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    Faculty of Biological Engineering Page 1

    INTRODUCTION

    CHAPTER 1

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    INTRODUCTION

    1.1 Arthritis

    Arthritis[1] is a form ofjoint disorder that involvesinflammation of one or more

    joints. There are over 100 different forms of arthritis. The mostcommonform,osteoarthritis (degenerative joint disease), is a result of trauma to the joint,

    infection of the joint, or age.

    Other arthritis forms arerheumatoid arthritis,psoriatic arthritis, and relatedautoimmune

    diseases.Septic arthritis is caused by jointinfection[1].Various drugs are used for the

    treatment of arthritis but a controlled drug delivery is closely associated with a polymer

    system that is degraded while the digestion and increases the effectiveness of the drug as

    such the drug Tinosporacordifolia, which is known by the common name Guduchi, is an

    herbaceousvine of the family Menispermaceae indigenous to the tropical areas of India,

    Myanmar andSri Lanka that cures the arthritis and Studies on induced oedema and arthritis

    and on human arthritis proved anti-inflammatorypotency of the water extract of plant.

    It also has antipyretic action. This drug relaxes the intestinal and uterine smooth muscles. It

    is proved effective in prevention of fibrosis and in stimulating regeneration in hepatic tissue.

    1.2 Drug Delivery

    Drug delivery[2] is the method or process of administering a pharmaceutical compound to

    achieve a therapeutic effect in humans or animals.Drug delivery technologies modify drugrelease profile, absorption, distribution and elimination for the benefit of improving product

    efficacy and safety, as well as patient convenience and compliance. Drug release is from:

    diffusion, degradation, swelling, and affinity-based mechanisms.Most common routes of

    administration include the preferred non-invasive peroral (through the mouth), topical

    (skin), transmucosal (nasal, buccal/sublingual, vaginal, ocular and rectal)[3] and inhalation

    routes. Many medications such as peptide and protein, antibody, vaccine and gene based

    drugs, in general may not be delivered using these routes because they might be susceptible

    to enzymatic degradation or can not be absorbed into the systemic circulation efficiently due

    to molecular size and charge issues to be therapeutically effective. For this reason many

    protein and peptide drugs have to be delivered by injection or a nanoneedle array. For

    example, many immunizations are based on the deliver it refers to approaches,

    formulations, technologies, and systems for transporting apharmaceutical compound in the

    body as needed to safely achieve its desiredtherapeutic effect.It may involve scientific site-

    targeting within the body, or it might involve facilitating systemic pharmacokinetics; in any

    case, it is typically concerned with both quantity and duration of drug presence. Drug

    delivery is often approached via a drug's chemical formulation, but it may also involve

    medical devices or drug-device combination products. Drug delivery is a concept heavily

    http://en.wikipedia.org/wiki/Arthropathyhttp://en.wikipedia.org/wiki/Inflammationhttp://en.wikipedia.org/wiki/Osteoarthritishttp://en.wikipedia.org/wiki/Rheumatoid_arthritishttp://en.wikipedia.org/wiki/Psoriatic_arthritishttp://en.wikipedia.org/wiki/Autoimmune_disorderhttp://en.wikipedia.org/wiki/Autoimmune_disorderhttp://en.wikipedia.org/wiki/Septic_arthritishttp://en.wikipedia.org/wiki/Infectionhttp://en.wikipedia.org/wiki/Infectionhttp://en.wikipedia.org/wiki/Herbaceoushttp://en.wikipedia.org/wiki/Menispermaceaehttp://en.wikipedia.org/wiki/Indiahttp://en.wikipedia.org/wiki/Myanmarhttp://en.wikipedia.org/wiki/Sri_Lankahttp://en.wikipedia.org/wiki/Pharmaceuticalhttp://en.wikipedia.org/wiki/Therapeutic_effecthttp://en.wikipedia.org/wiki/Therapeutic_effecthttp://en.wikipedia.org/wiki/Pharmaceuticalhttp://en.wikipedia.org/wiki/Sri_Lankahttp://en.wikipedia.org/wiki/Myanmarhttp://en.wikipedia.org/wiki/Indiahttp://en.wikipedia.org/wiki/Menispermaceaehttp://en.wikipedia.org/wiki/Herbaceoushttp://en.wikipedia.org/wiki/Herbaceoushttp://en.wikipedia.org/wiki/Infectionhttp://en.wikipedia.org/wiki/Septic_arthritishttp://en.wikipedia.org/wiki/Autoimmune_disorderhttp://en.wikipedia.org/wiki/Autoimmune_disorderhttp://en.wikipedia.org/wiki/Psoriatic_arthritishttp://en.wikipedia.org/wiki/Rheumatoid_arthritishttp://en.wikipedia.org/wiki/Osteoarthritishttp://en.wikipedia.org/wiki/Inflammationhttp://en.wikipedia.org/wiki/Arthropathy
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    integrated with dosage form and route of administration, the latter sometimes even being

    considered part of the definition.

    1.3 Ayurveda

    Ayurveda[4] is a complete or holistic system that integrates the mind, body and spirit.

    For a few centuries, the tradition of Ayurveda was dimmed due to the natural and human

    calamities and also by the invasion of foreign cultures into India. The sacred texts were

    either destroyed or stolen. However there were many vaidyas or doctors in India who

    managed to preserve some of the knowledge available in these Holy Scriptures. Divine

    plants that sustain long life and good health are now being rediscovered. Many renowned

    families of Vaidyas, who are specialized in certain branches of Ayurveda, have started

    functioning again in India. Today there is a revival of the ancient culture and traditions

    inherent to Ayurveda, which is a true gift of the ancient civilization to the modern world.

    The true history of Ayurveda starts from the time of the Holy books, the Vedas. Ancient

    mythology contends that the concept and essence of Ayurveda was revealed by the creator

    of the world himselfLord Brahma.

    There are four Vedas. They are

    Rigveda

    Yajurveda

    Samaveda

    Atharvaveda

    The Vedas date back to about five thousand years. They preach the philosophy of life.

    The Atharvaveda contains the principles of healing on which Ayurveda is based. 'Ayur'

    means 'life' in Sanskrit. Ayurveda is the most ancient science of healing which enhanceslongevity. It has influenced many of the older traditional methods of healing including

    Tibetan, Chinese and Greek medicine. Hence, Ayurveda is considered by many as the

    'mother of healing.'

    The practical tenets of Ayurveda are divided into eight sections or branches. These

    sections include:

    Internal medicine,

    Surgery,

    Organic medicine,

    Pediatrics,

    Toxicology,

    Rejuvenating remedy,

    Aphrodisiac remedies and

    Spiritual healing.

    These eight sections are called "Astanga Ayurveda".

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    1.4 Giloy( Tinospora Cordifolia)

    Tinosporacordifolia[5], which is known by the common name Guduchi, is an

    herbaceousvine of the family Menispermaceae indigenous to the tropical areas of India,

    Myanmar andSri Lanka.

    The plant is a glabrous climbing shrub found throughout India, typically growing indeciduous and dry forests. The leaves are heart shaped. The succulent bark is creamy white

    to grey in color, with deep clefts spotted withlenticels.It puts out long, slenderaerial roots,

    and is often grown on mango or neem trees Flowers are yellow, growing in lax racemes

    from nodes on old wood. Fruits aredrupes,turning red when ripe.

    Various name of giloy in India

    Hindi Name

    Giloya, Guduchi (Hindi)

    Bengali Name

    Gulancha/palo(Bengali),

    Telugu NameTippaatigo (Telugu)

    Tamil Name

    Shindilakodi (Tamil)

    Scientific classification

    Kingdom: Plantae

    Division: Magnoliophyta

    Class:Magnoliopsida

    Order: Ranunculales

    Family: Menispermaceae

    Genus: Tinospora

    Species: T. cordifolia

    Binomial name

    Tinosporacordifolia

    http://en.wikipedia.org/wiki/Herbaceoushttp://en.wikipedia.org/wiki/Menispermaceaehttp://en.wikipedia.org/wiki/Indiahttp://en.wikipedia.org/wiki/Myanmarhttp://en.wikipedia.org/wiki/Sri_Lankahttp://en.wikipedia.org/wiki/Glabroushttp://en.wikipedia.org/wiki/Lenticelhttp://en.wikipedia.org/wiki/Aerial_roothttp://en.wikipedia.org/wiki/Mangohttp://en.wikipedia.org/wiki/Neemhttp://en.wikipedia.org/wiki/Drupehttp://en.wikipedia.org/wiki/Biological_classificationhttp://en.wikipedia.org/wiki/Biological_classificationhttp://en.wikipedia.org/wiki/Planthttp://en.wikipedia.org/wiki/Planthttp://en.wikipedia.org/wiki/Flowering_planthttp://en.wikipedia.org/wiki/Flowering_planthttp://en.wikipedia.org/wiki/Magnoliopsidahttp://en.wikipedia.org/wiki/Magnoliopsidahttp://en.wikipedia.org/wiki/Ranunculaleshttp://en.wikipedia.org/wiki/Ranunculaleshttp://en.wikipedia.org/wiki/Menispermaceaehttp://en.wikipedia.org/wiki/Menispermaceaehttp://en.wikipedia.org/w/index.php?title=Tinospora&action=edit&redlink=1http://en.wikipedia.org/w/index.php?title=Tinospora&action=edit&redlink=1http://en.wikipedia.org/wiki/Binomial_nomenclaturehttp://en.wikipedia.org/wiki/Binomial_nomenclaturehttp://en.wikipedia.org/wiki/Binomial_nomenclaturehttp://en.wikipedia.org/w/index.php?title=Tinospora&action=edit&redlink=1http://en.wikipedia.org/wiki/Menispermaceaehttp://en.wikipedia.org/wiki/Ranunculaleshttp://en.wikipedia.org/wiki/Magnoliopsidahttp://en.wikipedia.org/wiki/Flowering_planthttp://en.wikipedia.org/wiki/Planthttp://en.wikipedia.org/wiki/Biological_classificationhttp://en.wikipedia.org/wiki/Drupehttp://en.wikipedia.org/wiki/Neemhttp://en.wikipedia.org/wiki/Mangohttp://en.wikipedia.org/wiki/Aerial_roothttp://en.wikipedia.org/wiki/Lenticelhttp://en.wikipedia.org/wiki/Glabroushttp://en.wikipedia.org/wiki/Sri_Lankahttp://en.wikipedia.org/wiki/Myanmarhttp://en.wikipedia.org/wiki/Indiahttp://en.wikipedia.org/wiki/Menispermaceaehttp://en.wikipedia.org/wiki/Herbaceoushttp://en.wikipedia.org/wiki/Herbaceous
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    Uses of Giloy:

    Immunity Disorders:

    Giloy or Amrita carries anti-inflammatory and antipyretic properties. This herb has been

    used in Ayurvedicrasayanas since centuries which is very helpful in building up the immune

    system and the body's confrontation against definite infecting organisms. In a scientific

    study conducted using human WBC (white blood corpuscles), the Ayurvedic herb helps in

    increasing the killing ability of macrophages, the resistant cell those are accountable for

    fighting foreign materials as well as microorganisms

    Stomach Ulcer:

    Giloy use for soothing inflamed and injured mucous membranes in the digestive tract. This

    herb protect the stomach and duodenum by increasing production of mucin, a substance that

    protects the lining of these organs against stomach acid and other harmful substances.

    Urinary disorder :

    This herb is used in urinary affections. The juice of the roots is very much effective in

    Urinary problems.

    Mental Disorder :

    The whole plant and the juice of the leaves is traditionally used in various mental disorders.

    This is regarded as one of the best psychotropic drugs in India.

    Fig-1.2

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    Other uses :

    There are many uses of Tinosporacordifolia. The stem is used in conditions like generalized

    weakness, dyspepsia, pyrexias of unknown origin (fevers), swine flu and many urinary tract

    infections. The bitter properties present in the drug show antiperiodic and antispasmodic

    properties which is again helpful in preventing swine flu.

    It is also used as an immune-modulator in immune-suppression of certain ailments like as

    obstructive jaundice, hepatic fibrosis, peritonitis and sepsis. The Tinosporaor Giloy or

    amrita has been exposed very effective in preventing fibrous changes and promoting

    regeneration of the liver against CCl4 induced hepatic toxicity.

    The herb is also used in stomach ulcer, urinary affections, vitalizer and remedy for diabetes

    and metabolic disorders.

    Studies on induced oedema and arthritis and on human arthritis proved anti-

    inflammatorypotency of the water extract of plant. It also has antipyretic action. This drugrelaxes the intestinal and uterine smooth muscles. It is proved effective in prevention of

    fibrosis and in stimulating regeneration in hepatic tissue.

    FIG-1.3 GILOY- HERB

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    Medicinal properties of giloy:

    External uses: Guduchi is very effective inskin disorders. It helps to boost resistance ofskin

    to microbes and is found effective in leprosy and ulcers formed due togout.This herb heals

    theherpes lesions and also accelerates the healing process of wounds in venereal diseases.

    Digestive system: Usage of Guduchi is indicated in conditions likeindigestion,excess thirst,

    acidity,irritable bowel syndrome (IBS),dysentery and vomiting. It is a good anti spasmodic

    and reduces stomach pain which arises due to intestinal spasms. Giloy is a very good

    hepatotonic .Preparations of this herb is very effective in jaundice, hepatic fibrosis and other

    diseases related to liver. This herb expels toxins accumulated in liver.

    Circulatory system: Texts of Ayurveda eulogize this herb as a cardiac tonic as it strengthens

    the heart. Guduchi helps to purifyblood and expel the toxins which are circulating in it.

    Because of this property it is useful ingout to control blood uric acid level.

    Respiratory system: It strengthens the lungs and reduces chronic cough

    Reproductive system: Few herbal aphrodisiac preparations contain guduchi as their

    ingredient. Due to aphrodisiac properties itincreases sperm count and sperm motility.Its

    adaptogen properties revitalize themale reproductive system and help in conditions like

    erectile dysfunction and premature ejaculation.

    Excretory System: Guduchi strengthens the urinary system and increases the resistance ofinner layers of bladder and urethra to fight repeated urinary tract infections.

    Giloy is very effective indiabetes and reduces fever. It boosts immunity and increases body

    energy level. Due to its anti microbial property the growth of micro organisms is inhibited in

    body. This boosts body resistance to diseases. Herbal preparations of this wonder herb help

    immensely in diseases of joints likearthritis and gout.

    1.5 Tamarind Seed Powder

    The tamarind seed powder is a complex mixture containing a galactoxyloglucan

    polysaccharides, proteins, lipids and certain minor constituents e.g. Fibers, Sugar etc. It

    forms a uniform solution on heating with water while stirring. It is used majorly in Paper

    Industry, Textile Industry, Dyeing and Printing Mills, Jute Mills, Card Board Industry and

    Explosive Plants.

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    REVIEW OF LITERATURE

    CHAPTER 2

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    REVIEW OF LITERATURE

    2.1 Pharmacological review

    PahadiyaS et al [7] has evaluated the radioprotective effect of an aqueous extract of

    Tinosporacordifolia (TC) against Co(60) gamma radiation in the dose of 5 mg/kg body

    wt to Swiss albino mice. It has shown significant protection in terms of survival

    percentage.

    Grover JKet al [8]has evaluated the extract of M. charantia (200 mg/kg), E. jambolana

    (200 mg/kg), M. pruriens (200 mg/kg) and T. cordifolia (400 mg/kg) was administered

    for 50 days in STZ induced diabetic mice, the plasma glucose concentration was reduced

    by 24.4, 20.84, 7.45 and 9.07% respectively.

    Mary NK et al [9] has evaluated T. cordifolia as antioxidant, anticoagulant, platelet

    antiaggregatory, lipoprotein lipase releasing, anti-inflammatory and hypolipidaemic

    activity in rats in the dose of 5 mg/kg. The extract has significantly (p

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    forming chemicals in the dose of 25 mg/kg b.wt, 10 days in mice. It has found extract of

    Tinosporacordifolia has been shown to inhibit the lipid peroxidation and superoxide and

    hydroxyl radicals in vitro.

    Singh SMet al[14]has evaluate influence of T. cordifolia on myeloid differentiation ofbone marrow progenitor cells and the recruitment of macrophages in response to tumor

    growth in situ. It has found extract of Tinosporacordifolia can influence the myeloid

    differentiation of bone marrow progenitor cells and the recruitment of macrophase in

    response to tumor growth in situ.

    Thatte UM et al[15] has evaluate the protective effects of Asparagus racemosus (AR)

    and Tinosporacordifolia (TC) against myelosuppression induced by single doses of

    cyclophosphamide (CP) 200 mg/kg. It has found that extract of Tinosporacordifolia is

    potent immunostimulant, with effects comparable to lithium and glucan.

    Chaudhary Ret al[16] Tinosporacordifolia (Guduchi), an Indian medicinal plant, was

    used to explore antitumor promoting activity in a two-stage skin carcinogenesis model at

    100 mg/kg body weight/day for 16 weeks. It has been observed that cumulative number

    of papillomas, tumor yield, tumor burden, and tumor weight showed significant

    reduction along with significant elevation of phase II detoxifying enzymes, and

    inhibition of lipid peroxidation in liver and skin in the animals administered with such

    plant extract concomitant to carcinogen exposure.

    Kar A et al [17] has evaluate hypoglycaemic activities of the experimental herbalsamples in the dose of 250 mg/kg . It has found that extract of Tinosporacordifolia

    shows the blood glucose lowering effect within 2 weeks in alloxan diabetic albino rats.

    Nemmani KV et al[18] has evaluate cell proliferation and natural killer cell activity by

    polyherbal formulation, Immu-21containing Ocimum sanctum, Withaniasomnifera,

    Emblicaofficinalis and Tinosporacordifolia in mice in the dose of 30 mg/kg once a day

    for 14 and 21 days. The results indicate that pretreatment with Immu-21 selectively

    increased the profileration of splenic leukocyte to B cell mitogen, Lipopolysaccharide

    (LPS), and cytotoxic activity against K 562 cells in mice.

    .Dhuley JN et al[19] The effect of Indian herbs namely, Asparagus racemosus,

    Tinosporacordifolia, Withaniasomnifera and Picrorhizakurrooa on the functions of

    macrophages obtained from mice treated with the carcinogen ochratoxin A (OTA) was

    investigated. The chemotactic activity of murine macrophages was significantly

    decreased by 17 weeks of treatment with OTA compared with controls. Production of

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    interleukin-1 (IL-1) and tumor necrosis factor (TNF) was also markedly reduced.

    Treatment with Asparagus racemosus, Tinosporacordifolia, Withaniasomnifera and

    Picrorhizakurrooa significantly inhibited OTA-induced suppression of chemotactic

    activity and production of IL-1 and TNF-alpha by macropahges. Moreover, we found

    that Withaniasomnifera treated macrophage chemotaxis and that Asparagus racemosusinduced excess production of TNF-alpha when compared with controls.

    Wadood Net al [20] The aqueous, alcoholic, and chloroform extracts of the leaves of

    Tinosporacordifolia were administered in doses of 50, 100, 150 and 200 mg/kg body

    weight to normal and alloxan-diabetic rabbits. The blood glucose and total lipid levels

    were estimated before and 2, 4, 6, and 8 hours after administration of the extract. The

    extract exerted a significant (P less than 0.5) hypoglycaemic effect in normal as well as

    in alloxan-treated rabbits

    Singh N et al[21] This article presents evidence to show that an alcoholic extract of

    Tinosporacordifolia (ALTC) enhances the differentiation of TAM to dendritic cells (DC)

    in response to granulocyte/macrophage-colony-stimulating factor, interleukin-4, and

    tumor necrosis factor. DC differentiated in vitro from TAM that were harvested from

    tumor-bearing mice after i.p. administration of ALTC (200 mg/kg body weight) 2 days

    post tumor transplantation shows an enhanced tumor cytotoxicity and production of

    tumoricidal soluble molecules like TNF, IL-1, and NO. Adoptive transfer of these TAM-

    derived DC to Dalton's lymphoma-bearing mice resulted in prolongation of survival of

    tumor-bearing mice. This is the first report regarding the differentiation and antitumor

    functions of TAM-derived DC obtained from tumor-bearing host administered with

    ALTC. The possible mechanisms involved also are discussed.

    Leyon PV et al [22] The antiangiogenic activity of Tinosporacordifolia was studied

    using in vivo as well as in vitro models. In vivo antiangiogenic activity was studied

    using B16F10 melanoma cell-induced capillary formation in animals. Intraperitoneal

    administration of the extract at a concentration of 20 mg/kg significantly inhibited the

    tumour directed capillary formation induced by melanoma cells. Analysis of the serum

    cytokine profile showed a drastic increase of proinflammatory cytokines such as IL-

    1beta, IL-6, TNF-alpha, granulocyte monocyte-colony stimulating factor (GM-CSF) and

    the direct endothelial cell proliferating agent vascular endothelial cell growth factor(VEGF) in the angiogenesis-induced control animals. Administration of Tinospora

    extract could differentially regulate these cytokine's elevation. The differential regulation

    is further evidenced by the increased production of antiangiogenic agents IL-2 and tissue

    inhibitor of metalloprotease-1 (TIMP-1) in the B16F10-injected, extract-treated animals.

    Moreover, using an in vitro rat aortic ring assay, it was observed that the extract at

    nontoxic concentrations inhibited the production of proangiogenic factors from B16F10

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    plants widely used in the Ayurvedic and Unani systems of medicine for treatment of

    chronic infections and immunological disorders. The effect of an ethanolic extract of

    each drug was studied on delayed type hypersensitivity, humoral responses to sheep red

    blood cells, skin allograft rejection, and phagocytic activity of the reticuloendothelial

    system in mice. Picrorhizakurroa was found to be a potent immunostimulant, stimulatingboth cell-mediated and humoral immunity. Tylophoraindica, Aconitum heterophyllum

    and Holarrhenaantidysenterica appeared to stimulate phagocytic function while

    inhibiting the humoral component of the immune system. Tinosporacordifolia and

    Ocimumgratissimum appeared to improve the phagocytic function without affecting the

    humoral or cell-mediated immune system. Hemidesmusindicus suppressed both the cell-

    mediated and humoral components of the immune system.

    StanelyMainzen, Prince P etal [26] We undertook the present study to evaluate the

    hypolipidaemic effect of an aqueous extract of Tinosporacordifolia roots, an indigenous

    plant used in Ayurvedic medicine in India. Administration of the extract of T. cordifolia

    roots (2.5 and 5.0 g/kg body weight) for 6 weeks resulted in a significant reduction in

    serum and tissue cholesterol, phospholipids and free fatty acids in alloxan diabetic rats.

    The root extract at a dose of 5.0 g/kg body weight showed highest hypolipidaemic effect.

    The effect of T. cordifolia roots at 2.5 and 5.0 g/kg body weight was better than

    glibenclamide. Insulin restored all the parameters to near normal values.

    2.2 Phytochemical Review

    Gangan VD et al [27] Several glycosides were isolated, as polyacetates, from the n-

    BuOH fraction of the Tinosporacordifolia stems. The structures of three new

    norditerpene furan glycosides cordifoliside A, B and C have been established by 1D and

    2D NMR spectroscopy.

    JahfarM et al [28] Polysaccharide from Tinosporacordifolia was isolated, purified,

    methylated, hydrolyzed, reduced and acetylated. The partially methylated alditol acetate

    (PMAA) derivative thus obtained was subjected to GC-MS studies. The following types

    of linkages were noticed: terminal-glucose, 4-xylose, 4-glucose, 4,6-glucose and 2,3,4,6-glucose.

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    AIM AND OBJECTIVES

    To design the tsp blended polymeric system for controlled drug delivery ofanti-arthritic drug giloy that can be used as a drug for curing arthritic disesases

    and studying the antimicrobial activitites of the drug.

    Several steps are to be carried out for estimating the release of the drug from

    the matrices at gastro-intestinal P.H for successful degradation of the

    polymeric substance and easy with quick adsorption of the drug at the mean

    time.[29]

    To determine the moisture contain analysis of giloy.

    To determine the total ash content in giloy.

    To determine the acid insoluble ash of the sample.

    To determine the acid insoluble ash of the sample.

    To determine the alcohol soluble extractive in giloy.

    To determine the soluble ash of giloy.

    To determine the percentage of sodium chloride content in giloy.

    To determine the percentage of borax content present in giloy

    To determine the mercury content in giloy. To determine the percentage of Iron content in giloy

    To perform the separation of two phases using TLC.

    Preparation of giloy films blended with TSP (tamarind seed powder) forcontrol drug delivery and anti- microbial studies.

    To perform Drug Loading and Dissolution experiments:

    To check anti-microbial activity of 5% composition giloy film blended

    with TSP.

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    MATERIALS AND METHODS

    CHAPTER-4

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    MATERIALS AND METHODS

    4.1 CHEMICAL PARAMETERS[30]

    4.1.1 MOISTURE CONTAIN & ANALYSIS

    AIM OF THE EXPERIMENT:-

    To determine the moisture contain analysis of giloy.

    APPARATUS REQUIRED:-

    Hot air oven

    Electronic balance

    Petri dish

    Spatula Desiccator

    Motor and pistle

    PRINCIPLE:-

    Excess of water in food , spices, medicine, will encourage the microbial growth of fungi or

    insect detoriation following hydrolysis.

    Limit of water contain should therefore be set for every given food or medicinal plant

    material. Although moisture in an important component of food and drugs it should be

    eliminate as per required. The objective of drying of fresh material are to add to their

    preservative to fix their constituents to check enzymatic &hydrolytic reaction that might

    effect that chemical composition of drugs, to reduce their bulk and weight. To facilate

    subsequent grinding into powder form. The test for loss or drying determines both water and

    volatile matter. Drying can be carried out by heating at a temp. 100-105C inside the hot air

    oven and then cooling for 30 min. inside the dessicator and wt. it.

    PROCEDURE:-

    5-10gms. of freshly prepared sample is taken (W) in pre weight petridish (W1) .

    The petridish containing the sample is placed inside the oven for 1-2 hour at a

    temp. between 100C-105C.

    The sample is taken out from oven and place inside the dessicator for 30 mins. for

    cooling.

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    Immediately take the weight (W2) in the balance and calculate the loss of wt. of

    the given sample.

    4.1.2 .TOTAL ASH

    AIM OF THE EXPERIMENT:-

    To determine the total ash content in giloy.

    APPRATUS REQUIRED:-

    Muffle furnance

    Desiccators

    Crucible,

    Tongs, Electronic balance.

    PROCEDURE:-

    A clean dry crucible of platinum, nickel or silica was taken.

    The blank weight of crucible measured (w1).

    Let sample weight be w ( 2-4 gms sample weigh accurately in the crucible) .

    Then ignite (incinerate) the sample with crucible inside the muffle furnace at 600-

    650C for 1 to 2 hour (until it is white).

    The crucible was taken out and cooled inside the desiccator and weight If the carbon free ash cannot be obtained in this manner then the crucible was cooled

    and moisture with 2ml of water over a saturated solution of ammonium nitrate.

    Then it was dried inside a water bath, then on hot plate.

    Ignition was done inside the muffle furnace up to a constant weight.

    Then the residue was allowed to cool inside the desiccators for30 minutes and then

    the weight was taken.

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    4.1.3 ACID INSOLUBLE ASH

    AIM OF THE EXPERIMENT:-

    To determine the acid insoluble ash of the sample.

    APPRATUS REQUIRED:-

    Muffle furnance

    Desicator

    Crucible

    Tong

    Hot plate

    Chemicals required:- 7% HCL.

    PROCEDURE:-

    Transfer the ash obtained above in 250 ml beaker without loss of a ash and 100 ml of

    dil.hydrochloric acid.

    Wash the crucible with 10 ml of acid and transfer the washings to the beaker.

    Heat the beaker till the liquid boils.

    Filter the solution and collect the insoluble matter on ashless-filter paper (Whatman

    filter paper No.4).

    Wash with hot water until the filtrate is neutral.

    Transfer the filter paper containing the insoluble matter to the original crucible.

    Dry on a hot plate and ignite at around 600C in a muffle furnace(until it become

    white ash).

    Allow the residue to cool in suitable desiccator for 30 min. and weight without delay.

    Repeated the process until the constant weight is obtained.

    Calculate the insoluble ash with reference to the air dried drug.

    4.1.4 WATER SOLUBLE EXTRACTIVE

    AIM OF THE EXPERIMENT:-

    To determine the water soluble extractive in giloy.

    APPARATUS REQUIRED:-

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    conical flask

    Hot air oven

    Water bath

    Desiccator

    Electronic balance

    PROCEDURE:-

    Macerate (powder) 5 grams of the air dried drug coarsely powdered with 100 ml of

    chloroform water in a closed flask (stpppered conical flask) for 24 hour.

    Shaking frequently during 6 hours and allowing to stand for 18 hours.

    Filter rapidly taking precautions against loss of solvent.

    Evaporate 25 ml of the filtrate to dryness in a tared flat bottom shallow

    dish(petridish) and dry at 105C to constant weight and weigh it.

    4.1.5 ALCOHOL SOLUBLE EXTRACTIVE

    AIM OF THE EXPERIMENT:-

    To determine the alcohol soluble extractive in giloy.

    APPARATUS REQUIRED:-

    conical flask

    Hot air oven

    Water bath

    Desicator

    Electronic balance

    PROCEDURE:-

    Macerate(powered form) 5 grams of the air dried drug coarsely powered with 100 ml

    of alcohol(90% ethyl alcohol) of the specified strength, in a closed flask for 24 hour.

    Shaking frequently during 6 hour and allowing to stand for 18 hour.

    And filter rapidly ,taking the precursions against the loss of solvent.

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    Evaporate 25 ml of the filtrate to dryness in a tarred flat bottom shallow

    disc(petridish) and then dry at 105C to constant weight and to weight.

    4.1.6 WATER SOLUBLE ASH

    AIM OF THE EXPERIMENT:-

    To determine the soluble ash of giloy.

    APPRATUS REQUIRED:-

    Muffle furnace

    Desicator

    Crucible

    PROCEDURE:-

    Prepare ash as given under total ash.

    Boil the total ash for 5 min. with 25 ml of water.

    Collect insoluble matter in a Gooch Crucible or on an ash-less filter paper.

    Wash with hot water ignite for 15 min. at a temperature not exceeding 600C.

    Subtract the weight of the insoluble matter from the weight of ash.

    The difference in weight represents the water soluble ash.

    Calculate the percentage of the water soluble ash with reference to the Air-dried

    drug.

    4.2 ASSAY

    4.2.1 SODIUM CHLORIDE ASSAY

    AIM OF THE EXPERIMENT:-To determine the percentage of sodium chloride content in

    giloy.

    APPRATUS REQUIRED:-

    Muffle furnace

    Silica crucible

    Volumetric flask

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    Filter paper

    Electronic balance

    Conical flask

    Titration stand

    CHEMICALS REQUIRED:-

    0.1AgNo

    KCrOIndicator

    PROCEDURE:-

    Dissolve about 2-3 grams of accurately weighed drug in 25 ml of purified water andleft for 30 minutes then filtered.

    Wash the filter paper completely with perfect water and the filtration made 100 ml in

    volumetric flask and make the solution homogeneous

    Titrate 25 ml of the solution with 0.1N AgNousing KCrOas indicator

    The end point shows the light brick red colour.

    Each ml of 0.1N AgNOis equivalent to 0.005845 gram NaCl.

    Dissolve about 2-3 grams of accqurately weigh drug sample and dissolve it in 25 ml

    of purified water and leave for 30 min and filter.

    Wash the filter paper completely with purified water and the filtrate is made 100 ml

    in volumetric flask and make the solution homogenous.

    Titrate 25 ml of this solution in 0.1N AgNo3 (silver Nitrite) as indicator.

    The end point shows light brick red colour.

    4.2.2 BORAX

    AIM OF THE EXPERIMENT:-To determine the percentage of borax content present in

    giloy

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    APPRATUS REQUIRED :-

    Muffle furnace

    Conical flask

    Filter paper

    Silica Crucible

    Desiccators

    CHEMICALS REQUIRED:-

    0.5N Hcl

    Methyl orange Indicator

    PROCEDURE :-

    Powder 5-6 grams of drugs i.e liv-52 and Ignited at 450c for 3 hours in muffle

    furnace to get it ash and after that put it on desiccator for cooling.

    Then 20 ml of distilled water put in ash and rest it for 15 minutes and filtration will

    be done

    Finally titration of sample will occur against 0.5N Hcl by using methyl orange

    Indicator.

    Each ml of 0.5N Hcl is equivalent to 0.09536 gram of NaBO10HO.

    4.2.3 MERCURY

    AIM OF THE EXPERIMENT:-To determine the mercury content in giloy.

    APPRATUS REQUIRED:-

    RB Bottle

    Dessicator

    Muffle furnace

    Condensor

    Conical flask

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    Electronic balance machine

    CHEMICALS REQUIRED:-

    Concentrated Hcl

    2) Nitric acid (HNO)

    3) 0.02N Potassium permanganate

    4) HO

    5) Thiocyanine

    6) Ferricallum indicator

    PROCEDURE:-

    Take 0.5 grams of sample in reflux bottle.

    Add 15 ml of Concentrated NOto the sample.

    Then reflux it at temperature (35-40c) for 1 hour and after reflux cool it for some

    time.

    After cooling add 50 ml of HNOthen again it is heated for colourless.

    Then 100 ml of destiled water put on the solution using condenser.

    Then add 0.02N potassium permanganate(KMNO) till the solution looks like pink

    colour.

    Then decolourised it by adding 6% HO

    Again add 3ml of Concentrated Nitric acid and titrate the sample against

    theocyanine by using ferricallum indicator.

    4.2.4 IRON

    AIM OF THE EXPERIMENT :-

    To determine the percentage of Iron content in giloy.

    APPRATUS REQUIRED:-

    Reflux

    Beaker

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    Silica crucible

    Muffle furnish

    Volumetric flask

    Titration strand

    Conical flask

    CHEMICALS REQUIRED:-

    Mercury Chloride

    Phosphoric acid

    Stannous Chloride

    Barium diphenyl amine sulphonate Indicator

    0.1N KCrO HSO

    PROCEDURE :-

    Take 2 grams of the Sample in a pre-weighed silica crucible.

    Heat in muffle furnish at 600c until the colour changing to white ash

    Reflux the ash with 100ml of 1:1 Hcl for 30 minutes.

    Cool and filtered the sample and make up the volume of filterd sample to 250 ml in avolumetric flask with the help of distilled water.

    Pipette out 25 ml aliquot of the solution and boil it.

    Add hot stannous chloride in drops in sample till the yellow colour disappears.

    Then cool it and add 10 ml of mercuric Chloride.

    Then the silver colour pipette occurs.

    Add 10 ml of Phosphoric acid-sulphuric acid mixture.

    Add 1ml of barium diphenyl amine sulphonate indicator to the sample.

    Finally Titrate the sample against 0.1N KCrOand the end point seems to be violet

    blue colour. Repeat titration for concordant values.

    Avoid excess of stannous chloride solution so, that solution does not become black.

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    Each ml of 1N KCrOis equivalent to 0.05585 grams of Fe

    4.3 TLC(THIN LAYER CHROMATOGRAPHY)

    AIM OF THE EXPERIMENT:-

    To perform the separation of two phases using TLC.

    APPRATUS REQUIRED:-

    Flat glass plates

    Aligning tray or flat surface

    Spreader

    Storage rack

    Developing chamber

    Graduated micro pipette

    Reagent sprayer

    U.V light

    REAGENTS/CHEMICALS REQUIRED:-

    Ninhydrin

    Ether

    Lime water

    Silica gel

    PROCEDURE:-

    a.Preparation of plates:-

    Prepare a suspension of the coating substance in a accordance with the instructions of the

    supplier and using the spreading device designed for the purpose.

    Spread a uniform layer of the suspension 0.25 0.30 mm thick on a flat glass plate 20 c.m

    long.

    Allow the coated plates to dry in air , heat at 100 - 105C for at least 1 hour.

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    Note- Except in the case of plates prepared with cellulose when heating for 10 minutes is

    normally sufficient and allowed to cool, protected from moisture , store the plates protected

    from moisture and use within three days of preparation .At the time of use dry the plates

    again if necessary as presented

    .

    SPREADER USED FOR TLC METHOD

    b Method:-

    Unless un-saturated condition are prescribed prepare the tank by lining the walls

    with sheets of filter paper , pour into the tank, saturating the filter paper in the

    process sufficient of the mobile phase to form a layer of solvent 5-10 mm deep [n-

    butanol:acetic acid : water(80:20:20)].

    Close the tank and allowed to stand for 1 hour at room temperature.

    c. Extraction:-

    Weigh 100 gram of solid sample or 10 ml of liquid sample.

    For solid sample add Ca(OH) 1 gram diturate with motor pistle and transfer thecontents to a 100 ml beaker, dilute it to 200 ml with lime water (5-10%) and shake

    well for 30 minutes.

    Filter it to collect in a separating funnel 80 ml of filtrate by filter paper on a Gooch

    crucible.

    It is twice washed with solvent ether 25 ml each for removing resin, glycosides etc.

    Fig2.1

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    The ether layer is washed with 5 ml of lime water and 5 ml of water.

    After washing remove the ether layer and collect the rest as sample.

    Add 2 gram of ammonium sulphate to the whole extract.

    First extraction is done with alcohol and chloroform in 1:1 dilution ratio.

    Second extraction is done with alcohol and chloroform in 1:2 ratio, during this add

    Barium hydroxide for clear distinction in layers. The whole mixture is evaporated at 100C using water bath for 30 minutes.

    After evaporation the residue is dissolved with 2 ml of alcohol and 25 ml of 0.02 N

    Sulphuric acid and it is warmed.

    It is cooled and titrated with 0.02 N NaOH with methyl orange/red as indicator, it

    gives red yellow colour end point.

    Remove a narrow strip of the coating substance or the stationary phase about 5 mm

    of the vertical sides of the lead.

    Apply the solution being examined in the form of circular spots about 2-6 mm in

    diameter or in the form of bands (10-20 mm 2-6 mm).On a line parallel with and

    20 mm from one end of the plate and not nearer than 20 mm of the sizes, the spots

    could be 50 mm apart.

    If necessary the solution may be applied in portions drying between applications.

    Mark the sides of the plates 15 cm or the distance specified in the monograph.

    Allow the solvent to evaporate and place the plate in tank, ensuring that it is nearly

    vertical as possible and that the spots or bands are above the level of the mobile

    phase.

    Close the tank and allow to stand at room temperature until the mobile phase has

    ascended to the marked line.

    Remove the plates and dry. Visualize as directed in monograph. Where as a spraying

    technique is prescribed. It is essential that the reagent be evenly applied as a fine

    spray. For 2-D chromatography dry the plate after the first development and carry out the

    second development in a direction perpendicular to the first.

    When the method prescribed in the monograph specifies protected from light or in

    subdued light it is intended that the entire procedure is carried out under this

    conditions.

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    SEPARATING FUNNEL USED FOR EXTRACTION METHOD

    d. Visualization:-

    The phrases U.V light (254 nm) and U.V light(365nm) indicates that the plates

    should be examined under an U.V light having a maximum out put at about 254 nmor at about 365 nm , as the case may be.

    The term secondary spot means any spot other than the principal spot, similarly a

    secondary band is any band other than principal band.Both the terms are applicable

    for 2-D chromatography.

    Fig-2.2

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    IDENTIFICATION OF PROTEIN

    4.4 SYNTHESIS OF TSP BLENDED WITH GILOY

    AIM OF THE EXPERIMENT:-

    Preparation of giloy films blended with TSP (tamarind seed powder) for control drug

    delivery and anti- microbial studies.

    MATERIAL REQUIRED:-

    Magnetic stirrer

    Beaker

    Hot air oven

    Petriplates

    PROCEDURE:-

    Fig-2.3

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    1 ) ONLY TSP FILM

    0.5gm of TSP was taken and made it soluble in25ml of distilled water at 500c with

    continuous stirring in magnetic stirrer.

    After about 31/2

    hours add 5-6 drops of glycerol.

    Then after 4 hours add the mixture onto a petriplate. Keep the petriplate in hot air oven at 50-55

    0c for turning into thin film.

    After about 24 hours a thin film of TSP is taken out from the petriplate.

    TSP FILM

    2) 1% COMPOSITION FILM

    0.5gm of TSP was taken and made it soluble in25ml of distilled water at 500c with

    continuous stirring in magnetic stirrer.

    Then add 1ml of the extract to the polymer.

    After that add 5-6 drops of glycerol to the mixture.Then after 4 hours of continuous

    stirring add the mixture onto a petriplate.Keep the petriplate in hot air oven at 50-

    550

    c for turning into thin film. After about 24 hours a thin film of TSP blended with 1ml of giloy extract is taken

    out from the petriplate.

    Fig-2.4

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    Fig-2.5- 1% COMPOSITION FILM

    3) 5% COMPOSITION

    0.5gm of TSP was taken and made it soluble in25ml of distilled water at 500c with

    continuous stirring in magnetic stirrer.

    Then add 5ml of the extract to the polymer.

    After that add 5-6 drops of glycerol to the mixture.

    Then after 4 hours of continuous stirring add the mixture onto a petriplate.

    Keep the petriplate in hot air oven at 50-550c for turning into thin film.

    After about 24 hours a thin film of TSP blended with 5ml of giloy extract is taken

    out from the petriplate.

    Fig-2.6- 5% COMPOSITION FILM

    4)7.5% COMPOSITION

    0.5gm of TSP was taken and made it soluble in25ml of distilled water at 500c with

    continuous stirring in magnetic stirrer.

    Then add 7.5ml of the extract to the polymer.

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    After that add 5-6 drops of glycerol to the mixture.

    Then after 4 hours of continuous stirring add the mixture onto a petriplate.

    Keep the petriplate in hot air oven at 50-550c for turning into thin film.

    After about 24 hours a thin film of TSP blended with 7.5ml of giloy extract is taken

    out from the petriplate.

    Fig-2.7- 7.5% COMPOSITION FILM

    5) 10% COMPOSITION

    0.5gm of TSP was taken and made it soluble in25ml of distilled water at 500c with

    continuous stirring in magnetic stirrer.

    Then add 10ml of the extract to the polymer.

    After that add 5-6 drops of glycerol to the mixture.

    Then after 4 hours of continuous stirring add the mixture onto a petriplate.

    Keep the petriplate in hot air oven at 50-550c for turning into thin film.

    After about 24 hours a thin film of TSP blended with 10ml of giloy extract is taken

    out from the petriplate.

    Fig-2.8 -10% COMPOSITION FILM

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    4.5 CONTROL DRUG DELIVERY[31]

    Drug Loading :

    Giloy blended with TSP(tamarind seed powder nanocomposites were prepared by

    emulsion/solvent evaporation method. In short, giloy of different loadings, i.e., 1 wt%, 5wt%, 7.5% and 10wt% were dissolved in glycerol. The formed solution was poured into a

    labeled Petri dish and allowed to evaporate the solvent overnight at room temperature. This

    compound was used for drug delivery purposes.

    Dissolution experiments:

    Dissolution experiments were performed at 37oC using the dissolution tester (Disso test,

    Lab India, Mumbai, India) equipped with six paddles at a paddle speed of 100 rpm. About

    900 ml of phosphatebuffer solution (pH 5.8,6 & 7) was used as the dissolution media to

    stimulate gastrointestinal tract (GIT) conditions. A 5 ml aliquot was used each time for

    analyzing the giloy content at a fixed time interval. The dissolution media was replenished

    with a fresh stock solution. The amount of giloy released was analyzed using a UV

    spectrophotometer (Systronics, India) at the max value of 380 nm.

    Drug release mechanism from matrices:

    From time to time, various authors have proposed several types of drug release mechanisms

    from matrices. It has been proposed that drug release from matrices usually implies water

    penetration in the matrix, hydration, swelling, diffusion of the dissolved drug (polymer

    hydro fusion), and/or the erosion of the gelatinous layer. Several kinetic models relating to

    the drug release from matrices, selected from the most important mathematical models, aredescribed over here.

    However, it is worth mention that the release mechanism of a drug would depend on the

    dosagefrom selected, pH, nature of the drug and, of course, the polymer used.

    (i) Zero - Order Kinetics [29].

    W = k1 t . (1)

    (ii) First - Order Kinetics [29,32].

    ln (100- W) = ln 100 - k2 t (2)

    (iii) Hixon-Crowels Cube- Root Equation (Erosin Model) [32].

    (100- W) 1/3 = 100 1/3k3 t .. (3)

    (iv) Higuchis Square Root of Time Equation (Diffusion Model) [30].W = k4 t . (4)

    (v) Power Law Equation (Diffusion/ Relaxation model) [31].

    Mt / M = k5 t n . (5)

    Mt / M is the fractional drug release into dissolution medium and k5 is a constant

    incorporating the structural and geometric characteristics of the drug.

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    Analysis:

    The term n is the diffusional constant that characterizes the drug release transport

    mechanism. When n = 0.5, the drug diffused and released from the polymeric matrix with a

    quasi-Fickian diffusion mechanism. For n =0.5, an anomalous, non-Fickian drug diffusion

    occurs. When n =1, a non-Fickian, case II or Zero- order release kinetics.

    4.6 ANTI-MICROBIAL ACTIVITY

    AIM OF THE EXPERIMENT:-

    To check anti-microbial activity of 5% composition giloy film blended with TSP.

    MATERIALS REQUIRED:-

    Hot plate Magnetic stirrer

    Auto clave

    Laminar air flow

    Hot air oven

    Filter paper

    Forceps

    Inoculating wire loop

    Beaker

    Conical flask

    Chemicals required for preparation of LB (Luria Media)

    100ml distilled water

    Peptone water-1.5gm

    Yeast dextrose agar-3.5gm

    Agar-agar-0.5gm

    PROCEDURE

    Water, peptone water, yeast dextrose water and agar-agar is taken in a conical flask

    and mixed well in a magnetic stirrer.

    0.5gm of 5% composition of the film is properly mixed with 7.5ml distilled water on

    a magnetic stirrer for about 1 hour.

    Conical flask with the LB media is wrapped with paper and kept in auto clave along

    with a petriplate wrapped in a paper inside a polythene.

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    After the sterilization is done in the petriplate the conical flask and the petriplate is

    kept in the laminar air flow.

    2 small 1cm filter paper is taken.

    One filter paper is kept in 90% ethyl alcohol and the other is kept in the film.

    Then the culture is taken in a wire loop from the sterile culture and a smear is madeon the LB media which is spread with a glass spreader all around the LB media.

    With the help of a forcep the filter paper which was in 90% ethyl alcohol is kept on

    one side of the petriplate and the other filter paper is kept on the other side.

    The petriplate is inverted and kept in the hot air oven.

    4.6.1 ANTI-FUNGAL ACTIVITY

    AIM OF THE EXPERIMENT:-To check anti-fungal activity of 5% composition giloy film blended with TSP.

    MATERIALS REQUIRED:-

    Hot plate

    Magnetic stirrer

    Auto clave

    Laminar air flow

    Hot air oven

    Filter paper

    Forceps

    Inoculating wire loop

    Beaker

    Conical flask

    PROCEDURE:

    Water, peptone water, yeast dextrose water and agar-agar is taken in a conical flask

    and mixed well in a magnetic stirrer.

    0.5gm of 5% composition of the film is properly mixed with 7.5ml distilled water on

    a magnetic stirrer for about 1 hour.

    Conical flask with the LB media is wrapped with paper and kept in auto clave

    alongwith a petriplate wrapped in a paper inside a polythene.

    After the sterilization is done in the petriplate the conical flask and the petriplate is

    kept in the laminar air flow.

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    2 small 1cm filter paper is taken.

    One filter paper is kept in 90% ethyl alcohol and the other is kept in the film.

    Then the culture is taken in a wire loop from the sterile

    culture and a smear is made on the LB media which is spread with a glass spreader

    all around the LB media. With the help of a forcep the filter paper which was in 90% ethyl alcohol is kept on

    one side of the petriplate and the other filter paper is kept on the other side.

    The petriplate is inverted and kept in the hot air oven.

    Anti fungal activity

    Fig-2.9

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    RESULTS

    CHAPTER 5

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    RESULT

    TOTAL MOISTURE

    Percentage of moisture= (Initial weight-final weight/sample weight)*100

    Weight of petriplate=44.02gm

    Weight of sample=6.55gm

    Initial weight(weight of petriplate + weight of sample)=50.57gm

    Final weight=50.41gm

    % of moisture=(50.57-50.41/6.55)*100

    =2.44%

    Therefore the total % of moisture =2.44%

    TOTAL ASH

    Volume of total ash= (final weight-blank weight/sample weight)*100

    Blank weight of crucible=22.41gm

    Sample weight=2.10gm

    Final weight=22.58gm

    Volume of total ash=(22.58-22.41/2.10)*100

    =8%

    Therefore, the total ash %=8%

    ACID I NSOLUBLE ASH

    Percentage of acid insoluble ash=(final weightblank weight/sample weight)*100

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    Blank weight of crucible=22.42gm

    Sample weight=3.04gm

    Final weight =22.53gm

    % of acid insoluble ash=(22.53-22.42/3.04)*100

    =3.6%

    Therefore, the total acid insoluble ash percentage=3.6%

    WATER SOLUBLE EXTRACTIVE

    Percentage of water soluble extractive= (final weight-blank weight/sample weight)*100

    Blank weight of petriplate=44.85gm

    Sample weight=2.5gm

    Final weight=45.01gm

    % water soluble extractive=(45.01-44.85/2.5)*100*1/4

    ( because Ive used 2.5gm sample and 25ml distilled water)

    =1.6%

    Therefore, the total % water soluble extractive=1.6%

    ACID SOLUBLE EXTRACTIVE

    Percentage of alcohol soluble extractive= (final weightblank weight/sample weight)*100

    Blank weight of petriplate=44.25gm

    Sample weight=2.5gm

    Final weight=44.37gm

    % alcohol soluble extractive=(44.37-44.25/2.5)*100*1/4

    =1.2%

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    Therefore, the total alcohol soluble extractive=1.2%

    WATER SOLUBLE ASH

    Percentage of water soluble ash={final weight of the total ash with crucible- final

    weight(water insoluble ash)/sample weight}*100

    Final weight of total ash=22.58gm

    Final weight of water insoluble ash=22.50gm

    Sample weight=2.10gm

    % water insoluble ash= (22.58-22.50/2.10)*100

    =3.8%

    Therefore, water soluble ash=3.8%

    Assay:

    SODIUM CHLORIDE

    Sample taken = 2.14gm

    Initial Burette reading=0

    Final burette reading=3ml

    Burette Reading =(3-0) = 3ml

    Each ml of 0.1N AgNois equivalent to 0.00584 gm of NaCl.

    Calculation:-

    1 ml of 0.1N AgNois to 0.005845 gmNacl

    3 ml of 0.1N AgNO3 is = 0.0058453=0.0175gmNaCl

    So,25 ml of sample solution contains 0.0175gm of Nacl

    100 ml sample solution contain =(0.01754)gmNacl = 0.07gm Nacl

    So,2.14gm of sample contain 0.07gm Nacl

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    Therefore % of Nacl=(Nacl content/sample wt)100

    =(0.07/2.14)100 =3.27% of Nacl

    From the above calculation It is found that giloy contains 3.27% of Nacl.

    BORAX

    Sample taken = 3.01gm

    Initial Burette Reading = 0

    Final Burette Reading = 4

    Burette Reading = (4-0) = 4 ml

    Each ml of 0.5N Hcl = 0.09536gm og NaBO10 HO

    Calculation:-

    Each ml of 0.5N Hcl = 0.09536gm NaBO10 HO

    Each ml of 0.094N Hcl = (0.09536gm of NaBO10 HO/0.50.094)gm of Borax

    =0.017927gm of Borax

    4ml of 0.094N Hcl contains Borax = (0.0179274)gm of Borax

    = 0.07170gm Borax

    % of Borax = (0.07170gm.Borax/3.01)100 = 2.38% of Borax

    From the above calculation it is found that giloy contain 2.38% of Borax

    MERCURY:-

    Sample taken = 0.5gm

    Initial Burette Reading = 0

    Final Burette Reading = 4 ml

    Burette Reading = 4-0 = 4ml

    Each ml of 0.1N NHScN = 0.01003g of H

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    Calculation:-

    0.1N NHScN = 0.01003gm of Hg

    4ml of 0.1N NHScN=(0.010034)gm of Hg =0.04012gm of Hg

    0.5gm of Sample Contain Hg=(0.04012/0.5)100 =8.024%

    So, from the above calculation it is found that giloy contain 8.024% 0f Hg.

    I RON ASSAY:-

    Sample taken = 2gm

    Initial Burette Reading = 0

    Final Burette Reading = 3.5 ml

    Burette Reading = 3.5-0 = 3.5ml

    Each ml of 1N KCrOis equivalent to 0.05585 gm of Iron

    Calculation:-

    Each ml of 1N KCrO= 0.05585gm of Fe

    Each ml of 0.1N KCrO= (0.055850.1)gm of Fe

    3.5ml of 0.1N KCrO= (0.05585gm Fe3.50.1)gm Fe.

    =0.01954gm of Fe

    So,250 ml of sample contain (0.01954 10)gm of Fe = 0.1954gm of Fe

    So,2gm of sample contain 0.1954gm of Fe.

    Therefore % of Fe =( Amount of Fe in 2gm of sample/sample wt)100

    =( 0.1954/2)100 = 9.77%

    So From the above calculation it is found that giloy contain 9.77% of Iron.

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    CONTROL DRUG DELIVERY

    As 4-5 ml of aliquot is used as a replenished medium so the below graphs estmiates the

    plot of time interval with the release of drug.The absorbance is recorded by U.Vspectrophotometer denoted by n[32].

    pH 5.8 of 1% giloy

    n=1.4205

    pH 6 of 1% giloy

    n=1.8148

    0

    0.5

    1

    1.5

    2

    2.5

    3

    3.5

    4

    0 0.2 0.4 0.6 0.8 1 1.2

    log(mt)

    log(t)

    pH 5.8 0f 1% giloy film

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    Fig-3.0

    pH 7 of 1% giloy

    n=1.7366

    Fig-3.1

    pH 5.8 of 5% giloy

    n=1.4544

    0

    0.5

    1

    1.5

    2

    2.5

    3

    3.5

    4

    0 0.2 0.4 0.6 0.8 1 1.2

    log(mt)

    log(t)

    pH 6 of 1% giloy film

    0

    0.5

    1

    1.5

    2

    2.5

    3

    3.5

    0 0.2 0.4 0.6 0.8 1 1.2

    log(mt)

    log(t)

    pH 7 of 1% giloy film

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    Fig-3.2

    pH 6 of 5% giloy

    n=1.7866(absorbance)

    Fig-3.3

    pH 7 of 5%

    n=1.66999(

    0

    0.5

    1

    1.5

    2

    2.5

    3

    3.5

    4

    0 0.2 0.4 0.6 0.8 1 1.2

    log(mt)

    log(t)

    pH 5.8 of 5% giloy film

    0

    0.5

    1

    1.5

    2

    2.5

    3

    3.5

    4

    0 0.2 0.4 0.6 0.8 1 1.2

    log(mt)

    log(t)

    pH 6 of 5% giloy film

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    Fig-3.4

    pH 5.8 of 7.5% giloy

    n=1.4829

    Fig-3.4

    pH 6 of 7.5% giloy

    n=1.7718

    0

    0.5

    1

    1.5

    2

    2.5

    3

    3.5

    0 0.2 0.4 0.6 0.8 1 1.2

    log(mt)

    log(t)

    pH 7 of 5% giloy film

    0

    0.5

    1

    1.5

    2

    2.5

    3

    3.5

    4

    0 0.2 0.4 0.6 0.8 1 1.2

    log(mt)

    log(t)

    pH 5.8 OF 7.5% GILOY FILM

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    Fig-3.5

    Fig-3.6

    pH 7 of 7.5% giloy

    n=2.5048

    0

    0.5

    1

    1.5

    2

    2.5

    3

    3.5

    0 0.2 0.4 0.6 0.8 1 1.2

    log(mt)

    log(t)

    pH 6 of 7.5% giloy film

    0

    0.5

    1

    1.5

    2

    2.5

    3

    3.5

    0 0.2 0.4 0.6 0.8 1 1.2

    log(mt)

    log(t)

    pH 7 of 7.5% giloy film

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    Fig-3.7

    pH 5.8 of 10% giloy

    n=1.4826

    pH 6 of 10% giloy

    n=1.7859

    0

    0.5

    1

    1.5

    2

    2.5

    3

    3.5

    4

    0 0.2 0.4 0.6 0.8 1 1.2

    log(mt)

    log(t)

    pH 5.8 OF 10% GILOY FILM

    -5000

    0

    5000

    10000

    15000

    20000

    25000

    30000

    35000

    0 0.2 0.4 0.6 0.8 1 1.2

    log(mt)

    log(t)

    pH 6 of 10% giloy film

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    Fig-3.5 pH 7 of 10% giloy

    n=1.9828

    Where n= absorbancy

    0

    0.5

    1

    1.5

    2

    2.5

    3

    3.5

    0 0.2 0.4 0.6 0.8 1 1.2

    log(mt)

    log(t)

    pH 7 of 10% giloy film

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    DISCUSSION AND CONCLUSION

    CHAPTER 6

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    DISCUSSION AND CONCLUSION

    Giloy( Tinospora cordifolia) was used for treatment of various skin dieases ,piles ,dibetes

    swine flu,malaria,various other fever conjunctivitis ,Immunity Disorders, StomachUlcerUrinary disorder Mental Disorder enzymes.

    So it was very important for human being. During the above experiments when the

    dissolution experiments were carried out and are observed with the absorbancy methods.the

    rates of dissolution that are termed at least possible P.H dissolved and disintegrated at a

    faster rate. The total ash value is an indicative of total amount of inorganic material after

    complete incineration and the acid insoluble ash value is an indicative of silicate impurities,

    which might have arisen due to improper washing of drug.The loss on drying value obtained

    is an indicative of amount of moisture content present in the drug. The extractive values

    names water soluble and alcohol soluble indicates the amount of active constituent in given

    amount of plant material when extracted with respective solvent, values obtained supports

    the fact that drug is unexhausted which is contrary to lower extractive value[6].

    In summary, a very simple, basic method was developed to fabricate drug-loaded

    biocompatible polymeric films by directly spreading polymer/drug solution on the

    nonsolvent surface. Tamarind polymer were used as the model polymer and drug,

    respectively.

    By controlling the weight ratio of giloy, different drug loading percentage films can be

    prepared. The drug release behaviors of the as-prepared products show their potential

    applications in a drug controlled delivery system. The release rate of drug(giloy) from these

    films into phosphate buffered solutions appeared to depend on a number of factors including

    drug loading content and the pH of the release mediums. This method can easily be scaled

    up and potentially extended to the fabrication of other drug-loaded composites for theapplication in drug delivery systems.Also giloy has shown anti-microbial activity that

    extends the purpose of being only a preparative medicine in various parts of world it is used

    extensively for curing illness by simply warming the tuber roots of the plant or chewing the

    stem and leaf of the plant.Present study simply enlightens the fact that the herbal drug can be

    formulated in different ways and can be pelleted by using different polymer systems and

    based on which the coating and pelleting can be done[32].

    As we are discussing the In-vitro release rate of sublingual tablets will becarried out using

    United State Pharmacopoeia (USP) XXIV dissolution testing apparatus (Paddle method). A

    aliquot sample of the solution is withdrawn from the dissolution apparatus. The samples are

    replaced with fresh dissolution medium of same quantity. The samples are filtered through

    Whatman filter paper No 40 and analyzed in UV spectrophotometer. The percentage drugrelease is calculated using an equation obtained from the calibration curve[5]

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    CHAPTER 7

    REFERNCES

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    REFERENCES

    [1].Wikipedia.The free Encyclopediahttp://en.wikipedia.org/wiki/Arthritis

    [2] Wikipedia.The free Encyclopedia

    http://en.wikipedia.org/wiki/Drug_delivery

    [3] An Overview on: Sublingual Route for Systemic Drug Delivery K. Patel

    Nibha1 and SS. Pancholi2*1Department of Pharmaceutics, BITS Institute of

    Pharmacy, Gujarat Technological university, Varnama, Vadodara, Gujarat, Ind.

    [4]Standardization and phytochemical evaluation of tinospora cordifolia(willd.) miers. (menispermaceae) shivani tanwar1*, jainendra jain1, sristi

    verma1, deepa solanki2

    [5]Quality control methods for herbal materials.Updated edition of Quality

    control methods for medicinal plant materials, 1998 1. Plants, Medicinal. 2.

    Medicine, Herbal. 3. Medicine, Traditional. 4. Quality control. 5. Manuals. I.

    World Health Organization.

    ISBN 978 92 4 150073 9

    [6]Evaluation and Standardization of Marketed Polyherbal Formulation

    Arogya Vati Yogendr Bahuguna*, Jaseem Saqib, Praveen Kumar, Ashutosh

    Badola

    [7].Alteration of lethal effects of gamma rays in Swiss albino mice by

    Tinospora cordifolia. Pahadiya S, Sharma J. Phytother Res. 2003

    May;17(5):552-4[8]Evaluation of the antineoplastic activity of guduchi (Tinospora cordifolia) in

    Ehrlich ascites carcinoma bearing mice. Grover J.K, Rao SK

    [9].Tinospora cordifolia induces enzymes of carcinogen/drug metabolism

    http://en.wikipedia.org/wiki/Arthritishttp://en.wikipedia.org/wiki/Arthritishttp://en.wikipedia.org/wiki/Arthritishttp://en.wikipedia.org/wiki/Drug_deliveryhttp://en.wikipedia.org/wiki/Drug_deliveryhttp://en.wikipedia.org/wiki/Drug_deliveryhttp://en.wikipedia.org/wiki/Arthritis
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    and antioxidant system, and inhibits lipid peroxidation in mice.Singh RP,

    Banerjee S, Kumar PV, Raveesha KA, Mary N.K. Phytomedicine. 2006

    Jan;13(1-2):74-84. Epub 2005 Jun 29

    [10]Sharma.A.Antifertility effect of Tinospora cordifolia (Willd.) stem extractin male rats. GuptaRS,IndianJExpBiol.2003Aug;41(8):885-9.

    [11]Radioprotective potential of an herbal extract of Tinospora cordifolia.

    Goel HC, Prasad J, Singh S, Sagar RK, Agrawala PK, Bala M, Sinha AK,

    Umamaheswari.S. J Radiat Res (Tokyo). 2004 Mar;45(1):61-8.

    [12]Anti-hyperglycemic effect of Eugenia jambolana and Tinospora cordifolia

    in experimental diabetes and their effects on key metabolic enzymes involved

    in carbohydrate metabolism.J K Grover,V Vats,S S Rathi

    [13] Restoration of antioxidant defence by ethanolic Tinospora cordifolia root

    extract in alloxan-induced diabetic liver and kidney.Prince PS, Padmanabhan

    M, Mathew S.

    [14] Effect of alcoholic extract of Ayurvedic herb Tinospora cordifolia on theproliferation and myeloid differentiation of bone marrow precursor cells in a

    tumor-bearing host.Singh SM, Singh N, Shrivastava P.

    [15] Immune stimulating properties of a novel polysaccharide from the

    medicinal plant Tinospora cordifolia. Nair PK, Rodriguez S, Ramachandran R,Alamo A, Thatte.U.L Int Immunopharmacol. 2004 Dec 15;4(13):1645-59.

    [16] Rubia cordifolia, Fagonia cretica linn and Tinospora cordifolia exertneuroprotection by modulating the antioxidant system in rat hippocampal slices

    subjected to oxygen glucose deprivation.Rawal AK, Muddeshwar MG,

    Chaudhry R.

    http://www.researchgate.net/publication/12236852_Anti-hyperglycemic_effect_of_Eugenia_jambolana_and_Tinospora_cordifolia_in_experimental_diabetes_and_their_effects_on_key_metabolic_enzymes_involved_in_carbohydrate_metabolismhttp://www.researchgate.net/publication/12236852_Anti-hyperglycemic_effect_of_Eugenia_jambolana_and_Tinospora_cordifolia_in_experimental_diabetes_and_their_effects_on_key_metabolic_enzymes_involved_in_carbohydrate_metabolismhttp://www.researchgate.net/publication/12236852_Anti-hyperglycemic_effect_of_Eugenia_jambolana_and_Tinospora_cordifolia_in_experimental_diabetes_and_their_effects_on_key_metabolic_enzymes_involved_in_carbohydrate_metabolismhttp://www.researchgate.net/researcher/39942265_V_Vatshttp://www.researchgate.net/researcher/39198693_S_S_Rathihttp://www.researchgate.net/researcher/39198693_S_S_Rathihttp://www.researchgate.net/researcher/39942265_V_Vatshttp://www.researchgate.net/publication/12236852_Anti-hyperglycemic_effect_of_Eugenia_jambolana_and_Tinospora_cordifolia_in_experimental_diabetes_and_their_effects_on_key_metabolic_enzymes_involved_in_carbohydrate_metabolismhttp://www.researchgate.net/publication/12236852_Anti-hyperglycemic_effect_of_Eugenia_jambolana_and_Tinospora_cordifolia_in_experimental_diabetes_and_their_effects_on_key_metabolic_enzymes_involved_in_carbohydrate_metabolismhttp://www.researchgate.net/publication/12236852_Anti-hyperglycemic_effect_of_Eugenia_jambolana_and_Tinospora_cordifolia_in_experimental_diabetes_and_their_effects_on_key_metabolic_enzymes_involved_in_carbohydrate_metabolismhttp://www.researchgate.net/publication/12236852_Anti-hyperglycemic_effect_of_Eugenia_jambolana_and_Tinospora_cordifolia_in_experimental_diabetes_and_their_effects_on_key_metabolic_enzymes_involved_in_carbohydrate_metabolism
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    [17] Comparative evaluation of hypoglycaemic activity of some Indian

    medicinal plants in alloxan diabetic rats.Ajit Kar,B K Choudhary,N G

    Bandyopadhyay Satsang Herbal Research and Analytical Laboratories, PO

    Satsang-814 116 Deoghar, India. Journal of Ethnopharmacology (Impact

    Factor: 2.76). 02/2003; 84(1):105-8.

    [18] Nemmani KV, Jena GB, Dey CS, Kaul CL, Ramarao P (2002) Cell

    proliferation and natural killer cell activity by poly herbal formulation, Immu-

    nity in mice. Indian J Exp. bioi. 40 (3): 282-287.

    [19] Antioxidant and Lipophilic Constituents of Tinospora crispa. Cavin A,Hostettmann K, Dyatmyko W, Dhuley J.N. Planta Med. 1998 Jun;64(5):393-6.

    [20]. Hepatoprotective and immunomodulatory properties of Tinospora

    cordifolia in CCl4 intoxicated mature albino rats. Bishayi B, Roychowdhury S,

    Ghosh S, Wadood N. J Toxicol Sci. 2002 Aug;27(3):139-46

    [21] Effect of Tinospora cordifolia on the antitumor activity of tumorassociated

    macrophages-derived dendritic cells.Singh N, Singh SM, Shrivastava P Int

    Immunopharmacol. 2004 Dec 15;4(13):1569-75.

    Effect of Tinospora cordifolia on the cytokine profile of angiogenesisinducedanimals.Leyon PV, Kuttan G.

    [22]Tinospora cordifolia induces enzymes of carcinogen/drug metabolism andantioxidant system, and inhibits lipid peroxidation in mice. Singh RP, Banerjee

    S, leyon PV, Raveesha KA, Rao AR.

    [23] Evaluation of the antineoplastic activity of guduchi (Tinospora cordifolia)

    in Ehrlich ascites carcinoma bearing mice.Jagetia GC, Rao SK.

    [24] Effect of Tinospora cordifolia on the antitumor activity of tumorassociated

    macrophages-derived dendritic cells.

    Singh N, Singh SM, Shrivastava P

    http://www.researchgate.net/researcher/60261206_Ajit_Karhttp://www.researchgate.net/researcher/8064041_B_K_Choudharyhttp://www.researchgate.net/researcher/8064042_N_G_Bandyopadhyayhttp://www.researchgate.net/researcher/8064042_N_G_Bandyopadhyayhttp://www.researchgate.net/journal/0378-8741_Journal_of_Ethnopharmacologyhttp://www.researchgate.net/journal/0378-8741_Journal_of_Ethnopharmacologyhttp://www.researchgate.net/researcher/8064042_N_G_Bandyopadhyayhttp://www.researchgate.net/researcher/8064042_N_G_Bandyopadhyayhttp://www.researchgate.net/researcher/8064041_B_K_Choudharyhttp://www.researchgate.net/researcher/60261206_Ajit_Kar
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    .[25]. A study of drug release from homogeneous PLGA microstructures. J

    Contr Release. Acharya G, Shin CS, Atal C,K, et al 2010;146:201206.

    [26] Restoration of antioxidants by ethanolic Tinospora cordifolia in

    alloxaninduced diabetic Wistar rats.Prince P, Kamalakkannan N, StanleyManzen.

    [27] The influence of ph on drug release from metformin hcl matrices

    containing different grades of hydroxypropyl methyl cellulose Masheer Ahmed

    Khan,Gangan V.D.

    [28] ] Effect of alcoholic extract of Ayurvedic herb Tinospora cordifolia on the

    proliferation and myeloid differentiation of bone marrow precursor cells in a

    tumor-bearing host.Singh SM, Singh N, Jaffer N.

    [29] Quality control methods for herbal materials.Updated edition of Quality

    control methods for medicinal plant materials, 1998 1. Plants, Medicinal. 2.

    Medicine, Herbal. 3. Medicine, Traditional. 4. Quality control. 5. Manuals. I.

    World Health Organization.

    ISBN 978 92 4 150073 9

    [30] Evaluation and Standardization of Marketed Polyherbal Formulation

    Arogya Vati Yogendr Bahuguna*, Jaseem Saqib, Praveen Kumar, AshutoshBadola

    [31] Standardization and phytochemical evaluation of tinospora cordifolia(willd.) miers. (menispermaceae) shivani tanwar1*, jainendra jain1, sristi

    verma1, deepa solanki2

    [32] Modeling and comparison of dissolution profilesPaulo Costa*,JoseManuel Sousa Lobo

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    APPENDIX

    REAGENTS AND SOLUTIONS

    1) 0.1N AgNo:-Dissolve 17gm of AgNoin sufficient amount of destiled water and make

    the volume 100 ml.

    Standardisation:- Titrate the above prepared solution with 0.1N Nacl using KCrO

    (potassium dichromate) as an indicator to slight and Red pipette is found.

    Calculation:- Actual Normality of AgNo= Normality of Nacl 10/Burette Reading.

    2).1N KCrO:- A 7% w/v solution of KCrOdissolve 4.9gm.of Potassium dichromate in

    sufficient water to produce 100 ml.

    Standardisation:- 20ml of 0.1N sodium thiosulphate add in 1gm.of potassium iodide and 7ml

    of 2M Hcl add in 250ml of water and titrate with above prepared solution using 3ml of

    starch indicator until the colour changes from blue to light green or green to blue.

    Calculation:-Actual Normality of KCro=

    Normality of NaSO20

    3) 0.1M EDTA:-Dissolve 37.225gm of disodium EDTA in a small amount of destiled water

    and make up to 1000ml of destiled water.

    Standardisation:-Weigh 200mg or 2gms of CaCoin a 250ml of conical flask and add 50 ml

    of HO and then 2ml of dilute Hcl added till solution dissolves then it is boiled on a hot plate

    till effervescence(CaCo gas) evolved, then cooled and 10ml of dilute ammonia solution

    along with 15ml of ammonia ammonium chloride buffer(NHClNH) was added to render

    the solution highly Alkalines then a pinch of 10ml of Erechrome black Tea indicator was

    added and immediately titrated above prepared EDTA solution.

    The end point sharp change in the colour mostly pink to blue or blue to pink.

    Calculation:-1ml of 0.01M EDTA = 1mg.CaCo

    Actual Normality of EDTA = The artificial Normality(0.02N)Mass of neutralizing

    sample(2gm CaCo)/Eqquivalence factor of sample(0.001gm)Actual burette reading

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    4) Ammonia Ammonium chloride buffer:-Dissolve 67.5gm.of Ammonium chloride in

    200ml of water and add 570ml of strong ammonia solution and dilute with HO to 1000ml.

    5) Ammonia Buffer(pH 10.0):-Dissolve 5.4 gm of Ammonium chloride in 20ml of HO, and

    35 ml of 10M Ammonia solution and dilute with water to 100ml.

    6 )0.1N NaSO:-Dissolve 24.82 or 25gm of NaSOin small amount of water along with

    0.1gm of sodium carbonate as preservative and make upto 1000 ml destiled water.

    Standardisation of NaSO:-weighy .2gm of Pottasium dichromate and add 100ml of

    destiled water then 7ml concentrated Hcl are added and again added 25ml of 10% of

    potassium iodide,finally titrate with the sodium theosulfate above prepared solution using

    starch indicator.

    Calculation:-Normality of NaSO= Weight of KCrO/.0490Burette reading.

    7) 0.1N NaOH:-Dissolve 4gm of pure NaOH in sufficient destiled water and make it volume

    1000ml.

    Standardisation:-Take 20 ml of above prepared solution in 200ml of conical flask and titrate

    with 0.1N Hcl using Phenopthaline as an indicator.

    Calculation:-Normality of NaoH = Normality of HclBurette Reading/20

    8)Methyl Orange Indicator:-Dissolve .1gm of methyl orange in some of HO and add

    sufficient ethanol(95%) to produced 100ml.

    9)Methyl Red Indicator:-Dissolve 50mg(0.05) of Methyle red in a mixture of 1.86ml of .1M

    NaOH and 50ml of ethanol(95%).Add sufficient amount of HO to produce 100ml.

    10)Ferricallum Indicator:-Prepare a saturated solution of Ferric ammonium sulfate in HO

    about 40% and add a few drops of 6N HNO.

    11)Phenopthaline Indicator:-Dissolve 1gm of purest grade phenopthaline in 50ml of Alcohol

    and dilute with water to 100ml.Add .1N NaOH solution until a faint pink colour

    appears.Remove this colour with ,1N Acid(HSo).

    12)Starch Indicator:-Dissolve 1gm of arrow root starch tapioca starch in small amount of

    water and add this content to 100ml of boiled distilled water contains stirring and then