PHYTOCHEMICAL AND BIOLOGICAL EVALUATION OF ANAGALLIS ARVENSIS MASTER OF PHILOSOPHY (Pharmaceutical Chemistry) By ZARMEENA YASMEEN Department of Pharmacy Faculty of Pharmacy & Alternative Medicine The Islamia University of Bahawalpur SESSION 2015-2017 List of Contents Sr. No. Contents Page 1 Acknowledgment I 2 List of tables II 3 List of figures III 4 Abbreviations IV 5 Abstract V Chapter 1 INTRODUCTION 1 Introduction 1 Historical sources and evolution of indigenous plants and 1.1 1 medicine system 1.2 Phytochemicals 4 1.3 The Family Primulaceae 15 1.4 Medicinal importance of Primulaceae 15 1.5 General description of Genus Anagallis 16 1.6 Botanical description of Genus Anagallis 16 1.7 Anagallis arvensis 16 1.8 Taxonomical classification of Anagallis arvensis 17 1.9 Synonym 17 1.10 Picture gallery of Anagallis arvensis 18 Chapter 2 LITERATURE SURVEY 2.1 Folk uses of Anagallis arvensis 20 Literature survey on pharmacological activities of Anagallis 2.2 20 arvensis 2.3 Literature survey on chemical constituents of Anagallis arvensis 26 Chapter 3 EXPERIMENTAL 3 Experimental 33 3.1 Plant material 34 3.2 Extraction 34 3.3 Preparation of reagents 34 3.4 Preparation of solutions 35 3.5 Procedures for qualitative tests of secondary metabolites 37 3.6 Procedures for biological evaluation 40 3.6.1 Antioxidant activity 41 3.6.2 Enzyme inhibitory assays 42 Chapter 4 RESULTS 4.1 Results of extraction 44 4.2 Results of phytochemical analysis of Anagallis arvensis 44 4.3 Results of biological screening 45 Chapter 5 DISCUSSION 5 Discussion 48 Chapter 6 REFERENCES 6 References 51 Abstract The main aim of this research work was to explore the hidden medicinal potentials of plant which is still not explored. The present research work emphasis on the phytochemical and biological evaluation of Anagallis arvensis (aerial and root parts) which belongs to family Primulaceae. The dichloromethane and methanol extracts of aerial and root parts were prepared. The extracts were subjected to a series of biological screening assays i. e antioxidant assay (2, 2-diphenyl-1-picryl hydrazyl and ferrous reducing power assay) and enzyme inhibition assays (Urease, Acetylcholinestrase and α- glucosidase inhibitory assays). The dichloromethane extracts from aerial and root parts showed significant antioxidant activity (71.5±0.3% and 73.5±0.3% respectively) while methanol extracts of aerial and root parts of the plant showed good antioxidant activity (65.3±0.3% and 64.4±0.2% respectively). When the extracts were screened for anti- urease activity significant (76.94±0.23% with IC50 142.53±0.19) and moderate (59.27±0.23% with IC50 187.41±0.19) results were found with methanol extracts of aerial parts and root parts respectively. The methanol extracts of root parts also showed moderate (51.82±0.15% with less than 500) acetylcholinestrase inhibitory activity. Dichloromethane extracts showed significant (97.62±0.13% with IC50 19.73±.03 and 89.26±0.19% with IC50 119.75±0.15) α-glucosidase inhibition respectively. Methanol extracts showed significant (91.54±0.17% with IC50 87.24±0.12) and good (62.53±0.21% with IC50 232.28±0.17) α-glucosidase inhibition. The plant powdered material was subjected to phytochemical investigations for presence of secondary metabolites, the results revealed the presence of saponins and flavonoids. V 1. INTRODUCTION Plants existence is universal which account for the variety of vast usages like beverages, medicines, dyeing, nutrition, fragrances, pharmaceutical and also for industrial purposes. Plants play a vital role in life and its growth and the importance cannot be ignored at any stage. Along with a vigorous and eco-friendly environment, plants produce food which is very important for our survival (Siddiqui et al., 2009). Humans have been gaining proteins, carbohydrates, fats and minerals from plants for millions of years and also build homes for their protection. Apart from primary ingredients, plants serve as a vital source of secondary metabolites which are used as agrochemicals, flavors, food additives, colorants, pharmaceuticals and bio-pesticides (Hassanpour et al., 2011). People have accepted the use of medical plants as alternative treatment since long time and the effects are less likely adverse. Old studies pour a light on enhancing immunity against various diseases using traditional systems. Also recent researches favor natural anti-oxidant compounds against the use of synthetic drugs (Subramanian et al., 2011). In traditional system of medications, plants have played a vital role in cure or prevention of different diseases. Treatments were not only limited to humans but animals as well have been cured with plant formulated medicines. This system has also been adopted by conventional Chinese system of medicines. Contemporary science also acknowledges the importance of plant based medicines and included it in pharmacotherapy as wide range of medications (Nasri, 2015). 1.1 Historical Sources and Evolution of Indigenous Plants and Medicine System There was a common practice in ancient times that medicinally valued plants were used for their specific actions against specific disease. For treating some particular ailment ancients use different parts of these plants as they were enriched with medicinal worth. Similarly Ayurvedic, Suddha and Unani medicinal indigenous system has been used and practiced from centuries and still exists (Bhattacharjee, 2004) (Gupta and Sharma, 2006). Healing and treatment of ailments with the aid of plants having medicinal importance is as old as human being is itself. The relationship between human beings and his efforts for the drugs search in nature evidenced from the far past. Although there are abundant evidences from numerous sources: preserved monuments, written documents. One of the 1 oldest evidence in written form is on usage of medicinal plants for preparation of different drugs for treatment of different diseases have been related to a Sumerian clay slab about five thousand years aged from Nagpur. It consists of twelve standard operating procedures for preparation of drugs relating to almost over 250 numerous medicinal plants, for example some medicinal plants were alkaloids as mandrake, poppy and henbane (Petrovska, 2012). For human survival it is very important to have a clear knowledge for the use of medicinally valued plants so it has been traditionally inherited to the generations. Plants had an abundant use for treating many ailments in the past and different books were compiled to provide the knowledge regarding information and the usage of medicinal plant in World‘s religious book named as ―Rigveda‖ and another book ―Soma‖ which served as a medicinal means. It was written by Indo-Aryans, in 4000 and 1600 B.C (Sher, 2004). Ayurveda is the medicinal indigenous system in the sub-continent of India which is known from old to 700 B.C. Ayurveda system is systemized by Sushruta and Charaka who have great attribution for citing nearly 700 plants of medicinal value. Medicinally important plants were used for different ailments. This is mentioned in the Indian old and ancient literature which provides intense help for herbal cure. Among them physostigma, digitalis and strophanthus have been used for different heart problems, quassia acts as a bitter tonic and the cinchona for malaria. ―Sushruta Samhita‖ is the name of a book which was compiled in 1000 B.C. This book consists of detailed chapters on therapeutically important herbs and herbal remedies and also includes the information for the use of such medicinal plants. Hippocrates (460 B.C.) known as a father of medicine started pharmacy. Greeks developed first ―Materia Medica‖. 500 drugs made by medicinal plants were enlisted and compiled in two bulky books named as history of plants and classes of the plants. These books were written by Jheophratus (287-370 B.C.) (Singh and Abrar, 1990). There are many references that show the narration of beneficial values of different plants like Nigella sativa and Crotalaria juncea by Our Prophet Muhammad (P.B.U.H). Hazrat Ayesha Siddiqa (R.A), wife of Muhammad (P.B.U.H) and his other associates had acknowledged of herbal remedies and are used by them for the treatment of various 2 diseases and injuries. All these show the significance and values of medicinal Plants. Additionally, herbal use of medicines was increased and it was considered as an aim in the science era of Islam (500 A.D.-1700 A.D.). For awareness of Europe countries Muslims provided the horizontal path line by the amendments of medicinal system of Greeks. Muslims also translated that entire medical task into Arabic, which was performed by Greeks and Indians. Among which opium a poppy plant has a unique significance in medicinal worth which has also been accredited with Arabs/Muslims into India (Saleem et al., 2013). In ancient times, different people used different techniques for the extraction of medicinal plants to get pharmaceutically active constituents that may serve as a source for treating different ailments. These extraction techniques may be maceration, decoction or infusion. Not only medicinally active plants get their significance in the era of pharmacy but also there is contribution of sources from animal origins too with the introduction of compound drugs in the sixteenth and eighteenth centuries. Hence people demanded such formulations which were the combination of both medicinally active plant as well as animal source. Such drugs were proved to be very effective and its extensive use was observed (Sewell and Rafieian-Kopaei, 2014). There has been a significant revival of interest in the study of natural products, which is based on the discovery of antibiotics and the importance which have been attained by some of the medicinally active constituents of plants in the treatment of cardiovascular diseases, mental disorders and certain forms of cancer. From the ancient times to present, plants have been used as a source of medicinal sources for the treatment of different ailments in all cultures throughout the world.