bicyclogermacren
(9,4%), β-caryophyllen (7,0%),
α-humulen (6,7%)
5 Piper carnibracteum
Leaves 0.22 42
α-pinen (28,1%), β-pinen
(17,1%), β-caryophyllen
(9,0%)
Stems 0.18 37
α-pinen (18,3%), β-pinen
(15,5%), germacren D
(12,2%), β-caryophyllen
(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
bicyclogermacren (11,8%),
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
bicyclogermacren (10,7%),
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 benzoat (29,4%),
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
(9,8%), β-caryophyllen (9,1%),
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
(10,4%), β-caryophyllen
(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
(10,0%), bicyclogermacren
(9,2%)
16 Piper pubicatulum
Leaves 0.25 39
bicycloelemen (11,9%),
bicyclogermacren (11,5%), β-
elemen (9,8%)
Stems 0.20 14
bicyclogermacren (11,9%), β-
caryophyllen (11,8%), β-
elemen (11,0%), α-humulen
(10,7%), bicycloelemen
(10,3%)
17 Piper retrofractum Leaves 0.20 65
benzyl benzoat (14,4%),
myrcen (14,4%),
bicycloelemen (9,9%),
bicyclogermacren, (7,0%) và
β-caryophyllen (5,3%)
18 Piper sarmentosum Leaves 0.20 19
benzyl benzoat (49,1%),
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.