Mar 2, 2017 - Studies on Spinacia oleracea L. , Coriandrum sativum L. , Mentha spicataâ has been independently carried out by me at Christ University, ...
A Comparative Phytochemical Studies on Spinacia oleracea L. , Coriandrum sativum L. Mentha spicata
A RESEARCH PROJECT REPORT
Submitted to Department of Life Sciences, Christ University for the Partial Fulfillment of the Requirements for the Degree of Bachelor of Science in Biotechnology, Chemistry and Botany
Under the Guidance of Fr. Jobi Xavier
By Prakruthi S 1415636
03/02/2017 Bengaluru -29
DECLARATION
I Prakruthi S, student of 6th semester Bachelor of Science in BCB (Biotechnology, Chemistry and Botany) at Christ University, Bengaluru, hereby declare that the project entitled “A Comparative Phytochemical Studies on Spinacia oleracea L. , Coriandrum sativum L. , Mentha spicata” has been independently carried out by me at Christ University, under the guidance of Rf. Jobi Xavier, Head of the Department Of Life Sciences, Christ University, Bengaluru. This work is submitted in partial fulfillment of the requirement for the award of degree of Bachelor of Science in BCB, from Christ University, during the academic year 2016-2017. This project work has not been submitted in part or fully, for award of any degree in this university or any other university previously.
Date: 03/02/2017
Prakruthi S
Place: Bangalore
1415636 6th SEMESTER BCB Christ University Bengaluru-29
CERTIFICATE
This is to certify that the project work entitled “A Comparative Phytochemical Studies on Spinacia oleracea L. , Coriandrum sativum L. , Mentha spicata” is a bonafide work of Prakruthi S 1415636 student of 6th semester, BCB, Christ University in completion of the requirement for the award of the degree of Bachelor of Science in BCB from Christ University during the
academic year
2016-2017 carried out under the guidance of Fr. Jobi Xavier, Head, Department Of Life Sciences, Christ University, Bengaluru -29.
Fr Jobi Xavier Head, Department of Life Sciences Christ University Bangaluru-29
CERTIFICATE
This is to certify that the project work entitled “A Comparative Phytochemical Studies on Spinacia oleracea L. , Coriandrum sativum L. , Mentha spicata” is a bonafide work of Prakruthi S 1415636 student of 6th semester, BCB, Christ University in completion of the requirement for the award of the degree of Bachelor of Science in BCB from Christ University during the academic year 2016-2017 carried out under my guidance for the partial fulfillment of the requirements for the award of the Degree of Bachelor of Science in BCB from Christ University, Bengaluru-29.
Fr. Jobi Xavier Head of the Department of Life Sciences Department of Life Sciences Christ University Bangaluru-29
TABLE OF CONTENT Chapter 1 ........................................................................................................................... INTRODUCTION .................................................................................................................. 1.1 Botanical Description of the plants ................................................................................... 1.2 Traditional uses of the leafy vegetables............................................................................ 1.3 Objectives .......................................................................................................................... CHAPTER 2 ......................................................................................................................... REVIEW OF LITERATURE ..................................................................................................... CHAPTER 3 ......................................................................................................................... MATERIALS AND METHOD .................................................................................................. Methods .................................................................................................................................. 3.1 Phytochemical Screening .............................................................................................. 3.2 Protein Estimation using Biuret Method ....................................................................... 3.3 TLC ................................................................................................................................. CHAPTER 4 ......................................................................................................................... RESULTS ............................................................................................................................. 4.1 Organoleptic characters ................................................................................................ 4.2 Phytochemical Screening .............................................................................................. 4.3 Protein Estimation ......................................................................................................... 4.4 Separation of Plant Pigment using TLC ......................................................................... 4.5 Images ........................................................................................................................... CHAPTER 5 ......................................................................................................................... DISCUSSION ....................................................................................................................... CHAPTER 6 ......................................................................................................................... CONCLUSION ...................................................................................................................... REFERENCES .......................................................................................................................
ABSTRACT
The study is based on the comparative pharmacological activity of Spinacia oleracea, Coriadrum sativum and Mentha spicata and their protein estimation. The study includes the estimation of secondary metabolites along with the plant pigment separation using Thin Layer Chromatography. Biuret method was used for the estimation of protein and it was found that spinach had 0.078g of protein followed by mint with 0.093g and least was coriander with 0.031g. Spinach increases iron content in blood . It was also found that spinach had most of the secondary metabolites present in it which has significant therapeutic activity. Spinach also showed high amount of Plant pigments and thus had higher Rf value of 0.078g. The active constituents present in spinach impart unique characteristics to it.
CHAPTER 1 INTRODUCTION The study is a comparison between the leafy vegetables of Spinacia oleracea L., Coriandrum sativum L. and Mentha spicata L belonging to different families. These leafy vegetables are used for cooking purpose yet have much dissimilarity in all. This study includes phytochemical analysis, organoleptic analysis, thin layer chromatography and protein estimation. Medicinal plants are of significant importance to the health of individuals. The medicinal value of plant lies in some of their chemical constituents that create a distinct physiological action on the human body. The major chemical constituents of plants are alkaloids, tannins, flavanoids, phenolic compound etc. Most of these plants are used as spices and in food to give it a flavour. Plant and plant based medicines are the basis of many of the pharmaceuticals we use today for our various afflictions. The progress in the plant derived medicine has been depended upon firstly, the efficacy of the medicine in human being and secondly, the major active chemical constituents present in plant. Herbs and spices have been used during the middle ages for flavoring, food preservation, and/or medicinal purposes. In India thousands of species are known to have medicinal values and the use of different parts of several medicinal plants to cure specific disease has been in vogue since ancient times. North-East India is regarded as a hot spot of biodiversity and hence different types of herbal and medicinal plants are available there. These medicinal plants have been used traditionally by its local people for a long time to cure diseases. Acknowledge of the chemical constituent is desirable for the discovery of therapeutic agents as well as discovering new source of economic materials, such as herbals, tannins, oil, gums and precursors for the synthesis of complex substances. Medicinal plants will continuously provide a source for generating novel drug compounds. Plants may become the base for the development of a new medicine or they may be used as phyto-medicine for the treatment of diseases. The compounds are synthesized by primary or rather secondary metabolism of living organisms. Secondary metabolites are chemically and taxonomically extremely diverse compounds with obscure function. They are widely used in the human therapy, veterinary, agriculture, scientific research and countless other areas. [1]
1.1 Botanical Description of the plants
Spinacia oleracea
Figure.1 Spinach leaves Scientific classification based on Bentham and Hooker: Kingdom:
Plantae
Division:
Angiosperms
Class:
Dicotyledonae
Sub class:
Polypetalae
Order:
Caryophyllales
Family: Amaranthaceae/ Chenopodiaceae Genus:
Spinacia
Species:
S. oleracea
Spinach (Spinacia oleracea) is an edible flowering plant in the family Amaranthaceae native to central and western Asia. It is an annual plant, which grows up to 30 cm tall. Spinach may survive over winter in temperate regions. The leaves are alternate, simple, ovate to triangular, and very variable in size from about 2–30 cm long and 1–15 cm broad, with larger leaves at the base of the plant and small leaves higher on the flowering stem. The flowers are inconspicuous, yellow-green, 3–4mm in diameter, maturing into a small, hard, dry, lumpy fruit cluster 5–10mm across containing several seeds.
Common spinach, S. oleracea, was long considered to be in the family Chenopodiaceae, but in 2003, that family was merged into the family Amaranthaceae in the order Caryophyllales. Within the family Amaranthaceae, Amaranthoideae and Chenopodioideae are now subfamilies, for the amaranths and the chenopods, respectively [2].
Coraiandrum Staivum
Figure.2 Coriander leaves Kingdom:
Plantae
Division:
Angiosperms
Class:
Dicotyledonae
Sub class:
Polypetalae
Order:
Apiales
Family:
Apiaceae
Genus:
Coriandrum
Species:
C. sativum
Coriander (Coriandrum sativum) also known as cilantro Chinese parsley or dhania is an annual herb in the family Apiaceae. Coriander is native to regions spanning from southern Europe and northern Africa to southwestern Asia. It is a soft plant growing to 50 cm (20 in) tall. The leaves are variable in shape, broadly lobed at the base of the plant, and slender and feathery higher on the flowering stems. The flowers are borne in small umbels, white or very pale pink, asymmetrical, with the petals pointing away from the center of the
umbel longer (5–6 mm or 0.20–0.24 in) than those pointing toward it (only 1–3 mm or 0.039–0.118 in long). The fruit is a globular, dry schizocarp 3–5 mm (0.12–0.20 in) in diameter. The seeds are generally used as a spice or an added ingredient in other foods or recipes, although sometimes they are eaten alone [3].
Mentha Spicata
Figure.3 Mint leaves
: Kingdom:
Plantae
Division:
Angiosperms
Class:
Dicotyledonae
Sub class:
Polypetalae
Order:
Lamiales
Family:
Lamiaceae
Genus:
Mentha
Mentha (also known as mint, from Greek míntha, Linear B mi-ta) is a genus of plants in the family Lamiaceae (mint family). The species are not clearly distinct, and estimates of their number vary from 13 to 18. Hybridization between some of the species occurs naturally. Many other hybrids, as well as numerous cultivars, are known.
The genus has a sub cosmopolitan distribution across Europe, Africa, Asia, Australia, and North America. Mints are aromatic, almost exclusively perennial, rarely annual, herbs. They have wide-spreading underground and over ground solons and erect, square, branched stems. The leaves are arranged in opposite pairs, from oblong to lanceolate, often downy, and with a serrated margin. Leaf colors range from dark green and gray-green to purple, blue, and sometimes pale yellow. The flowers are white to purple and produced in false whorls called verticillasters. The corolla is two-lipped with four sub equal lobes, the upper lobe usually the largest. The fruit is a nut let, containing one to four seeds [4].
1.2 Traditional uses these leafy vegetables The leaves of spinach, coriander and mint are mostly used for its health benefits, although all parts of the plant including root, stem etc., have an important and significant role in medicine. Different parts of plants are used in Ayurveda and Siddha System of Medicine. A few of the uses are mentioned below. Chewing of leaves also cures ulcers and infections of mouth and stomach issues. Research indicates that the leaves of spinach has a very high safety margin with exceptionally low toxicity, providing general beneficial effects at doses without adverse reactions or other undesirable side effects. Recently when a few experiments were carried out it was determined that these 3 leaves protects and reduces stress, enhances stamina, increases the body’s efficiency to use Oxygen, boosts the immune system, reduces inflammation, protects against radiation damage, lessens aging, Gastrial ailments, Antiinflammatory action, reduces blood Sugar levels, prevents ulcers, Lowers fever, Cold and cough and also provides rich supply of antioxidants. It is found that flavonoid (methylenedioxyflavonol glucuronides) and carotenoid content of spinach helps fight cancer and functions as an antioxidant. Spinach is also effective in the treatment of diabetes. If taken together with alfalfa, spinach helps lower the blood sugar levels of the body. Therefore it is said as one of the best green leafy vegetable which has Anti-cancerous, Anti- inflammatory and Anti-Diuretic activity. Mint is said to be rich in many antioxidant vitamins, including vitamin A, beta carotene, vitamin-C and vitamin E. The leaves of mint also contain many important B-complex vitamins like folates, riboflavin and pyridoxine (vitamin B-6); and the herb is an excellent source of vitamin-K. It also has Anti-oxidant
properties, helps in stomach disorders, good for Asthma patients. All three leafy vegetables are essential source of Vitamin, Protein, Minerals and provide immunity to fight against many chronic diseases [5]. Spinach has a very good medicinal value. It is packed with vitamins such as vitamin C, vitamin A and vitamin E and minerals like magnesium, manganese, iron, calcium and folic acid. Spinach is also a good source of chlorophyll, which is known to aid in digestion. Spinach is also rich in the carotenoids betacarotene and lutein.
1.3 Objectives 1. To do the comparative phytochemical studies on Spinacia oleracea, Coriandrum sativum , Mentha spicata. 2. To find the protein content present in Spinacia oleracea, Coriandrum sativum , Mentha spicata. 3. To Perform phytochemical screening to find out major secondary metabolites present. 4. Detection of the plant pigments present in Spinacia oleracea, Coriandrum sativum , Mentha spicata using TLC.
CHAPTER 2
REVIEW OF LITERATURE Saxena et al. (2013) in their study described all about the importance of different phytochemical analysis of medicinal plant extracts. They also demonstrated all the procedure for determination of all the secondary metabolites. According to Saxena et al. (2013) Phytochemicals (from the Greek word phyto, meaning plant) are biologically active, naturally occurring chemical compounds found in plants, which provide health benefits for humans further than those attributed to macronutrients and micronutrients. They protect plants from disease and damage and contribute to the plant’s colour, aroma and flavour. Some of the phytochemicals and their importance: The term “phenolic acids”, in general, designates phenols that possess one carboxylic acid functional group. Naturally occurring phenolic acids contain two distinctive carbon frameworks: the hydroxycinnamic and hydroxybenzoi structures. Phenolic acid compounds and functions have been the subject of a great number of agricultural, biological, chemical and medical studies. Phenolic compounds in many plants are polymerized into larger molecules such as the proanthocyanidins (PA; condensed tannins) and lignins. Moreover, phenolic acids may arise in food plants as glycosides or esters with other natural compounds such as sterols, alcohols, glucosides and hydroxyfatty acids. Varied biological activities of phenolic acids were reported. Increases bile secretion, reduces blood cholesterol and lipid levels and antimicrobial activity against some strains of bacteria such as staphylococcus aureus are some of biological activities of phenolic acids [6]. Phenolics acid possesses diverse biological activities, for instance, antiulcer, anti- inflammatory, antioxidant, cytotoxic and antitumor, antispasmodic, and antidepressant activities Flavonoids are polyphenolic compounds that are ubiquitous in nature. More than 4,000 flavonoids have been recognised, many of which occur in vegetables, fruits and beverages like tea, coffee and fruit drinks. Flavonoids have been reported to exert multiple biological property including antimicrobial, cytotoxicity, anti-inflammatory as well as antitumor activities but the best-described property of almost every group of flavonoids is their capacity to act as powerful antioxidants which can protect the human body from free radicals and reactive oxygen species.
Marmount et al., (1967) in this journal, described about the effects of alkaloids of oleracea. The mechanism by which the spinacia oleraceae alkaloids effect cellular division consists chiefly a blockade of mitosis in metaphase, which is the result of the inactivation of mitotic spindle. These property
account not only for the for the introduction of the stathmokinetic method for evaluating the proliferating activity of blood cells but also for the universally adopted methods of colchicine blockade for chromosomal study. According to the result by Marmount et al. (1967) collected during this experiment it has been speculated that mitotic arrest is not the only operative mechanism of these alkaloids, and this hypothesis appeared supported by the demonstration that metaphasic blockade occurred at more than 100 times the concentration necessary to cause 50% inhibition of growth as estimated by protein determination or cell count [7].
Inamdar et al. in his work described about the anatomy of pathological leaves of Coriandrum sativum. It is noticed that the affected leaves are pale green to pale yellow membranous, and reduced in size compared to the normal ones. It has also been noticed that the reduction in the general surface results in the reduction of the internal tissue. The frequency of stomata, stomatal index, guard cells, number of epidermal cells is all affected. The stomatal abnormalities are induced artificially by growth regulators as there was no report found for the abnormalities of stomata due to mycoplasma infection. Inamdar et al. thought that these abnormalities were caused due to the mycoplasma infection [8].
The study by Zhou et al. (2011) shows how Soybean transcription factor GmMYBZ2 represses catharanthine biosynthesis in hairy roots of Coriandrum sativum. Coriandrum sativum (L.) G. Don is a plant species known for its production of a variety of terpenoid indole alkaloids, many of which have pharmacological activities. Production of catharanthine in cell cultures or in hairy roots established by transformation with Agrobacterium rhizogenes is of interest because catharanthine can be chemically coupled to the abundant leaf alkaloid vindoline to form the valuable anticancer drug vinblastine. Here, it observed a high amount of catharanthine in hairy roots of C. sativum, established by infecting leaf explants with the A. rhizogenes >agropine-type A4 strain carrying plasmid pRi. In dicots, hairy roots can be induced after infection with A. rhizogenes, a soil-borne pathogenic bacterium responsible for the development of the hairy roots disease. A. rhizogenes-induced hairy root cultures exhibit stable and fast growth rates, genetic and biochemical stability, and increased production of secondary compounds compared to cell suspension cultures. Moreover, transformed hairy roots can also be successfully cultured in large-scale bioreactors. It was noticed that the hairy roots from the AR lines grew slowly, were thin and straight, and had slender, dense, and white branches with thin tips, whereas hairy roots from the Z lines grew slowly with thick and fewer branches. Zhou et al. (2011) observed that along with
the generation of brown pigments in some of the Z lines, most likely due to the accumulation of phenolic compounds [9]
Phatat et al, (2014) studied about the herb Mentha spicata. Herb Mentha spicata is grown as weeds in the tropical countries like India. It has a wide range of active constituents which has potent medicinal properties. The main objective was to overview the total antioxidant capacity of the herb Mentha spicata. Antioxidant activities were evaluated in terms of total phenolics content, total antioxidant activity, and reducing power. Different studies were carried out by comparing Mintha spicata extract with antioxidant references such as gallic acid; ascorbic acid. All these antioxidant activities increased with increasing concentrations in a dose dependent manner. It was found to be significant and valuable. Hydroalcoholic extract of Mentha spicata was optimized in ratio of methanol to water (1:1,v/v) which may be superior to its ethanolic extract to observe the exact antioxidant nature of the herb Mentha spicata. The result concluded that this herb rich in alkaloids, triterpenes, glycosides, flavonoids, cardenolides, steroids, bufadienolides and lipids, pharmacognostic studies, pharmacological activities such as antileishmanial, hepatoprotective, nephroprotective, neuropharmacological, antimutagenic, anti-ulcer,
antibacterial,
antidiabetic,
immunosuppressive,
antihypertensive,
analgesic,
anti-
inflammatory, wound healing, uterine contractility, insecticidal, fungitoxic, phytotoxic activities and other in-vitro activities [10]
Gowswami et al. (2013) described the microscopy, macroscopy, standardization for purity and strength as per WHO and preliminary phytochemical screening of the leaf of Spinacia oleracea. Coriandrum sativum belongs to the family Apocyanaceae, is a bushy perennial herb and evergreen shrub. More than 100 alkaloids and related compounds have so far been isolated and characterized from the plant. Main alkaloids are vincristine and vinblastine which are responsible for anticancerous activity. Gowswami et al. (2013) subjected the dried leaves of Periwinkle for different standardization parameters like Total ash (0.4%), Acid insoluble ash (0.68%), Water soluble ash (1.68%), Sulphated ash (4.12%), Water soluble extractive (6.34%), Alcohol soluble extractive (4.8%), Moisture content (10.09%), Loss on drying (5.01%), Foaming index (0.9cm height) and Swelling index (0.8g) for their purity and strength as per WHO guideline. The Preliminary phytochemical screening report reveals the presence of Alkaloids, Glycosides, Terpenoids, Flavonoids, Phenols, Tannins, Carbohydrates, Saponins, Phytosterols, Protein and amino acids in ethanolic extract of the leaf of Coriandrum sativum. The transverse section of the plant exhibits
small layer of rectangular cells covered with thick cuticle, uni-cellular covering trichome and vascular bundle present in the middle of midrib region and cruciferous stomata [11]
Ghos et al. (2001) described about the effect of extract of Mentha spicata in the treatment of alloxan diabetics in male albino rat. Administration of aqueous extract of Mentha flowers and leaves has found to regulate the blood sugar level in alloxan diabetic male albino rat. This therapy not only produced blood glucose homeostatis but also reversed changes in carbohydrate, protein, lipid metabolisms and metabolic and pathological changes that took place in pancreatic islet cells, liver and kidney following a single dose of allaxon monohydrate were irreversible until death after 6 days of alloxan administration and the B cells secretory activity resumed near normalcy [12].
Verma et al. (2008) discussed and worked on the current and future status of herbal plants. Nature always stands as a golden mark to exemplify the outstanding phenomena of symbiosis. As the people are becoming aware of the potency and side effect of synthetic drugs, there is an increasing interest in the natural product remedies with a basic approach towards the nature. Throughout the history of mankind, many infectious diseases have been treated with herbals. A number of scientific investigations have highlighted the importance and the contribution of many plant families i.e. Asteraceae, Liliaceae, Apocynaceae, Solanaceae, Caesalpinaceae, Rutaceae, Piperaceae, Sapotaceae used as medicinal plants. Medicinal plants play a vital role for the development of new drugs. The bioactive extract should be standardized on the basis of active compound. The bioactive extract should undergo safety studies. Almost, 70% modern medicines in India are derived from natural products. Medicinal plants play a central role not only as traditional medicines but also as trade commodities, meeting the demand of distant markets. India has a very small share (1.6%) of this ever-growing global market. To compete with the growing market, there is urgency to expeditiously utilize and scientifically validate more medicinally useful plants. Medicinal herbs as potential source of therapeutics aids has attained a significant role in health system all over the world for both humans and animals not only in the diseased condition but also as potential material for maintaining proper health. A major factor impeding the development of the medicinal plant based industries in developing countries has been the lack of information on the social and economic benefits that could be derived from the industrial utilization of medicinal plants. Except for the use of these plants for local health care needs, not much information has been available on their market potential and trading possibilities. As a result, the governments or entrepreneurs have not exploited the real potential of these plants [13]
It has been reported by Cutts, Beer, and Xoble (1960) that extracts of the plant Spinacia oleracea, when injected into rats, led to a marked fall in circulating leukocytes and to a depression of the bone marrow. Subsequently a new alkaloid, Vincaleukoblastine (VLB), was isolated from such ex tracts; it caused marked hemopoietic effects and affected the growth of experimental tumours. It has been reported by Cutts, Beer, and Xoble that extracts of the plant Vinca rosea, when injected into rats, led to a marked fall in circulating leukocytes and to a depression of the bone marrow. Subsequently a new alkaloid, Vincaleukoblastine (VLB), was isolated from such extracts; it caused marked hemopoietic effects and affected the growth of experimental tumours. VLB has also been reported to have carcinostatic activity. This has been demonstrated with L1210 and P-1534 leukemia transplanted in BDF hybrid mice, AKr leukemia transplanted in AK mice, and Sarcoma 180 in Swiss mice [14]
Nascimento et al (2000) described in his work about biological effects of a new alkaloid, Vincaleukoblastine (VLB). In mice bearing the transplantable leukemias LI~10, P1534, and AKr or Ehrliehascites tumour, treatment with VLB effectively prolonged survival. The effectiveness of treatment depended on the amount and spacing of dosage and the time of the institution of treatment after transplantation. A single injection of VLB was highly active, but hourly doses were probably the most effective. VLB was also active when incubated with tumour cells. Transplanted or spontaneous mammary tumours in C3H mice also showed a reduced growth during treatment with VLB. Mice "cured" of AKr or P1534 transplanted leukemia by treatment with VLB were subsequently resistant to repeated challenges by the tumour. Fischer rats bearing transplanted IRC 741 leukemia also responded to VLB treatment Circulating tumour cells were particularly affected. VLB did not inhibit regeneration of the liver of the partially hepatectomized rat. Bone marrow depression was striking, although megakaryoeytes were little affected. Intestinal lesions were absent. The mouse tolerated greater doses of VLB than did the rat or the guinea pig [15]
Mahlberg et al. (1971) evaluated the antimicrobial activity of plant extracts and phytochemicals with antibiotic susceptible and resistant microorganisms. In addition, the possible synergistic effects when associated with antibiotics were studied. Extracts from the following plants were utilized: Achillea millifolium (yarrow), Caryophyllus aromaticus (clove), Melissa offficinalis (lemon-balm), Ocimun basilucum (basil), Psidium guajava (guava), Punica granatum (pomegranate), Rosmarinus officinalis (rosemary), Salvia officinalis (sage), Syzygyum joabolanum (jambolan) and Thymus vulgaris (thyme). The
phytochemicals benzoic acid, cinnamic acid, eugenol and farnesol were also utilized. The highest antimicrobial potentials were observed for the extracts of Caryophyllus aromaticus and Syzygyum joabolanum, which inhibited 64.2 and 57.1% of the tested microorganisms, respectively, with higher activity against antibiotic-resistant bacteria (83.3%). Sage and yarrow extracts did not present any antimicrobial activity. Association of antibiotics and plant extracts showed synergistic antibacterial activity against antibiotic-resistant bacteria. The results obtained with Pseudomonas aeruginosa was particularly interesting, since it was inhibited by clove, jambolan, pomegranate and thyme extracts. This inhibition was observed with the individual extracts and when they were used in lower concentrations with ineffective antibiotics. The extracts from basil, clove, guava, jambolan, lemon balm, pomegranate, rosemary and thyme presented antimicrobial activity to at least one of the tested microorganisms. The extracts from clove and jambolan presented the highest activities, i.e. e., they were able to inhibit 9 (64.2 %) and 8 (57.1 %) types of microorganisms of interest, respectively. Moreover, they also had the highest activity rate against antibiotic resistant bacteria, which was 83.3 %. On the other hand, the extracts from sage and yarrow did not show any anti-microbial activity [16]
Boke et al (1949) experimented on the crown gall tissue of the plant Catharanthus and found out that Crown gall tissue was a promising experimental material for the study of the metabolism and mode of action of cytokinins because it contains high levels of these compounds, which may be responsible for its unregulated growth. The tissue most studied to date has been a culture of Vinca rosea L. Crown gall, which has been found to contain a range of glycosides of zeatin (Z): the side chain O-glucoside (ZOG), the 9-riboside (ZR) and its O-glucoside (ZROG), and the 9-glucoside (Z9G). The accurate quantitative analysis of these cytokinins is essential for an understanding of their bio chemistry in this tissue. The range of zeatin glycosides found in crown gall tissue of Vinca rosea L. has been quantified using a mass spectrometric isotope dilution procedure. Problems in the quantitative analysis of cytokinins in plant extracts are discussed. The accuracy of bioassay techniques for cytokinin quantification is limited by their variability, susceptibility to interference, and lack of chemical specificity. Although recentlydeveloped HPLC methods are capable of resolving all the natural cytokinins, we have found that current detection methods are not sufficiently selective to permit confident identification of plant extract components. Quantitative mass spectrometry is the most reliable technique available at present. The labeled and natural compounds can be distinguished in the analysis because the m/e values of their mass spectral fragments are displaced by four units. A known quantity of the 15N-labelled analogue is added to the initial tissue extract, and after purification the molar ratio of natural cytokinin to labelled
internal standard in the extract is determined by GC/MS in the selected ion monitoring (SIM) mode. Hence the quantity of natural compound in the initial extract may be calculated [17]
Braun et al (1962) found out through his study that the plasma membrane in immature cells is often irregular in contour. Some irregularities become conspicuous folds that continue to enlarge in to the cytoplasm. These invaginations may continue to increase in size and typically expand into the central vacuole. Sections taken by Braun et al (1962) showed two closely parallel membranes in areas where the invagination projects into the vacuole. A narrow layer of cytoplasm may traverse the intermembrane zone between the membranes. The interior of an invagination may lack obvious content, or may be occupied by a fibrous material, or vesicular and tubular structures. The small vesicles bound by a single membrane appear to be derived from a projection formed most frequently near the orifice of the invagination. The origin of large vesicles and tubules possessing one membrane is not certain although they may arise by the fusion of several small vesicles. Alternatively, tubules, once formed, often possess constrictions along their length which suggest that these structures may become divided into a series of smaller vesicles. There is some evidence that cytoplasmic vesicles may fold into the membrane of the invagination and subsequently be pinched off into the interior of the sac. These vesicles are bounded by two membranes. Vesicles frequently contain an electron-dense content more or less homogeneous in composition and unlike the typical ribosomal character of the cytoplasm. Plasma-membrane- derived invaginations are a common developmental feature in immature cells and are present in protodermal and ground meristem cells in several parts of the shoot system. They appear to undergo enlargement and expansion into the vacuole and may occupy a considerable volume of the cell. Evidence to support an interpretation that they are real and not fixation artefacts, is derived from recent comparative studies of living hair cells of Trade-scantia virginiana which possessed membranous structures comparable in dimensions to those present in ultrastructural views [18].
The morphological interpretation of floral organs has been a controversial subject for many years. According to the classical theory supported by Eames (1931), the flower is a determinate axis the appendages of which are the homologues of leaves. The classical theory has been attacked by Thomas (1931, 1934, 1935), Thompson (1935, 1936), Gretgoire (1938) and others. The literature on floral interpretation is already voluminous, and is cited in the several excellent reviews on the subject (Arber, 1937; Bancroft, 1935; Kozo-Poljanski, 1936; Wilson and Just, 1939). The study by Ethalsa et al (2014) is the last of a series of three on floral initiation and development in Vinca rosea L. This project was
presented cy Ethalsa et al (2014) with the hope that the results of a detailed ontgenetic study may provide further information which will be of use in interpreting the stamen and carpel in flowering plants. The problem of the direction of procambial differentiation recently has been receiving considerable attention. Investigators agree that in most vascular plants the procambium is related to foliar primordia. With respect to the place of origin of the procambial strand associated with a given primordium, there is, however, a divergence of opinion [19]
Govindswamy et al (2012) in his study found that as a result of the transition from a normal plant cell to a fully autonomous rapidly growing crown-gall tumour cell, a series of quite distinct but well defined biosynthetic systems, which represent the entire area of metabolism concerned with cell growth and division, become progressively and permanently activated. This leads to the permanently increased synthesis by such tumour cell types of the nucleic acids, mitotic proteins, and other substances concerned specifically with growth accompanied by cell division.2' 3 Included among the biosynthetic systems shown to be activated in the plant tumour cell are those involved in the synthesis of two growth-regulating substances one of which, an auxin, is concerned with cell enlargement while the other, a kinin, is mitogenic and acts svnergistically with the first to promote growth accompanied by cell division. In addition, the systems concerned with the synthesis of the vitamin myo-inositol, glutamine, asparagine or aspartic acid as well as with purines and pyrimidines are permanently activated in the tumour cell types. The degree of activation of these biosynthetic systems appears, moreover, to determine the rate of growth of the plant tumour cells. These systems are precisely regulated in normal cell types [20]
Gajalakshmi et al (2013) described that plants are a very important source for medicines; documentation of different uses of plants in medicine originates from ancient times. Plant has been a rich resource for natural drug research and development. Medicinal plant-based drugs have higher advantage of being simple, effective and they offer a broad spectrum of activity with greater emphasis on preventive action.
The aim study by Gajalakshmin et al (2013) was to determine the antibacterial activity of crude extracts from root parts of Coriandrum sativum against several bacterial species of clinical significance. Root part of C. roseus was extracted in appropriate solvent followed by evaluation of antibacterial activity by agar well diffusion assay against a total of eight bacterial stains. Further, minimum inhibitory concentration(s)
was evaluated for the crude extracts. Among all the extracts, the chloroform extract was found to be most active against almost all the bacterial species tested. Gram-positive bacteria were found more sensitive than Gram-negative bacteria. Other focuses included the determination of antioxidant activity using DPPH assay and IC50 (Inhibitory concentration) values were also determined using broth dilution assay. Preliminary phytochemical screening of the crude chloroform extract revealed the presence of alkaloids, flavonoids, tannins, saponins, proteins and phenolics. The study promises an interesting future for designing potentially active antibacterial agents from Coriandrum sativum [21].
CHAPTER 3 MATERIALS AND METHOD
Chemicals required: Wagner’s reagent, Dragendroff’s reagent, Mayer’s reagent, 1% lead acetate, 10% lead acetate, 2N HCl, Concentrated H2SO4, 10% NaOH, Ammonia solution, Chloroform, Acetic anhydride, Distilled water, Ethanol, standard protein solution, biuret reagent, sodium potassium tartrate, copper sulphate, Potassium iodide, sodium phosphate (Monobasic and diabasic). Methods 3.1 Phytochemical Screening
Preparation of Powder: The samples were over dried at 50oC for about half an hour and then crushed using a motar and pestle into fine particles.
Preparation Of extract: 1 g of each powdered leaves were placed in conical flask and 10ml of methanol was added. The powder material was extracted with methanol for 24 hours at room temperature with continuous stirring. After 24 hours the supernatant was collected by filtration and the solvent was evaporated to make the crude extract. The residues obtained were stored in airtight bottles in a refrigerator for further use.
Preliminary Phytochemical Screening: The methanolic extracts of following plants was subjected to different chemical tests for the detection of different phytoconstituents using standard procedures
1. Test for Alkaloids: The aqueous extract is evaporated in a test tube. To the residue , dilute HCl is added, shaken well and filtered. With the filtrate tests is performed. 1) Wagner’s Test :To the filtrate, add Wagner’s Reagent- Red ppt is formed 2) Mayer’s reagent: To the filtrate add 2-4 drops of Mayer’s Reagent.This Reagent gives cream colour ppt with alkaloids. 3) Dargendorff’s reagents: To the filtrate add Dargendroff’s Reagent.It gives yellow ppt ppt with alkaloids. 2. Test for flavonoids 1) With lead acetate: To the small quantity of aqueous extract 10% of lead acetate solution was added. Formation of yellow precipitate 2) With Sodium Hydroxide: On addition of an increasing amount of sodium hydroxide, Shows yellow coloration 3. Test for Saponins 1) Foam Test- The extract is diluted with distilled water and made up to 20 ml. The suspension is shaken in a graduated cylinder for 15 minutes. A layer of foam is formed. 4. Test for Tanin :For 2 ml of extract add few drops of 1% lead acetate. Yellowish ppt seen. 5. Test for Carbohydrates: Extract were dissolved individually in 5ml of distilled water and filtered. Use the filterate 1) Molisch’s Test- Filterate+ 2 drops of alcoholic α-napthol solution and add conc.H2SO4. Violet ring formed at the junction of two colours
2) Fehling’s Test- Filterate was hydrolyzed with dil.HCl, neutralized with Fehling A and Fehling B solution. Put it on water bath for 10 minutes. Formation of a Red ppt. 6. Detection of phytosterols: 1) Salkowski’s test: extract was treated with Chloroform & filtered. The filtrates was treated with few drops of conc. H2SO4, shaken & allowed to stand for 5 min. Golden yellow color ppt formed 2) Liebermann’s Burchard test: extracts were cooled. Treated with chloroform & filtered. Filtrates were treated with few drops of acetic anhydride boiled & conc. H2SO4 was added. Formation of brown ring at junction. 7. Detection of phenols: extracts were treated with few drops of ferric Chloride solution. Bluish black ppt 8. Detection of proteins 1) Xanthoproteic test: extract was treated with concentrated HNO3. Yellow color PPT 9. Detection of Terpenoids: 1) Copper Acetate test: extract was dissolved in water and treated with 4 drops of copper acetate solution. Emerald green ppt – Diterpenoids 10. Test for Courmarin: 2ml extract + acl. NaOH. Yellow color ppt. 3.2 Protein Estimation using Biuret Method
Sample extract: 20g of sample was soaked in distilled water overnight and grinded into fine paste and then made up to the volume to 100 ml with phosphate buffer. PART A: Extraction of Protein: Extraction of protein was carried out in ice cold conditions. 1. 5ml of the respective leaves extract was taken in a beaker and diluted to 10ml using ice cold phosphate buffer. 2. Transferred into a centrifuge tube and centrifuged at 3000 rpm for 5 minutes at 4°C to remove suspended particles. 3. The supernatant was collected into a beaker and 8g of ammonium sulphate was added carefully in small amounts with constant stirring on ice to make 45% saturation with ammonium sulphate and allow standing for 5 minutes. 4. Transfer this into a centrifuge tube and centrifuge at 3000 rpm for 10 minutes at 4°C.
5. The supernatant was discarded and suspend the pellet in 5 ml of distilled water. 6. 5ml of 10% TCA was added and mixed well and centrifuged at 3000 rpm for 10 minutes at 4°C. 7. The supernatant was discarded and re-suspend the pellet in 5 ml of distilled water. This is the protein test sample. . PART B: Estimation of Extracted Protein: Procedure: 1. 0.0, 0.5, 1.0, 1.5, and 2.0 ml of working standard in to the series of labeled test tubes taken out. 2. 0.5, 1.0 and 1.5 ml of the extracted protein sample from plant into another three test tube and label it. 3. The volume to 2.0 ml was made in all the test tubes with distilled water. A tube with 2.0 ml of distilled water serves as the blank. 4. 4 ml of biuret reagent was added to all the test tubes including the test tubes labeled 'blank' and 'unknown'. 5. The contents of the tubes were mixed by vortexing / shaking the tubes and incubate at room temperature for 10 minutes 6. The absorbance was readed at 520 nm against blank. 7. Plot the standard curve by taking amount of Protein along X-axis and absorbance at 520 nm along Yaxis. 8. From the standard curve calculate the concentration of protein in the given. 3.3 TLC Thin layer chromatography is one of the chromatographic methods where it is used to separate nonvolatile compounds.For the running of TLC mobile phase is essential. 1. Mobile phase used was petroleum ether:acetone:chloroform. 2. TLC plate was prepared using silica and water. 3. After the preparation of silica gel on the glass plate, It is kept inside the hot air oven for 1 hour. 4. A dot was made using the extract on the plate and kept it in the glass jar having the mobile phase. 5. Wait till the components were separated on the plate. 6. compound was estimated with the help of Rf value
7. Rf value= Distance travelled by the component/distance travelled by the solvent.
CHAPTER 4 RESULTS 4.1 Organoleptic characters The leaf of plant Spinacia oleracea was found to be dark green in color with a characteristic taste having bitter smell. The shape of the leaf is found to be ovate; the flowers are inconspicuous, yellow in color. The leaf of herb Coriandrum sativum was found to be light green in color, having aromatic smell. The shape of the leaf is sub globular; the flowers are white and produced in false whorls called verticillaster. The leaf of Mentha spicata was found to be dark green in color with minty cool taste having aromatic smell. The leaves are wide to narrow with blunt tip [Table. 1]. 4.2 Phytochemical Screening The aliquot extract of these leaves were prepared using methanol and tested for different secondary metabolites. Spinacia oleracea showed positive results for almost all the test like protein, carbohydrates, phenols, flavonoids etc., whereas Coriandrum sativum showed two test as negative that is iodide and saponins., least results were showed by Mentha spicata [Table. 2] 4.3 Protein Estimation Estimation of protein was done using biuret method. Biuret method can be used for both qualitative and quantitative estimation of protein. This method is based on the fact that under alkaline conditions substances containing two or more peptide bonds form a purple complex with cupric ions in the reagent and can be measured at 540nm. The color is stable, but all readings should be taken within 10 min. of
each other. Proteins with an abnormally high or low percentage of amino acids with aromatic side groups will give high or low readings, respectively. According to the study it was found that Spinach has more amount of protein content followed by coriander and least is mint [Table 3 to 5] 4.4 Separation of Plant Pigment using TLC According to the study the Rf value was found to be more in Spinach leave followed by Coriander leaf and the less Rf value was found in Mint leaves [Table 6]
Characteristics
Spinacia oleracea
Coriandrum sativum
Mentha spicata
Color
Bright to dark green
Light green to yellow
Dark green or purple
Taste
Characteristics
Characteristics
Minty or cool
Odor
Bitter
Aromatic
Aromatic
Shape
Ovate to triangular
Sub globular
Wide to narrow with a blunt tip
Extra Feature
The
flowers
are Flowers are white to Wide
spread
inconspicuous, yellow– purple and produce in overground green, the fruits are false lumpy
whorls
called underground
verticillasters
square
stolons, branched
stems.
Table.1 Organoleptic characters of Spinacia oleracea, Coriandrum sativum and Mentha spicata
Test
Spinacia oleracea Coriandrum satiyum
Mentha spicata
Protein
Positive
Positive
Positive
Carbohydrate
Positive
Positive
Positive
Iodine
Negative
Negative
Negative
Phenols
Positive
Positive
Positive
Glycosides
Positive
Positive
Negative
Alkaloids
Positive
Positive
Positive
Flavinoids
Positive
Positive
Positive
Saponins
Positive
Negative
Positive
Tannins
Positive
Positive
Positive
Cumarins
Positive
Positive
Negative
Table.2 Phytochemical screening of Spinacia oleracea, Coriandrum sativum and Mentha spicata
Test tube
Volume
of Volume
of Volume
protein(ml) water (ml)
of
Amount of Absorbence
biuret
protein
reagent
(mg)
520(nm)
(ml) 1
0.0
2.0
4
Keep
2
0.5
1.5
4
room
0.5
0.02
3
1.0
1.0
4
temperatur 1.0
0.04
4
1.5
0.5
4
e
10 1.5
0.06
5
2.0
0.0
4
minutes
2.0
0.08
6
0.1(UK1)
1.9
4
-
0.05
7
0.2(UK2)
1.8
4
-
0.06
for
at 0
0.0
Table.3 Protein estimation of Spinacia oleracea using Biuret method Unkown 1 of spinach: 0.1 ml of protein sample has = 1.25mg of proteins
5 ml of protein sample has = 31.25mg of proteins 5ml of protein solution was obtained from 2ml of stock. So, 2ml of stock has = 31.25mg The 2ml of stock solution was taken from 100ml of stock solution. Therefore 100 ml of stock solution has = 1.562g/ 1562mg of proteins This 100 ml of stock solution was made from 20 g of spinach leaves. Therefore, 20g contains 1562mg of proteins. 1 g of sample will have = 78.125mg/ 0.078g of proteins. Unknown 2 of spinach: 0.2 ml of protein sample has = 1.50mg of proteins 5 ml of protein sample has = 37.5mg of proteins 5ml of protein solution was obtained from 2ml of stock. So, 2ml of stock has = 37.5mg The 2ml of stock solution was taken from 100ml of stock solution. Therefore 100 ml of stock solution has = 1.8725g/ 1875mg of proteins This 100 ml of stock solution was made from 20 g of spinach leaves. Therefore, 20g contains 1875mg of proteins. 1 g of sample will have = 93.75mg/ 0.093g of proteins. Test tube
Volume
of Volume
of Volume
protein(ml) water (ml)
of
Amount of Absorbence
biuret
protein
reagent
(mg)
520(nm)
(ml) 1
0.0
2.0
4
Keep
at 0.0
0.00
2
0.5
1.5
4
room
0.5
0.02
3
1.0
1.0
4
temperatur 1.0
0.04
4
1.5
0.5
4
e
10 1.5
0.06
5
2.0
0.0
4
minutes
2.0
0.08
6
0.1(UK1)
1.9
4
-
0.01
7
0.2(UK2)
1.8
4
-
0.02
for
Table.4 Protein estimation of Coriandrum sativum using Biuret method
Unkown 1 of Coriander: 0.1 ml of protein sample has = 0.25mg of proteins 5 ml of protein sample has = 12.5mg of proteins 5ml of protein solution was obtained from 2ml of stock. So, 2ml of stock has = 12.5mg The 2ml of stock solution was taken from 100ml of stock solution. Therefore 100 ml of stock solution has = 0.625g/ 625mg of proteins This 100 ml of stock solution was made from 20 g of spinach leaves. Therefore, 20g contains 625mg of proteins. Ig of sample will have = 31.25mg/ 0.031g of proteins. Unknown 2 of coriander: 0.2 ml of protein sample has = 0.25mg of proteins 5 ml of protein sample has = 12.5mg of proteins 5ml of protein solution was obtained from 2ml of stock. So, 2ml of stock has = 12.5mg The 2ml of stock solution was taken from 100ml of stock solution. Therefore 100 ml of stock solution has = 0.625g/ 625mg of proteins This 100 ml of stock solution was made from 20 g of spinach leaves. Therefore, 20g contains 625mg of proteins. 1g of sample will have = 31.25mg/ 0.031g of proteins. Test tube
Volume
of Volume
protein(ml) water (ml)
of Volume
of
Amount of Absorbence
biuret
protein (mg) 520(nm)
reagent (ml) 1
0.00
2.0
4
0.0
0.00
2
0.5
1.5
4
Keep
at 0.5
0.02
3
1.0
1.0
4
room
1.0
0.04
4
1.5
0.5
4
temperatur 1.5
0.06
5
2.0
0.9
4
e
10 2.0
0.08
6
0.1(UK1)
1.9
4
minutes
-
0.03
7
0.2(UK2)
2.0
4
-
0.05
for
Table.5 Protein estimation of Mentha spicata using Biuret method Unkown 1 of mint: 0.1 ml of protein sample has = 0.75mg of proteins 5 ml of protein sample has = 37.5mg of proteins 5ml of protein solution was obtained from 2ml of stock. So, 2ml of stock has = 37.5mg The 2ml of stock solution was taken from 100ml of stock solution. Therefore 100 ml of stock solution has = 1.875g/ 1875mg of proteins This 100 ml of stock solution was made from 20 g of spinach leaves. Therefore, 20g contains 1875mg of proteins. Ig of sample will have = 93.75mg/ 0.093g of proteins. Unknown 2 of mint: 0.2 ml of protein sample has = 1.25mg of proteins 5 ml of protein sample has = 31.25mg of proteins 5ml of protein solution was obtained from 2ml of stock. So, 2ml of stock has = 31.25mg The 2ml of stock solution was taken from 100ml of stock solution. Therefore 100 ml of stock solution has = 1.562g/ 1562mg of proteins This 100 ml of stock solution was made from 20 g of spinach leaves. Therefore, 20g contains 1562mg of proteins. 1g of sample will have = 78.125mg/ 0.078g of proteins. Distance travelled by solvent and different plant pigments present in spinach, coriander and mint leaves.
Spinacia oleracea
Coriandrum sativum
Mentha spicata
Distance travelled by solvent Distance travelled by solvent Distance travelled by solvent =12cm
= 10.5cm
=10cm
Chlorophyll a = 9.7cm
Chlorophyll a = 7.2cm
Chlorophyll a = 6.3cm
Chlorophyll b = 7.4cm
Chlorophyll b = 6cm
Chlorophyll b = 5.9cm
Carotene = 5.4cm
Carotene = 3.4cm
Carotene = 3.2cm
Xanthophyll = 2.5cm
Xanthophyll = 2cm
Xanthophyll = 1cm
Table.6 Distance travelled by plant pigments in thin layer chromatography plate.
Plant pigment
Spinacia oleracea
Coriandrum sativum
Mentha spicata
Chlorophyll a
0.78
0.69
0.63
Chlorophyll b
0.62
0.58
0.59
Carotene
0.45
0.37
0.32
Xanthophyll
0.21
0.18
0.10
4.5 Images Photochemical test for Spinacia oleracea
Ninhydrin
Phenols and flavanoids
Carbohydrates
Glycosides
Tannins and Cumerins
Photochemical test for Coriandrum sativum
Glycosides
Phenols
Wagners
Ninhydrin
Flavanoids
Flavanoids
Saponins
Photochemical test for Mentha spicata:
Phenols Ninhydrin and Carbohydrates
Wagner’s
Thin Layer Chromatography:
Spinach Coriander
Mint
CHAPTER 5 DISCUSSION Plant and plant based medicines are the basis of many of the pharmaceuticals we use today for our various afflictions. The progress in the plant derived medicine has been depended upon firstly, the efficacy of the medicine in human being and secondly, the major active chemical constituents present in plant. Herbs and spices have been used during the middle ages for flavoring, food preservation, and/or medicinal purposes. In India thousands of species are known to have medicinal values and the use of different parts of several medicinal plants to cure specific disease has been in vogue since ancient times. North-East India is regarded as a hot spot of biodiversity and hence different types of herbal and medicinal plants are available there. These medicinal plants have been used traditionally by its local people for a long time to cure diseases. It was found that Spinacia oleracea gave almost all positive phytochemical tests for alkaloids, amino acid, carbohydrates, phenols etc. According to Gaikwad Priyanka Subhash “Spinacia oleracae pharmacological and pharmacognostic overview” it is found that flavonoid (methylenedioxyflavonol glucuronides) and carotenoid content of spinach helps fight cancer and functions as an antioxidant. Spinach is also effective in the treatment of diabetes. If taken together with alfalfa, spinach helps lower the blood sugar levels of the body. Therefore it is said as one of the best green leafy vegetable which has Anti-cancerous, Anti- inflammatory and Anti-Diuretic activity. Coriandrum sativum comes next in phytochemical analysis test, which gave around 8 positive tests for amino acid, carohydrates, alkaloids, flavanoids etc. From “Importance of Coriander (Coriandrum Sativum L.) Between the Medicinal and Aromatic Plants by K Maroufi et al”, talks about the secondary metabolites which plays a major role in controlling Blood sugar, Cholesterol and shows Anti-oxidant property too. It also help in treating digestive disorders, gastric, skin diseases etc.
The last is Mentha spicata which shows around 6 positive phytochemical test such as amino acid, carbohydrates, tannins, saponins etc.“From USDA national nutrition database”: Mint is said to be rich in many antioxidant vitamins, including vitamin A, beta carotene, vitamin-C and vitamin E. The leaves of mint also contain many important B-complex vitamins like folates, riboflavin and pyridoxine (vitamin B6); and the herb is an excellent source of vitamin-K. It also has Anti-oxidant properties, helps in stomach disorders and good for Asthma patients. All three leafy vegetables are essential source of vitamin, protein, minerals and provide immunity to fight against many chronic diseases. The leaves of spinach, coriander and mint are mostly used for its health benefits, although all parts of the plant including root, stem etc., have an important and significant role in medicine. Different parts of plants are used in Ayurveda and Siddha System of Medicine. A few of the uses are mentioned below. Chewing of leaves also cures ulcers and infections of mouth and stomach issues. Research indicates that the leaves of spinach has a very high safety margin with exceptionally low toxicity, providing general beneficial effects at doses without adverse reactions or other undesirable side effects.
CHAPTER 6
CONCLUSION
Through Comparative Quantitative and Qualitative study on Spinacia oleracea, Coriandrum sativum, Mentha spicata the following conclusion were drawn: Spinach Showed more positive result for phytochemical screening then that of coriander and mint [table2]. It also had high amount of protein content than coriander and mint [table 3 t0 5] and even showed high Rf values.We can infer that spinach has a very good medicinal value. It is packed with vitamins such as vitamin C, vitamin A and vitamin E and minerals like magnesium, manganese, iron, calcium and folic acid. Spinach is also a good source of chlorophyll, which is known to aid in digestion. Spinach is also rich in the carotenoids betacarotene and lutein. It is a good source of the bioflavonoid quercetin with many other flavonoids which exhibits anti-oxidant, antiproliferative, antiinfammatory, antihistaminic, CNS depressant, protection against gamma radiation, hepatoprotective properties in addition to its many other benefits.
CHAPTER 7
REFERENCES
1.
Saxena, Mamta, Jyoti Saxena, Rajeev Nema, Dharmendra Singh, and Abhishek Gupta. "Journal of Pharmacognosy and Phytochemistry." Vol 2, pp 29-35, 2013.
2.
Marmont, Alberto M., and Eugenio E. Damasio, "The effects of two alkaloids derived from Spinacia oleracea on the malignant cells of Hodgkin’s disease, lymphosarcoma and acute leukemia in vivo." Blood 29, vol 1, pp 1-21, 1967.
3.
Zhou, Mei-Liang, Hong-Li Hou, Xue-Mei Zhu, Ji-Rong Shao, Yan-Min Wu, and Yi-Xiong Tang. "Soybean transcription factor GmMYBZ2 represses catharanthine biosynthesis in hairy roots of Coriandrum sativum." Applied microbiology and biotechnology 91, vol 4 pp 1095-1105, 2011.
4.
Phatak, Rohan Sharadanand, and Anup Subhash Hendre. "Total antioxidant capacity (TAC) of fresh leaves of Kalanchoe pinnata." Journal of Pharmacognosy and Phytochemistry 2, vol 5 pp 3235, 2014.
5.
Goswami, Shambaditya. "Preliminary Phytochemical Screening and Standardisation of Leaves of Coriandrum sativum (L.) G. Don." Indian Journal of Research in Pharmacy and Biotechnology 1, vol 1 pp 21, 2013.
6.
Ghosh, Sumana, and S. A. Suryawanshi. "Effect of Mintha spicata extracts in treatment of alloxan diabetes in male albino rats." Indian Journal of Experimental Biology 39, vol 8 pp 748-759 2001.
7.
Verma, Sheetal, and S. P. Singh. "Current and future status of herbal medicines." Veterinary world 1, vol 11 pp 347-350, 2008.
8.
Warwick, O. H., J. M. M. Darte, T. C. Brown, C. T. Beer, J. H. Cutts, and R. L. Noble, "Some biological effects of Vincaleukoblastine, an alkaloid in Spinacia oleracea Linn in patients with malignant disease." Cancer research 20, vol 7 Part 1 pp 1032-1040, 1960.
9.
Gaikwad S. et. al, “Spinacia oleraceae Linn: A Pharmocognostic and pharmacological overview”, International Journal of Research in Ayurveda & Pharmacy, Vol 1, Issue 1, Sep-Oct, pp 7884,2010.
10.
Maroufi K. et. al, “Importance of Coriander (Coriandrum Sativum L.) Between the Medicinal and Aromatic Plants”, Advances in Environmental Biology, 4(3): pp 433-436, 2009.
11.
Gopi G. et.al, “Concise Review on Mentha spicata” International Journal of Phytopharmacy Research, 3(1), pp16-19, 2011.
12.
Kiranami P. et.al, “Toxicological Evaluation of Chloroform fraction of Spinacia oleraceae L. on mice”, International Journal of Drug Development and Research, vol 1(1), pp161-162.
13.
Gajalakshmi, S., S. Vijayalakshmi, and RAJESWARI V. Devi. "Pharmacological activities of Mintha spicata: a perspective review." Int J Pharm Bio Sci 4, vol 2, pp 431-439, 2013.
14.
Bisla, Gita, Shailza Choudhary, and Vijeta Chaudhary. "Evaluation of the nutritive and organoleptic values of food products developed by incorporated Mitha spicta (Sadabahar) fresh leaves explore their hypoglycemic potential." Vol 6(2), pp 198-220.
15.
Svoboda, Gordon H., Norbert Neuss, and Marvin Gorman. "Alkaloids of Coriander Linn .(Coriander sativum G. Don.) V. Preparation and characterization of alkaloids." Journal of the American Pharmaceutical Association 48, vol 11, pp 659-666, (1959).
16.
Tan, Puay Yok, and Thomas Whitlow. "Physiological responses of Mintha spicata (pudina) to ash yellows phytoplasmal infection." New Phytologist 150, vol. 3 pp 757-769, 2001.
17.
C.O.A.Z.S.S. Ballabh B, “Traditional medicinal plants of cold desert Ladakh- used against kidney and urinary disorders”, Journal of Ethno Pharmacology, Vol 118, pp 331-339, 2008.
18.
S.S. S. S. K. V. D. P. M. R. Palit G. “Evaluation of anti ulcer genic and ulcer-healing properties of Spinacia oleracea Lian.,” Journal of Ethnophamacology. Vol 93. pp. 197-206, 2004.
19.
Lewington A, “Medicinal plants and plant extracts. A review of their importation into Europe”, Traffic International, Vol – 4, 2014.
20.
Bamoniri A et. al., “Antioxidant and antimicrobial activity evaluation and essential oil analysis of Mentha sp. from Iran” Food Chem Vol- 122, p. 553–558, 2010.
21.
Flagan S F, “Utilization of carotenoid and vanillylamine as growth substrates by Capsicum (hot pepper) associated bacteria” Environ. Microbiol., Vol - 8, p. 560–565, 2006.
