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) 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 2018Department 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 0Department 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) . 1H 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