TECHNOCRATS INSTITUTE OF TECHNOLOGY PHARMACY�

PY504

PHARMACOGNOSY

Model Question Paper

1-Q(a)WHAT IS PRIMARY METABOLITES?

Ans :- Primary metabolites:- A primary metabolite is a type of metabolite that is directly involved in normal growth, development, and reproduction. It generally performs a physiological function in the organism (i.e. an essential function). A primary metabolite is typically present in many organisms or cells. It is also referred to as a central metabolite, which has an even more restricted meaning (present in any initially growing cell or organism)

Examples: ethanol, lactic acid, sugar, fatty acid & amino acids

Q(b)WHAT IS SECONDRY METABOLITES?

Ans :- Secondary metabolites are organic compounds that are not directly involved in the normal growth, development, or reproduction of an organism. Unlike primary metabolites, they represent chemical adaptations to environmental stresses, or serve as defensive, protective or offensive chemicals against microorganisms, insects and higher herbivorous predators. They are sometimes considered as waste or secretary products of plant metabolism and are of pharmaceutical importance.

Examples of secondary metabolites include:- alkaloids, glycosides, flavonoids, volatile oils

Q (c) WRITE A NOTE ON SHIKMIC ACID PATHWAY?

Ans :- This appears to be an important route from carbohydrate for the biosynthesis of the C6-C3 units (phenylpropane derivatives) , of which phenylalanine & tyrosine are both examples. A scheme of biogenesis for these aromatic amino acids, as elucidated in various organism for higher plants, the presence of the enzyme system responsible for the synthesis of shikimic acid has been confirmed.

An important branching point arises at chorismic acid ; anthranilate synthase uses chorismic acid as a substrate to give anthranilic acid which is a precursor of tryptophan. The synthesis is controlled by the latter acting as a feedback inhibitor ; chorismate mutase converts chorismic acid to prephenic acid the preculsor of phenylalanine and tyrosine and a variety of control mechanisms appear to activate at the branching point.

Q(d) Write a brief description of Alkaloid?

Ans :- INTRODUCTION:-The term alkaloids (or alkali-like) was first and foremost proposed by the pharmacist, W. Meissner, in 1819, for the basic nitrogen-containing compounds of plant origin.

Ladenburg defined alkaloids,as naturally occurring plant compounds having a basic character and�� containing at least one nitrogen in a heterocyclic ring. With the advent of recent advanced knowledge in� the chemistry of various alkaloids two more inevitable characteristic features were logically and justifiably added to the definition of alkaloids, namely:-

(a) Complex molecular structure, and (b) Significant pharmacological activity.

Furthermore, it was broadly observed that the basic properties of the alkaloids is solely by virtue of the presence of N-atom embedded into the five-or six- membered ring. Therefore, the alkaloids are now generally defined as,physiologically active basic compounds of plant origin, in which at least one�� nitrogen atom forms part of a cyclic system. Even this definition has a few anomalies as stated below,� namely:

(i) Cholines and Betaines: These two substances have the N-atom in the side chain and not in the aromatic ring as shown below: HOCH2CH2N+ (CH3)3

Choline

(CH3)3N+CH2 COO Betaine� �

The cholines and betaines are regarded as simple alkylamines and not classified as alkaloids. They are designated by some school of thoughts as biological-amines or protoalkaloids.� � � �

(ii) Ephedrine: It has the N-atom only in the side chain and not embedded in the aromatic ring as given below:

Nomenclature:-

The major characteristic of the nomenclature of alkaloids is the lack of any agreed systematic prevailing system. Therefore, by a general agreement, the chemical rules designate that the names of all alkaloids must end with the suffix (ine). The latin names end with (ina). Thus, the names of the alkaloids are� � usually obtained in a number of ways, namely:

(a) From the generic name of the plant producing them: Examples: Atropine from Atropa belladona Linn., (Solanaceae); and Hydrastine from Hydrastis canadenisis L. (Ranunculaceae).

(b) From the specific name of the plant yielding them: Examples: Belladonine from Atropa belladona L. (Solanaceae); and Cocaine from Erythroxylum coca Lam. (Erythroxylaceae).

(c) From the common name of the drug producing them: Example: Ergotamine from Claviceps purpurea (Er.) Tul. (Hypocreales) commonly known as ergot.

(d) From their specific physiological activity: Examples: Emetine from Hedera helix L. (Araliaceae) called Ivy; Narcotine from Papaver somniferum L. (Papaveraceae) known as Opium Poppy; and Morphine from P. somniferum L.

(e) From the name of the discoverer: Example: Pelletierine from the barks of Puniea granatum Linn., (Punicaceae).

(f) From their physical property: Example: Hygrine from the roots of Withania somniferum (L.) Dunal (Solanaceae) called Ashwagandh.

Classification of Alkaloid:-

Hegnauer* (1963) conveniently classified alkaloids into six important groups, corresponding to the six amino-acids legitimately considered as the starting points for their biosynthesis, such as: anthranilic acid, histidine, lysine, ornithine phenylalanine and tryptophan. Price* (1963) further took a leading clue from the earlier observation and considered in details the alkaloids present in one of the families, (Rutaceae) and logically placed them in the following nine chemical-structural categories, namely: acridines, amides, amines, benzylisoquinolines, canthinones, imidazoles, indolquinazolines, furoquinolines, and quinazolines. Another school of thought classifies alkaloids in the following four heads, namely:

(a) Biosynthetic Classification In this particular instance the significance solely lies to the precursor from which the alkaloids in question are produced in the plant biosynthetically. Therefore, it is quite convenient and also logical to group together all alkaloids having been derived from the same precursor but possessing different taxonomic distribution and pharmacological activities. Examples (i) Indole alkaloids derived from tryptophan.

(ii) Piperidine alkaloids derived from lysine.

(iii) Pyrrolidine alkaloids derived from ornithine.

(iv) Phenylethylamine alkaloids derived from tyrosine.

(v) Imidazole alkaloids derived from histidine.

(b) Chemical Classification It is probably the most widely accepted and common mode of classification of alkaloids for which the main criterion is the presence of the basic heterocyclic nucleus (i.e., the chemical entity).

Examples (i) Pyrrolidine alkaloids e.g., Hygrine;

(ii) Piperidine alkaloids e.g., Lobeline;

(iii) Pyrrolizidine alkaloids e.g., Senecionine;

(iv) Tropane alkaloids e.g., Atropine;

(v) Quinoline alkaloids e.g., Quinine;

(vi) Isoquinoline alkaloids e.g., Morphine;

(vii) Aporphine alkaloids e.g., Boldine;

(viii) Indole alkaloids e.g., Ergometrine

; (ix) Imidazole alkaloids e.g., Pilocarpine;

(x) Diazocin alkaloids e.g., Lupanine;

(xi) Purine alkaloids e.g., Caffeine;

(xii) Steroidal alkaloids e.g., Solanidine;

(xiii) Amino alkaloids e.g., Ephedrine;

(xiv) Diterpene alkaloids e.g., Aconitine.

(c) Pharmacological Classification Interestingly, the alkaloids exhibit a broad range of very specific pharmacological characteristics. Perhaps this might also be used as a strong basis for the general classification of the wide-spectrum of alkaloids derived from the plant kingdom, such as:

analgesics, cardio-vascular drugs, CNS-stimulants and depressants, dilation of pupil of eye, mydriatics, anticholinergics, sympathomimetics, antimalarials, purgatives, and the like. However, such a classification is not quite common and broadly known.

Examples (i) Morphine as Narcotic analgesic;

(ii) Quinine as Antimalarial;

(iii) Strychnine as Reflex excitability;

(iv) Lobeline as Respiratory stimulant;

(v) Boldine as Choleretics and laxatives;

(vi) Aconitine as Neuralgia;

(vii) Pilocarpine as Antiglaucoma agent and miotic;

(viii) Ergonovine as Oxytocic;

(ix) Ephedrine as Bronchodilator;

(x) Narceine as Analgesic (narcotic) and antitussive.

(d) Taxonomic Classification This particular classification essentially deals with the Taxon i.e., the� � taxonomic category. The most common taxa are the genus, subgenus, species, subspecies, and variety. Therefore, the taxonomic classification encompasses the plethora of alkaloids exclusively based on their respective distribution in a variety of Plant Families, sometimes also referred to as the Natural order. A� � few typical examples of plant families and the various species associated with them are stated below, namely:

(i) Cannabinaceous Alkaloids: e.g., Cannabis sativa Linn., (Hemp, Marijuana).

(ii) Rubiaceous Alkaloids: e.g., Cinchona Sp. (Quinine); Mitragyna speciosa Korth (Katum, Kratum, Kutum); Pausinystalia johimbe (K. Schum) (Yohimbe).

Characteristic of Alkaloid:-

Atropine sulphate and morphine hydrochloride are much more soluble in water than their corresponding bases i.e., atropine and morphine. However, there are a few exceptions to the above stated generalizations, namely:

(i) Certain alkaloid bases are water soluble, but these may be solely regarded as exceptions rather than any specific rule, such as: ephedrine, colchicine, pilocarpine; the quaternary alkaloid-base like berberine and tubocurarine; caffeine-base readily extracted from tea with water.

(ii) Narceine and pilocarpine are insoluble in organic solvents, whereas morphine is sparingly soluble in organic solvents viz., solubility in either 1:5000.

(iii) Certain alkaloidal salts, for instance: lobeline hydrochloride and apoatropine hydrochloride are found to be soluble in organic solvent like chloroform.

(iv) Some alkaloidal salts are sparingly soluble in water whereas others are extremely watersoluble, such as: Quinine sulphate-soluble in 1:1000 parts of water, Quinine hydrochloridesoluble in 1:1 part of water

Silent feature:-

1. The weaker bases, i.e., alkaloids having low pKa values, shall require a more acidic medium to form their respective salts with the corresponding acid.

2. The strongly basic alkaloids i.e., those possessing high pKa values, shall require comparatively low acidic medium to form their respective salts with the acid. Note: In a medium at a weakly acidic pH certain strongly basic alkaloids would be easily converted to their respective salt by interaction with the corresponding acid, whereas the alkaloids which are relatively weaker bases having lower pKa values shall still remain in their free-base form. Such a critical situation is skillfully exploited for the separation of a specific alkaloid or a group of alkaloids having closely identical pKa values, from other alkaloids that essentially possess either very low or very high pKa values.

3. The alkaloids are usually neutrallized with acids to form salts that may be converted to the corresponding free-base by the cautious addition of selective weak bases, such as, ammonia, calcium hydroxide or sodium carbonate. The usage of either NaOH or KOH solutions must be avoided so as to prevent the decomposition or destruction of highly sensitive alkaloids.

4. Amphoteric alkaloids: There are some alkaloids which are amphoteric in nature i.e., they are neither acidic nor basic in character; this is due to the presence of phenolic (OH) moiety in Morphine, or the� presence of carboxylic (COOH) function in Narceine, �

Isolation of Alkaloid:-

To isolate alkaloids from plants, the dried and powdered plant material is extracted with pet ether (or hexane, colemans etc.) first. This removes fats, oils, terpenes, waxes etc. This extract is discarded.

The material is now subjected to an alcohol extraction, eg with methanol or ethanol. The extract is evaporated to leave the crude alkaloids mixture.

This extract is partitioned between an diluted aq. tartaric acid solution and ethyl acetate. Other acids like citric acid can be used, and other solvents may substitute here. The ethyl acetate layer contains neutral and weakly basic alkaloids. Evaporate the solvent to isolate them.

The aq. layer is neutralised with NH3 or Na2CO3 and again extracted with ethyl acetate. The organic layer now contains basic alkaloids, while the aq. layer contains quarternary ammonium ions.

Many alkaloids can be isolated directly from the alkoholic extract by chromatographic methods. This is a separation which works well for tropane alkaloids (atropine, cocaine, scopolamine).

The alcohol extract is fractionated by column chromatography on SiO2, solvent chloroform followed by chloroform with rising methanol content. This separates lipids and terpenes from the crude alkaloid fraction. The alkaloid fraction is again chromatographed (SiO2; CHCl3 : MeOH = 10:1) to isolate the pure alkaloids.

CHEMICAL TEST FOR ALKALOID:-

Dragendorff reagent(Potassium-bismuth-iodide solution)

Mayer reagent (Potassium-mercuric-iodide solution)

Wagner reagent (iodine-potassium-iodide solution)

Hager reagent (Saturated solution of picric acid)

(a)Mayer's Test:-

Filtrates were treated with Mayers reagent (Potassium Mercuric Iodide).Formation of a yellow coloured� precipitate indicates the presence of alkaloids.

(b)Wagner,s Test:-

Filtrates were treated with Wagners reagent (Iodine in Potassium Iodide). Formation�

of brown/reddish precipitate indicates the presence of alkaloids.

Or

Q(e).WEITE A NOTES ON PHYTOCHEMICAL SCREENING?

Ans :- INTRODUCTION:-Phytochemical screening refers to the extraction, screening and identification of the medicinally active substances found in plants.

Some of the bioactive substances that can be derived from plants are flavonoids, alkaloids, carotenoids, tannin, antioxidants and phenolic compounds. Although the knowledge of how these substances provide medicinal value to humans reflects a relatively recent scientific understanding, the use of plants and plant extracts to heal, relieve pain and promote good health.

In phytochemical evaluation the powdered leaves were subjected to

phytochemical screening for the detection of various plant constituents, characterized for their possible bioactive compounds, which have been separated and subjected to detailed structural analysis.

PREPRATION OF EXTRACT:-

Preparation of the Extracts About 2 kg of air-dried plant material was extracted in soxhlet

assembly successively with n-hexane, chloroform, ethyl acetate and methanol (order of increasing polarity). Each time before extracting with the next solvent, the powdered material was dried.

Each extract was concentrated by using rotary vacuum evaporator. The extract obtained with each solvent was weighed and the percentage yield was calculated in terms of dried weight of the plant material. The colour and consistency of the extract were also noted. All the solvents used for this entire work were of analytical reagent grade

PHYTOCHEMICAL SCREENING:-

examinations were carried out for all the extracts as per the standard methods

(1)DETECTION OF ALKALOID:-

A small portion of the extract was stirred separately with a few

drops of dilute hydrochloric acid and filtered. The filtrate was carefully tested with various alkaloidal reagents such as :-

(a)Mayer's Test:-

Filtrates were treated with Mayers reagent (Potassium Mercuric Iodide).Formation of a yellow coloured� precipitate indicates the presence of alkaloids.

(b)Wagner,s Test:-

Filtrates were treated with Wagners reagent (Iodine in Potassium Iodide). Formation�

of brown/reddish precipitate indicates the presence of alkaloids.

(c)Dragendroff's Test:-

Filtrates were treated with Dragendroffs reagent (solution of Potassium Bismuth Iodide). Formation of� red precipitate indicates the presence of alkaloids.

(d)Hager's Test:-

Filtrates were treated with Hagers reagent (saturated picric acid solution). Presence of alkaloids� confirmed by the formation of yellow coloured precipitate..

(2)DETECTION OF CARBOHYDRATES:-

The minimum amount of the extracts were dissolved in 5ml of distilled water and filtered. The filtrate was subjected to test for carbohydrates.

(a)Molisch Test:-

Filtrates were treated with 2 drops of alcoholic α-naphthol solution in a test tube. Formation of the violet ring at the junction indicates the presence of Carbohydrate

(b)Benedict'sTest

Filtrates were treated with Benedict’s reagent and heated gently. Orange red precipitate indicates the presence of reducing sugars.

(c)Fehling's Test

Filtrates were hydrolysed with dil. HCl, neutralized with alkali and heated with Fehlings A & B solutions.� Formation of red precipitate indicates the presence of reducing sugars.

(3)DETECTION OF GLYCOSIDES:-

Extracts were hydrolysed with dil. HCl, and then subjected to test for glycosides.

(a)Modified borntrager's Test:-

Extracts were treated with Ferric Chloride solution and immersed in boiling water for about 5 minutes. The mixture was cooled and

extracted with equal volumes of benzene. The benzene layer was separated and treated with ammonia solution. Formation of rose-pink colour in the ammonical layer indicates the presence of anthranol glycosides.

(b)Legal's Test:- Extracts were treated with sodium nitropruside in pyridine and sodium hydroxide. Formation of pink to blood red colour indicates the presence of cardiac glycosides.

(4)DETECTION OF SAPONIN:-

(a)Froth Test:- Extracts were diluted with distilled water to 20ml and this was shaken in a graduated cylinder for 15 minutes. Formation of 1 cm layer of foam indicates the presence of saponins.

(b)Foam Test:-: 0.5 gm of extract was shaken with 2 ml of water. If foam produced persists for ten minutes it indicates the presence of saponins.

(5)DETECTION OF PHYTOSTEROL:-

(a)Salkowski's Test:-Extracts were treated with chloroform and filtered. The filtrates were treated with few drops of Conc. Sulphuric acid, shaken and allowed to stand. Appearance of golden yellow colour indicates the presence of triterpenes.

(2)Libermann Burchard'sTest:-Extracts were treated with chloroform and filtered. The filtrates were treated with few drops of acetic anhydride, boiled and cooled. Conc. Sulphuric acid was added. Formation of12brown ring at the junction indicates the presence of phytosterols.

(6)DETECTION OF PHENOL:-

Ferric Chloride Test: Extracts were treated with 3-4 drops of ferric chloride solution. Formation of bluish black colour indicates the presence of phenols.

(7)DETECTION OF TANNIN:-

(a)Gelatin Test:-

To the extract, 1% gelatin solution containing sodium chloride was added. Formation of white precipitate indicates the presence of tannins.

(8)DETECTION OF FLAVONOIDS:-

(a)Alkaline Reagent Test:-

Extracts were treated with few drops of sodium hydroxide solution. Formation of intense yellow colour, which becomes colourless on addition of dilute acid, indicates the presence of flavonoids.

(a)Lead acetate Test:-

Extracts were treated with few drops of lead acetate solution. Formation of yellow colour precipitate indicates the presence of flavonoids.

(9)DETECTION OF PROTEIN:-

(a)Xanthoproteic Test:-

The extracts were treated with few drops of conc. Nitric acid. Formation of yellow colour indicates the presence of proteins.

(b) Ninhydrin Test:-

To the extract, 0.25% w/v ninhydrin reagent was added and boiled for few minutes. Formation of blue colour indicates the presence of amino acid.

(10) DETECTION OF DITERPENES:-

Copper acetate Test: Extracts were dissolved in water and treated with 3-4 drops of copper acetate solution. Formation of emerald green colour indicates the presence of diterpenes

(11) DETECTION OF FIXED OIL AND FAT:-

(a) Small quantity of various extracts was separately pressed between two filter papers. Appearance of oil stain on the paper indicates the presence of fixed oil.

(b) Few drops of 0.5N alcoholic potassium hydroxide was added to a small quantity of various extracts along with a drop of phenolphthalein. The mixture was heated on a water bath for 1-2hrs. Formation of soap or partial neutralization of alkali indicates the presence of fixed oil and fats.

THIN LAYER CHROMATOGRAPHY; TLCinvolve various methods of separating and isolating plant constituents, thin layer chromatography (TLC) is one of the most powerful technique used for the separation, identification and estimation of single or mixture of components present in various extracts.

Mechanism employed in this reliable technique is adsorption in which solute adsorbs on the stationary phase according to its affinity. Substances are separated by differential migration that occurs when a solvent flows along the thin layer of stationary phase. The substance which is having more affinity towards mobile phase moves faster when compared to the substance which has less affinity leading to the separation of the compounds.

Tlc plate:-Precoated silica gel on aluminium plates were used as a stationary phase.

Sample application:-The extracts to be analysed were diluted with respective solvents and then spotted with help of capillary tube just 2 cm above its bottom.

Selection of mobile phase:-

Solvent mixture was selected on the basis of the phyto constituents present in each extract. Solvents were analysed as its order of increasing polarity. Several mobile phases were tried for the separation of maximum components. After several trials, the best solvent system was selected which showed good separation with maximum number of components.

Solvent system:-

- Methanol :Chloroform(9:1)

Chloroform extract - Methanol :Chloroform(9:1) :

Ethyl acetate: Hexane: Acetic acid (2:7:1: 0.5)

Methanol extract - Ethyl acetate: Water ( 6:3:1)

Rf values were noted down for each selected extracts after elution

by using different detecting agents such as Dragendroffs, Ninhydrin, Libermann Burchard, concentrated� sulphuric acid and ferric chloride.

2- Q(a)DEFINE PHYTOCONSTITUTE?

Ans :- Definition of phytoconstituents:- Phytoconstituents are chemical compounds that take place naturally in plants (phyto means "plant" in Greek). Some are responsible for colour and other organoleptic properties, such as the deep purple of blueberries and the smell of garlic. The term is generally used to refer biologically significant chemicals, for example antioxidants, but are not established as essential nutrients. Scientists estimate[ that there may be more than 10,000 different phytoconstituents having the potential to affect diseases such as cancer, stroke or metabolic syndrome.

Q(b).DEFINE ALKALOID?

Ans :- The term alkaloids (or alkali-like) was first and foremost proposed by the pharmacist, W. Meissner, in 1819, for the basic nitrogen-containing compounds of plant origin.

Ladenburg defined alkaloids,as naturally occurring plant compounds having a basic character and�� containing at least one nitrogen in a heterocyclic ring. With the advent of recent advanced knowledge in� the chemistry of various alkaloids two more inevitable characteristic features were logically and justifiably added to the definition of alkaloids, namely.

Q(c)WRITE PHARMACOGNOSY OF RAUWOLFIA?

Ans :- Synonym:- Indian Snakeroot Or Sarpagandha

Biological source:- Rauwolfia consists of the dried rhizome and roots of Rauwolfia serpentina

Family:- apocynaceae

Macrosopical characters:-

Colour of root- The outer surface is greyish-yellow, light brown or brown with slight wrinkles (young pieces) or longitudinal ridges (older pieces)Odour- slight odour

Taste- BitterChemical Constituents:-

Rauwolfia contains at least 30 alkaloids. Which total some 0.7- 2.4%. Other substances present include phytosterols, Fatty acids, unsaturated alcohols & sugars.

It also contains a number of chemicals, including ajmaline, aricine, corynanthine,

deserpidine,lankanescine rauwolscine, rescinnamine, reserpine, reserpiline, isoreserpine, isoreserpiline, serpentinine, and yohimbine.

Reserpine yohimbine

usesNausea & vomiting Weight gainDryness of mouth

Q(d).WHAT IS RADIO TRACER TECHNOLOGY?

Ans :- A radioactive tracer, or radioactive label, is a chemical compound in which one or more atoms

have been replaced by a radioisotope so by virtue of its radioactive decay it can be used to explore the mechanism of chemical reactions by tracing the path that the radioisotope follows from reactants to products. Radiolabeling is thus the radioactive form of isotopic labeling.

PRINCIPLE:-

It is based on the principle that a stable isotope is replaced by a radioisotope.The radioisotope is capable of emitting radiations that can be detected andanalysed.

It is powerful than chemical reactions as they can be detected even inlower concentration as seen inside a cell.

In general, isotopes of hydrogen,carbon, phosphorus, sulphur, and iodine have been used extensively to trace thepath of biochemical reactions.

ISOTOPES:-

Isotopes of a chemical element differ only in the mass number. For example, the isotopes of hydrogen can be written as 1H, 2H and 3H, with the mass number at top left. When the atomic nucleus of an isotope is unstable, compounds containing this isotope are radioactive.

USE OF ISOTOPES:-

Ca-Studying cellular function & bone formation

Carbon14- Metabolism

Cesium137 -Cancer treatment

Chromium51- RBC studies

Cobalt57 -Diagnosis of pernicious anemia

Cobalt60 -Sterilization of surgical instruments

Copper67 -Treat cancer

ADVANTAGE:-

the advantages of using radiotracers. Some of them are

(1) the radiation emitted by radiotracers is generally easy to detect and measure with high precision

(2) the radiation emitted is independent of pressure, temperature, chemical and physical state,

(3) radiotracers do not affect the system and can be used in nondestructive techniques, and

(4) the radiation intensities measured furnish direct information concerning the amount of the labeled species and no special models are required to draw quantitative conclusions.

The radioactive isotopes are chemically identical to stable isotopes of the same element.

An important assumption in the use of the tracer technique is that the raelement

emitted does not affect the chemical and physical properties of the system under investigation, and the decay product following the radioactive disintegration does not have any influence on the behavior of the system. In case of radiotracers with short half-life, the changes caused by the decay of the radioisotope are taken into consideration.

REQUIREMENT:-

There are two main ways in which radioactive tracers are used

(1)Preparation of labelled compound:-

When a labeled chemical compound undergoes chemical reactions one or more of the products will contain the radioactive label. Analysis of what happens to the radioactive isotope provides detailed information on the mechanism of the chemical reaction.

In metabolism research, Tritium(3H) and 14C-labeled glucose are commonlyused to measure rates of glucose uptake, fatty acid synthesis, and othermetabolic processes.

2. In medicine, tracers are applied in a number of tests, such as 99mTc inautoradiography and nuclear medicine, including single photon emissioncomputed tomography (SPECT), positron emission tomography (PET) andscintigraphy. 3. The urea breath test for helicobacter pylori commonly used a dose of 14Clabeled urea to detect H. pylori infection.

(2)introduce labelled compound:-

A radioactive compound is introduced into a living ororganism by

Route feeding

Stem feeding

Direct injection

Flitting method

and the radio-isotope provides a means to construct an image showing the way in which that compound and its reaction products are distributed around the organism.

Criteria for Radio tracer technology:-

Starting concentration of tracer should be sufficient in amount.

Physical and chemical nature of compound should be well known.

1/2 life should be sufficient.

Radio labelled should not be damaged.

Asumption of Radio tracer technology:-

use of radiotracers is dependent on certain basic assumptions being fulfilled.

The first assumption, mentioned above, is that the radioactive isotopes of a given element behave identically as the stable isotopes of the same element. Actually this assumption is not exactly true.

The second basic assumption is that the radioactivity does not change the chemical and physical properties of the experimental system.

A third basic assumption, for biological studies, is that there is no deviation from the normal physiological state.

A fourth basic assumption is that the chemical and physical form of the radionuclide-labeled compound is the same as the unlabeled variety.

The fifth basic assumption is that only the labeled atoms are traced. Never assume that the appearance of the radioactive label in a given sample indicates the presence of the

Methods for radio tracer techniques-

Precursor-product sequence:- These a common method for the elucidation(identification) of the biosynthetic pathway, in these method presumed precursor of constituents under investigation. in a labelled form, is fed into the plant and after a suitable time the constituent its isolated and purified and its radioactivity is determined.

If specific atoms of the precursor are labelled, it may be possible to degrade the isolated metabolite and ascertain whether the distribution of radioactivity within the molecule is in accordance with the hypothesis under test.

The radio activity of isolated compound alone is not usually sufficient evidence because particular compound may enter in general metabolic pathway & form randomly distributed through the whole plant , so the double and triple-labelling experiment with different isotopes by one or two position are used.

Competitive feeding: - When be admitted in precursor a radio labelled compound there is two way of kindness-

Main pathway

Subsidiary pathway

May be obtained as a result of typically availability of admitted compound

This method value for two possible intermediates is normally used by the plant & distinguishable. The inactive B & B' fed with labelled A to separate group of plant.

If the incorporation of the activity of C is inhibited in plant receiving B but unaffected in group B' ; then conclusion is a possible pathway from A to C is possible

figure- competitive feeding

Sequential analysis:- In this method 14C is used .

The plant grow in 14C atmosphere and analysis the plant in given time interval to obtain the sequence , in which various related compound become labelled , so some routes are selected and other rejected.

Uses:- This method is generally used for elucidation of photosynthesis and also for determining the sequential formation of opium and tobacco.

Or

Q(e) WRITE A BRIEF DESCRIPTION OF ATROPINE?

Ans :- Biological source:-Atropine is found in many members of the Solanaceae family. The most commonly found sources are Atropa belladonna, Datura inoxia, D. metel, and D. stramonium

Family:-solanacea

Chemistry:-

The most common atropine compound used in medicine is atropine sulfate (monohydrate) (C17H23NO3)2·H2SO4·H2O,

the full chemical name is 1α H, 5α H-Tropan-3-α ol (±)-tropate(ester), sulfate monohydrate

structure

Pharmacological activity:-

Eyes:Topical atropine is used as a cycloplegic, to temporarily paralyze the accommodation reflex, and as a mydriatic, to dilate the pupils.

Heart:Injections of atropine are used in the treatment of bradycardia (a heart rate < 60 beats per minute).

Secretions:Atropine's actions on the parasympathetic nervous system inhibit salivary and mucus glands. The drug may also inhibit sweating via the sympathetic nervous system.

Poisonings:Atropine is not an actual antidote for organophosphate poisoning. However, by blocking the action of acetylcholine at muscarinic receptors, atropine also serves as a treatment for poisoning by organophosphate insecticides and nerve gases, such as tabun (GA), sarin (GB), soman (GD) and VX.

Contraindicated:Atropine is contraindicated in patients pre-disposed to narrow angle glaucoma.

Side effect :-Adverse reactions to atropine include ventricular fibrillation, supraventricular or ventricular tachycardia, dizziness, nausea, blurred vision, loss of balance, dilated pupils, photophobia, dry mouth

and potentially extreme confusion, dissociative hallucinations and excitation especially amongst the elderly.

Biosynthesis of pathway:-

Atropine can be synthesized by the reaction of tropine with tropic acid in the presence of hydrochloric acid.

Biosynthesis of atropine starting from L-Phenylalanine

The biosynthesis of atropine starting from l-phenylalanine first undergoes a transamination forming phenylpyruvic acid which is then reduced to phenyl-lactic acid.Coenzyme A then couples phenyl-lactic

acid with tropine forming littorine, which then undergoes a radical rearrangement initiated with a P450 enzyme forming hyoscyamine aldehyde. A dehydrogenase then reduces the aldehyde to a primary alcohol making (−)-hyoscyamine, which upon racemization forms atropine.

3-Q(a)WHAT IS STEREOISOMERS?

Ans :- When isomer is made by different arrangement of atom or group in 3d space these isomer is

called stereoisomer & the phenomenon is known as stereoisomerism.In other words it have same� �

structural formula but different in arrangement of atom in 3d space �

Figure :- Naproxen and its Stereoisomer

Q(b)DEFINE THE TERM PHYTOCHEMICAL SCREENING?

Ans :- Definition- phytochemical investigation means to investigate the plant material in terms of its active constituents.it involves the isolation of active constituents and to identify them qualitatively.

it is the primary sequential screening of various plant material metabolites / product which provide identification of secondary metabolites. Phytochemical screening is followed by a successive solvent extraction.

Q(c) GIVE THE CLASSIFICATION OF ALKALOID?

Answer � The term Alkaloid (Alkali-like) was first & foremost introduced by Pharmacist W. Meissner in 1819 used for the basic nitrogen containing compound of plant origin.

Classification Of Alkaloids:-

(a) Biosynthetic Classification:- In this particular occurrence the significance exclusively lies to the precursor from which the alkaloids in query are produced in the plant biosynthetically. Therefore, it is relatively suitable and also logical to group together all alkaloids having been derived from the same precursor but possessing diverse taxonomic distribution and pharmacological activities.

Examples

(i) Indole alkaloids derived from tryptophan.

(ii) Piperidine alkaloids derived from lysine.

(iii) Pyrrolidine alkaloids derived from ornithine.

(iv) Phenylethylamine alkaloids derived from tyrosine.

(v) Imidazole alkaloids derived from histidine

(b) Chemical Classification :- It is probably the most widely established and common mode of classification of alkaloids for which the main principle is the presence of the basic heterocyclic nucleus (i.e., the chemical entity).

Examples

(i) Pyrrolidine alkaloids e.g., Hygrine;

(ii) Piperidine alkaloids e.g., Lobeline;

(iii) Pyrrolizidine alkaloids e.g., Senecionine;

(iv) Tropane alkaloids e.g., Atropine;

(v) Quinoline alkaloids e.g., Quinine;

(vi) Isoquinoline alkaloids e.g., Morphine;

(vii) Aporphine alkaloids e.g., Boldine;

(viii) Indole alkaloids e.g., Ergometrine;

(ix) Imidazole alkaloids e.g., Pilocarpine;

(x) Diazocin alkaloids e.g., Lupanine;

(xi) Purine alkaloids e.g., Caffeine;

(xii) Steroidal alkaloids e.g., Solanidine;

(xiii) Amino alkaloids e.g., Ephedrine;

(xiv) Diterpene alkaloids e.g., Aconitine.

(c) Pharmacological Classification:- fascinatingly, the alkaloids exhibit a broad range of very precise pharmacological characteristics. possibly this might also be used as a strong basis for the general classification of the wide-spectrum of alkaloids derived from the plant kingdom, such as: analgesics, cardio-vascular drugs, CNS-stimulants and depressants, dilation of pupil of eye, mydriatics, anticholinergics, sympathomimetics, antimalarials, purgatives, and the like. However, such a classification is not quite common and largely known.

Examples

(i) Morphine as Narcotic analgesic;

(ii) Quinine as Antimalarial;

(iii) Strychnine as Reflex excitability;

(iv) Lobeline as Respiratory stimulant;

(v) Boldine as Choleretics and laxatives;

(vi) Aconitine as Neuralgia;

(vii) Pilocarpine as Antiglaucoma agent and miotic;

(viii) Ergonovine as Oxytocic;

(ix) Ephedrine as Bronchodilator;

(x) Narceine as Analgesic (narcotic) and antitussive.

Q(d)WRITE A BRIEF DESCRIPTION OF EPHEDRINE?

Ans:- It is obtained from the dried tender stems of the Chinese wonder drug Ma Huang which is being used in the Chinese systems of Medicine for more than five thousand years. It occurs in Ephedra vulgaris Hook. F. (E. gerardiana Wall); Ephedra sinica Stapf. (1-3%); Ephedra equisetina Bunge. (2%) belonging to the natural order Gentaceae; and several other Ephedra species. Besides, it is also found in the roots of Aconitum napellus L. (Ranunculaceae) (Aconite, Monkshood, Blue Rocket); and Ephedra nevadensis S. Wats. (Ephedraceae) (Mormon Tea, Nevada Jointfir).

structure

Chemistry:-

Ephedrine exhibits optical isomerism and has two chiral centres, giving rise to four stereoisomers. By convention, the pair of enantiomers with the stereochemistry (1R,2S) and (1S,2R) is designated ephedrine, while the pair of enantiomers with the stereochemistry (1R,2R) and (1S,2S) is called pseudoephedrine. Ephedrine is a substituted amphetamine and a structural methamphetamine analogue. It differs from methamphetamine only by the presence of a hydroxyl group (OH).�

The isomer which is marketed is (1R,2S)-()-ephedrine.[24]�

Ephedrine hydrochloride has a melting point of 187−188 °C.[25]

In the outdated D/L system (+)-ephedrine is also referred to as L-ephedrine and (−)-ephedrine as D-ephedrine (in the Fisher projection, then the phenyl ring is drawn at bottom).[24][26]

Often, the D/L system (with small caps) and the d/l system (with lower-case) are confused. The result is that the levorotary l-ephedrine is wrongly named L-ephedrine and the dextrorotary d-pseudoephedrine (the diastereomer) wrongly D-pseudoephedrine.

The IUPAC names of the two enantiomers are (1R,2S)- respectively (1S,2R)-2-methylamino-1-phenylpropan-1-ol. A synonym is erythro-ephedrine

Identification test:-

1. Dissolve 0.01 g of ephedrine in 1 ml water by adding a few drops of dilute HCl. To this add two drops of CuSO4 solution (5% w/v) followed by a few-drops of NaOH solution when a reddish colour is developed. Now, add 2-3 ml ether and shake the contents thoroughly; the ethereal layer turns purple while the lower aqueous layer becomes blue.

2. Dissolve 0.2 g of ephedrine in 30 ml of chloroform in a stoppered flask and shake the contents vigorously. Allow the mixture to stand for at least 12 hours at room temperature and then remove the chloroform over an electric water bath. The crystals of ephedrine hydrochloride separate out.

3. Triturate 0.05 g of ephedrine with a few crystals of [K3Fe(CN)6] i.e., potassium ferricyanide, followed by a few drops of water and heat on a water bath slowly when a distinct odour of benzaldehyde (i.e., similar to the odour of bitter almonds) in given out.

Pharmacological activity:-

l-Ehedrine is used extensively as a bronchodilator.

2. It also exerts excitatory action on the CNS and produces noticeable effects on skeletal muscles.

3. It is also employed as nasal decongestant.

Contraindicated:-

Ephedrine should not be used in conjunction with certain antidepressants, namely norepinephrine-dopamine reuptake inhibitors (NDRIs), as this increases the risk of symptoms due to excessive serum levels of norepinephrine.

Side effect :-

Cardiovascular: tachycardia, cardiac arrhythmias, angina pectoris, vasoconstriction with hypertension

Dermatological: flushing, sweating, acne vulgaris

Gastrointestinal: nausea

Genitourinary: decreased urination due to vasoconstriction of renal arteries, difficulty urinating is not uncommon, as alpha-agonists such as ephedrine constrict the internal urethral sphincter, mimicking the effects of sympathetic nervous system stimulation

Nervous system: restlessness, confusion, insomnia, mild euphoria, mania/hallucinations (rare except in previously existing psychiatric conditions), delusions, formication (may be possible, but lacks documented evidence) paranoia, hostility, panic, agitation

Respiratory: dyspnea, pulmonary edema

Biosynthesis of pathway:-

Interestingly, phenylalanine and ephedrine not only have the same carbon and nitrogen atoms but also have the same arrangement of C and N-atoms i.e., the skeleton of atoms. Noticeably, L-phenylalanine is a precursor, possessing only seven carbons, a C6C1 fragment, gets actually incorporated. It has been observed that phenylalanine undergoes metabolism, probably via cinnamic acid to benzoic acid; and this perhaps in the form of its coenzymeA ester, which is acylated with pyruvic acid and undergoes� decarboxylation during the addition

A thiamine PP-mediated mechanism is put forward for the formation of the diketone, and a transamination reaction shall give rise to cathinone. Further reduction of the carbonyl moiety from either face yields the diastereomeric norephedrine or norpseudoephedrine (Cathine). Ultimately,

N-methylation would give rise to ephedrine or pseudoephedrine

Or

Qe)WRITE A BRIEF DESCRIPTION OF ERGOMETRINE?

Ans :- synonym:-

Ergometrine; Ergobasine; Ergotocine; Ergostetrine; Ergotrate; Ergoklinine; Syntometrine

Biological source:

Biological Sources It is obtained from the seeds of Ipomea violaceae Linn. (Ipomea tricolor Cav.) belonging to family Convolvulaceae (Morning glory, Tlitliltzen, Ololiuqui); and also from the dried seeds of Rivea corymposa Hall. F. (Convolvulaceae) (Snakeplant).

structure

Chemistry:-

Chemically it is know as [8β (s)]-9, 10-Diadehydro-N-(2-hydroxy-1-methylethyl)-6-methylergoline-8-carboxamide; (C19H23N3O2).

Characteristic features:-

1. Ergonovine is obtained as tetrahedral crystals from ethyl acetate, and as fine needles from benzene. It tends to form solvated crystals having mp 162°C.

2. It has specific optical rotation [α]20D + 90° (in water).

3. Its dissociation constant is pKa 6.8.

4. It is found to be freely soluble in lower alcohols, acetone and ethyl acetate; more soluble in water than the other principal alkaloids of ergot; and slightly soluble in chloroform.

Identification Test:-

Identification Tests As per se the ergot alkaloids may be identified either by general precipitation and colour reactions or by preparing their derivatives as stated below

a)Precipitation reaction:-

(i) The ergot alkaloids are readily precipitated by the alkaloidal reagents. However, Mayers reagent is regarded to be the most sensitive test whereby on opalescence in dilutions of 1 ppm can be obtained.

(ii) Iodine solution in KI also gives an instant precipitate with very dilute solutions of ergot alkaloids.

b)colour Test:-

The most vital colour tests are given as under:

(i) Keller's Test:

To a solution of the alkaloid in glacial acetic acid add a few mg of solid FeCl3 and then add 1-2 ml of concentrated sulphuric acid along the side of the tube. The appearance of an intense blue colouration is accomplished at the junction of the two layers.

(ii) Van Urk Test:

When a solution containing an ergot alkaloid is mixed with Van Urk Reagent**, it gives rise to a characteristic deep blue colouration.

Note: (a) Van Urk Reagent may also be used in spraying developed paper chromatograms of the ergot alkaloids, and for this purpose 10% (v/v) HCl is used instead of H2SO4.

(b) The spectrophotometric assay for total ergot alkaloids is also based on the blue colour given with Van Urk Reagent.

(iii) Glyoxylic Acid Reagent Test:

Ergot alkaloids gives a blue colouration with the addition of Glyoxylic acid reagent and a few drops of concentrated H2SO4.

(iv) Fluorescence Test: The aqueous solution of the salts of ergot alkaloids produce a distinct blue fluorescence

Pharmacological activity:-

1.Ergonovine is used as an oxytocic.

2. Ergonovine maleate also acts as an oxytocic and produces much faster stimulation of the uterine muscles as compared to other ergot alkaloids.

3. Methylergonovine meleate is observed to act as an oxytocic whose actions are slightly more active and longer acting than ergonovine.

Contraindicated:-

The drug is contraindicated in pregnancy, vascular disease, and psychosis.

Side effect :-

Possible side effects include nausea, vomiting, abdominal pain, diarrhea, headache, dizziness, tinnitus, chest pain, palpitation, bradycardia, transient hypertension and other cardiac arrhythmias, dyspnea, rashes, and shock.[10] An overdose produces a characteristic poisoning, ergotism or "St. Anthony's fire": prolonged vasospasm resulting in gangrene and amputations; hallucinations and dementia; and abortions. Gastrointestinal disturbances such as diarrhea, nausea, and vomiting, are common.

Biosynthesis of pathway:-

4- Q(a)GIVE THE BIOLOGICAL SOURCE OF KURCHI?

Ans :- Synonyms:- Holarrhena Eng.; �

Kura Kurchi- Hindi;

Kutaha- Sans;

Botanical Source:- Kurchi consists of the dried stem bark of Holarrhena antidysenterica Wall.

Family:- Apocynaceae

Q(b) WHAT IS THE USE OF RAUWOLFIA?

It is used under antihypertensive category as per the I.P.(Dose- 500µg daily).It produces the vasodilator effect.It is used as sedative in psycho category (depress the mental activity). It is also used in mild anxiety condition.

Q(c) GIVE A NOTE ON ISOLATION OF TERPENOID?

Ans :- Terpenoids are present in volatile oils in the form of mixture. These terpenoids are present either in the form of hydrocarbon or their oxygenated derivative (alcohol, aldehyde, ketone etc.). These are separated usually by two methods: Physical method, and Chemical method.

1. Physical method: In physical method different chromatographic methods and fractional distillation is applied for separation of constituent terpenoids.

2. Chemical method:

a. Separation of terpenoid hydrocarbon: These are separated by using Tilden reagent composed of solution of Nitrosyl chloride (NOCl) in chloroform. The terpenoid hydrocarbons on treatment with Tilden reagent forms crystalline adduct having sharp m.p., which is separated from volatile oil followed by hydrolysis or decomposed to get back the terpenoid hydrocarbon.

b. Separation of terpenoid alcohol: Terpenoid alcohols on reaction with thallic anhydride forms di-ester, which precipitate out from volatile oil. These di-esters on treatment with NaHCO3 in presence KOH, yields back terpene alcohol and thallic acid.

c. Separation of terpenoid aldehyde and ketone: Terpenoid aldehydes and ketones forms crystalline adduct on reaction with NaHSO3 and phenyl hydrazines etc. These crystalline adducts can be hydrolyzed to get back carbonyl compounds.

Q(d)WRITE A BRIEF DESCRIPTION OF MORPHINE?

Ans :- B.s:-Morphine is the most abundant opiate found in opium, the dried latex extracted by shallowly scoring the unripe seedpods of the Papaver somniferum poppy.

Morphine is generally 814% of the dry weight of opium�

Family:- papaveracea

Chemistry:-

Moleculer formula:- C17H19NO3H2O

Systematic (IUPAC) name:- (4R,4aR,7S,7aR,12bS)-3-methyl-2,3,4,4a,7,7a- hexahydro-1H-4,12-

methanobenzofuro[3,2-e]isoquinoline-7,9-diol

Morphine is a benzylisoquinoline alkaloid with two additional ring closures. It has:

A rigid pentacyclic structure consisting of a benzene ring (A), two partially unsaturated cyclohexane rings (B and C), a piperidine ring (D) and a tetrahydrofuran ring (E). Rings A, B and C are the phenanthrene ring system. This ring system has little conformational flexibility.

Two hydroxyl functional groups: a C3-phenolic OH (pKa 9.9) and a C6-allylic OH,

An ether linkage between C4 and C5,

Unsaturation between C7 and C8,

A basic, tertiary amine function at position 17,

5 centers of chirality (C5, C6, C9, C13 and C14) with morphine exhibiting a high degree of stereoselectivity of analgesic action.

Most of the licit morphine produced is used to make codeine by methylation. It is also a precursor for many drugs including heroin (3,6-diacetylmorphine), hydromorphone (dihydromorphinone), and oxymorphone (14-hydroxydihydromorphinone); many morphine derivatives can also be manufactured using thebaine or codeine as a starting material. Replacement of the N-methyl group of morphine with an N-phenylethyl group results in a product that is 18 times more powerful than morphine in its opiate agonist potency. Combining this modification with the replacement of the 6-hydroxyl with a 6-methylene group produces a compound some 1,443 times more potent than morphine, stronger than the Bentley compounds such as etorphine (M99, the Immobilon tranquilliser dart) by some measures.

structure:-

Pharmacological activity:-

Pain:Morphine is used primarily to treat both acute and chronic severe pain. It is also used for pain due to myocardial infarction and for labor pains.Its duration of analgesia is about three to seven hours.

However, concerns exist that morphine may increase mortality in the setting of non ST elevation myocardial infarction. Morphine has also traditionally been used in the treatment of acute pulmonary edema.

Shortness of breath:Immediate-release morphine is beneficial in reducing the symptom of shortness of breath due to both cancer and noncancer causes.In the setting of breathlessness at rest or on minimal exertion from conditions such as advanced cancer or end-stage cardior

Morphine is also available as a slow-release formulation for opiate substitution therapy (OST) in Austria, Bulgaria, and Slovenia, for addicts who cannot tolerate either methadone or buprenorphine.

contraindicated:Relative contraindications to morphine include: respiratory depression when appropriate equipment is not available

Although it has previously been thought that morphine was contraindicated in acute pancreatitis, a review of the literature shows no evidence for this.

Biosynthesis of pathway:-

Morphine is biosynthesized in the opium poppy from the tetrahydroisoquinoline reticuline. It is converted into salutaridine, thebaine, and oripavine. The enzymes involved in this process are the salutaridine synthase, salutaridine:NADPH 7-oxidoreductase and the codeinone reductase.

Bio synthesis pathway in human body:-

Morphine Can endogenous opioid in humans that can be synthesized and released by white blood cells.[7] CYP2D6, a cytochrome P450 isoenzyme, catalyzes the biosynthesis of morphine from codeine and dopamine from tyramine along the biosynthetic pathway of morphine in humans.[7][72] The morphine biosynthetic pathway in humans occurs as follows:[7]

L-tyrosine → para-tyramine or L-DOPA → dopamine → (S)-norlaudanosoline → (S)-reticuline → 1,2-dehydroretinulinium → (R)-reticuline → salutaridine → salutaridinol → thebaine → neopinone → codeinone → codeine → morphine

(S)-Norlaudanosoline (also known as tetrahydropapaveroline) can also be synthesized from 3,4-dihydroxyphenylacetaldehyde (DOPAL), a metabolite of L-DOPA and dopamine.[7] Urinary concentrations of endogenous codeine and morphine have been found to significantly increase in individuals taking L-DOPA for the treatment of Parkinson's disease.

Or

Q(e)WRITE A BRIEF DESCRIPTION OF QUININE?

Ans:- B.s:-cinchona species (Rubiaceae) specifically contains quinine in the bark upto 16% (mostly 6-10%) in a variety of its species, namely: Cinchona calisaya Wedd.; C. ledgeriana Moens ex Trimen; C. officinalis Linn. f.; C. robusta How.; and C. succirubra Pavon ex Klotzsch. The representative samples of dried cinchona, cinchona bark or peruvian bark is found to contain nearly 0.4 to 4% quinine.

structure:-

Chemistry:-

Formula:-C20H24N2O2

IUPAC name: (2-ethenyl-4-azabicyclo[2.2.2]oct-5-yl)- (6-methoxyqu

Characteristic Features:-

1. It is obtained as triboluminescent, orthorhombic needles from absolute ethanol having mp 177° (with some decomposition).

2. It sublimes in high vacuum at 170-180°C.

3. Its specific optical rotations are: [α]15D - 169° (C = 2 in 97% ethanol); [α]17D - 117° (C = 1.5 in chloroform); [α]15D - 285° (C = 0.4 M in 0.1 N H2SO4).

4. Its dissociation constants are: pK1 (18°) 5.07; and pK2 9.7.

5. The pH of its saturated solution in 8.8.

6. It gives a distinct and characteristic blue fluorescence which is especially strong in dilute sulphuric acid.

7. Solubility Profile: 1 g dissolves in 1900 ml water; 760 ml boiling water; 0.8 ml ethanol; 80 ml benzene; 18 ml benzene at 50°; 1.2 ml chloroform; 250 ml by ether; 20 ml glycerol; 1900 ml of 10% ammonia water; and almost insoluble in petroleum ether.

Identification Test:-

Quinine may be identified either by a series of Colour Tests or by the formation of several known derivatives having characteristic features; and these shall be discussed separately as under:

(a)Colour Test

1. Oxygenated Acids:

Oxygenated acids, such as: sulphuric acid or acetic acid gives a strong blue fluorescence with

2. Herpathite Test:

To a boiling mixture of quinine (0.3g) in 7.5 ml glacial acetic acid, 3 ml ethanol (90% v/v) and 5 drops of concentrated H2SO4, add 3.5 ml of I2 solution (1% w/v) in ethanol, crystals of iodosulphate of quinine or Herpathite* separates out on cooling. The crystals thus obtained exhibit metallic lustre, appears dark in reflected light and alive-green in transmitted light.

3. Thalleioquin Test:

When a few drops of bromine water are added to 2 or 3 ml of a weakly acidic solution of quinine salt, followed by the addition of 0.5-1.0 ml of strong ammonia solution, it produces a distinct characteristic emerald green colouration. It is an extremely sensitive colour test which may detect quinine even upto a strength as low as 0.005% (w/v). The end coloured product is known as thalleioquin for which the exact chemical composition is not yet known.

Note: (a) This test is given by quinidine and also by other Remijia alkaloids e.g., cupreine.

(b) Both cinchonine and cinchonidine do not respond to the Thalleioquin Test.

4. Erythroquinine Test (or Rosequin Test):

Dissolve a few mg of quinine in dilute acetic acid, add to it a few drops of bromine water (freshly prepared), followed by a drop of a 10% (w/v) solution of potassium ferrocyanide [K4Fe(CN)6]. Now, the addition of a drop of concentrated NH4OH solution gives rise to a red colouration instantly. If shaken quickly with 1-2 ml of chloroform, the red colouration is taken up by the lower chloroform-layer.

salt test:These are as follows:

1. Quinine Trihydrate:

It is obtained as a microcrystalline powder having mp 57°C. It effloresces and loses one mol of water in air, two moles of water over H2SO4, and becomes anhydrous at 125°C.

2. Quinine Bisulphate Heptahydrate (C20H24N2O2.H2SO4.7H2O) [Synonyms: Quinbisan, Dentojel, Biquinate): It is obtained as very bitter crystals or crystalline powder. It effloresces on exposure to air and darkens on exposure to light. 1 g dissolves in 9 ml water, 0.7 ml boiling water, 23 ml ethanol, 0.7 ml ethanol at 60°C, 625 ml chloroform, 2500 ml ether, 15 ml glycerol and having a pH 3.5.

3. Quinine Dihydrochloride

1. Oxiate): It is obtained as a powder or crystals having a very bitter taste. 1g dissolves in about 0.6 ml water, 12 ml ethanol; slightly soluble in chloroform; and very slightly soluble in ether. The aqueous solutions are found to be strongly acidic to litmus paper (pH about 2.6).

4. Quinine Hydrochloride Dihydrate (C20H24N2O2.HCl.2H2O): It is obtained as silky needles having a bitter taste. It effloresces on exposure to warm air. It does not lose all its water below 120°C. 1 g dissolves in 16 ml water, in 0.5 ml boiling water, 1.0 ml ethanol, 7.0 ml glycerol, 1 ml chloroform, and in 350 ml ether. A 1% (w/v) aqueous solution shows a pH 6.0-7.0.

5. Quinine Sulphate Dihydrate [(C20H24N2O2)2.H2SO4.2H2O] (Synonyms: Quinamm; Quinsan; Quine, Quinate): It is obtained as dull needles or rods, making a light and readily compressible mass. It loses its water of crystallization at about 110 °C. It becomes brownish on exposure to light. Optical rotation [α]15D - 220° (5% solution in about 0.5 N . HCl). 1g dissolves in 810 ml water, 32 ml boiling water, 120 ml ethanol, 10 ml ethanol at 78°C; slightly soluble in ether and chloroform, but freely soluble in a mixture of 2 vols. chloroform and 1 vol. absolute ethanol. Its aqueous solutions are neutral to litmus. The pH of a saturated solution in 6.2.

Pharmacological activity:-

1. It is frequently employed as a flavour in carbonated beverages.

2. It is used as an antimalarial agent.

3. It is also employed as a skeletal muscle relaxant.

4. It has been used to treat hemorrhoids and varicose veins.

5. Quinine is also used as a oxytocic agent.

6. Quinine is supposed to be prophylactic for flu.

contraindication :-

Quinine can cause abnormal heart rhythms, and should be avoided if possible in patients with atrial fibrillation, conduction defects, or heart block. Quinine can cause hemolysis in G6PD deficiency (an inherited deficiency), but this risk is small and the physician should not hesitate to use quinine in patients with G6PD deficiency when there is no alternative.[23]

Biosynthesis of quinine:-

A survey of literature reveals that the intrinsic details of the biosynthetic pathways are lacking; however, an assumed biogenetic process essentially involving the followingsteps:

1. L-Tryptophan and secologanin yields strictosidine, which upon hydrolysis and decarboxylation produces coryantheal.

2. Coryantheal undergoes intramolecular changes, first-by cleavage of C-N bond (via iminium), and secondly-by formation of an altogether new C-N bond (again via iminium). This gives rise to an intermediate.

3. The resulting intermediate undergoes further intramolecular changes to yield cinchoninone having a quinoline nucleus.

4. Cinchoninone in the presence of NADPH* reduces the carbony function and generates quinine:

5-Q(a) WRITE PHARMACOLOGICAL ACTITY OF VINCA?

Ans :- The alkaloid is used for the treatment of a wide variety of neoplasms. It is also recommended for generated Hodgkins disease, lymphocytic lymphoma, hystiocytic hymphoma, mycosis fungoides,� advanced testicular carcinoma, Kaposi's sarcoma, and choriocarcinoma and lastly the breast cancer unresponsive to other therapies.

It is effective as a single entity, however, it is normally given along with other neoplastic agents in combination therapy for the increased therapeutic effect without any noticeable additive toxicity.

It arrests mitosis at the metaphase.

It is aound to be effective in the acute leukemia of children.

Q (b) DEFINE TERPENOID?

Answer Terpenoids:- � The terpenoids sometimes called isoprenoids, are a large and various class of naturally occurring organic chemicals like to terpenes , derived from five-carbon isoprene units assembled and adapted in thousands of ways. Most are multicyclic structures that vary from one another not only in functional groups but also in their basic carbon skeletons. These lipids can be found in all classes of living things, and are the largest group of natural products. About 60% of known natural products are terpenoids

Terpenoid is the mixture of isoprene unit� �

Isoprene Unit:- (CH2=C(CH3)-CH=CH2)n

Q (c)WRITE ABOUT T.S OF DATURA LEAF?

Ans :- Biological Source:- Datura is a genus of poisonous vespertine flowering plant known as Datura stramonium

Family:- Solanaceae

Microscopical characters:-

Lamina: Dorsiventral

Upper epidermis:They are single layered, cells rectangular with cuticularized outer walls. Trichomes, both covering and glandular are seen. Covering trichomes are uniseriate, multicellular, warty and blunt at the apex. Glandular trichomes are made up to a stalk of one cell and a 2 to 4 celled glandular head.

Mesophyll:It is differentiated into palisade and spongy parenchyma.

Palisade:It is a single layered, compact and cells radially elongated

Spongy parenchyma: They are many layered, loosely arranged with intercellular spaces. Sphaeraphides, microsphenoidal crystals and vascular strands are found in the upper layers of spongy parenchyma.

Lower epidermis:It is identical to upper epidermis. Stomata and numerous trichomes are seen on the lower epidermis.

Midrib:The epidermis layers of lamina are continuous in the midrib region also. Strips of collenchymas appear below the upper and above the lower epidermis. This is followed by cortical parenchyma containing prisms of calcium oxalate and microsphenoidal crystals. Embedded in the central region of the cortical parenchyma is a bicollateral bundle.

Q(d)WRITE A BRIEF DESCRIPTION OF TAXOL?

Ans :- B.s:-It is obtained from the bark of the Pacific Yew tree, Taxus brevifolia Nutt belonging to the family Taxaceae

structure:-

Chemistry:

Systematic (IUPAC) name:-

(2α,4α,5β,7β,10β,13α)-4,10-Bis(acetyloxy)-13-{[(2R,3S)-3-(benzoylamino)-2-hydroxy-3-phenylpropanoyl]oxy}-1,7-dihydroxy-9-oxo-5,20-epoxytax-11-en-2-yl benzoate

Formul:-C47H51NO14)

(a) It has a taxane ring system,

(b) It has a four membered octane ring

(c) An ester side chain at C-13 of the taxane ring is a prime requirement for taxols cytotoxic activity, and�

(d) The presence of an accessible hydroxyl moiety at C-2 of the ester side chain renders an appreciable enhancement of the cytotoxic activity.

Pharmacological activity:-

1. Taxol is primarily employed in the treatment and management of metastatic carcinoma of the ovarian glands after the failure of follow-up chemotherapy.

2. It is also used in the treatment of breast cancer usually after the observed failure of combination chemotherapy for metastatic disease.

3. Because of its hydrophobic nature the injectable concentrate of taxol formulation meant for

intravenous infusion is normally solubilized duly in polyoxyethylated caster oil. However, before

injection it should be appropriately diluted in normal saline or dextrose solution or combination thereof.

Side effect :-

slow heart rate;

seizure (black-out or convulsions);

pale skin, easy bruising or bleeding, unusual weakness;

fever, chills, body aches, flu symptoms;

white patches or sores inside your mouth or on your lips;

numbness, tingling, or burning pain in your hands or feet;

increased blood pressure (severe headache, blurred vision, buzzing in your ears, anxiety, confusion, chest pain, shortness of breath, uneven heartbeats); or

warmth or redness under your skin.

Less serious side effects may include:

pain, swelling, redness, or skin color changes where the Taxol was injected;

joint or muscle pain;

mild nausea, vomiting, diarrhea; or

hair loss.

Biosynthesis of pathway:-

The biosynthetic pathway to paclitaxel has been investigated and consists of approximately 20 enzymatic steps. The complete scheme is still unavailable. The segments that are known are very different from the synthetic pathways tried thus far (Scheme 1). The starting compound is geranylgeranyl diphosphate 2 [which is a dimer of geraniol . This compound already contains all the required 20 carbon atoms for the paclitaxel skeleton. More ring closing through intermediate 3 (taxadiene) leads to taxusin . The two main reasons why this type of synthesis is not feasible in the laboratory is that nature does a much better job controlling stereochemistry and a much better job activating a hydrocarbon skeleton with oxygen substituents for which cytochrome P450 is responsible in some of the oxygenations. Intermediate 5 is called 10-deacetylbaccatin III.

Or

Q(e)WRITE A BRIEF DESCRIPTION OF RUTIN?

Ans :- Introduction:-

Synonyms :-Melin; Phytomelin; Eldrin; Ilixanthin; Sophorin; Globularicitrin; Paliuroside; Osyritrin; Osyritin; Myrticolorin; Violaquercitrin; Birutan; Rutabion; Rutozyd; Tanrutin.

Biological Source:-Rutin is found in many plants, especially the buckwheat plant (Fagopyrum esculentum Moench; family: Polygonaceae); in forsythia [Forsythia suspensa (Thunb).) Vahl ver. Fortunei (Lindl). Rehd., family Oleaceae]; in hydrangea (Hydrangea paniculata Sieb., family: Saxifragaceae); in pansies (Viola sp. Violaceae); from leaves of Eucalyptus macroryncha F.v. Muell., family : Myrtacea); in Fagopyrum tartaricum Gaertn: family: Polygonaceae); in Ruta graveolens L., (family: Rutaceae); in buds of Sophora japonica L., (family: Leguminoseae); in fresh leaves of tobacco plants, Nicotiana tabacum L., (family : Solanaceae); in cotton seed Gossypium hirsutum;(family Malvaceae); in Viola tricolor, (family : Violaceae).

structure

Chemistry:-

The iupac name is 2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-3-{[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-({[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}methyl)oxan-2-yl]oxy}-4H-chromen-4-one

Chemical formula:- C27H30O16

Pharmacological activity:-

1.Rutin is used to decrease the capillary fragility (i.e., to enhance the tensile strength of capillary walls), reduce capillary permeability by tissue injury, and minimise the destruction of epinephrine in body tissues.

2. It has been mostly used in certain disease condition to reduce capillary bleeding promptly.

3. It is found to be useful in the treatment of retinal harmorrhages

Chemical Test:-

1. It gives a distinct yellow precipitate with basic lead acetate.

2. It yields a greenish brown colour with ferric chloride.

3. It produces a silver mirror with ammonical silver nitrate solution (Tollens Reagent)�

Biosynthesis of pathway:-

This pathway mostly describes the metabolic situation found in buckwheat species and represents a branch of the phenylpropanoid biosynthesis (see also flavonoid biosynthesis , flavonol biosynthesis and quercetin glycoside biosynthesis (Arabidopsis) ). This has been demonstrated through monitoring the differential expression of involved genes during developmental stages of buckwheat which are characterized by the accumulation of rutin as one of the most abundant flavonol glycoside in this plant [Gupta11 ] [Li10a ]. The increased production of rutin after overexpressing the flavonol-specific transcription factor AtMYB12 in Fagopyrum esculentum and observing the expression of genes presumably involved in rutin biosynthesis shows that key enzymes of the phenylpropanoid and flavonoid biosynthetic pathways up to phenylpropanoid biosynthesis, initial reactions are channeled towards rutin formation [Park12b ].

An important metabolic step in this pathway is carried out by the flavonol synthase catalyzing the 2-oxoglutarate dependent oxygenation of (+)-taxifolin to form quercetin . The corresponding gene has been cloned, heterologously expressed and characterized with regard to protein structure and catalytic properties [Li12a ]. The following conversion in the pathway forming the immediate precursor for rutin, quercetin-3-glucoside (isoquercetin) is realized by the flavonol 3-O-glucosyltransferase [Suzuki05 ] [Lucci09 ] (compare also quercetin glycoside biosynthesis (Arabidopsis) ). The enzyme catalyzing the final step in the pathway, i.e. flavonol-3-O-glucoside L-rhamnosyltransferase has been partially purified from mung bean [Barber62 ] and Dimorphandra mollis (fava d'anta) [Lucci09 ]. The enzyme catalyzes the transfer of a rhamnosyl group from UDP-β-L-rhamnose to quercetin 3-O-glucoside forming rutin. Although dTDP-rhamnose had been considered as the primary donor [Barber62 ] it has been demonstrated that UDP-L-rhamnose is the native donor for the transfer of L-rhamnose [Barber91 ] [Barber63 ].

It had long been a pending question how exactly UDP-L-rhamnose was biosynthesized in plants. It is clear now that UDP-L-rhamnose can be catalyzed from UDP-D-glucose (see UDP-L-rhamnose biosynthesis ) [Barber91 ] [Barber63 ] which is an ubiquitous activated nucleotide sugar readily available for metabolic processes in plants.