CHEMISTRY AND INSECTICIDAL POTENTIAL OF PARTHENIN AND ITS TRANSFORMATION ...

81 Pages·2012·0.89 MB·English

CHEMISTRY AND INSECTICIDAL POTENTIAL OF
PARTHENIN AND ITS TRANSFORMATION REACTION
PRODUCTS AGAINST Tribolium castaneum (Herbst).

Thesis

Submitted to the Punjab Agricultural University
in partial fulfillment of the requirements
for the degree of

MASTER OF SCIENCE
in
CHEMISTRY
(Minor Subject: Biochemistry)

By

Ramandeep Kaur
(L-2010-BS-196-M)

Department of Chemistry
College of Basic Sciences and Humanities
© PUNJAB AGRICULTURAL UNIVERSITY
LUDHIANA – 141 004

2012
CERTIFICATE I

This is to certify that the thesis entitled Chemistry and insecticidal potential of
parthenin and its transformation reaction products against Tribolium castaneum
(Herbst) submitted for the degree of M.Sc., in the subject of Chemistry (Minor subject:
Biochemistry) of the Punjab Agricultural University, Ludhiana, is a bonafide research work
carried out by Ramandeep Kaur (L-2010-BS-196-M) under my supervision and that no part
of this thesis has been submitted for any other degree.
The assistance and help received during the course of investigation have been fully
acknowledged.

_________________________
MajorAdvisor
Dr. (Mrs.) K. K. Chahal
Professor-cum-Head
Department of Chemistry
Punjab Agricultural University
Ludhiana - 141004
CERTIFICATE II

This is to certify that the thesis entitled, Chemistry and insecticidal potential of
parthenin and its transformation reaction products against Tribolium castaneum
(Herbst) submitted by Ramandeep Kaur (Admn. No. L-2010-BS-196-M) to the Punjab
Agricultural University, Ludhiana, in partial fulfillment of the requirements for the degree of
M.Sc. in the subject of Chemistry (Minor subject: Biochemistry) has been approved by the
Student’s Advisory Committee along with Head of the Department after an oral examination
on the same.

_ _
{Dr. (Mrs.) K. K. Chahal} {Dr. (Mrs.) K. K. Chahal}
Head of the Department Major Advisor

______________________
(Dr. Gursharan Singh)
Dean Postgraduate Studies
Acknowledgements
Firstly, I bow my head with utmost reverence before the Almighty whose eternal
blessing has enabled me to accomplish this noble effort.
It gives me immense pleasure to record my thanks and sense of profound gratitude
to my Major Advisor Dr. (Mrs.) K. K. Chahal, Professor-cum-Head, Department of
Chemistry, Punjab Agricultural University, Ludhiana, for her expert guidance,
encouragement, inspiration and advice throughout my research work. It was my privilege
to be guided by a person of calibre, whose blessings bring best in every one of my
endeavours.
Special thanks are accorded to Dr. B. R. Chhabra, Professor Adjunct for his
excellent technical guidance and support.
I owe my unpayable debt to the other esteemed members of my advisory committee.
Dr. (Mrs.) Manpreet Kaur, Assistant Professor, Department of Chemistry, Dr. (Mrs.)
Bavita Asthir Senior Biochemist, Department of Biochemistry, Dr. (Mrs.) B.K. Kang,
Associate Professor, Department of Entomology, Dr. (Mrs.) S.K. Uppal ,Senior
Biochemist-cum-Incharge of Sugarcane Section, Department of Plant Breeding and
Genetics, for their able guidance , constructive suggestions and continuous support.
I am indebted to my respected family for their constant words of encouragement,
deep affection and heartful blessings that enabled me to this stage of career.
Friends are always a moral support which is extremely important when one is
feeling low. I take great pleasure in thanking my friends Ajay, Dalvir, Amanpal and Amit
for giving me moral support, sharing the burden of my work and making things smooth.
My sincerest thanks to Mr. Banjit Singh, Mr. Mukesh Kumar and Mr. Raj Singh
for their invaluable and generous help in the laboratory. I feel proud to be a part of PAU,
Ludhiana where I learnt a lot and spent some unforgettable moments of my life.
I thankful to Punjab Agricultural University for providing merit fellowship
during final year of my M.Sc.
Last but not least, I duly acknowledge my sincere thanks to all who love and care
for me.

_____________
(Ramandeep Kaur)
Title of the Thesis : Chemistry and insecticidal potential of parthenin and
its transformation reaction products against Tribolium
castaneum (Herbst)
Name of the Student : Ramandeep Kaur
and Admission No. L-2010-BS-196-M

Major Subject : Chemistry

Minor Subject : Biochemistry

Name and Designation : Dr. (Mrs.) K. K. Chahal
of Major Advisor Professor- cum-Head

Degree to be Awarded : M.Sc.

Year of Award of Degree : 2012

Total Pages in Thesis : 73 + VITA

Name of University : Punjab Agricultural University,
Ludhiana-141 04
ABSTRACT
The present investigation deals with Chemistry and insecticidal potential of
parthenin and its transformation reaction products against Tribolium castaneum
(Herbst).The shade dried and powdered leaves of Parthenium hysterophorous were extracted
in chloroform using Soxhlet extraction method. Parthenin was isolated by column
chromatography using chloroform:acetone (5%) solution as the eluent. Parthenin was
subjected to reaction with diazoester which resulted into the formation of two compounds-
pyrolysis product and diazoester adduct. Parthenin on reactions with dry hydrochloric acid
gas and formic acid gets converted into anhydroparthenin. Parthenin on irradiation with
microwave gets converted into anhydroparthenin. Parthenin and its derivatives were
1
characterised on the basis of melting point, TLC, FT-IR and H NMR. Parthenin and its
derivatives were tested for their bioefficacy against adults of Tribolium castaneum (Herbst)
by releasing them in wheat grains spiked with various concentrations of test compounds viz.
-1
1,000, 2,000, 4,000, 5,000, 10,000 and 20,000 μg g of wheat respectively. The observations
of mortality were noted every 24 hours till complete or constant mortality was obtained. The
corrected per cent mortality was calculated using Abbott’s formula. All the compounds
-1
exhibited complete mortality at the spiking level of 10,000 and 20,000μg g . Parthenin was
found to be most potent followed by anhydroparthenin, pyrolysis product and diazoester
adduct.

Key words: Parthenium hyseterophorous, Bioefficacy, Tribolium castaneum, Soxhlet
extraction.

_ _
Signature of Major Advisor Signature of the Student
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CONTENTS

CHAPTER TOPIC PAGE
I. INTRODUCTION 1-4
I. REVIEW OF LITERATURE 5-24
I. MATERIAL AND METHODS 25-32
IV. RESULTS AND DISCUSION 3-57
V. SUMMARY 58-60
REFERENCES 61-73
VITA
CHAPTER – I
INTRODUCTION
Wheat (Triticum aestivum L.) of Family Gramineae is an important staple foodstuff
in North India. Post-harvest losses of wheat are due to biotic (insects, molds, rodents and
birds) as well as abiotic (temperature, relative humidity and moisture content of the grains)
factors. Among the biotic processes, insect pests are the major agent, which cause
considerable losses in terms of quality and quantity of food grains. The damage caused by
insect pests to wheat grain has been estimated at 10 to 20 percent (Ramzan et al 1991, Khan
et al 2010). Tribolium castaneum has been found to be one of major insect pest of wheat
according to surveys conducted (Mahmood et al 1996, Ghizdavu and Deac 1994, Khalil and
Irshad 1994, Desimpelaere 1996, Bandyopadhyay and Ghosh 1999).
Red flour beetle, T. castaneum (Herbst) is one of the major insect pests of stored
grains with cosmopolitan distribution (Ghizdavu and Deac 1994, Desimpelaere 1996, Abro
1996, Wong et al 1996 and Hulasare et al 2003). Although, T.castaneum is considered a pest
of flour and other milled cereal products and is also considered as a secondary pest in stored
wheat (Le Cato 1975, Hamed and Khattak1985, Irshad and Talpur 1993). A single larva can
attack 88 grains during its life time which leads to a considerable loss of quality and viability
of grain (Atanasov 1978). Apart from loss of weight and quality of food grains, insects of
genus Tribolium secrete a variety of toxic quinones which are known to be carcinogenic.
Presence of Tribolium species in the food grains give pungent smell and infested flour
becomes dirty yellow in colour (El-Mofty et al 1989) which affect baking quality of flour
(Flogliazza and Pagani 2003).
To prevent the loss during storage, farmers usually rely on synthetic chemical
insecticides. Methyl bromide was used as fumigant in past. Its use has been restricted due to
ozone layer depletion (Zhang and Van Epenhuijsen 2004).These problems lead to
increasingly stringent environment regulation of pesticides (Isman 2006, Pavela 2007). At
present there is an urgent need to develop safer, more environmentally friendly and efficient
alternatives that have the potential to replace synthetic pesticides.
Among the various alternatives, use of natural plant products called allelochemicals
offer a new approach for the management of noxious weeds and pests in sustainable manner
(Macias et al 2001).Their toxicities as well as repellent effects on the pests were of special
interest during the last decade. Plant essential oils are alternative to synthetic pesticides
possess insecticidal, ovicidal, repellent and ovipositional activities against various stored
product insects (Chiasson et al 2004, Tripathi and Kumar 2007,Tripathi et al 2009, Aboua et
al 2010).
Secondary metabolites are known to exhibit a broad spectrum of biological activities.
Among them, sesquiterpene lactones are most widely distributed in the members of family
Compositae. Sesquiterpenes containing α-methylene-γ-lactone moiety have attracted a great
deal of interest to explore their role as cytotoxic agents (Liu et al 2008), anticancer agents
(Zhang et al 2005), anti-inflammatory agents (Hall et al 1979), antioxidants (Jung et al 2004)
and plant growth regulators (Chhabra et al 1998). Due to their ability to undergo a Michael
reaction with biological nucleophiles, α-methylene-γ-lactone has been reported to possess
biological activity (Macias et al 1996). Due to their biologically active nature, these
compounds have been investigated the most for their chemistry, mechanistic pathways,
chemical transformations and synthesis (Rodriguez et al 1976). The possibility that they can
provide a lead in the search for new plant growth regulators have helped in isolation and
partial synthesis of some of the most potent compounds in which structural features other
than α-methylene-γ-lactone moiety are significant which keep the field open for further
chemical studies.
Parthenium hysterophorous L. is rich source of α-methylene-γ-lactone containing
sesquiterpenoids. This annual or biennial herbaceous plant originated from tropical America
but has spread throughout the world’s tropical areas. It can grow and reproduce itself any
time of the year. During a favorable growing season, four or five successive generations of
seedlings can emerge at the same site. Low temperature considerably reduces plant growth,
mainly flowering and seed production by reducing leaf area index, relative growth rate, net
assimilation rate, and leaf area duration (Pandey et al 2003). The weed grows fast and
comfortably on alkaline to neutral clay soils. However, its growth is slow and less prolific on
a wide range of other soil types (Rezene et al 2005). It has become a serious problem in
many parts of the world due to its threat to agricultural activities, biodiversity and human
health and has also been labelled as a useless weed. The weed is particularly problematic in
India and Australia where it was first noticed in the 1950s and has continued to spread at an
alarming rate (Navie et al 1996). In South Africa, the weed first invaded the warmer and
wetter eastern parts of the country in the 1880s, and is currently spreading to several
prominent game reserves, including Hluhluwe-I Mfolozi and the Kruger National Park
(KNP) (Strathie et al 2005). P. hysterophorus is a known invader of disturbed areas such as
roadsides, agricultural fields and wastelands (Navie et al 1996). Characteristics that make the
weed such an effective invader include tolerance of a wide range of ecological and climatic
conditions, a fast growth rate, high fecundity and efficient utilization of resources (Hedge
and Patil 1982). The plant, however, is a folk remedy (Towers et al 1977) against various
afflictions:
2
SOURCE USES
Barbados Flowers and leaves used for inflammation eczema, skin rashes.
Cuba Common name Escobar Amarga, medicinal plant, febrifuge, bitter and
corroborant (Roig 1953).
Gaudeloupe Febrifuge, used for herpes and rheumatic pains.
Guadeloupe and Common names Absinthe batard, herbe, a pian, metricaire allude to
Martinique cure of female ailments (Duss 1972).
Guyana Used for skin eruptions.
Jamaica Supposed to be used in resolutive baths and infusions and for treatment
of wounds. Country people use it to prepare a decoction for colds and to
make a bath for fleas on dogs. The plant is said to contain bitter
glucosides. The plant is still used for ‘bush baths’ in the Kingston area
and perhaps, elsewhere as well.
Trinidad Used, along with other herbs, in the preparation of bush baths for
cleansing the skin.
Mexico Analgesic properties, particularly in muscular rheumatism. Used by
aztecs as remedy against headache and ulcerated sores (Herz et al
1962).
U. S. Virgin Used for muscular strains, analgesic, vermifuge and heart trouble.
Islands
United states In Montgomery, Ala, it was purposed to be efficacious as a skin tonic
by older people.
The aqueous extracts of this allelopathic weed are important as potential agents to be
manipulated for biological control of pathogenic fungi, only when these extracts are used at
lower concentrations; whereas, at higher concentrations, a potent increase in biomass
production may prove to be beneficial for mass production of mycoherbicides to control the
weeds of economically important crops.
Parthenin has been reported to be located in various plant parts with especially high
concentrations occurring in trichomes on the leaves (Kanchan 1975, McFadyen 1995,
Reinhardt et al 2004). On the molecular level, sesquiterpene lactone biosynthesis is regulated
at the transcriptional level, and these compounds generally originate from the mevalonic acid
pathway. It has been suggested that all terpenes originate from the common precursor,
isopentenyldiphosphate. Reinhardt et al (2004) determined that one trichome type in
3