Department of Pharmaceutical Analysis and Quality Assurance

Department of Pharmaceutical Analysis and Quality Assurance, AUCOP 1 | P a g e GC – MS ANALYSIS OF PHYTOCHEMICAL
COMPOUNDS PRESENT IN TARENNA ASIATICA
LEA VES EXTRACT

Dissertation submitted to the
Jawaharlal Nehru Technological University,
Hyderabad, T.S.
In the partial fulfilment of requirement of degree
of Master of pharmacy
By
UROOJ HASHMI
(16S61S0407)

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Under the guidance of

Dr. SEEMA FIRDOUSE, M.Pharm, Ph.D
Associate professor
Department of Pharmaceutical Analysis & Q.A

Anwarul Uloom College of Pharmacy, Mallepally,
Hyderabad, T.S. Approved by AICTE
SEPTEMBER 2018

Department of Pharmaceutical Analysis and Quality Assurance, AUCOP 2 | P a g e CONTENTS

Serial number
Title
Page number

1
Abstract
3

2
Introduction
4

3
Literature review
6

4
Aim and objective
8

5
Materials and methods
10

6
Results and discussion
2 9

7
Summary
4 8

8
Conclusion
49

9
Bibliography
5 0

Department of Pharmaceutical Analysis and Quality Assurance, AUCOP 3 | P a g e 1. ABSTRACT
Tarenna a siatica is a thornless evergreen shrub growing to a height of about 1 – 6m. These are
profoundly located in Western Ghats. GC – MS analysis was carried out on the leaf extract of
Tarenna a siatica to determine the phytoconstituents (compounds) present. GC – MS along
with high performan ce thin – layer chromatography (HPTLC) Fourier transform infrared
radiation (FTIR) and nuclear magnetic resonance spectroscopy (NMR) was employed for the
research.
Sample preparation for analysis was done by Soxhlet apparatus while ethanol was employed
as s olvent of choice. Next step involved phytochemical screening of the extract and chemical
constituents recognised were alkaloids, carbohydrates, volatile oils, flavonoids, glycosides,
tannins, steroids and triterpenoids . HPTLC helped in separation of the co nstituents and peaks
obtained on FTIR spectrum lead to the determination of functional groups such as alkanes,
alkenes, aromatic alkenes, halides, alcohols, Si – OR, S – OR, C – Cl, C – Br, S – S (disulphides) . 1
H
NMR was employed to determine the nature of proton s in the sample which in turn gives
knowledge or an idea about the structural elucidation of components . GC – MS technique was
employed for determination and identification of the phytoconstituents present in the
ethanolic plant extract which were found to be as follows., benzaldehyde, glycerine,
benzofuran – 2,3 – dihydro – , propane – 1,3 – diol 4 – methyl – benzeneboronate, n – hexadecanoic acid,
phytol, D – mannitol, propylene glycol mono o leate and squalene . This development opens
pathways for future research that can be un dertaken to explore various other benefits and uses
of Tarenna a siatica leaves in the field of medicine.

Department of Pharmaceutical Analysis and Quality Assurance, AUCOP 4 | P a g e 2. INTRODUCTION
2.1: HERBAL MEDICINE 1
Herbal medicine is also referred to as Herbalism or Botanical medicine and is the oldest form
of healthcare known to mankind. It can primarily be defined as the use of herbs for its
therapeutic or medicinal values. “Herb” is a part of plant (such as roots, stem, leaves, flowers,
stigma, fruits etc.,) or the whole plant that has significant value in the field of medicine or
favoured for its aromatic or savoury qualities. These herbs produce a variety of chemical
compounds (also known as phytoconstituents) th at act upon the body. World H ealth
O rganization (WHO) states that about 4 billion i.e., approximately 80% of world population
presently use herbal medicine for some aspect of their primary healthcare.

2.2: THE HISTORY OF HERBAL MEDICINE 2
Herbs or medic inal plants have been used since a very long time. Ancient Chinese and
Egyptian papyrus writings signify their purpose. Ayurvedic and homeopathic medicine was
relied upon before allopathy came into practice.
In the early 19 th
century, chemical analyst firs t began extracting and modifying the active
ingredient from plants. This lead to synthe sizing of these ingredients resulting in the
development of synthetic pharmaceuticals. Overtime, the use of pharmaceuticals was
favoured which lead to decline in the use of herbal medicines.

2.3: HERBAL MEDICINE TODAY
In the last two decades, a steep decline in the use of synthetic pharmaceuticals has been
recorded. This is mainly due to increasing public dissatisfaction with the cost, safety, efficacy
and potential side effects of prescription drugs.
This has resulted in herbal medicine making a comeback with herbal plants being harvested
on large scale to compensate for its increasing public demand. These botanical medicine,
even though possess slow therapeutic action b ut are characterized by little to no side – effects.

Department of Pharmaceutical Analysis and Quality Assurance, AUCOP 5 | P a g e 2.4: IMPORTANCE OF SOME HERBS WITH THEIR MEDICINAL
VALUE 3
? Black pepper, cinnamon, sandalwood, ginseng and saffron are used to heal wounds,
sores and boils
? Basil, fennel, mint, rosemary etc., are put in to various uses for their aromatic and
savoury properties
? Turmeric is used for its antibiotic property in wound healing
? Sandalwood and cinnamon are astringent in nature
? Ginger and cloves are used in certain cough syrups
? Foxglove plant leaves are used as ca rdiac stimulant
? Piperine along with rifampicin act as anti – TB drug
? Ephedra treats asthma and other respiratory problems.

2.5: QUALITY ? SIGNIFICANT FOR EFFICACY
Quality of the herb is of foremost importance as the safety, purity and efficacy of the drug
d epends on it. Reproducibility of the therapeutic action in each dosage unit is also to be
considered. The environmental stressors such as cl imatic conditions, soil changes and
geographical changes have an ardent impact on the plant nutritional value. There fore, it also
affects its pharmacological activity.
To maintain the quality, manufacturers/farmers need to supervise the biological source,
selection of herbs and other conditions and changes while harvesting plant herbs. Thus,
standardization of herb is e ssential which includes authentication, purity and assay of herbs
which is a lengthy and tiring process. Parameters for standardization that are to be followed
are outlined by Ayurvedic pharmacopoeia of India and even World Health Organization
standards su ch as ash value, volatile matter, microscopic examination, organoleptic properties
etc., are to be taken into consideration for attaining the quality of not only the raw material
but also of finished herbal product.

Department of Pharmaceutical Analysis and Quality Assurance, AUCOP 6 | P a g e 3. LITERATURE REVIEW
? The present investigations find, out of an ethanolic extract of Tarenna a siatica fruits have
a potent of cytotoxic activity against MCF – 7 cells. The results obtained from the in – vitro
studies performed using the MCF – 7 cell lines reveals that the ethanolic extract of fruits of
Tarenna a siatica has a higher anticancer activity. There wa s increase in the cell growth
inhibition when concentration of samples was incre ased; The IC50 value was
237.08 µg/ml for the cell line studies as shown by the MTT assay method. The upshot of
this study encourages to carrying out further studies to be exten ded for other cell lines and
in vivo cytotoxicity investigations are required to identify anticancer activity. 4

? The leaf – bud exudate of Tarenna a siatica (Rubiaceae: Ixoroideae, Pavetteae) is
investigated for its biological activity. The crude benzene extract and corymbosin (pure
compound isolated) were screened for antiviral activity by using ELISA and PCR
methods against animal (blue tongue and chikungunya) and plant (papaya ring spot,
sesban ia mosaic and common bean mosaic) viruses. Both corymbosin and benzene
extract showed significant antiviral activity though corymbosin was found relatively more
potent against the animal and plant viruses tested. This is the first report of antiviral
activ ity for the gumresin of T arenna a siatica , so also for the compound corymbosin,
against the plant viruses. 5

? The study was aimed to analyze the phytochemical, antioxidant and antibacterial
potentials of T arenna a siatica . Methods: The leaves, barks and flowers extracts were
analyzed for total phenolic and flavonoid contents. Antioxidant activities were evaluated
using DPPH scavenging, ABTS+ scavenging, FRAP, phosphomolybdenum reduction,
metal chelating, nitric oxide radical scavenging, superoxide radical scavenging, hydroxyl
radical scavenging and lipid peroxidation assays. Antibacterial activity was examined
using agar well diffusion method against pathogenic microorganisms. Results: The total
phenolic and flavonoid contents were found to be higher in a c etone extract of leaves
(57.21 g GA E/100g and 619.67 mg RE/g respectively). Antioxidant assays revealed that
leaves acetone extract possesses significant (p