Module 2 Pharmacognosy

Module 2 Pharmacognosy Plant Chemistry, Biochemistry Kathreen Mae D. Cascabel BSPharmacy

INTRODUCTION TO PHARMACOGNOSY Pharmacognosy – knowledge of drugs and pharmaceuticals | Greek: pharmakon (drugs) and gnosis (knowledge) BRIEF HISTORY Babylonians – made clay models of man Egyptians – very apt in the practice of embalming Greeks Pedanios Dioscorides – De materia medica libre cinque (five volumes), 600 plants (Rome) Galen – described the method of preparing formulas containing plant and animal drugs and the term ”Galenical pharmacy” originated (Cold cream – Galen’s cerate) Germans C.A. Seydler – coined the term pharmacognosy in his dissertation in 1815 (title: Anaclectica pharmacognosthica) Fluckiger –most comprehensive definition of pharmacognosy : “simultaneous application of various scientific disciplines with the object of acquiring knowledge of drugs from every point of view” J.A. Schmidt – Lerbuch der Materia medica Pharmacognosy – “an applied science that deals with the biologic, biochemical and economic features of natural drugs and their constituents that originate in the plant and animal kingdom” General Process of Extraction Removes substances that can be dissolved by the solvent/menstruum The undissolved portion of the drug is known as the marc Product of extraction process: Percolate/Extractive Crude Drugs Natural Substances Derivatives/ Extractives Geographic source & Habitat Indigenous Naturalized

Appropriate Solvents: 1. Fats 2. Resins 3. Chlorophyll 4. Chrysarobin

- Hexane - Alcohol - Acetone - Hot benzene

– undergone only collection and drying – have not had changes made in their molecular structure as found in nature – chief constituents obtained by extraction – region in which the plant or animal yielding the drug grows – growing in their native countries – growing in foreign land

PREPARATION OF DRUGS FOR THE COMMERCIAL MARKET 1. Collection – (small scale) collection time: part of the plant that constitutes the drug is highest in its content of AI 2.

Harvesting – (large scale) Mechanical devices : pickers, mowers, binders, swath, steel Manual – for potent constituents

3.

Drying (Air Drying and Artificial Air Drying) • remove moisture • fixes the constituents • prevent molding • facilitate grinding and milling • prevent action of bacteria • prevent environmental hydrolysis

Collection time Pectin – citrus fruits Unripe fruits - protopectin Just ripe - pectin Overripe - pectic acid Marijuana Young leaves - cannabidiol Mature leaves - cannabinol (THC)

4.

Garbling – final stage; removal of extraneous matter (other parts of the plant, dirt, added adulterants)

5.

Packaging, Storage and Preservation To avoid insect attacks: a. SIMPLEST METHOD: Expose the drug to 65°C b. Fumigation with methyl bromide c. Add adrop of chloroform or carbon tetrachloride

Examples of Plants in De Materia Medica Aloe Belladonna (beautiful lady) Tamoxifen Colchicum Allopurinol Ergot (“fingers” Claviceps purpurea) Opium (stone of immortality) Premarin Digoxin (Digitalis lanata) Digitoxin (Digitalis purpurea)

Emollient, tx for burns Atropine – mydriatic (SE: cycloplegia) For breast CA (SE: endometrial CA) Acute gout Chronic gout (Xanthine Oxidase inhibitor) Ergotamine (migraine), Ergonovine (oxytocic), LSD (hallucinogen) Estrogen derivative from MARE

Bioassay Digoxin - pigeon Atropine - cats Glucagon - cats Oxytocin - chicken (F) Head drop test - rabbits/rats PTH - dogs Heparin - sheep plasma Insulin - rabbit

Lipid soluble, Long half life

Street names of illegal drugs 1. Shabu Ice, Meth 2. Ecstacy Sky 3. Marijuana Hashish, MJ/Maryjane, Pot, Grass, Weed, Cannabis, Indian hemp EVALUATION OF DRUGS (determine the quality (intrinsic property) and purity (absence of adulterants) of drugs) 1. Organoleptic - by means of the organs of sense; macroscopic appearance 2. Microscopic - essential in the study of adulterant in powdered plants (identity and purity) 3. Biologic - pharmacologic activity of certain drugs has been applied to their evaluation or standardization 4. Chemical - assay of AC using chemicals (titration); best method of determining official potency 5. Physical - physical constants (solubility, spec grav, optical rotation, congealing point, refractive index, MP) CLASSIFICATION OF DRUGS Morphology Taxonomy Pharmacologic or Therapeutic Chemical classification

– forms – phylogeny (natural relationship or among plants and animals) – based on effect on the body – based on active constituents; Preferred Method of Classification

CHEMISTRY OF NATURAL DRUG PRODUCTS 1. Pharmaceutically active – cause chemical changes in the preparation 2. Pharmacologically active – responsible for the therapeutic activity 3. Inert constituents – no definite pharmacologic activity 4. Secondary plant substance – extracted, crystallized and purified; presence of inert matter which may modify or prevent absorbability or potency of AC Drug biosynthesis/Biogenesis – study of the biochemical pathways leading to formation of secondary constituents (drugs) Primary metabolites

to

Secondary metabolites

Carbohydrates Proteins Fats

Glycosides BIOGENESIS OR DRUG BIOSYNTHESIS

Tannins Volatile oils or Fixed oils

3 Principal factors that influenced the secondary constituents of drug plants 1. heredity (genetic composition) 2. ontogeny (stage of development) 3. environment (soil, climate, associated flora and method of cultivation)

CARBOHYDRATES AND RELATED COMPOUNDS polyhydroxyaldehydes or polyhydroxyketones containing C, H and O; same ratio as in H2O (2:1) 1st products of photosynthesis (glucose) General formula: CnH2nOn | Cn(H2O)n | (CH2O)n

CHO 4 cal/g CHON 4 cal/g Fats 9 cal/g

MONOSACCHARIDES Cannot be hydrolyzed to simple sugar substances that is ketonic or aldehydic substitution product of a polydroxy alcohol C 2 3 4 5 6 7 8

Name Diose Triose Tetrose Pentose Hexose Heptose Octulose

Aldose Hydroxyacetaldehyde Glyceraldehyde Erythrose Arabinose, Xylose, Ribose Glucose, Galactose Sedoheptose D-glycero-D-mannoctulose

Ketose Dihydroxyacetone Erythrulose Ribulose Fructose

HEXOSES (most important monosaccharides found in plants) Glucose (aldohexose)

Fructose (ketohexose)

Other names D-glucose Dextrose Blood sugar Grape sugar Physiologic sugar Fruit sugar Levulose (sweetest monosaccharide)

Uses Nutrient (parenteral), Excipient Ca gluconate/gluceptate/levulinate – Ca replenisher Ferrous gluconate – hematinic

Identification Test Moore’s Test (caramel)

Food for diabetics Ingredient in infant feeding formula Ingredient in fructose injection, nutrient

Seliwanoff’s Test

PENTOSES Xylose (D-xylose, Wood Sugar) obtained by boiling corn cobs, straw or similar materials with dilute acid to hydrolyze the xylan polymer diagnostic aid in intestinal malabsorption

OLIGOSACCHARIDES DISACCHARIDES Sucrose Glu + Fru (non-reducing)

Maltose Glu + Glu Lactose Glu + Gal (reducing)

Other N Saccharum Cane sugar Beet sugar Table sugar Malt sugar

Origin Saccharum officinarum (cane) Beta vulgaris (beets) Acer saccharum (maple)

Milk sugar

Cow’s milk (Bos Taurus)

Uses Demulcent, Sweetening, Coating, Preservative, Production of syrups, Retards oxidation

From germination of barley

Lactulose (Alkaline lactose) lactic & acetic acid (irritation, catharsis) Dec serum NH3 in encephalopathy

Isolation of Lactose Butter – cream, united fat globules Buttermilk – liquid separating from butter Skimmed milk – left after cream separates Cheese – coagulum, skimmed m. + rennin Whey – liquid separated from coagulum

POLYSACCHARIDES Polysaccharides are polymers of monosaccharides (sugar) linked together through glycosidic(ether) linkages Temporary storage form of photosynthetic products systematic nomenclature are given the ending “an”, glycan is another word

Amylose Linear 250-300 glucose units 25% α-1,4 glucosidic bonds Soluble Blue

Amylopectin Branched 1000 or more glucose units 75-80% α -1,4 and α-1,6 (every 25 glucose) Insoluble (paste forming) Blue-violet or purple

HOMOGLYCANS Starch (Glucosan) Zea mays (corn) Triticum aestivum (wheat) Solanum tuberosum (potato)

Use Tablet filler, binder, and disintegrant Antidote for iodine poisoning Permanent reserve food material in plants

Enzymes that break down starches: β amylase – pancreatic juice and saliva α amylase – hydrolyzes it to maltose Starch

Gluten Pregelatinized Starch Na Starch Glycolate Hetastarch

– tachy proteins – binder – disintegrant – plasma expander

Soluble starch Dextrin Maltose

Inulin (Fructosan) Dextran (Glucosan) - plasma expander Cellulose

2 glucose Ingredient in culture media Used in evaluation of renal function dextran sucrase (from Leuconostoc mesenteroides) Sucrose Dextran Purified/Absorbent Cotton – from the hair of the seeds of Gossypium hirsutum Use: mechanical protection against bacteria; absorb mucus, pus, and blood Soluble guncotton/Pyroxylin – formed by action of nitric acid & sulfuric acid on cotton Other ingredients: Castor oil (flexibility) | Camphor (proof) Use: topical protectant Other derivatives: Cellulose Acetate Pthalate (CAP) – film coating, Methyl/Ethylcellulose

HETEROGLYCANS (yield more than one type of monosaccharide units upon hydrolysis) Gums and Mucilages Natural plant hydrocolloids that may be classified as anionic or non-ionic polysaccharides Produced by plants as a protective after injury Upon hydrolysis, they yield arabinose, galactose, glucose, mannose, xylose, and other uronic acid derivatives USES: suspending agents, dental adhesives, laxatives

PLANT EXUDATES Tragacanth

Astragalus gummifer

Bassorin (swells) Tragacanthin (thickener)

Arabin

Best mucilage (resistant to acid hydrolysis)

Acacia

Gum arabic

Acacia senegal

Ghatti

Indian Gum

Anogeissus latifolia

Karaya

Sterculia Gum

Sterculia urens

D-galacturonic acid D-glucoronic acid

(fetid odor)

Suspending agent, Emulsifier, Adhesive, Demulcent, Emollient 5-6% dispersion in H2O Incompatible with OH 35-36% dispersion Acacia substitute Laxative, Suspending agent, Adhesive

MARINE GUMS Agar Algin

Japanese Gelidium cartilagineum Insinglas/Gelatin (Red algae) Macrocystis pyrifera

Agarose Agaropectin Sodium alginate

(brown seaweed)

Carageenan

Irish Moss, Chrondus

Chondrus crispus Gigartina mamillosa

Danish agar

Furcerallan

Furcellaria fastigiata

Kappa- & Iotacarageenan (swelling) Lambda-carageenan (nonswelling) k-carrageenan

Laxative, tablet diluents/disintegrant Suspending agent, tablet binder, thickener Demulcent, bulk laxative

Suspending agent

SEED GUMS Plantago Cydonium

Psyllium, Plantain Seed Quince seed

Plantago psyllium (Spanish) P. ovate (Blonde/Indian) Cydonia vulgaris

Guar Gum

Guaran

Cymopsis tetragonolobus

Locust Bean Gum

Carob pulp St. John’s bread

Ceratonia siliqua

Bulk laxative

1,4 linked Dmannopyranosyl

Laxative, thickener, binder Chocolate substitute

MICROBIAL GUM Xanthan Gum

Xanthomonas campestris

Peudoplastic activity which enables toothpaste and ointment both to hold shape and spread readily

PECTIN – purified carbohydrate product obtained from the dilute acid extract of inner portion of the rind of citrus fruits or from apple pomace Active Constituent : linear 1,4 linked D-galacturonan Uses : protectant, s.a., ingredient in many antidiarrheal formulations

[O]

PLANT ACIDS a. Cherry Juice or Succus cerasi Ripe fruit of Prunus cerasus Contains pectin (incompatible with OH) Constituent: Malic acid (present in cherry, apple, pear) Use: preparation of cherry syrup b. Plant Acids Uses: Acidulants in effervescent formulations, Component of buffer systems a. Citric acid - (2-hydroxy 1,2,3, propane trioic acid) isolated by Scheele from lemon juice in 1784 b. Lactic acid - (alpha-hydroxy propionic acid) acidulant in infant feeding formula c. Tartaric acid - (1,2,3, dihydroxy butane dioic acid) by product of wine industry d. Fe fumarate - hematinic c. Alcohol or Ethanol 95% ethanol by volume @15.56°C Product of fermentation Undergo the process of distillation to concentrate the alcohol content to 40-55% brandy - distilled wine whiskey - malted grain rum - molasses wine - mild stimulant and tonic

[H]

SUGAR ALCOHOLS Mannitol (D-mannitol) From Manna, Fraxinus ornus Osmotic diuretic and Osmotic laxative Sorbitol (D-glucitol) From the ripe berries of mountain ash, Sorbus aucuparia half as sweet as sucrose has humectant properties (ingredient in toothpaste, chewing gums and various dietetic products) not absorbed on oral ingestion that is why it is used in the manufacture of chewing gums

TESTS FOR CARBOHYDRATES Molisch’s Fehling’s Benedict’s Barfoed’s Tollen’s Picric acid Nylander’s Osazone/Kowarsky Seliwanoff’s Keller-Killiani Test Moore’s Mucic acid Test Bial’s

Test for General for carbohydrates Reducing sugars Reducing sugars Reducing sugars; Monosaccharides Reducing sugars (Aldehydes) Reducing sugars Reducing sugars Mannose Ketohexose (Fructose) Deoxysugars (cardiac glycosides) Reducing sugars (Glucose) Galactose Pentoses

Composition α-naphthol A – CuSO4 B – K Na tartrate NaOH + CuSO4 Cupric acetate + acetic acid Silver + Ammonia Picric acid Na K tartrate + K/NaOH + Bi Phenylhydrazine Resorcinol Acetic acid, Fe chloride, Sulfuric acid Heat with alkali Nitric acid Resorcinol + Fe

Positive result Purple/Red Brick red ppt Brick red ppt Brick red ppt Silver mirror Mahogany red Brown Yellow ppt (ozone) Red Reddish brown Caramel Insoluble ppt Blue green

GLYCOSIDES Heteroglycans (yield one ore more sugars among the products of hydrolysis); simple glycosides: sugar ethers The most frequently occurring sugar is Beta-D-glucose (plants) Aglycone or genin – nonsugar component, Glycone – sugar component

CARDIOACTIVE OR CARDIAC GLYCOSIDES MOA: inhibits Na/K/ATPase pump increased influx of Ca2+ ions (+) inotropy 2 types of steroidal aglycones or genin: Cardenolides - most prevalent in nature Bufadienolides - are homologs of the cardenolides and an unsaturated 6-membered lactone ring DOC: control rapid ventricular rate in px with atrial fibrillation/atrial flutter Keller-Killiani Test - Detects presence of 2-deoxysugars Other Names Grecian Foxglove Foxglove Convallaria Adonis Strophanthus Apocynum Oleander/Adelfa Squill Cactus Grandiflorus Black Hellebore

Origin Digitalis lanata Digitalis purpurea Convallaria majalis Adonis vernalis Strophanthus kombe Apocynum cannabinum

Active Constituent Digoxin Digitoxin (lipid soluble, long t½) Convallatoxin Adonitoxin k-strophanthoside/Strophanthidin Cymarin Oleandrin Scillaren A

Night blooming cereus

Nerium oleander Urginea maritime Selenicereus grandiflorus

Christmas rose

Hellebborus niger

hellebrin

Lily-of-the-Valley Pheasant’s eye Black Indian Hemp Dog bane, Canadian Hemp Rose bay

ANTHRAQUINONE GLYCOSIDES Cathartics (except chrysarobin) Aglycone: Anthracene ID test: Borntrager’s test

Cascara Sagrada

Sacred bark

Origin Rhamnus purchianus

Active Constituent D glycosides – based on emodin C glycosides – based on aloin Cascarosides A&B – barbaloin isomers Cascarosides C&D – chrysaloin isomers Lane's Pill

Casanthranol – purified mixture of Present in Movicol = Karaya + Frangula anthranol glycosides from cascara sagrada

Frangula Aloe

Buckthorn bark

Rhubarb

Rheum/Chinese Rhubarb

Senna

Cultivated on wet lands resembling rice paddies

Chrysarobin

Rhamnus frangula Aloe barbadensis/Aloe vera (Curacao) A. ferox + africana + spicata (Cape) Rheum officinale (Chinese) medicinal R. emodi, R. webbianum (Indian/Himalayan) Cassia angustifolia (Tinnevelly) Cassia acutifolia (Alexandria) Andira araroba (Goa powder; hot benzene)

Frangulin A & B Barbaloin from the latex Aloe-emodin Rhein anthrones ornamental

Sennosides A, B, C, and D Derivatives of Senna: Blue Coloration

*More potent than cascara Keratolytic agent

SAPONIN GLYCOSIDES β-hemolytic substances – destroys RBC by hemolysis and are toxic to cold blooded animals (used as fish poison) Soap-like (foam upon shaking) o Steroidal in nature (cyclopentanoperhydrophenanthrene/CPPP nucleus) o Sapotoxin – toxic aglycone Sternutatory - irritating the mucous membrane; people normally sneeze o Sapogenin – aglycone portion, 2 types: Steroid (neutral saponin) and Triterpenoid (acid saponin) Identification tests: 1. Froth test - (+) honeycomb froth that persists for 10 mins 2. Capillary test 3. BAM (Blood Agar Media) Test or Hemolysis Test α hemolysis – partial hemolysis β hemolysis – complete hemolysis γ hemolysis – no hemolysis

Glycirrhiza Licorice root

Ginseng • fave remedy in Chinese medicine

Dioscorea Mexican Yam

Origin Glycyrrhiza glabra

Panax quinquefolius (American) Panax ginseng (Asian/Korean) D. floribunda – best source of steroid D. spiculiflora – contains diosgenin

Active Constituent Glycyrrhizin/glycyrrhic acid (50x as sweet as sugar)

Use Peptic ulcer and Addison’s Increases foaminess of beer.

Glycyrrhetic acid (anti-inflam) Ginsenosides Panaxosides Chikusetsusaponins Botogenin Diosgenin

CI: cardiac problems & HTN – ↑ Na retention and ↓K Aphrodisiac Adaptogen (antistress) Cortisone precursor Costisol

CYANOPHORE OR CYANOGENIC GLYCOSIDES (Bound Poisons) yeilds hydrocyanic acid as one of the products found in Rosaceae plants amygdalin is the most widely distributed derivatives of mandelonitrile (benzaldehyde-cyanohydrin) Cyanide-containing compounds: Prunasin, Laetrile/Vitamin B17, Amygdalin Guinards Test: Brick Red / Red Coloration

Amygdalin Amygdalase Mandelonitrile glucose Prunase Mandelonitrile + Glucose

*Amygdalase + Prunase = EMULSIN

Benzaldehyde + HCN

Bitter Almonds Wild Cherry prunus virginiana, wild black cherry tree Apricot Pits Cassava

Origin Amygdalus communis Prunus serotina

Active Constituent Amygdalin Prunasin

Prunus armeniaca, Rosaseae Manihot esculenta

Laetrile/Vit. B17 Mannihotoxin

Use sedative expectorant

controversial treatment for sickle cell anemia (antiCA)

GLUCOSINATES/ISOTHIOCYANATES (From family Cruciferae/Brassicaceae) Black Mustard White mustard Sinapis nigra Sinapis alba Brassica nigra Brassica alba Sinigrin Sinalbin myrosin myrosin Allyl isothiocyanate Acrinyl isothiocyanate Local irritant and emetic, Rubefacient, vesicant, and condiment

Synonym Origin Active Constituent Uses

FLAVONOL

Rutin &Hisperidin Hesperitin & Naringen

(Vitamin P or Permeability Factors)

Tx of capillary bleeding secondary to capillary fragility Tx of symptoms of common colds

ALCOHOL

Salicin (Salix and Populus)

Salix purpurea Salix fragilis

Saligenin (salicyl alcohol) - antirheumatic (closely resembles salicylic acid)

ALDEHYDE

Vanilla

Vanilla plantifolia

Vanillin (4-hydroxy-3- methoxy benzaldehyde or methylprocatechuic aldehyde)

Other sources of Vanillin: Coniferin - glycoside present in cambium sap of pine trees Eugenol - phenol present in clove oil Lignin - by product of the pulp industry Gluco-vanillin (avenein) and glucovanillic alcohol hydrolyzed into glucose and vanillin and glucovanillic alcohol which is oxidized to vanillic aldehyde

PHENOL

Uva Ursi (Bearberry)

Arctotaphylos uvaursi

Arbutin – antiseptic and astringent

LACTONE GLYCOSIDES Coumarin

Cantharides

Psoralens (photosensitizing furocoumarins)

Origin Dicumarol, Dipteryx odorata Bishydroxycoumarin (tonka beans) Spanish flies, Russian flies, Blistering flies

Cantharis vesicatoria

Ammi majus

Use Anticoagulant

AC : Cantharidin Effects: Blisters – irritant, vesicant, rubefacient Priapism (used as aphrodisiac) Methoxsalen – repigmentation in vitiligo Trioxsalen

TANNINS A group of complex phenolic compounds capable of combining with proteins Polypeptides that are difficult to separate because they do not crystallize Precipitates proteins from solutions and can combine with proteins, rendering resistant to proteolytic enzymes Medicinal use : astringent Industrial use : manufacture of leather tannin Skin and Hides Leather Tanning of leather CHEMICAL CLASSES OF TANNINS Hydrolyzable

Heating

Gallic acid

Pyrogallol

Hydrolyzation Boil with HCl FeCl3 Test Br Water Lead Acetate Leather Type

Phenolic acids and Sugar Bluish black precipitate No precipitate Soluble Bloom

Tests for presence of Tannins: 1. Goldbeater test 2. Gelatin test Characteristics of Tannins 1. Acidic 2. Sharp puckering taste 3. Precipitates alkaloids 4. Precipitates proteins (tanning industry) 5. Astringents – GI and skin 6. Treatment of burns

Hamemelis Leaf Hazel leaves

Nutgall

Origin Hammemelis virginiana

Nonhydrolyzable/Condensed/Proanthocyan idin (from condensation of catechin and leucocyanidin) Catechol

Polymerization Plobaphenes (insoluble, red colored products) Greenish black precipitate Precipitate Tanner’s red

Tannins can precipitate: Proteins Alkaloids Glycosides Tannic acid – antidote for alkaloidal poisoning Universal antidote : Tannic acid - precipitant Activated charcoal - adsorbent Magnesium Oxide - cathartic

Active Constituent Hamamelitannin

Excrescence from the young twigs Tannic acid/Gallotannic acid of Quercus infectoria, Fagaceae Insect: Cynips tinctoria

Use Astringent Incorporated in hemorrhoidal preparations Tx: insect bites and stings Used in tanning industry Astringent Manufacture of inks

LIPIDS ester of long chain of fatty acids and alcohols Uses: emollients vehicles for IM injection – Corn Cottonseed Peanut Sesame cathartic – castor oil soap production increase caloric value – TPN

CLASSES OF LIPIDS 1. Fixed Oils - Long chain fatty acids + Glycerol o From vegetables o Liquid at room temperature (except theobroma) 2. Fats - Long chain fatty acids + Glycerol o From animals o Solid at room temperature (except cod liver oil)

SATURATED FATTY ACIDS 10C Capric 12C Lauric 14C Myristic 16C Palmitic (most abundant FA) 18C Stearic 20C Arachidic UNSATURATED FATTY ACIDS 18:1 Oleic 18:2 Linoleic 18:3 Linolenic (dietary precursor of PGE) 18:4 Arachidonic (precursor of PGE) Vitamin F (essential FAs): Oleic, Linoleic, Linolenic

3. Waxes – long chain fatty acids + High MW alcohol such as cetyl or stearylalcohol 4. Sterols – alcohols containing cyclopentanophenanthrene nucleus (cholesterol) 5. Phospholipids (Phosphatides) – Ester consisting of glycerol in combination with fatty acids, phosphoric acid and nitrogenous compounds (Lecithin) 6. Glycolipids (Cerebrosides) – Substances isolated from the brain and various sources which on hydrolysis yield fatty acids, galactose and nitrogenous compounds (Phrenosin and kerasin)

Tests or constants based on the chemical constitution of the fatty acids: 1. Acid value or acid number # of mg. of KOH required to neutralize the free fatty acids in 1 g of substance Indicates the presence of free fatty acids 2. Saponification value , saponification number or Koettsdorfer number # of mg. of KOH required to neutralize the free acids and saponify the ester contained in 1 g. of substance serves to detect the presence of the glycerides of acids 3. Iodine number # of grams of iodine absorbed by 100g of the substance indicates the degree of unsaturation of the fatty acids Classification of Fixed oils base on Iodine Value 1 drying oils >120 Linseed, cod liver oil 2 semidrying 100-120 Cottonseed, sesame 3 nondrying oils < 100 Olive, almond

FIXED OILS Origin

Active Constituent

Use

Coconut Oil

Cocos nucifera

Lauric & Myristic acid

Solvent for IM injection

Palm Oil

Elaeis guincenss

Shortening, frying, soap & candle manu

Castor Oil

Ricinus communis

Ricin (toxic) Ricinine (alkaloid) Azeleic acid (anti-acne)

Stimulant cathartic (ricinoleic acid) from Tricinolein hydrolysis by lipase (duodenum) CI: Pregnancy Olive oil grades (extracted with): 1. First Grade Virgin – less pressure 2. Second Grade Virgin – more P 3. Technical Oil – hot water 4. Sulfur Grade – CS2; low quality 5. Tournant Oil – fallen, decomp fruits

Olive Oil Sweet oil

Olea europaea

Oleic acid

Peanut Oil Arachis Oil

Arachis hypogaea

Oleic acid, Linoleic acid

Solvent for IM injection

Rapeseed Oil Canola Oil

Brassica campestris Brassica napus

Erucic acid

Salad oil Shortening

Soybean Oil

Glycine soja

Linolenic Acid

Lecithin – useful in controlling deranged lipid and cholesterol metabolism. Stigmasterol – from lipid fraction of soybean; steroid hormone precursor

Cottonseed Oil

Gossypium hirsutum

Linoleic, Oleic, Palmitic Myristic, Arachidonic

Make substitues for lard, soap

Sesame Oil Teel oil, Benne oil

Sesamum indicum

Oleic acid Linoleic acid

Sesamol – phenolic constituent which owes the excellent stability of the oil produced by the hydrolysis of sesamolin

Prunus amygdalus

Oleic, Linoleic Palmitic, Stearic

Emollient

Almond Oil Persic Oil Apricot/Peach kernel oil

Prunus armeniaca Prunus persica

Corn Oil

Zea mays embryo

Safflower Oil

Carthamus tinctoria

Sunflower Oil

Helianthus annus

Linseed Oil Flaxseed oil

Linum usitatissimum

Theobroma Oil Food for the Gods

Theobroma cacao

Cocoa syrup, Beverage Cocoa butter – supp base

Ethiodized Oil Injection

Iodine addition product of ethyl ester of the fatty acids of poppy seed

Diagnostic aid

Solvent for IM injection Linoleic acid

Dietary supp (False claim: ↓ cholesterol) Alternative to corn oil and safflower oil

Linileic acid – inedible (drying oil in paint & varnish)

Linum means thread; usitatissimum means most useful

FATS Lanolin

Origin Wool of Ovis Aries

Cod Liver Oil - liquid animal fat

Gadus morrhua (fresh livers)

Undecylenic Acid

Pyrolysis of Ricinoleic acid from castor oil

Active Constituent Hydrous Lanolin: 25% water Anhydrous lanolin/wool fat: 0.25% water Myristic acid Palmitic acid

Use H2O absorbable ointment base Emollient Sodium morrhuate - sclerosing agent (varicose) Source of Vit. A and D Antifungal

WAXES – stiffening agents Spermaceti Jojoba Oil Beeswax Yellow wax Carnauba wax

Origin head of sperm whale, Physeter macrocephalus Simmondsia chinensis Honeycomb of Apis mellifera Copernicia prunifera

Active Constituent Synthetic Spermaceti (Cetyl Ester Wax)

Use Emollient, Cold cream

Eicosenoic acid Myricyl palmitate

Emollient Stiffening agent

Myricyl cerotate

Candles, wax varnishes

Prostaglandins – class of natural products with potent and diverse biological activities Lipid metabolites formed in the body from essential, unsaturated fatty acids from the diet. Linoleic – most important dietary precursor of the prostaglandins – converted by elongation and desaturation in the liver to arachidonic acid

Prostanoic acid – parent compound of all natural prostaglandins Elcosonoids – arachidonic acids and most of the metabolites contain 20 carbon atoms 3 Main Classes of Major Prostaglandins 1 Prostaglandin A 2 Prostaglandin E 3 Prostaglandin F

VOLATILE OILS (Ethereal oils/Essences) Physical Characteristics Possess characterisctic odor Evaporate when exposed to air High refractive index Optically active Immiscible in water but they can be sufficiently soluble to impart their odor in water May act as insect repellants or insect attractants May occur in specialized secretory structures: glandular hairs modified parenchymal cells oil tubes called vittae lysigenous or schizogenous passages Formed by:

– Labiatae Lamiaceae – Piperaceae – Umbelliferae Apiaceae – Pinaceae, Rutaceae

Decomposition of resigenous layer of cell wall Hydrolysis of certain glycosides

2 classes: Terpene derivatives Aromatic compounds

- via acetate-mevalonic acid pathway - via schikimic acid-phenylpropanoid route

Medicinal Uses: Carminative, Antiseptic , Antipruritic (camphor), Counterirritant (methyl salicylate) Fixed Oil Ester of Glycerol + Fatty Acid Can be saponified Permanent spot on brown paper Cannot be isolated by distillation Become rancid

METHODS OF OBTAINING VOLATILE OILS 1. Distillation a. Water distillation b. Water and steam distillation c. Steam distillation d. Destructive distillation

Volatile Oil Made up of terpenes Cannot be saponified No permanent spot on brown paper Can be distilled from natural source Resinify on long standing

- for those not injured by boiling (turpentine oil) - for those injured by boiling (clove oil, cinnamon oil) - for fresh plant drugs (peppermint, spearmint) - empyreumatic oils (heat without the access of air)

2. Enzymatic Actions – mustard oil 3. Expression a. Ecuelle b. Enfleurage

- rolling the fruit over a trough lined with sharp projections (citrus oils) - expression with the use of cold fat (perfume)

4. Extraction – using solvent systems based on volatile solvents (petroleum ether/benzene)

MEDICINAL AND COMMERCIAL USES 1. Fabrication of Perfumes

Top Notes Middle Notes Base Notes aka Fixatives

Definition Most volatile products Leave the skin readily Intermediate tenacity and volatility Low volatility and high tenacity

Examples Lemon oil, Lavender oil, Anise oil Thyme oil, Neroli oil, Rose oil Musk Dried secretion from the preputial follicles of the male musk deer of Asia (Moschus spp) Civet A glandular secretion appearing in an outwardly discharging pockets of Civet cats (Paradoxurus hermaphroditus) Ambergris Most valuable material Pathologic product formed in the stomach of spermwhale when it feeds on squid or cattlefish

Volatile oils generally consist of: Eloptene – hydrocarbon portion, liquid Stearoptenes – oxidized hydrocarbon portions, solid (generally obtained by freezing the oil)

CONSTITUENTS OF VOLATILE OILS Terpenes – natural products whose structures may be divided into isoprene units C5H8 Monoterpenes – most abundant in nature Sesquiterpenes Diterpenes Triterpenes

2 isoprene units 3 isoprene units 4 isoprene units 6 isoprene units

C10H16 C15H24 C20H32 C20H48

Phenylpropanoids – contain C6 phenyl ring with an attached C3 propane side chain – those in volatile oils are phenol or phenol ethers

BIOSYNTHESIS OF VOLATILE OIL CONSTITUENTS Isoprene units – building blocks for terpenes Synthetically active isoprene units: isopentyl pyrophosphate dimethylallyl pyrophosphate Geranyl pyrophosphate – C-10 precursor of terpenes Principal precursors of phenylpropanoid compounds: cinnamic acid p-hydroxycinnamic acid (p-coumaric acid) These compounds arise from phenylalanine and thyrosine, which in turn synthesized via shikimic acid pathway.

HYDROCARBON 1. Turpentine Oil (Spirit of Turpentine) BO : Pinus palustris and from other species of Pinnus Linne Use : Disinfectant, Deodorant, Counterirritant in Vicks Vaporub Rectified turpentine oil - rectified by distillation from an aqueous solution of NaOH Terpin hydrate/Terpinol/cis-p-menthane-1,8-diol hydrate - formed by action of nitric acid on rectified turpentine oil in the presence of alcohol - expectorant

ALCOHOL Peppermint Oil

Mentha piperita

Menthol

Japanese Peppermint Oil Cardamom Oil

Mentha arvensis Elettaria cardamomum

High menthol concentration than M. piperita but it has an inferior flavor Flavor, carminative

Coriander Oil

Coriandum sativum

Flavor, carminative

Rose Oil Otto of Rose Orange Flower Oil Neroli Oil Juniper Oil

Rosa gallica, R. damascena, R. alba, R. centifolia Citrus aurantium

Perfume

Juniperus communis

Flavor, diuretic in the OTC drug, Odrinil®

Cinnamomum loureirii

Flavoring, carminative, pungent aromatic. Antiseptic

Perfume, flavor

ALDEHYDE Cinnamon Oil Cassia Oil

Cinnamaldehyde Natural: cassia oil Synthetic: benzaldehyde + acetaldehyde (action of NaOH) Lemon Peel

Citrus limon

Limonene & Citral (neral & geranial)

Lemon Balm

Melissa officinalis

Citral

Orange Oil

Citrus sinensis

Limonene

Citronella Oil

Cymbopogon winterianus, C. nardus

Perfume, insect repellant

Hamamelis Water

Hamamelis virginia

Astringent in the OTC hemorrhoid preparations

KETONE Camphor

Cinnamomun camphora Carvone

Antipruritic

Synthetic: racemic Natural: dextrorotatory (+)Carvone has the same odor as that of caraway

Flavor, carminative Gums, toothpaste, mouthwash

Spearmint

Mentha spicata

Caraway Oil

Carum carvi

(+)Carvone

Flavor, carminative

Buchu

Barosma betulina, B. Crenulata B. Serratifolia

*Diosphenol

Diuretic in OTC drugs for menstrual problems, Fluidex, Odrinil

Wormwood Oil Absinthe Oil

Artemisia absinthium

Thujone

Counterirritant in Absorbine Jr.

Cedar Leaf Oil

Thuja accidantalis

Thujone, Fenchone

Counterirritant in Vicks Vaporub

Clove Oil

Eugenia caryophyllus

Dental analgesic

Thymol

Thymus vulgaris

CREOSOTE Beachwood creosote

Fagus grandiflora

Eugenol (4-allyl-2methoxyphenol) Thymol - synthetically from m-cresol or p-cymene Gualacol, Creosol

JUNIPER TAR Cade oil

Pinus palustris

Myrcia oil

Pimenta racemosa

PHENOL VOLATILE OILS Antifungal Antibacterial disinfectant in Dewitt’s Toothache drops, expectorant in Creomulsin Antieczematic and anti-psoriasis (Packer’s Pine Tar, Polytar), Expectorant

PHENOLIC-ETHER VOLATILE OILS Biosynthesis of Phenolic Ethers – takes place from phenylalanine (schikimic acid-phenylpropanoid pathway) Methionine – methyl donor for methoxylation reaction Nutmeg Myristica Oil Fennel Oil

Myristica fragrans

Anise Oil

Pimpinella anisum (anise or aniseed), Illicium varum (chinese anise or star anise)

Foeniculum vulgare

Safrole Myristicin Anethole

Flavoring, Carminative

Anethole

Flavoring, Carminative

OXIDE VOLATILE OILS 1. Eucalyptus Oil/Cineole BO : Eucalyptus globulus AC : Cineole/Eucalyptol/Cajuputol Use : flavor, antiseptic, diaphoretic, expectorant 2. Ascaridol

Flavoring, Carminative

ESTER VOLATILE OILS Biosynthesis of Esters - terpene esters are generally formed from the respective alcohols by reaction with aliphatic acid moieties (commonly acetic acid) Gaultheria Oil Wintergreen/Betula/ Sweet birch

Gaultheria procumbens Betula lenta

Lavender Oil

Lavandula angustifolia

Perfume

Pine Needle Oil Dwarf pine needle oil

Pinus mugo

Perfume

Mustard Oil

Brassica nigra

SESQUITERPENES 1. Chamomile 2. Artemisinin Quinghaosu 3. Valerian 4. Feverfew DITERPENES 1. Taxol 2. Ginkgo

Methylsalicylate

Allyl isothiocyanate Acrinyl isothiocyanate

Flavor (aromatic cascara sagrada fluidextract local irritant, antiseptic, antirheumatic

Rubefacient in Musterolew

Matricaria chamomila Artemisia annua

Carminative, spasmolytic Antimalarial

Valeriana officinalis Tanasetum parthenium

Sedative and anxiolytic Antipyretic, migraine tx

Taxus brevifolia Ginkgo biloba

Antineoplastic Increases blood fluidity

TRITERPENES Quassinoids – pediculocide 1. Neem Tree Azadirachta indica

Antifeedant properties (mosquito repellant)

RESIN AND RESIN COMBINATIONS Amorphous products with complex chemical nature Hard transparent or translucent and when heated they soften and melt Resin Combinations: 1. Oleoresins 2. Oleo-gum-resins 3. Balsams 4. Glucoresins or glycoresins

– resins + volatile oils – resins + volatile oils + gum – contain cinnamic acid, benzoic acid, or both – glycosidal combinations (ex. jalap and podophyllum)

Resin Acids (abietic acid, copaivic & oxycopaivic acid, commiphoric acid) • contain large proportion of oxyacids, usually combining the properties of carboxylic acids and phenols • resinates – metallic salts of resin acids (some are used in soaps and varnishes) Resin Alcohols (benzoresinol, storesinol) • do not give a tannin reaction with iron salts • occur in free state and as esters in combination with simple aromatic acids Resinotannols (aloeresinotannol, peruresinotannol, siaresinotannol and sumaresinotannol, and toluresinotannol) • gives a tannin reaction with iron salts Resenes – B-complex natural substances devoid of characteristic chemical properties Glycoresins – yields sugars and complex resin acids on hydrolysis Pharmaceutic Resins are usually obtained by: - extracting with alcohol & precipitating the resins in water (jalap & podophyllum) - separating the oil from oleoresin by distillation (copaivic resin) - collecting the natural product throught natural or artificial punctures (mastic)

RESINS Other Name

Origin

Rosin

Colophony

Podophyllum

Mayapple Mandrake

Eriodictyon

Yerba santa

Jalap

Jalap root

Exogonium purga

Mastic

Mastiche, Mastich

Pistacia lentiscus

Kava

Kava-kava

Piper methysticum

Cannabis

Indian hemp Marihuana, Pot

Cannabis sativa

Constituent

Pinus Palustris Podophyllum peltatum Indian Podophyllum – rhizome of P. emodi Eriodictyon californicum

Use Stiffening agent

Podophyllin

Drastic purgative Antimitotic (papilomas) Disguise Quinine bitterness Stimulating expectorant (1g)

Jalap resin

Hydragogue cathartic Dental varnish to seal cavities

Hashish

Centrally acting muscle relaxant Antipyretic, local anaesthetic THC – euphoric agent Cannabidiol – manuf of rope

OLEORESINS Turpentine

Other Name Gum turpentine Gum thus

Origin

Constituent

Pinus palustris

Counterirritant

Capsicum

Cayenne pepper

Capsicum frutescens

Ginger

Zingiber

Zingiber officinale

White Pine Copaiba

Pinus srobus Balsam of Copaiba

Copaifera spp.

Use

Capsaicin

Coniferin

Irritant, carminative, Rubefacient, Stimulant, Condiment Flavor, condiment, stimulant, carminative, ginger ale Expectorant Genitourinary disinfectant, Diuretic, Expectorant, Laxative

OLEO-GUM-RESINS 1. Myrrh/Gum myrrh 2. Asafetida/Devil’s Dung

Commiphora molmol

Astringent, Embalming

BALSAMS Storax Liquid Storax Styrax Peru Balsam Tolu Balsam

Origin Liquidambar orientalis

Use Pharmaceutic aid for compound benzoin tincture Stimulant, Expectorant, Antiseptic

Myroxylon pereirae Myroxylon balsamum

Local protectant and rubefacient, Antiseptic Pharmaceutic aid in compound benzoin tincture Expectorant, Flavoring in syrups, confectionery, chewing gum, perfumery Antiseptic Compound Benzoin Tincture (benzoin, aloe, storax, tolu) Benzoic acid (synthetic product; first obtained by sublimation from Sumatra benzoin; antifungal)

occurs as plastic solid that gradually hardens

Benzoin

Styrax benzoin

ALKALOIDS Bitter, basic nitrogenous compounds, sparingly soluble in water Most of them end in –ine All are solids except: o Coniine o Nicotine o Sparteine Form double salts with compounds of mercury, gold, platinum, and other heavy metals

Alkaloidal reagents/precipitants: 1. Mayer’s Mercuric potassium iodide 2. Marmers Potassium-cadmium iodide 3. Dragendorff’s Potassium-bismuth iodide 4. Wagner’s Iodine TS 5. Scheibler’s Phosphotungstic acid 6. AuCl 7. Valser’s KI and mercuric iodide 8. Tannic acid 9. Hager’s Picric acid 10. Sonnenscheins Phosphomolybdic acid

White or slightly yellow ppt. White ppt. Orange Reddish brown or red brown ppt.

White ppt. Yellow

CHARACTERISTICS OF ALKALOIDS 1. 2. 3. 4. 5. 6. 7. 8.

Free alkaloids - sparingly soluble in H2O + acids --> salts + salts of heavy metals --> highly insoluble precipitates nitrogenous, physiologically active alkaloidal salt + Na2CO3 --> free base Solid alkaloid - white, soluble or slightly soluble in H2O, soluble in organic solvents Pure alkaloids - insoluble in water, soluble in organic solvents alkaloids + acids = alkaloidal salts (soluble in water)

Alkaloid yielding plants: 1. Leguminosae 2. Solanaceae 3. Rubiaceae 4. Ranunculaceae 5. Papaveraceae 6. Berberidaceae

Pharmacologic Action of Alkaloids 1. Analgesic and narcotics 2. Central stimulant 3. Mydriatics 4. Miotics 5. Cause a rise in B.P 6. Excessive hypertension

-morphine, codeine - strychnine, brucine - atropine - physostigmine, pilocarpine - ephedrine - reserpine

The names of the alkaloids are obtained in various ways: 1. generic name (Hydrastine, Atropine) 2. specific name (Coccaine, Belladona) 3. common name (Ergotamine) 4. physiologic activity (Emetine, morphine) 5. discoverer (Pelletietine) FUNCTIONS OF ALKALOIDS IN PLANTS 1. poisonous agents – protect plants againts insects & herbivores 2. end products of detox representing metabolic locking up of compounds otherwise harmful to the plant 3. Regulatory growth factor 4. Reserve substance capable of supplying nitrogen or others

PYRIDINE-PIPERIDINE Origin Nicotine

Use

Nicotiana tabacum

Nicotine (smoking deterent)

Areca

Betel nut

Areca Catechu

Arecoline Anthelminthic Tannin content causes esophageal CA

Lobelia

Indian tobacco

Lobelia inflata

Lobeline (smoking deterent)

Origin Atropa belladonna Atropos – fate that cuts the thread of life

Use Parasympathetic depressant Adjunct in tx of peptic ulcer

TROPANE/SOLANACEOUS

Belladonna

Deadly nightshade

Hyoscyamus

Henbane

Hyoscyamus niger

Hog + Bean

Stramonium

Jimson Weed Jamestown Weed

Datura stramonium D. metel (Talumpunay/Thorn apple)

Treatment of asthma (vapor)

Mandragora

European mandrake

Mandragora officinarum

Has folkloric uses

Cocaine

Coca

Erythroxylum coca

Psychomotor stimulant with a strong abuse potential, vasoconstrictor

toxic to swine

Principal Solanaceous Alkaloids: 1. Atropine o Antispasmodic o Antisialogogue – decrease production of secretion o Mydriatic and cycloplegic o Antidote for cholinesterase inhibitors 2. Scopolamine or Hyoscine o Hyoscyamus niger o Treatment of motion sickness o Trans-scoop or transderm Scop – 4 hours before anti-emetic effect

QUINOLINE Cuprea o Remijia purdieana Cinchona or Peruvian bark o Cinchona succirubra (Red cinchona) Cinchona calisaya (Yellow cinchona) o Quinidine - anti-arrthythmic Quinine - antimalarial o Identification of the quinine /quinidine by Thalleioquin test

ISOQUINOLINE – emerald green color of thalleioquin (Joseph Caventou with Pelltier) Other Name Ipecac

Origin

Constituent

Cephalis ipecacuanha

Emetine

Use Ipecac syrup – emetic Fluidextract – 14x more potent Ipecac + Opium - diaphoretic

Sanguinaria

Bloodroot

Sanguinaria canadensis

Sanguinarine

Expectorant & emetic

Tubocurarine

Curare

Strychnus toxifera

Tubocurarine

Skeletal muscle relaxant

Hydrastis

Golden seal

Hydrastis canadensis

Hydrastine

Astringents in inflammation of the mucous membrane

Opium

Stone of immortality

Papaver somniferum

Alkaloids of Opium 1. morphine 2. codeine methyl morphine 3. heroin diacetylmorphine 4. Papaverine 5. Hydromorphine 6. Hydrocodone 7. Apomorphine 8. Narcotine/Noscapine 9. Paregoric 10. Dover’s powder

Stimulate and depresses CNS

- most abundant, most important - most widely used; antitussive - hallucinogen - relief of ischemia & arrhythmias

- emetic - ANARCOTINE, antitussive - camphorated opium tincture - Ipecac + opium

IMIDAZOLE Pilocarpine (Pilocarpus, Jaborandi) BO : Pilocarpus jaborandi Use : Cholinergic drug used in the Tx of glaucoma; miotic

STEROIDAL Veratum Viride (American or Green hellebore) BO : Veratrum viride Uses : hypotension, cardiac depressant, Black hellebore – cardiac stimulant White Hellebore (European hellebore) BO : Veratrum album Use : Hypotensive, Insecticide (crude drug)

INDOLE Other Name Rauwolfia serpentina

Catharanthus

Vinca Chichirica

Nux Vomixa

Constituent Reserpine

Catharanthus roseus

Rescinnamine Deserpidine (IIdesmethoxyreserpine) Yohimbine - TX: impotence (Serpasil) Vincristine and Vinblastine antineoplastic/oncolytic Strychnine – toxic central stimulant Brucine – alcohol denaturant Physostigmine/Eserine - Tx glaucoma, miotic

Strychnos nux-vomica

Physostigmine Ergot

Origin Rauvolfia serpentina

Calabar bean Ordeal bean Rye ergot Secale cornutum

Physostigma venenosum Claviceps purpurea

Use Snake bites Insanity (Psychotherapy) Hypotensive Sedative, Tranquilizing

Ergonovine – oxytocic Ergotamine – migrane Lysergic acid Diethylamide (LSD) most specific and most active hallucinogen discovered by A. Hoffman in 1943

ALKALOIDAL AMINES Constituent Ephedrine

Ephedra, Mahuang

Colchicine

Use

Ephedra sinica

Sympathomimetic

Colchicum autumnale

Plant genetics: doubles the chromosomes Gout suppressant: Acute

Khat

Abyssinian tea

Catha edulsis

Cathinone - alleviate sensation of hunger and fatigue

Peyote

Mescal buttons

Lophophora williamsii

Mescaline (3,4,5 trimethoxy-ẞ-phenylethylamine) Hallucinogen and euphoric

PURINE OR METHYLXANTHINES 1. Caffeine 1,3,7 trimethylxanthine o Does not react with or ppt with alkaloidal reagnents o Kola (dried cotyledon of Cola nitida) o Cofee bean (dried seeds of Coffea arabica) o Guarana (Paullinia cupana) o Mate/Paraguay tea (Ilex paraguariensis)

Caffeine is detected by means of a Murexide test purple color

2. Theophylline 1,3 dimethylxanthine o Camella sinensis o Treatment of bronchial asthma, Tocolytic agent o Aminophylline – semisynthetic theophylline derivative 3. Theobromine 3,7 dimethyl xanthine o Theobroma cacao o Diuretic in cardiac & pulmonary edema (preferred over caffeine - little action on the CNS)

BIOLOGICS Any product derived from a living plant or animal source Any virus, therapeutic serum, toxin, antitoxin or analogous product Not chemotherapeutic nor an antibiotic Uses of biologics: o Prophylactic or Preventive Therapy o Therapeutic or Curative Measures

1. 2.

Types of Immunity Natural (Innate) Immunity Acquired Immunity Active Immunity a. Naturally Acquired Active b. Artificially Acquired Active Passive Immunity a. Naturally Acquired Passive b. Artificially Acquired Passive

2 GENERAL CATEGORIES Antigen provokes the immune response Chemically: Antigens are usually protein Biologically: causes formation of antibodies when introduced to tissue Physically: H molecular weight (more than 10,000 daltons) 2 properties: Immunogenicity – capacity to induce Ab formation Specificity Antibody predominant in the serum fraction of the blood 4 fractions obtained when serum proteins are separated by electrophoresis: 1. serum albumin 2. alpha globulin 3. beta globulin 4. gamma globulin Immunoglobulins/Antibodies/Serum Globulin Ab that occur predominately in the gamma globulin fraction Attacks antigens 5 subclasses of Ig: IgA External secretions (tears, saliva, respiratory, GI) IgD Found in surface of B lymphocytes (no fxn yet) IgE Allergy IgG Smallest (passes through placenta) IgM Largest First Ig to be formed during infection (newborn) Macrophages and Lymphocytes B cells T cells

Epitope simplest form of antigenic determinant present on a complex antigenic molecule Haptens compounds with m.w lower than 10,000 daltons partial antigens lack immunogenicity but can attach to host proteins to form a complete antigen

4 subsets of T cells

Cell-mediated immunity: TC cells – cytotoxic, attack cell membranes bearing their specific antigen TD cells – delayed hypersensitivity cells – releases lymphokines, which recruit & activate macrophages Immunoregulatory: TH (helper) cells – interaction with B cells is essential for optimal humoral immunity TS (supressor) cells – diminish activity of B cells

Major cellular components of immune system Orgin of serum antibodies (humoral immunity) originate in bone marrow but depend on thymus gland for differentiation cellular immunity, delayed hypersensitivity sentinels of immune surveilance against cancer and mediators of graft rejection

B memory cells

some progeny of the selected clones that remain as circulating B cells serve as immune system’s memory

Plasma cells

other member of selected B-cell clones grow larger, stop reproduction, & continuously secrete large quantities of Ab

VACCINES

AE: fever, malaise, soreness at injection site rubella vaccine: arthralgia and arthritis pertussis vaccine: convulsions “swine flu”, 1976: Guillain-Barre syndrome

living, attenuated, or killed viruses, killed rickettsiae, or attenuated or killed bacteria must be induced prior to infection (prophylactic) – exception: rabies vaccination Nonliving vaccines provide protection for a limited time: typhoid fever, cholera, plague, typhus With living agents (single vaccination): measles, rubella, or mumps vaccine Multiple immunizations: polio CI: immunosupressed patients

VIRAL VACCINES

• cultures: chick embryo, monkey kidney, or human diploid cells Description Grown on Smallpox Vaccine (Dr. Edward Jenner) Living virus of vaccinia (cowpox) Skin of a vaccinated bovine calf Rabies Vaccine Sterile lyophilized preparation of: Human diploid embryo lung tissue Human Diploid Cell Rabies Vaccine (whole virion) Whole virion (Wistar rabies (HDCV) Human diploid cell cultures (subvirion) virus) ® ® (Imovax , WYVAC ) Subvirion rabies virus (Pasteur- *Brain tissue – formerly used for preparing the vaccine; myelin Pasteur was able to “fix” rabies derived Pitman-Moore virus) (“paralytic factor”) virus Yellow Fever Vaccine Attenuated strain of living yellow Living embryo of domestic fowl (YF-VAX®) fever virus (Vector: Aedes mosq) (Gallus domesticus) Influenza Virus Vaccine Sterile, aqueous solution of Extra-embryonic fluid of influenza (Fluzone®, Fluogen®) suitably inactivated influenza virus virus-infected chick embryo types A and B high strain specificity and genetic instability (reevaluated regularly) Poliovirus Vaccine Inactivated Sterile suspension of inactivated Rhesus monkey kidney tissues IPV, Salk poliomyelitis virus types 1, 2 and 3 Type 1 (Brunhilde) poliovirus – responsible for major epidemics Type 2 (Lansing) poliovirus – sporadic Type 3 (Leon) poliovirus – less frequent epidemics Improved strains: Type 1 Mahoney strain Type 2 MEF-1 strain Type 3 Saukett strain

Poliovirus Vaccine Live Oral (Orimune®) Trivalent Oral Polio Vaccine (TOPV)

Landsteiner and Popper – first transmitted poliovirus John F. Enders – originated a method of cultivating poliovirus Jonas Salk – perfected the roller-tissue method, as well as the detoxified form of vaccine

Preparation of one or a combination of the 3 types of live, attenuated polioviruses

Monkey kidney tissue

Measles virus vaccine live Rubeola vaccine (Attenuvax®) Rubella virus vaccine live (Meruvax II®) - For German measles, CI:Pregnancy

Attenuated viruses derived from the original Edmonston B strain Wistar Institute RA 27/3 strain

Chick embryo tissue

Mumps Virus Vaccine Live (Mumpsvax®) Hepatitis B Vaccine (Heptavax-B®)

B-level Jeryl Lynn strain chemically inactivated hepatitis B surface antigens (HbsAg) particles

Chicken embryo tissue plasma of healthy chronic HbsAg carriers by plasmaphresis

Albert Sabin, Harold Cox, Hilary Koprowski – developed procedures for manufacturing the vaccine (separately)

Human diploid cell tissue

Combination Virus Vaccines o Live measles and rubella (M-R-Vax II), Live rubella and mumps (Biavax II), Live measles, mumps, and rubella (M-M-R II)

RICKETTSIAL VACCINES G : chick embryos or monkey kidney tissue cultures Use : murine typhus, tsutsugamushi fever, rickettsial diseases, epidemic typhus

BACTERIAL VACCINES suspensions of attenuated, more commonly, killed pathogenic bacteria in isotonic NaCl soln “S”/smooth strains are more antigenic than “R”/rough strains Typhoid Vaccine (Enteric vaccine)

Killed Salmonella typhi (Type 2)

Cholera Vaccine

Killed Vibrio cholerae (Inaba & Ogawa strains)

Plague Vaccine

Killed Yersinia pestis (Reservoir: rats | Vector: fleas)

Pertussis Vaccine

Killed Bordetella pertussis Adsorbed Pertussis Vaccine – pertussis vaccine + of AlOH3 or Al2(PO4)3

Tuberculosis Vaccine (BCG Vaccine)

Dried, living M. Tuberculosis(Calmette-Guerin strain)

Meningitis Vaccine

Capsular polyssacharides for Neisseria meningitidis (A, C, Y, and W-135)

Pneumococcal Vaccine Polyvalent

Polysaccharide antigens from Streptococcus pneumoniae

Haemophilus b Polysaccharide Vaccine

Capsular polyvalent polysaccharide of Haemophilus influenzae type b (Hib)

TOXINS AND TOXOIDS o o o o o

Toxins – (exotoxins) bacterial waste products that are considered poisonous to the animal body Toxoid – toxin whose toxicity has been inactivated or suppressed either by chemical (formalin) or heat Act as antigens because they stimulate certain cells of the body to produce antitoxins Culture medium: beef broth medium Clostridium botulinum – source of the most poisonous poison Fluid Toxoids - formaldehyde (reduces or eliminates toxic properties w/o affecting antigenicity) - induces artificial active immunity Adsorbed Toxoid - precipitated or adsorbed with alum, aluminum hydroxide, or aluminum phosphate - slower release of antigen, subsequent production of higher & more prolonged Ab titers - more prone to produce local reactions at the injection site

ANTITOXINS (From blood of toxin-immunized animals, usually horses) o o

Standardized in terms of “antitoxin units” Admin: prepare a syringe containing epinephrine injection and torniquet to counter anaphylactic reactions

Diphtheria Antitoxin Tetanus Antitoxin Botulism Antitoxin type A, B, E strains

Use Passive immunity, curative Treatment & prophylaxis of tetanus if Tetanus Ig is not available (passive immunity) (multivalent) treat all cases of toxemia caused by types of botulinus bacteria used

VENOMS AND ANTIVENINS (From blood of venom-immunized animals, usually horses) o o

Venoms – poisonous excretions produced by animals; equivalent to exotoxins in bacteria Snake venins or venoms – obtained by holding a poisonous snake over a conical glass container covered with a sheet of thin rubber

Antivenin (Crotalidae) Polyvalent Antivenin Spider-Bite Antivenin

Synonym North and South American antisnakebite serum

North American coral snake antivenin Antivenin (Latrodectus mactans) Black widow spider antivenin

Against venom of Crotalus atrox (Western diamondback) C. adamanteus (Florida diamondback) C. durissis terrificus (S. American rattlesnake) Bothrops atrox (S. Amarican fer-de-lance) Micrurus fulvius (eastern coral snake) M. fulvius tenere (Texas coral snake) Latrodectus mactans (black widow spider)

ANTISERUMS (From blood of bacteria/virus-immunized animals, usually horses) Antirabies Serum (more effective if administered with rabies vaccine) D : sterile, nonpyrogenic solution containing antiviral substances From : blood serum or plasma of a healthy horse immunized against rabies by vaccination

IMMUNE GLOBULINS (From immune humans (survived an attack or immunized) Immune Globulin Immune serum globulin Immune globulin intramuscular Gamma globulin

Use Prophylactic in chickenpox, hepatitis A, and other diseases Immune Globulin Intravenous/IGIV provides immediate Ab levels tx of immunodeficiency syndrome

Pertussis Immune Globulin

Prophylaxis and treatment of pertussis

Tetanus Immune Globulin

Passive immunization against tetanus in individuals with wounds that may have been contaminated

Rabies Immune Globulin

Used with rabies vaccine (best postexposure prophylaxis)

Hepatitis B Immune Globulin Varicella-Zoster Immune Globulin

Restricted to immunodeficient individuals

RH0 (D) Immune Globulin

For unsensitized Rh-negative women who give birth to RH0 (D)- or Dupositive infant

Lymphocyte Immune Globulin

Lymphocyte selective immunosuppresant

DIAGNOSTIC ANTIGENS positive response: localized, well-defined wheal accompanied by erythema From Human or bovine strains of tubercle bacillus

Tuberculin Old Tuberculin Purified Protein Derivative of Tuberculin Histoplasmin Coccidioidin Diphtheria Toxin Mumps Skin Test Antigen

Positive Response redness or inflammation

M. tuberculosis

inflammation and palpable induration or edema (5 mm)

M. tuberculosis (free from protein)

palpable induration (10 mm or more)

Histoplasma capsulatum (mycelial phase) Fungus Coccidioides immitis Corynebacterium diphtheriae (nlt 400 MLD/min lethal doses) Formaldehyde-inactivated mumps virus grown on extra-embryonic fluids of the mumps virus-infected chicken embryo

circumscribed area of redness (≥1 cm) erythema (at least 1.5 cm)

BIOLOGICS RELATED TO HUMAN BLOOD Whole Blood (human) Red Blood Cells Plasma Protein Fraction (human) Thrombin

From donor (+ citrate/heparin as anticoagulant) Whole blood minus plasma Selected proteins derived by fractionating material (source blood plasma, or serum) Sterile protein from prothrombin (bovine) + thromboplastin in the presence of calcium

Blood replenisher Blood replenisher in anemia Blood volume restoration (in shock) Hemostatic

Antihemophilic Derivatives – control of 2 types of hemophilia Antihemophilic factor

Antihemophilic factor IX complex Albumin Human

sterile, freeze-dried concentrate of human antihemophilic factor (prepared from the factor VIII-rich cryoprotein fraction of human venous plasma) dried plasma fraction comprising coagulation factors IX, II, VII, X sterile, nonpyrogenic preparation of serum albumin obtained by fractionating material (source blood plasma, serum, or placentas) from healthy human donors

Hemophilia A (classic hemophilia) Prevent dangerous bleeding or to perform surgery blood-volume supporter

Radio-Iodinated Serum Albumins contains serum albumin iodinated using mild conditions with either 125I or 131I sterile, buffered, isotonic solutions prepared to contain nlt 10 mg of radio-iodinated normal human albumin per ml & adjusted to provide nmt 1 millicurie of radioactivity per ml Iodinated 125I albumin injection and Iodinated 131I albumin injection Iodinated 131I aggregated injection

diagnostic aid to determine blood volume and cardiac output diagnostic aid for determination of pulmonary clearance

HERBS AND “HEALTH” FOODS Natural – identifies only a source, not necessarily degree of superiority or inferiority Organic – the produce was grown under conditions utilizing only natural fertilizers, no pesticides Alfalfa Aloe Angelica Apricot Pits Arnica Asafetida Aveloz Black Cohosh Blessed Thistle Blue Vervain Boneset Borage Bran Broom Buchu Burdock Butcher’s-Broom Calamus Canaigre Catnip Chamomile Chaparral Chicory Cholecystokinin Cocillana Collinsonia Coltsfoot Comfrey Cucurbita Seed Damiana Devil’s Claw Echinacea Eleuthero Evening Primrose Eyebright Fenugreek Feverfew Fo-ti-tieng Garlic Onion Gentian ( Ginseng Glucomannan Glycyrrhiza Goldenseal Gotu Kola

lucerne

gum asafetida, devil’s dung killwart

wild hyssop common borage Scotch broom lappa box holly sweet flag wild red American ginseng

creosote bush CCK stoneroot farfara

Cone/purple cone flower Siberian ginseng

Vitamin X

gentian root Konjac mannan licorice Hydrocotyle, Indian pennywort

Medicago sativa A. barbadensis(Aloe vera) or Curacao, A. spicata (Cape aloe) Angelica archangelica Prunus armeniaca Arnica montana Ferula assafoetida, F. rubricaulis, F. Foetida Euphorbia heterodoxa Cimicifuga racemosa Cnicus benedictus Verbena hastata Eupatorium perfoliatum Borago officinalis Triticum aestivum Cytisus scoparius Barosma betulina, B. crenulata, B. Serratifolia Arctium lappa Ruscus aculeatus Acorus calamus Rumex hymenosepalus Nepeta cataria Matricaria chamomilla (German), Anthemis nobilis (Roman) Larrea tridentata Cichorium intybus secreted by the duodenal mucosa Guarea rusbyi Collinsonia canadensis Tussilago farfara Symphytum officinale Cucurbita pepo, C. maxima, C. moschata Turnera diffusa Harpagophytum procumbens Echinacea angustifolia Acanthopanax senticosus/Eleutherococcus s. Oenothera biennis Euphasia officinalis Trigonella foenumgraecum Chrysanthemum parthenium Centella asiatica, Gillena trifoliata (meadowsweet), C. nitida Allium sativum Allium cepa Gentiana lutea Panax quinquefolius, P. Pseudoginseng Amorphophallus rivieri Hydrastis canadensis Centella asiatica

Hawthorn Hedge Hyssop Hibiscus Honey Hops Horehound Horse Chestnut Horsetail Hydrangea Hyssop Kelp Lecithin Life Root Linden Flowers Lovage Marigold Mistletoe Mormon Tea Muira Puama Oregano Pangamic Acid Parsley Passiflora Pau d’ Arco Pennyroyal Poke Root Pollen Propolis Red Bush Tea Rose Hips Royal Jelly Sarsaparilla Sassafras Schisandra Senega Snakeroot Senna Spirulina Super Oxide Dismutase Tamarind Tarragon Tienchi Tonka Beans L-Tryptophan Turmeric Valerian Wild Lettuce Wormwood Yarrow Yohimbe Yucca

Sudanese tea mel homulus

golden ragwort

American or European mistletoe Mexican/teamster/squaw tea, popotillo potency wood wild marjoram Ca pangamate, pangamic acid, vit B15

taheebo, ipe roxo

bee glue rooibos tea

sassafras bark schizandra senna leaves SOD

titnchi-ginseng, sanchi

midfoil

Crataegus monogyna Gratiola officinalis Hibiscus sabdariffa Apis mellifera Humulus lupulus Marrubium vulgare Aesculus hippocastanum Equisetium arvense Hydrangea arborescens Hyssopus officinale Macrosystis, Nereocystis, and Laminaria spp. Phosphatide mixture that yield α- or β-glycerophosphoric acid, fatty acids, and choline on hydrolysis Senecio aureus Tilia species Levisticum officinale Calendula officinalis Phoradendron tomentosum, P. serotinum, flavescens Ephedra nevadensis Ptychopetalum olacoides, P. uncinatum Origanum vulgare blood abnormalites, asthma, emphysema, alcoholism Petroselinum crispum Passiflora incarnata Tabebuia impetiginosa Phytolacca americana microspores of seed-bearing plants natural antibiotic Aspalathus linearis Rosa canina secretion from the paired salivary glands of Apis mellifera Smilax aristolochiaefolia, S. regelii, S. Febrifuga Sassafras albidum Schisandra chinensis Polygala senega Cassia spp. Leaflets Spirulina maxima metal lipoprotein from the liver or RBC of ox, Bos taurus Tamarindus indica Artemisia dranunculus Panax notoginseng Dipteryx odorata sleep-onset insomnia Curcuma longa Valeriana officinalis Lactuca virosa Artemisia absinthium Achillea millefolium Pausinystalia yohimbe Yucca spp.

BIOCHEMISTRY/MOLECULAR BIOLOGY Parts of a Cell

basic functional and structural unit of life

1. CELL MEMBRANE 1. Phospholipds 2. Glycoproteins and Glycolipids 3. Cholesterol 4. Arachidonic Acid

CELL WALL Plants Cellulose Bacteria Peptidoglycan Fungi Chitin

2. ORGANELLES 1. Mitochondrion - powerhouse of the cell; produces ATP (energy)

2. Ribosomes

- protein synthesis Prokaryote: 30s, 50s = 70s Eukaryote: 40s, 60s = 80s

3. Lysosomes

- suicide bag of the cell - has enzymes and free radicals (when released, will cause APOPTOSIS) Red blood cells – 120 days *Hemolytic anemia – 120 (linseed, fish) Semidrying 100-120 (cottonseed, sesame) Nondrying