Documente Academic
Documente Profesional
Documente Cultură
Contact Toxicity of Ginger (Zingiber officinale) Essential Oil and Coconut (Cocos
nucifera) Shell Liquid Smoke against mango stem borer larvae
April 2019
Abstract
Seeing as the mango stem borer is a very problematic pest, the study aims to
produce an eco-friendly and cost-friendly larvicide by testing its contact toxicity against
the stem borer. Ginger essential oil (GEO) and coconut liquid smoke (CLS) were the two
primary treatments as larvicide, because of its abundance, and its cheapness. The study
tested the contact toxicity of ginger essential oil, coconut liquid smoke and mixture of
both formulations (GEO and CLS). Tested on 4 treatment sets with 10 larvae each,
including ethanol treatment as the control group. The larvae were checked in 1 hour
intervals, checking for activity and responses when tapped, and also the time it took for
the larvae to die. It was seen that the mixed treatment was the most effective in killing
larvae, killing 9/10 larvae, with the first larva dying within the 1st hour, followed by the
coconut liquid smoke, which killed 8/10 larvae, with the first larva also dying within the
1st hour. It is then followed by the ethanol control group, which killed 4/10 larvae, and
killing the first larva at 4th hour, and then the ginger essential oil, which only killed 3/10
Approval Sheet
This Research Paper is accepted in partial fulfilment of the requirements for the
Science Research II:
_______________________
Date
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE iv
Acknowledgements
First and foremost, we would like to thank God Almighty for giving us the
We would also like to thank our research adviser, Mr. Ivan Hubert C. Juan, for his
limitless patience, encouragement and guidance, which helped us get through this study.
Our sincerest gratitude to the thesis panel for giving us their insights on our
To our classmates in 10 – Queen of All Saints, our class adviser Mr. Christian
Felix, for giving us the support and motivation to finish these dreams of ours, and
To our family and relatives who have given us support through this journey,
To Mr. Manny Correa, for giving us a safe journey throughout our research
appointments.
To the Chemicals & Energy Division of DOST-ITDI and its staff, Mr. Romulio
Estrella, Mr. Peter Rubio, and Ms. Evelyn Manongsong, who assisted in and facilitated
the extraction of the ginger essential oil, giving way for our study,
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE v
To the people in the Forest Products Research Development Institute, Engr. Gino
S. Pasco, Engr. George, Mr. Rodelio and Dr. Jennifer Tamato, who also guided us in our
study,
And Mater Dei Academy, for providing the education and the curriculum they
administered. It is because of our Alma Mater that we were taught on how to conduct the
We would also like to thank anyone and everyone who have directly or indirectly
List of Tables
5 Result of mixed GEO and CLS to one set of larvae for 24 hours ..........................38
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE vii
List of Figures
4 Pyrolysis .................................................................................................................21
larvae ......................................................................................................................22
8 (Top) Machine used for the extraction; (Bottom) Extracted ginger essential oil...39
9 (Top) Signing of forms for the extraction of ginger essential oil in ITDI-CED
DOST; (Bottom) Researchers after getting the ginger essential oil .......................40
11 (Top, Left) Apparatus used in Pyrolysis process; (Top, Right) Produced coconut
shells liquid smoke; (Bottom) Researchers with the staffs of FPRDI. ..................42
List of Appendices
E Photo Documentation.............................................................................................39
G Receipts ..................................................................................................................46
H Work Plan...............................................................................................................49
Table of Contents
Abstract ............................................................................................................................... ii
Acknowledgements ..............................................................................................................iv
CHAPTER
1 INTRODUCTION......................................................................................1
Hypotheses ...................................................................................................3
Definition of Terms......................................................................................5
species……………………….. ..................................................................10
from Three Plants from the Ginger Family (Zingiberaceae) and their
castaneum ...................................................................................................15
larvae….. ....................................................................................................19
Extraction .......................................................................................20
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE xi
Pyrolysis .........................................................................................20
Results ........................................................................................................24
Discussion ..................................................................................................24
Conclusion .....................................................................................................
Recommendations ..........................................................................................
REFERENCES .................................................................................................................27
APPENDICES.. ............................................................................................................…31
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 1
Chapter 1
Introduction
The larvae of stem borer, causes widespread damage against mango trees. This is
mango. The country has established itself as a leading producer and exporter of processed
mangos, particularly in the dried mango category (Duke CGGC, 2017). Locally, mango is
the third most important fruit both in terms of quantity and value of production after
bananas and pineapples. Thus, the mango industry plays an important role in the
economy of the country, providing a source of livelihood to about 2.5 million farmers
(DOST-PCAARRD, 2011).
Mango stem borer is one of the major pests of mango orchards. The mango stem
borer feeds internally and thus becomes difficult to control it once the larvae enter the
mango stem/shoot. This polyphagous pest attacks both living and dead trees, wherein
severe infestation affects the whole shoots and tree looks like burned trees and causes
simply ginger, is widely used as a spice or a folk medicine. Previous studies were
conducted about ginger in which many pharmacological properties of ginger have been
identified. This includes essential oil, zingiberol, zingiberone, zingiberene, and pungent
components such as gingerol and shogaol (Goto et al., 1990; Shoji et al., 1982; Connell
and Sutherland, 1969). The essential oil of Zingiber officinale (GEO) is proven that it has
larvicidal activity against Cx. Tritaeniorhynchus and An. subpictus mosquito species
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 2
(Govindarajan, 2011). It was also found to have larvicidal activity against Cx.
Coconut shell is a common wood waste material in Philippines. For the reason
that the Philippines is also known as the world’s second largest producer of coconuts
according to World Atlas. Coconut shell composition is composed of lignin and cellulose,
which is burned with limited air at high temperatures (Yulistiani, 1997). It contains many
compounds that are formed due to the pyrolysis of three components of wood,
including phenol, carbonyl, acids, furans, alcohols, lactones, hydrocarbons, and polycylic
aromatic that are anti-microbial and toxic to insect pests (Thamrin, 2007). The study will
use coconut shells and ginger essential oil to produce a liquid smoke larvicide targeting
The study investigated on what is more applicable solution against stem borers
and the accuracy of the quantity of killed pests with each solution (with GEO only, with
CLS only, and with mixed solution of GEO and CLS.) The following questions were
answered:
1. Which formulation of larvicide (GEO, CLS, GEO and CLS) will have the fastest
reaction against larvae in terms of kill time in killing larvae of each set(10 larvae)
after 24 hours?
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 3
2. Which formulation of larvicide (GEO, CLS, GEO and CLS) will have the most
Research Hypotheses
1. The mixed formulation of GEO and CLS would have the fastest reaction against
larvae in terms of kill time in killing larvae of each set(10 larvae) after 24 hours.
2. The mixed formulation of GEO and CLS would have the most number of killed
The stem borer is the most damaging pest because the larvae of the pest live for
long time (about 1 Year) and hibernate in winter inside the dry shoot, when climate
warms up they activate and pupate inside then adults emerge and start egg laying from
July-August. When the larvae matured it starts to bore too deep in the stem/shoot
It can destroy one of the abundant crop in our country at any stage of the plant for
the grub is damaging stage and damages by cutting and chewing of newly twigs and
shoots. Once grub enters in the shoot creates tunnel inside the stem and damages the stem
The study aims to eliminate these stem borer larvae by targeting it in its larval
stage. By addressing the problem of stem borers against mango trees, liquid smoke can be
The study targets to deal with the use of organic materials abundant in our country
and liquid smoke concentrations as a natural larvicide that is effective for killing stem
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 4
borers in mango trees. It also aims on recycling coconut shells, for it is one of the major
wood wastes that has not been utilized properly and will make use of ginger essential oil
Also, one of the objectives of the study is to investigate in which solutions and
The study can benefit the community by accomplishing the following goals:
address problems regarding stem borers against mango trees and the use of liquid smoke
as an alternative to synthetic pesticides, since the synthetic ones can cause negative
pollution); and raising environmental awareness by recycling wood waste material into
organic larvicide.
environmental friendly. The information of larvicide that this study will create can
contribute to the elimination of stem borers, one of mainly harmful pests in plants.
officinale essential oil and Cocos nucifera shell liquid smoke towards stem borers.
The study also investigated what solutions (GEO: CLS: 1:1 GEO and CLS) has
the best results in terms of the time it took to kill a group of stem borer larvae and in
The study was limited to the analysis that the said larvicide has the same
effectiveness in terms of the quantity of killed larvae and how fast it kills larvae against
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 5
other mango tree pest. The effects of other limitations such as effectiveness of the
larvicide on other species aside from stem borer, and effects of it in mango trees was not
Definition of Terms
Larvicide. An insecticide that targets larvae in breeding habitat before they can mature
and wood preservative due to the uniqueness of its complex structure and
chemical composition.
oxygen. The products of biomass include biochar, bio-oil and gases including
Emulsion. Refers to the mixture of two or more liquids that are normally immiscible.
Stem borer. Any insert larva, or arthropod, that bores into plant stems. However the term
most frequently refers among the Coleoptera to the larva of certain longhorn
beetles such as Dorysthenes buqueti and those of the genus Oberea, and among
Shoot. A young branch or sucker springing from the main stock of a tree or other plant.
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 6
Chapter 2
king of fruits, asking of fruits for its sweetness, excellent flavor, delicious taste and high
nutritive value. This important tropical fruit is being grown in more than 100 countries
(Sauco, 1997). It is also valuable environmental and shade tree, which contributes to the
protection of soil against erosion and different medicinal virtues (D Almeida, 1995). The
Philippines holds a relatively significant position in the mango GVC (Global Value
Chain). The country has been an important player in the global market since the 1980,
The Life Cycle study was done at RARS, Tarahara (Horticulture research unit) in
borer infested Amrapali mango tree. The standing plant was covered by cloth net
(mosquito net) and no any treatment was applied upon plant. The observation was taken
at 15 days interval. Study was started from July 2011 to October 2012. The study showed
that the mango stem borer adult starts egg laying upon the bark and dead shoots of mango
tree from July – August and egg hatching takes place within 10- 15 days. Newly hatched
larvae of the pest start feeding upon the newly flushed leaves and start boring onward.
Once larva enters the shoot, it starts cutting and boring the shoots.
The mango stem borer (Batocera rufomaculata) a species of beetle in the family
Cerambycidae. The adult starts egg laying upon the bark and dead shoots of mango tree
from July –August and egg hatching takes place within 10- 15 days. Newly hatched
larvae of the pest start feeding upon the newly flushed leaves and start boring onward.
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 7
Once larva enters the shoot, it starts cutting and boring the shoots. This pest damages the
young shoots of mango turning dry and no flowering and fruiting. It is also because that
dry shoot does not carry any type of pesticides towards the dry shoot where larva remains
(Upadhyay, 2013).
Nepal
indica L.) a member of family Anacardiaceae is known as king of fruits, asking of fruits
for its sweetness, excellent flavor, delicious taste and high nutritive value. This important
tropical fruit is being grown in more than 100 countries (Sauco, 1997). It is also valuable
environmental and shade tree, which contributes to the protection of soil against erosion
and different medicinal virtues (D Almeida, 1995). Mango stem borer is one of the major
pests of mango orchards. The farmers are destroying the orchards in ETR due to the
heavy infestation of this pest. A survey in Siraha and Saptari districts reported mango
fruit fly, mango hopper and mango fruit borer as prevailing insect pests of mango (Regmi
et al., 2004).
The survey and field observation of the orchards indicated that Mango stem borer
is a serious pest of mango in Saptari district and also shifting to Siraha and Udaipur
districts respectively. A detailed farm and farmers' survey was conducted among 30
farmers in each district in ETR of Nepal during 2010-11 by using pre-tested semi-
structured questionnaire. The survey assessed the information on insect pest incidence,
their nature and extent of damage (Symptoms, plant-parts infested, time of attack and
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 8
extent of damage due to insect pests) in different varieties of mango fruit and available
25% WG and Trizophos 40% SL performed best in management of mango stem borer.
Among the treatments, Imidacloprid 17.8% SL, Thiamethoxame 25% WG was found
In this study presented by Chowdhury, The aim of this present study is to focus
yield loss of mango fruit by different mango insect pests and their effect on economic
condition of mada peoples as well as national and international trading system. Northern,
eastern, western and southern regions of India are recognized as distinct mango growing
regions, though some cultivars are grown over wider areas (Yadav and Rajan, 1993). The
insects along with their immature and mature stages were collected by traditional
methods of hand picking. The adults and nymphs were collected and preserved in 75%
alcohol in vials for later identification in Laboratory. Grubs feed inside the stem boring
upward making irregular tunnels which results in interruption of nutrient and water
transport in the tissue. Drying of terminal shoot in early stages and severe symptoms
important element is the ADH system, which degrades 90% of the total ethanol in
Drosophila melanogaster (Heinstra et al., 1987; Geer et al., 1993). ADH action per se is
enough to convert a toxic exogenous substance (ethanol) into a common endogenous one
(acetate) (Kapoun et al., 1990; Chakir et al., 1993; Geer et al., 1993). Third instar larvae
were exposed to ethanol. The exposure was carried out in Petridishes sealed with PVC
film, each with a cellulose sponge soaked in a solution containing ethanol at different
concentrations, 0.15 M NaCl (to maintain the osmotic equilibrium) and 1% glucose (to
exposure experiment and after the first 12 h of exposure, the larvae were transferred to
new Petri dishes with fresh solutions at the same ethanol concentration. The
concentration required to kill half of the larvae exposed during a given time was called
the Lethal Concentration 50 (LC 50). The survival of larvae exposed to ethanol (whose
toxicity was placed in evidence by our model) was dependent on several factors.
Regarding the effects of the genetic background, data from LC analysis suggest that there
was hybrid superiority, thus characterizing a heterosis effect, although there was no clear
statistical significance for this statement. Through this rare and informative model, in
traits, is made possible, we demonstrated that the rearegenetic factors acting on the
Larvicidal activity of Tagetes patula essential oil against three mosquito species
In this study that was conducted by Dharmaggada, Naik, Mittal and Vasudevan,
Mosquitoes are well known vectors of several disease causing pathogens. The synthetic
insecticides are toxic and adversely affect the environment by contaminating soil, water
and air. There is a need to find alternatives to these synthetic pesticides. Botanical
pesticides are promising in that they are effective, environment – friendly, easily
biodegradable and also inexpensive. The steam carried the volatile organic compounds
(essential oil) present in the plant material into the vapor phase. Condensation decreased
the solubility of essential oil constituents in the vapor and the essential oil was easily
Fourth instar larvae, which is the fourth larval stage in the life cycle of mosquito
after which the larva finally becomes a pupa, were exposed to sub lethal concentrations
of 6.25, 12.5, 25, 50, 100 and 150 ppm of the oil in distilled water for 24 h according to
standard WHO procedure (1981). As the essential oil does not dissolve in water it was
first dissolved in ethanol (99.8%). The test medium was prepared by adding 1 ml of
appropriate dilution of essential oil in ethanol and mixed with 249 ml of water to make up
250 ml of test solution. The oil – ethanol – water solution was stirred for 30 s with a glass
road. After about 15 min 25 larvae taken on a strainer with fine mesh were transferred
gently to the test medium by tapping. Two replicates, with 25 larvae in each, were taken
for each mosquito species in each solution and the control of ethanol – water mixture.
The dead larvae were counted after every hour and percentage mortality is reported from
the average for the two replicates taken together. The diviation between the replicates is
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 11
±10 e. Observations were also made on the larval behavior, morphological changes and
adult emergence.
smoke
distillation on character of resulted liquid smoke has been done. Objective of the research
is to identify effect of coconut shell pyrolisis temperature and liquid smoke distillation on
affected by raw material type, duration of burning and burning temperature. Liquid
smoke is used commonly by agent providing aroma, texture and taste of food product,
such as mead, fish and cheese. Washed coconut shell with its dimension is reduced was
put into pyrolisis reactor and closed well. Reactor was heated in high temperature (150-
450°C) with heating rate of 30°C per minutes. After reaching desired temperature, it was
let 2 hours and smoke flow through evaporation pipe, cooled through cooling media.
Then its liquid was discharged through pipe placed in container. After there is no liquid,
the heating is stopped and reactor is cooled. Condensate and charcoal was weighted to
concentration was 28.75% (m/m). Characteristic (pH, acid level and phenol level) of
liquid smoke before and after purification process and distillation are various. Highest pH
(6.2) was D-I, while the lowest (4.1) at D-VI. The highest acid level (58.40%) was at D-
VI, while the lowest (1.86%) was at D-I. at D-I liquid smoke does not contain phenol,
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 12
while the highest phenol content (3.85%) was in D-VI and the lowest was liquid smoke
residue (1.93%).
In this study conducted by F.X. Wagiman, Arik Ardiansyah and Witjaksono, they
used liquid smoke as a form of pesticide to the rice brown planthopper. Since coconut
shell composition is composed of lignin and cellulose and is suitable for pyrolysis
process, they used it as a chip in making the liquid smoke pesticide. Laboratory trial
proved that the genuine of coconut-shell liquid-smoke grade II was very toxic against the
rice brown planthopper (Nilaparvata lugens) and phytotoxic to the rice plants.
The genuine liquid smoke is very acid (pH 1.2). It could be neutralized (pH 6, 46) by
adding it with calcium oxide at rate of 7 grams per 100 ml. LC50 of the neutral coconut-
shell liquid-smoke grade II against N. lugens at 24, 48, and 72 hours after treatment were
12.89, 11, and 9.94%, respectively. The concentration rate of 12.5% was feasible to be
developed into application dosage at which this concentration rate of neutral liquid smoke
Insecticidal properties of Zingiber officinale and Piper cubeba essential oils against
Essential oils are the complex mixture of volatile organic compounds produced as
secondary metabolites whose functions are other than nutrition. Essential oils and its
constituents are known to have repellent and insecticidal activities against several stored
grain insect pests. In the present study essential oils from Zingiber officinale rhizome and
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 13
Piper cubeba berries have been isolated and its insecticidal activities against red flour
beetle Tribolium castaneum have been explored. Repellency assay was carried out in
glass petri dishes (diameter 8.5 cm and height 1.2 cm). Test solutions of serial dilution of
Z. officinale and P. cubeba essential oils were prepared in acetone. Whatmann filter paper
was cut into two equal halves and each essential oil solution was applied to filter paper
half as uniform as possible using micropipette. The other half of filter paper was treated
Twenty adults of T. castaneum were released at the centre of filter paper disc and
then petri dishes were covered and kept in dark. Four replicates were set for each
concentration of essential oil solution. Number of insects on both treated and untreated
halves was recorded after 4 h in mild light. Fumigant toxicity of Z. officinale and P.
cubeba essential oils was tested against larvae and adults of T. castaneum. Ten
adults/larvae taken from the laboratory culture were placed with 2 g of wheat flour in
petri dishes (diameter 8.5 cm and height 1.2 cm). A filter paper strip (4 cm2) treated with
essential oil solutions of different concentration prepared in acetone, was pasted on the
under cover of petri dishes. Oviposition inhibitory activity of Z. officinale and P. cubeba
According to results, Z. officinale and P. cubeba essential oils are repellant and
toxic to growing larvae and adults of T. castaneum. These essential oils repel adult
cantonensis
the roots of ginger (Zingiber officinale), for the parasite Angiostrongylus cantonensis.
This study found that the above constituents killed A. cantonensis larvae or reduced their
mebendazole and albendazole. These constituents of ginger showed effects against DPPH
and peroxyl radical under larvicidal effect. Together, these findings suggest that these
officinale
dehydrogingerdione (2) and (6)-dihydrogingerdione (3); the latter has not previously been
reported from Z. officinale. The structures were established from infrared (IR), ultraviolet
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 15
(UV), 1H-nuclear magnetic resonance (NMR), 13C-NMR and mass spectral data.
instar larvae of A. aegypti (LC50 4.25, 9.80, 18.20 ppm) and C. quinquefasciatus (LC50
5.52, 7.66, 27.24 ppm), respectively. The results show that the most effective compound
was 4-gingerol.
Contact Toxicity, Feeding Reduction, and Repellency of Essential Oils from Three
Plants from the Ginger Family (Zingiberaceae) and their Major Components
The essential oils from rhizomes of Alpinia conchigera Griff, Zingiber zerumbet
Smitt, Curcuma zedoaria (Berg.) Roscoe; their major compounds (camphene, camphor,
essential oils comprised of mixtures of major pure compounds in the same ratios as the
extracted essential oils were tested for contact, feeding reduction, and repellency against
Several essential oils are also toxic to stored product insects (Huangetal. 2000
Negahban and Moharramipour 2007 Rajendran and Sriranjini 2008). Therefore, the
essential oils could also be used as alternative sources to control stored-product insects,
due to their low toxicity to warm-blooded mammals (Shaaya et al. 1991, 1997; Kim et al.
2003). The objective of this study is to determine whether the essential oils from three
Thai plants from the Zingiberaceae-A. conchigera, Z. zerumbet, and C. zedoaria – are
active against two stored-product insects, S. zeamais and T. castaneum. Pure Compounds
and Synthetic Essential Oils. Chemical constituents of the essential oils were determined
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 16
ml of three essential oils, eight pure compounds or three synthetic oils at different
applied topically onto the pronotum of S. zeamais (average weight of insect, 2.305 mg) or
The commercial sugarcane variety Ja 60-5 was obtained from the germplasm
methodology reported by Payan et al. Briefly, meristematic tissue from sugarcane was
excised, disinfected and placed on callus induction medium (P) containing MS salt, 100
mg/ml myo-inositol, 0.8 mg/l thiamine-HCl, 500 mg/l casein hydrolysate, 20 g/l
sucrose,4 mg/l 2,4-D, 7 g/l agar-agar, pH 5.6. The culture was maintained in the dark at
25 C for one month. Subsequently, friable embryogenic calli were transferred to fresh
SCSB colonies were established using semi-artificial diet: 33 g/l maize flour, 45
g/l wheat germ powder, 16.5 g/l sugarcane leaf powder, 42.5 g/l dry yeast,4.3 g/l ascorbic
acid, 1.32 g/l boric acid, 3 g/l methyl parabene, 10% formaldehyde. Soluble proteins from
E.coliex pressing the tcryIA(b)gene were assayed for lethality to neonate SCSB larvae by
The evaluation of larvicidal activity was carried out against 1-day old, third-instar SCSB
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 17
larvae by putting them onto the artificial diet. Ten larvae were used for each toxin dose. A
diet, supplemented with lysates obtained from pUC 19-transformed E. coli culture, was
used as negative control. Mortality was scored after 7 days and LC50 was calculated.
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 18
Chapter 3
This chapter presents the research processes that are going to be used, equipment
that were utilized, as well as the techniques that we were used in the treatment of data
gathered.
Research Design
Fig. 1. Schematic Diagram. Rectangles on the left demote major phases of the study,
Research Methodology
Phase 1 – Gathering of materials (ginger and coconut shells) and collection of stem
borer larvae. The first phase of the study entails the gathering of raw materials,
consisting of rhizome of ginger (30 kilograms) and coconut shells (six sacks) that were
gathered in wet markets of Pulong Buhangin and Poblacion in Santa Maria, Bulacan and
Also, the proponents went to mango farms in Santa Maria and Norzagaray,
Fig. 2. Gathering of materials and collection of mango stem borer larvae. 36-kilogram
sample of ginger (top, left), gathered six sacks of coconut shells (top, right), collected
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 20
mango stem borer in the stem (bottom, left), and the mango stem borer removed from the
Phase 2 – Extraction of ginger essential oil and production of coconut shell liquid
smoke (pyrolysis)
Technology Development Institute (CED – DOST Chemicals and Energy Division) for
Fig. 3. Extraction of ginger essential oil. Sliced ginger (top, left), Oil pressing machine
Pyrolysis. The collected six sacks of coconut shells (crushed) were passed to Forests
The coconut shells were placed to the combustion tube with a temperature of
350°C. Every hour for 4 hours, the combustion tube was refilled with new set of coconut
shells.
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 21
Fig.4. Pyrolysis. The combustion tube and tank condenser used for Pyrolysis (left), and
Phase 3 – Emulsion Process (Mixing of GEO, Ethanol, and distilled water) and
(Mixing of GEO, CLS, and distilled water). The extracted ginger essential oil and
produced coconut shell liquid smoke were mixed with ethanol and distilled water. For the
formulation of GEO only, 37.5µL of ginger essential oil, 5055µL of ethanol and distilled
water. For the formulation of mixed GEO and CLS, same values used in GEO and
Fig. 5. Emulsion Process. The mixed GEO with Ethanol ( left), the Ethanol as control
sample (center), and the mixed GEO and CLS with Ethanol ( right).
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 22
Fig. 6. Investigation of Contact toxicity of each formulation against mango stem borer.
The produced formulations used to kill stem borer larvae (left and right).
borer larvae
The parameter that was investigated is the formulation of GEO against mango
stem borer larvae. The investigation was done through spraying method (100 sprays) to
the set of larvae (10 larvae) and observing the time of death of each larva within time
ranges. Then, the study got the number of killed larva/larvae per hour and the number of
The parameter that was investigated is the formulation of CLS against mango
stem borer larvae. The investigation was done by spraying 10 mL (100 sprays) of CLS to
the set of larvae (10 larvae) and was observed for 24 hours. Then, the study got the
number of killed larva/larvae per hour and the number of killed larva/larvae in each set
The parameter was investigated is mixed formulation was done by spraying the
mixed concentrations of GEO (37.5 µL) and CLS emulsified in 5,055 µL of ethanol with
a total amount of 250 ml (100 sprays) to the set of larvae (10 larvae). The parameter that
was investigated was observed for 24 hours. Then, the study got the number of killed
larva/larvae per hour and the number of killed larva/larvae in each set under the
The parameter that was investigated is the control sample for the study, ethanol
against mango stem borer larvae. The investigation was done through spraying method to
the set of larvae (10 larvae) using 10mL (100 sprays) of ethanol and observing the time of
death of each larva within time ranges using 1 hour time intervals. Then, the study got the
number of killed larva/larvae per hour and the number of killed larva/larvae in each set
Chapter 4
Results
Table 1. Shows the results of four different formulations sprayed on 4 different sets
Discussion
The result of contact toxicity of Ethanol being the control sample against mango
stem borer was during the fourth hour of observation, the control sample started to affect
the larvae, killing one larva. From the seventh hour until twelfth hour, there were another
larvae killed. In 13th hour until 16th, a total of three larvae were killed. On the next hour
(17th hour) until 24th hour there was another larva killed resulting to 4 killed larvae out of
For the result of contact toxicity of GEO against mango stem borer, during the
12th hour after the GEO was sprayed to larvae, 1 out of 10 larvae was killed. For the next
4 hours (sixteenth hour), another two larvae were killed. In twenty-third hour up to the
last hour of observation, another larva was killed having a total of 3 killed larvae out of
10 larvae alive in the formulation of GEO. To summarize, for the whole 24-hour
observation of larvae with sprayed GEO, there were 3 out of 10 larvae killed.
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 25
For the contact toxicity of CLS against mango stem borer, for the first hour of
observation, there were already six larvae killed and this was the observation until
seventh hour. For the eighth hour up to ninth hour, another larva was killed and finally,
for the tenth hour up to the twenty-fourth hour of observation, another larva was killed
again. Over the 24-hour observation, there were 8 out of 10 larvae that were killed.
For the last observation of contact toxicity against mango stem borer with the
formulation of mixed GEO and CLS, during the first hour of observation, there were
already five dead larvae. During the third hour, another larva was killed. In the fourth
hour until tenth hour of observation, a total of seven were killed larvae. In 11th hour up to
17th hour, another larva was killed as well as in the 18th hour until 24th hour having a total
Fig. 7. The larvae sprayed with different formulation. Ethanol (top, left), mixed GEO and
CLS (top, right), GEO (bottom, left), and CLS (bottom, right).
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 26
Chapter 5
Conclusions and Recommendations
Conclusions
The researchers have concluded that the mixed formulation of CLS and GEO
have the highest contact toxicity among the other formulation (GEO and CLS) for it has
the fastest reaction against mango stem borer larvae. It had killed five larvae after the first
hour (larvae does not respond after tapping; straight in body shape but have minimal dark
spots). Also, it has the most number of killed larvae in set after 24 hours (9 over 10)
Recommendations
application of the formulations (Mixed, GEO, and CLS) against Batocera rufomaculata.
Also, for them to try different concentrations of the said formulations, to determine things
such as:
If ginger essential oil has really low contact toxicity to mango stem borer larvae,
The result to the ratio of mixed formulation such as (1:1), (1:2), and (2:1).
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 27
References
Abbott, WS. (1925). A Method of Computing the Effectiveness of an Insecticide. Journal
Agarwal, M., Walia, S., Dhingra, S., Khambay, B. (2001). Insect growth inhibition,
officinale Roscoe (ginger) rhizomes. Pest Management Science Vol. 57 (3), pp.
Bandong, J.P., Litsinger, J.A. (2005). Rice crop stage susceptibility to the rice yellow
International Journal of Pest Management Vol. 51 (1), pp. 37-43. DOI: 10.
1080/09670870400028276.
Burton, J. (2018). The world leaders in Coconut procudtion. WorldAtlas. Retrieved from:
https://www.worldatlas.com/articles/the-world-leaders-ini-coconut-
production.html
Butterworth, J.H., Morgan, E.D. (1968). Isolation of a substance that suppress feeding in
https://www.worldatlas.com/articles/the-countries-producing-the-most-rice-in-
the-world.html
species and their hybrids." Genetics and Molecular Biology 32, no. 1 (2009), 177-
185. doi:10.1590/s1415-47572009005000012.
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 28
Catling, H., Islam, Z. (1995). Studies on the ecology of the yellow stem borer
DOI: 10.1016/j.biotechadv.2009.09.003.
Dharmagadda, V.S.S., S.N. Naik, P.K. Mittal, and P. Vasudevan. "Larvicidal activity of
Ghosh, A., Chowdhury, N., Chandra, G. (2012). Plant extracts as potential mosquito
larvicides. The Indian Journal of Medical Research. Vol 135 (5), pp. 581-598.
doi:10.1016/s1995-7645(11)60047-3
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 29
Huesing, J., English, L. (2004). The impact of Bt crops on the developing world.
http://hdl.handle.net/10355
Lin, R., Chen, C., Chung, L., & Yen, C. (2010). Larvicidal activities of ginger (Zingiber
doi:10.1016/j.actatropica.2009.12.007
(PDF) Transgenic sugarcane plants resistant to stem borer attack. (2008, June 28).
Retrieved from
https://www.researchgate.net/publication/226582992_Transgenic_sugarcane_plan
ts_resistant_to_stem_borer_attack
Prabhakar, K., Maheshwari, R., & Vimal, O. (1986). Pyrolysis of coconut shell and its
4607(86)90140-x
Rahuman, A. A., Gopalakrishnan, G., Venkatesan, P., Geetha, K., & Bagavan, A. (2008).
Salim M, Masir R. (1987). Efficacy of insecticides against rice stem borer at NARC,
Senapati, B., Panda, SK. (1990). Rice stem borers in insect pests of cereals and their
http;//www.knowledgebank.irri.org/
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 30
Shirin, A. P., & Jamuna, P. (2010). Chemical composition and antioxidant properties of
2674-2679. doi:10.5897/jmpr09.464
Toxicity, Feeding Reduction, and Repellency of Essential Oils From Three Plants
From the Ginger Family (Zingiberaceae) and Their Major Components Against
Thamrin, M. (2007). Efek asap cair cangkang kelapa sawit terdahap jamur ganoderma sp.
Pada Kayu Kelapa. Jurnal Sains Kimia Vol. 11 (1), pp. 9-14.
Yulistiani, R., Purnama, D., Harmayani, E. (1997). The inhibition ability of liquid smoke
against growth pathogens and blight on cow tongue. Master Thesis. Study
Appendix A
Appendix B
Memorandum of Agreement
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 33
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 34
Appendix C
Proof of Guidance
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 35
Appendix D
Experimental Results
Table C1. Shows the effects of the control sample (Ethanol) to 10 larvae for 24 hours.
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 36
Table C2. Shows the effects of the GEO to 10 larvae for 24 hours.
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 37
Table C3. Shows the effects of the CLS to 10 larvae for 24 hours.
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 38
Table C4. Shows the effects of the mixed GEO and CLS to 10 larvae for 24 hours.
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 39
Appendix E
Photo Documentation
Fig. D. Machine used for the extraction (top). The extracted ginger essential oil (bottom).
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 40
Fig. D2. Signing of forms for the Extraction of ginger essential oil in ITDI-CED DOST
(top). The researchers after getting the ginger essential oil (bottom).
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 41
Fig. D4. The apparatus used in Pyrolysis process (top, left).The produced coconut shells
liquid smoke (top, right). The researchers with the staffs of FPRDI (bottom).
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 43
Fig. D5. The emulsification process of GEO and Ethanol (top, left); GEO and CLS with
Appendix F
Abbott’s Formula
Formula
Then x-y=the percentage killed by the treatment and the percentage killed by the
treatment (x-y) divided by the percentage living in the check (x) gives the control or
expressed by an equation:
𝑥−𝑦
(100) = percentage control
𝑥
Ethanol:
4
10
CLS:
6−2
(100) = 66.67%
6
GEO:
6−7
(100) = −16.67%
6
Mixed:
6−1
(100) = 83.3%
6
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 46
Appendix G
Receipts
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 47
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 48
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 49
Appendix H
Work Plan
CONTACT TOXICITY OF GINGER OIL AND LIQUID SMOKE 50
Appendix I
Budget Plan