MINISTRY OF EDUCATION AND TRAININGVINH UNIVERSITY LÊ DUY LINH RESEARCH ON THE COMPOSITION OF ESSENTIAL BEARING PLANTS SPECIES IN VU QUANG NATIONAL PARK, OIL-HA TINH PROVINCE Major: Botan
Trang 1MINISTRY OF EDUCATION AND TRAINING
VINH UNIVERSITY
LÊ DUY LINH
RESEARCH ON THE COMPOSITION OF ESSENTIAL BEARING PLANTS SPECIES IN VU QUANG NATIONAL PARK,
OIL-HA TINH PROVINCE
Major: Botany Code: 9420111
ABSTRACT OF A THESIS DOCTOR OF PHILOSOPHY IN BIOLOGY
VINH, 2020
Trang 2This thesis was fulfilled at Vinh University
Supervisors: 1 Assoc Prof Dr Pham Hong Ban
2 Assoc Prof Dr Tran Minh Hoi
Reviewer 1:
Reviewer 2:
Reviewer 3:
The thesis will be defended at Vinh University
The thesis can be found at: The National Library of Vietnam
Library of Nguyen Thuc Hao
Trang 31 Rationale
Located in the tropical monsoon region, Vietnam is ranked inthe sixteenth in the world in terms of biodiversity According to therecord and description in the “Indochinese General Plant” (FloreGénérale de l’Indochine) and its subsequent collections, there areover 240 families with over 7,000 vascular plant species In recentyears, many botanists have predicted that the figure could reach over12,000 species Currently, there have been about 10,500 species ofvascular plants, of which about 657 species belonging to 357 generaand 114 families containing essential oils (accounting for about 6.3%
of total species; 15.8% of total genera and 37.8% of total families).Along with the development of the society, the need to learn and
to use compounds of natural origin is increasing Among the plantsthat are considered as an important resource, those plants containingessential oils play an important part This is an essential source ofmany industries such as cosmetics, food and pharmaceuticals, etc.Most essential oil-bearing plants are in the two branches of Pinus(Pinophyta) and the Magnolia (Magnoliophyta) The family whichhave many species of essential oil plants include Cúc (Asteraceae),Cam (Rutaceae), Na (Annonaceae), Long não (Lauraceae), Bạc hà(Lamiaceae), Hoa tán (Apiaceae), Hoa hồng (Rosaceae), Dầu(Dipterocarpaceae), Nhài (Oleaceae), Thông (Pinaceae), Hoàng đàn(Cupressaceae)
Today, essential oils are widely used in many industries, especially the food,pharmaceutical and cosmetic industries Especially in the field of cosmetics,essential oil is an indispensable material Although there is an increase in syntheticaromatics, these artificial products cannot replace natural oils, because they do notcreate a cool aroma and especially synthetic fragrances are often too expensiveand highly toxic
Vu Quang National Park was established under the Prime Minister’sDecision No 102/2002 / QD-TTg dated July 30, 2002, with a total area of about
Trang 454,000 ha, including 76% of the natural forest area with 5 types of main forestdivided by different elevations This is the place of growth for many valuablespecies: Dalbergia oliveri (Cẩm lai), Chukrasia tabularis (Lát hoa), Erythrophleumfordii (Lim), Magnolia hypolampra (Giổi), Aquilaria crassna (Trầm hương), etcand many precious medicinal plants There have been a number of studies onvascular plant diversity in Vu Quang National Park, but there is a lack of in-depthstudies on plant resources Meanwhile, there exists a huge potential of essential oil
resources in there Therefore, the author selected the topic: “Research on the composition of essential oil-bearing plant species in Vu Quang National Park, Ha Tinh Province” as the content of the thesis.
2 Objectives
Assess the diversity of essential oil-bearing plants and thechemical composition of essential oils extracted from some oil-bearing species in Vu Quang National Park, Ha Tinh province
3 Scientific and practical significance
The research results of the thesis are the survey data that arebasic researches on the diversity of essential oil species in Vu QuangNational Park, Ha Tinh province, and provide new data on Thechemical composition of some essential oils of some families in theflora of Vu Quang National Park
4 New findings
- Providing new data including 366 species of essential oilplants in Vu Quang National Park This is a relatively comprehensiveand systematic data on essential oil plant resources in Vu QuangNational Park, Ha Tinh Province
- For the first time to provide data on essential oils of 9 species
(Bời lời lá thuôn (Litsea elongata (Wall Ex Nees) Hook.f.), Bời lời phiến lá thon (Litsea lancilimba Merr.), Bời lời biến thiên (Litsea variabilis Hemsl.), Kháo vàng thơm (Machilus bonii Lecomte), Kháo nhậm (Machilus odoratissima Nees), Nô vàng(Neolitsea aurata (Hayata) Koidz.), Nô bui san (Neolitsea buisanensis Yam & Kam.),
Trang 5Vàng tâm (Manglietia dandyi (Gagnep.) Dandy), Sa nhân lông hung (Amomum velutinum X E Ye, Skprnick & N H Xia)
5 The structure of the thesis
The thesis consists of 128 pages, 30 tables, 7 pictures, 39 photos structured intothe following main sections: Introduction (3 pages); Chapter 1: Literature review (28pages); Chapter 2: Objects, content and research methods (8 pages); Chapter 3:Research findings (86 pages); Conclusions and recommendations (2 pages); List ofpublished works related to the thesis (2 pages); References (14 pages) and appendix(including 3 appendices, 114 photos)
CHAPTER 1 LITERATURE REVIEW
1.1 Some concepts of essential oils
1.1.1 General concept of an essential oil-bearing plant
there was no exact definition of an essential oil-bearing plantbefore When some oil compounds were found that included in thebody of all organisms (animals’ bile acids, carotene in most plants,etc) As such, there is no clear boundary between essential oil-bearingplants and other plants From this perspective, Nicolaev (1968)defined: “Essential oil-bearing plants are those different from otherplants in that their essential oils can be obtained”
Later, when studying the structure and functioning of secretoryorgans, it became clear about the difference in the nature of essentialoil-bearing plants From that, it is possible to define that Essential oil-bearing plants are plants which contain specialized structures for thesecretion and accumulation of essential oils
1.1.2 Concept of essential oils
Trang 6The French Standard (1987), gives out the definition of essentialoils as follows: “Products obtained from plant-derived materials, by
steam distillation or by mechanical methods for fruit peels of Citrus
genus Essential oils are separated from the water by physicalmethods ” This definition has the limitation that is it excludesproducts obtained by solvent extraction as well as products obtained
by other methods
Essential oils are understood as blends of organic compounds,complex molecular structures and different physical and chemicalproperties Essential oils have several properties:
- All essential oils are liquid, viscous, with optical properties, causing therotation of light
- Most essential oils have less density than water (d<1), some have a greaterdensity than water (d>1), insoluble or very rarely soluble in water, but soluble inorganic solvents
- Essential oils have scent since the composition of essential oil is composedfrom free-form constituents
- Essential oils are evaporable
Based on the chemical molecular structure of essential oils, they areorganized into 4 main groups:
- Aliphatic compounds
- Terpenes and their derivatives
- Benzene derivatives
- Other ingredients
1.2 Studying essential oil-bearing plants
1.2.1 Research on essential oil-bearing plants in the world
So far there are not enough documents to visualize the history ofstudies on essential oil-bearing plants in the world The earliest
Trang 7available document is the “Medicinal Plants” found in Japan, written
in 890 In this document, there are nearly 100 species of essential bearing plants, and it describes the process of producing and usingthese plants
oil-However, research on essential oils and essential oils plantsstarted to attract a lot of attention from researchers in the earlytwentieth century The most remarkable works are those published byCharabot and his students in 1903, 1904, and 1907 At later times,there was a boom in the number of researches varying in many fields.According to Brian M Lawrence in “Progress in essential oils”(1992-1994) and “Essential oils” (1995-2005), the author hasestimated about 1,000 species of essential oil-bearing plants that havebeen analyzed for chemical composition in the world According toLPA Oyen and Nguyen Xuan Dung (1999) in the project “Essentialoil plants in South-East Asia”, in Southeast Asian countries, over 70species of essential oil plants have been analyzed for chemicalcomposition, in which about 30 species have been studied quitecomprehensively from biological characteristics, ecology,distribution, ability to grow, develop, use, yield and be harm by pest,traded for commercial purposes
1.2.2 Research on essential oil plants in Vietnam
The research works on essential oil in Vietnam started after
1956 During this time a series of researches on peppermint,lemongrass, etc were published According to La Dinh Mo and LuuDam Cu (2001), until now, we have exploited from nature andplanted about 20 species of essential oil-bearing plants out of over
600 known species (only 3% of the total) The aforementionedspecies are commonly planted species such as Lemongrass,Peppermint, Fragrance, Camphor, Melaleuca, Cinnamon, Basil,Anise, Patchouli, etc
1.3 Research on chemical composition of essential oil in the world and in Vietnam
Trang 81.3.1 Research on the chemical composition of essential oil in the world
1.3.1.1 Camphor family (Lauraceae)
Research on essential oils in the Camphor family (Lauraceae) has typical works such as XD Ji et al (1991), L Zhu et al (1993), Choudhury, S N et al (1995), Sriramavaratharajan, V et al (2019), etc
1.3.1.2 Magnoliaceae
Many scientific works have been studied on the chemical
composition of essential oils of Magnolia genus such as Theo Oyen
L.P.A., Nguyen Xuan Dung (1999), Grag, SN & Sushil Kumar(1999);Zheng et al (2015), Scharf et al (2016)
1.3.2 Research on chemical composition of essential oil in Vietnam
1.3.2.1 Study on chemical composition of essential oil of Camphor
(Lauraceae)
Research on Camphor’s essential oil (Lauraceae) focuses mainly
on the genera Cinnamomum, Litsea, Machilus, Phoebe There are
some typical works such as: Nguyen Xuan Dung (1996), Tran DinhThang et al (2005), Le Cong Son (2013), Do Ngoc Dai et al (2019)
1.3.2.2 Study on chemical composition of Magnoliaceae family
Many studies have been done on essential oil-bearing plants ofMagnoliaceae family by Dung et al (1997), Lesueur et al (2007), Do
et al (2016a), Bui Van Huong et al (2014), Do et al (2016b),
Trang 91.3.2.3 Study on chemical composition of essential oil-bearing plants of Ginger family (Zingiberaceae)
In Vietnam, the study of the chemical composition of theiressential oil-bearing plants of ginger family (Zingberaceae) is typical
of the works of Nguyen Xuan Dung et al (1996), Nguyen Xuan Dung
et al (2005), Phan Minh Giang et al (2007), Le Thi My Chau et al(2015) , Le Thi Huong (2016), Le Thi Huong, et al (2019), NguyenThanh Hung et al (2018), etc
1.4 The natural, socio-economic conditions of Vu Quang National Park
This section outlines geographical location, terrain, climaticcharacteristics, socioeconomic characteristics, flora and fauna of theresearch area
CHAPTER 2 OBJECTIVES AND RESEARCH METHODS 2.1 Subjects and Research locations
The subject of research is essential oil-bearing plant speciesdistributed in Vu Quang National Park, Ha Tinh
2.2 Time conducted for the research
The thesis was conducted from 2015 to 2019
Each year, the sample is collected into 2 batch, each happens inthe interval of 7-10 days
Research was carried out at Vu Quang National Park, Ha TinhProvince
The total number of samples obtained was 1,200 samples A total
of 1061 templates is collected and stored in the Plant Exhibitionroom, at the Center of Laboratory Practice, Vinh University
Trang 10The chemical composition of essential oils is analyzed at theChemical Institute of Natural Compounds, the Academy of Scienceand Technology of Vietnam.
oil-+ Diversity in terms of gene sources that are rare and precious,
as well as, conservation status
- Analyze the chemical composition of essential oils of some important plants of the families
2.4 Research methodology
2.4.1 Researching from secondary survey
Data on natural and socio-economic conditions in the studyarea, plant samples kept in domestic and foreign museums, publishedworks related to the topic
2.4.2 Methodology of field investigations and surveys
Based on the map, some main areas were chosen to survey.There are 05 main typical areas which are representative for VuQuang National Park The first route is Park Center to Sao La Station– Po Mu The second route is Park Center – Co station (Trạm Cò) –Phan fortress (Thành Cụ Phan) – De slope (Dốc Dẻ) The third route
Trang 11is Son Kim I The fourth route is Son Kim II.The fith route is HuongKhe The process and fieldwork survey were applied following the
method introduced by Nguyen Nghia Thin in “Plant research methods” (2007).
To identify essential oil-bearing plants, it is often necessary tolook up published documents about the chemical composition ofessential oils and rely on photosensitivity (using nose to smell forfragrance) during field investigation
2.4.3 Methods of sampling and classification
Sampling: For each type of tree, 1-3 specimens are usually
collected from the same place After collecting the sample, the sample
is then numbered When collecting samples, the researchers make arecord of some characteristics that are easily faded away when thesamples are dried or soaked in alcohol or hormones such as colors,latex, properties of flowers, leaves, etc Also photos of trees are takenwith Canon digital cameras
Classification: The research method used to classify is a
comparative morphological method This is a traditional researchmethod that is being used today This method is done based onstudying some characteristics of vegetative organs and reproductiveorgans Among the two, reproductive organs are the focus, such asinflorescence position, flower structure (characteristics of bracts,calyx, corolla, stamens, fruits, seeds, etc)
In the process of identifying scientific names of species, theresearch relies on the classified work about Vietnamese plants (PhamHoang Ho, 1999-2000)
The following documents are also used: A handbook forsearching and identifying families of angiosperms in Vietnam (NguyenTien Ban, 1997); Vietnamese timber trees (Forest Inventory andPlanning Institute, Ministry of Forestry) (1971-1989); IndochineseGeneral journal plants (1907); Chinese plantology (1994-2002);Common plants found in Vietnam; Vietnamese plantology: Annonaceae
Trang 12family, Mint Family (Lamiaceae), Ginger family (Zingiberaceae),Camphor family (Lauraceae)], Asteraceae and some other specializeddocuments.
Rectification of scientific names and list development:
Revision of scientific names according to Vietnam’s Plant SpeciesList, RK Brummitt (1992) and http://www.theplantlist.org
2.4.4 Methods to assess the diversity of the flora
-Diversity of taxon of flora: According to “Handbook of
biodiversity research” by Nguyen Nghia Thin (1997):
- Diversity of the trunk types: Based on the records of field
surveys as well as related documents (Specifically, the names ofVietnamese forest trees to collect and evaluate the different forms oftrunks of essential oil-bearing trees
- Diversity in the application of the flora: The application
value of essential oil bearning plants is explored from documents such as: “Dictionary of Vietnamese medicinal plants” (Vo Van Chi, 2012), “1900 species of useful plants in Vietnam” (Tran Dinh Ly et
al, 1993), “List of Vietnamese plant species” (Nguyen Tien Ban (Editor) et al, 2003, 2005), “External forest products Vietnamese timber “(Trieu Van Hung et al, 2007),” Vietnamese medicinal plants and herbs “(Do Tat Loi, 2003),” Vietnamese medicinal plants, how to grow, pick, process, and treat” (Le Tran Duc, 1997), “Medicinal plants and animals in Vietnam” (Do Huy Bich et al, 2004), etc
- Geographical diversity
Establish the spectrum of geographical factors, and apply thedivision of the authors Póc Tamás (1965), Le Tran Chan (1999) andNguyen Nghia Thin (2007)
- Diversity of rare and precious plant species and conservation issues: Based on the Vietnam Red Book (2007), including extinct
species (EX), extinct species in nature (EW), critically endangered
Trang 13species (CR), endangered species (EN), vulnerable species (VU), lowrisk species (LR).
2.4.5 Collect samples and distill essential oil
Specimens for distillation for essential oils are separate parts ofplants (leaves, branches, bark, stems, rhizomes, flowers, fruits) Eachsample collected are of between 0.5-3 kg in fresh condition Thesample is inscribed with the serial number (this number coincideswith the model number for classification) and the time of collection.After collection, the sample was chopped and distilled by steamextraction method with reflux in Clevengert equipment for 2-4 hours
at normal pressure according to Vietnam Pharmacopoeia IV standard(2009)
2.4.6 Methods of quantifying essential oils
Essential oils of various parts are quantified according tomethod I of Vietnam Pharmacopoeia II (2002) The content of freshessential oil is calculated by the formula
Or follow the formula
X(%) = (when d>1)
In which: a is the volume of essential oil, measured in ml
b is the mass of the sample, measured in grams
Essential oils are dried with anhydrous Na2SO4, stored instandard sealed jars, stored at 0-5oC before being analyzed
a x 0.9
a
Trang 142.4.7 Methods of analyzing chemical composition of essential oils
Preparation of analytical sample for gas chromatography:Dissolve 1.5 mg of dried essential oil with anhydrous Na2SO4 in 1 ml
of pure hexane of the type used for chromatographic analysis
+ Gas chromatography (GC) with FID probe: Made on Agilent
Technologies HP 7890A Plus with FID detector, HP-5MS capillarycolumn at length of 30 m, inner diameter (ID) = 0.25 mm, a layer of0.25m-film with hydrogen as carrier gas The injection chambertemperature is 250 ° C Detector temperature is 260oC Program thetemperature of 60 ° C (2 min), increase by 4 ° C / min until 220 ° C, stop
at this temperature for 10 minutes
+ Gas chromatography (GC / MS): This is performed on Agilent
Technologies HP 7890A / HP 5973 MSD combined gas chromatography system with separation column and chromatographic operating conditions as mentioned above and with Helium as carrier gas
2.4.8 Method of data processing
Data are processed using Microsoft Office Excel 2007 software
CHAPTER 3 RESEARCH FINDINGS AND DISCUSSION
3.1 Diversity of essential oil plants in Vu Quang National Park
3.1.1 Diversity in phyla and classes
Results of surveys and identification of essential oil-containingplants in Vu Quang National Park, Ha Tinh There are 366 species,
145 genera and 45 families of 2 vascular plant phyla (Table 3.1) thathave been identified
Trang 15Table 3.1 Distribution of taxon with essential oils in some phyla of flora in Vu
Quang National Park
Percent age %
Numbe
r of species
Percent age %
359 species, accounting for 98.09% of the total number of species,
139 genera, accounting for 95.86% and 41 families, accounting for91.11% of the total number of families; Pinophyta with 7 species,accounting for 1.91%; 6 genera, accounting for 4.14% and 4families, accounting for 8.89% of the total number of families Thus,the essential oil-bearing species concentrate in the phylum of NgocLan with over 90% of genera and species, which is quite reasonablecompared to the evolution of plants because the Magnoliophyta is anphylum dominating vascular plants
The analysis of the taxons of two classes in the Magnoliophytaphylum (Table 3.2), there are also some differences Magnoliopsidahas a dominant taxon number of over 80% of the total number offamilies, genera and species; The class of Onions (Liliopsida) accountsfor a low rate with 7 families (accounting for 16.67%); 19 genera(accounting for 13.67%) and 55 species (accounting for 15.32%)
Trang 16Table 3.2 The ratio of two classes in the Magnolia phylum (Magnoliophyta)
Percentag
e %
Numbe
r of genera
Percentag
e %
Numbe
r of species
of the total families) with the number of species from 6 to 10 species,with the total of 40 species accounting for 10.93% There are 4 families(accounting for 8.89% of the total families) of 11-14 species, with a total
of 51 species accounting for 13.93% There are 4 families vvith morethan 21 species with 206 species, acounting for 56.28%
3.1.3 Diversity of genera
Essential oil-bearing plant species in Vu Quang National Park belong to 145genera In particular, the number of species distributed in the genera is uneven Thereare 17 genera accounting for 11.72% of the total number of genera, but there are 164species accounting for 44.81% of the total species The number of genera with thenumber of species less than 5 is 128 genera (accounting for 88.28%), with a total of
202 species accounting for 55.19% The genera with species which produce more
essential oils are Litsea with 25 species, Cinnamomum with 21 species, followed by Alpinia – 13 species, Piper and Fissistigma with 11 species and Syzygium – 10 species The remaining genera have the number of species ranging from 5 to 9.
Trang 17Further analysis of the genera having essential oil-bearingspecies in Vu Quang National Park showed that 17 genera have morethan 5 species belonging to 7 families: Camphor (Lauraceae), Ginger(Zingiberaceae), Annonaceae, Pepper (Piperaceae), Myrtaceae,Magnoliaceae, Euphorbiaceae Of which, the Camphor family has 7genera (accounting for 41.17%) with 81 species, accounting for49.39% The results of this analysis show that in Vu Quang NationalPark, the Camphor family is very diverse and abundant This is alsoperfectly reasonable, because camphor trees are adapted to the localclimatic conditions, … while Vu Quang National Park is home totropical monsoon climate, with both low and high belts, therefore, it
is very convenient for camphor tree species to grow and develop
Ginger family has 3 genera: Alpinia with 13 species, Amomum with 8 species, Zingiber with 7 species The total number of species of the
three genera of the Ginger family is 28, accounting for 17.07% of the
total species The Annonaceae family also has 3 genera: Fissistigma
with 11 species, Desmos and Uvaria with 5 species The total number
of species of 3 genera is 21 species, accounting for 12.80%
3.1.4 Comparing composition of essential oil plants in Vu Quang National Park with that of Pu Mat National Park
- Compare with Pu Mat National Park
To see the diversity of essential oil-bearing plants in Vu Quang National Park,
Ha Tinh Province, the results were compared with essential oil species in Pu MatNational Park (2016) by Nguyen Viet Hung (Table 3.5)
Table 3.5: Comparison essential oil plants in Vu Quang with Pu Mat
of species
Proportion (%)
Number of species
Proportion (%)
Trang 18of species of Magnoliaceae in Vu Quang is higher than that of Pu Mat
at 111.23% (359 compared to 355) The difference is not significant.Meanwhile, the area of Vu Quang accounts for 59.57% compared tothat of Pu Mat Thus, the above results show that the number ofspecies of essential oil-bearing plants in Vu Quang National Park ishigher than that of Pu Mat even though the area of Pu Mat is 1.8times higher The number of vascular plant species currently known
in Vu Quang National Park is 1.5 times higher than that in Pu MatNational Park The results show that the species of plants bearingessential oils do not depend on the area and the number of species,but investigation
3.1.5 Diversity in trunk form
Through surveys which record the trunk form of essential bearing plants in Vu Quang National Park, and also based on "Names ofVietnamese forest trees", there are five main types of trees whose trunktypes having been classified: trees whose trunk is large, trees whosetrunk is small, shrubs, liana and herbaceous plants (Table 3.7)
oil-Table 3.7 Trunk forms of some essential oil-bearing plants in Vu Quang National