Dr. Areej M. Al-Taweel Pharmacognosy Department Pharmacognosy Department.
Analytical Pharmacognosy
Transcript of Analytical Pharmacognosy
SUKRASNO SITI KUSMARDIYANI
-- explain --
Tekelan, Epatorium reptansKumis kucing, Orthosiphon stamineus
Authenticity ---explain
KUSMARDIYANI
BOTANIC IDENTITY TEST Morphological, Microscopical, Microchemical,
and Genetic Methods
© SK
“SIMPLISIA”
Ver. 22SK005
CRUDE DRUGS
DriedIntactSliced
Morphological Method
THE ORGAN COMPOSING CRUDE DRUGS:– Leaves and tops (herbs)– Barks– Woods– Leaves or leaflets– Fruits– Seeds– Subterranean Organs
– Aerial stem• Dimension,
• shape,
• colour,
• herbaceous or woody,
• up right or creeping,
• smooth or ridged,
• Hairs
• Position and arrangement of leaves– Radical or cauline
– Adnation
– Alternate
– Opposite
– Decussate
– Whorled
Morphological Method
LEAVES AND TOPS
Alternate = Placed singly along the stem or axis, not opposite or whorled.Distichous = Arranged in two vertical ranks.Opposite leaves = With two leaves at a node, one on each side of the stem or axis.Deccusate = In opposite pairs, each pair at right angles to the next.
LEAF ARRANGEMENTS
ALTERNATE ALTERNATE, DISTICHOUS
OPPOSITE OPPOSITE, DECUSSATE
Get more types of leaf arrangements and give examples
Cambridge Glossary of Botanical Terms
Wallis, 573.
TERMS FOR LEAF DESCRIPTION
• Origin of preparation– Trunk, branches, roots.– Whole, inner bark
• Inner surface– Colour, striations, furrows
• Size and shape– Lichens, mosses, lenticels, cracks or furrows,
colour before and after scrapping.• Fracture
– Short, fibrous, splintery, granular• Transverse surface
– Smoothed transverse surface after staining with fluoroglucinol and HCl
Morphological Method
BARKS / CORTEX
© SK
Cinnamomi cortex
Litseae cortex
Cinchonae cortex
Alstoniae cortex
• Size and colour– after and before staining
• Relative density– Guaiacum 1.33 and Poplar 0.38
• Hardness and behavior when split• Transverse surface
– Distribution of wood fibers and wood parenchyma, annual ring (true or false)– Distance between medullary rays and annual rings
• Longitudinal surfaces– Height of medullary rays.
Morphological Method
WOODS
Ligustrinae lignum
Sappan lignum
• A: primary structure• B: development of a
complete cambial ring• C: beginning of secondary
growth• D: stem after a number of
seasons of growth
• Types of vascular bundle• E: collateral• F: bicollateral• G: amphivasal• H: amphicribal
Trease, 517.Stem structure of dicotyledons
STEM
• Duration – Deciduous or evergreen
• Leave base– Stipulate or exstipulate
• Petiole– Petiolate or sessile– Shape, colour, hair
• Lamina– Composition, incision, shape, venation,
margin, apex, base, surface, texture
Morphological MethodLeaves or Leaflets
Morphology of leavesA. Shape
1. acicular 2. elliptical 3. oval 4. oblong 5. round
6. linear 7. lanceolate 8. ovate
9. obovate 10. subulate 11. spatulate 12. diamond shape 13. cuneate
14. cordate 15. auriculate 16. lyrate 17. reniform
Trease, 519.
Morphology of leavesB. Composition and Incision
1. pinnatifid 2.pinnatripartie 3. pinnatisect, 4.palmatifid 5. imparipinnate
Trease, 519.
Morphology of leaves
D. Margin
E. Base
C. Apex
Trease, 519.
• Type of inflorescences– Racemose, cymose or mixed
• Axis or receptacle of influorescence• Type of flower• Receptacle of the flower• Calyx• Corolla (number of petals, venation, oil glands• Androecium• Gynaecium
Morphology of Flowers
Inflorescences
Actinomorphic Radially symmetrical, a line draw through the middle of the structure along any plane will produce a mirror image on either side
ZygomorphicBilaterally symmetrical, a line draw through the middle of the structure along only one plane will produce a mirror image on either side
Irregular Bilaterally symmetrical, a flower in which all parts are not similar in size and arragemnent on the receptacle
FLOWER TYPES
INFLORESCENCE
Trease, 521.
PERIANTH FORMS
Plant Identification Terminology, 170-171.
PERIANTH FORMS
Plant Identification Terminology, 170-171.
• Classification:– Simple, Aggregate, Collective
• Simple, dry, indehiscent fruits– Achene, nut, caryopis
• Simple, dry, dehiscent fruits– Legume, follicle, capsules
• Schizocarpic or splitting fruits
• Succulent fruits (druppe, berry)
Morphological MethodFRUITS
• Shape and dimensions– Adhesion
– Dehiscence
– Pericarp
– Placentation
– Seeds
Morphological Method
FRUITS
Foeniculi fructus
Coriandri fructus
Capsici fructusCardamomi fructus
© SKPiperis nigri fructus
Trease, 522.
FRUITSFruits of the Solanaceae:
A – B capsulesA ripe fruit of Datura stramoniumB pyxidia of Hyoscyamus niger
with upper fruit showing calyx partly removed
C berries of Atropa belladonaD berries of Capsicum sp
• Size, shape and colour
• Funicle
• Hilum and micropyle
• Seed coats
• Perisperm
• Endosperm
• Embryo
Morphological Method
SEEDS
Coffeae arabicae semen
Coffeae robustae semen
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Myristicae semen
Piperis albi semen
• Morphological nature– Root, rhizome
• Condition – Fresh, dry, whole or sliced, peeled or un peeled
• Subaerial stems • Part of the crude drug or adulteration
• Subterranean stems• Size and shape• Direction of growth and branching• Surface characters• Fracture and texture• Transverse section
• Roots• Kind• Size and shape• Surface characters• Fracture and texture• Transverse section
Morphological MethodSUBTERRANEAN ORGANS
Languatis rhizoma
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Calami rhizoma
Zingiberis rhizoma
Rauwolfiae radix
Curcumaedomesticae rhizoma
Fixed oils, fats, waxes, volatile oils, resins, oleoresins, oleo-gum-resins, balsam
Dried juices
Latices
Extracts
Physical state– Solid– Liquid
Odour and taste
Chemical tests
Unorganized Drugs
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MicroscopicalAnalysis
Trease, 520.
Trease, 528.
Anomocytic Anisocytic Diacytic Paracytic Actinocytic Bidiacytic
Ver. 22SK005
Give description for each type of stomata.
BENTUK DAN KONFIGURASI STOMATA yang berlainan dapatdigunakan sebagai patokan analisis mikroskopik serbuk simplisia.
Stahl, 54
Tipe stomata khusus:
1. Tipe Ranunculaceae= tipe anomositik
2. Tipe Cruciferae= tipe aninositik
3. Tipe Caryophyllaceae= tipe diasitik
4. Tipe Rubiaceae= tipe parasitik
Ver. 22SK005
Trease, 529.
EPIDERMISA. Lower epidermis of Digitalis purpureaB. Lower epidermis of Hyoscyamus nigerC. Upper epidermis of Atropa belladonnaD. Lower epidermis of Cassia angustifoliaE. Lower epidermis of Rosmarinus officinalisF. Lower epidermis of Mentha piperitaG. Lower epidermis of Pilocarpus jaborandiH. Upper epidermis of Lobelia inflataI. Lower epidermis of Digitalis lanataJ. Lower epidermis of Erythroxylum coca
A
B and C
D
E and F
G
Type of stomata: anomocyticanisocytic
paracytic
actinocyticdiacytic
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Microscopical Analysis
• Epidermal trichomes (hairs)Leaves, stems, flowers, fruits and seeds may have trichomes.
Trichomes– Non glandular (clothing)
• Unicellular, multicellular
– Glandular• Unicellular or multicellular stalk• Unicellular or multicellular head
EPIDERMAL TRICHOMES
Trease, 530.
A Papillae of lower epidermis of Coca leafB – G Unicellular hairsB Papillae epidermal cell with cystolith from leaf
of CannabisC Cystolith clothing hair from floral bract of
CannabisD Lobelia inflata leafE Senna leafF Lignified hair of AilanthusG ComfreyH Group of unicellular hairs from Hamamelis
leafI T-shaped hair of Artemisia absinthium
Trease, 531.
EPIDERMAL TRICHOMES
A – H Uniseriate clothing hairsI Multicellular branched hairJ Biseriate hair
A Datura metelB Datura stramoniumC Mentha piperitaD Thymus vulgarisE Plantago lanceolataF Hyoscyamus nigerG Digitalis purpureaH Xanthium strumariumI Verbascum thapsusJ Calendula officinalis
GLANDULAR HAIRS
Trease, 532.
A – B Atropa belladonnaC Datura stramoniumD Digitalis purpureaE Multicellular labiatae
glandular hairF Hyoscyamus nigerG – H Primula vulgarisI Digitalis luteaJ Cannabis sativaK Artemisia maritima
Phloem
A.sieve tubes with companion cells,
B and C longitudinal and transversal views of sieve tube,
D. sieve plate in winter condition,
E. radial longitudinal view of laticifers
A. tracheidB. fibre tracheidC. xylem fibreD. septate fibreE. annular F. spiral vesselG. scalariform vesselH. reticulate vesselI. vessel with round bordered
pits and simple perforation pits
J. hexagonal pitsK. vessel segmentL.M.N. bordered pitO.half bordered pitP. transverse section vessel
Xylem
Trease, 532.
Microscopical Analysis
• Secretory cells– Oil cells– Secretary cavities or sacs
• Schizogenous• Lysigenous• Schizolysigenous
– Vittae• Schizogenous oleoresin canals
– Latex• Cells or tissue containing fluid with milky appearance
SECRETORY CELLS AND DUCTS
Trease, 535.
Ergastic Cell Contents
• Microscopically, physically or chemically identified non-living particles.
– Carbohydrate, protein, fixed oils, fat, glycoside, volatile oil, gum, silica, etc.
– Crystal • Calcium oxalate• Calcium carbonate
ERGASTIC CELL CONTENT
Trease, 537.
Calcium oxalate crystalsA-D tetragonal systemE-I monoclinic systemA1-A3 rosette crystals of
tetragonal systemD a tetragonal prismE a monoclinic prismG raphidesH a single needle crystalI a sphaerocrystal
MICROCHEMICAL ANALYSISDistribution of tissue
– Phloroglucinol and HCl• Lignified cells: red
– Chlor-zinc-iodine solution • Cellulose walls: blue or violet• Lignified or suberized walls: yellow or brown• Starch grains: blue
Granules of wheat starch, stained with iodine, photographed through a light microscope - Wikipedia -
Cinchona bark, moistured with phloroglucinol followed by adding hydrochloric acid - Kusmardiyani-
Clearing, defatting and bleaching• Chloral hydrate
– Dissolve chlorophyll, protein, starch, resin, volatile oil, but not sodium oxalate
• Solution of potash– Dissolve aleurone, starch, protein, swelling cell walls.
• Ether-ethanol– Defatting
• NaOCl– Bleaching dark colour sections.
Disintegration and isolation of tissue• Potassium chlorate and nitric acid• Chromic acid and nitric acid or sulphuric acid• Solution of potash or soda
• Ethanol• Alkanna tinctures• Chloral hydrate and glycerin• Chloral hydrate with iodine• Clove oil (clearing agent for oily powder)• Chlor-zinc-iodine solution (Schultze’s solution)• Copper oxide, ammoniacal solution (swelling cellulose)• Corallin, alkaline solution (stain callose, gums and mucilages)• Ferric chloride (tannin)• Glycerin (mountant)• Iodine
Reagents
• Lactophenol (clearing)• Mercury-nitric acid (Millon’s Reagent) for protein containing material• Nitric acid (crude fibre)• Picric acid (aleurone and animal fibres)• Potassium cupri-tartrate (Fehling’s solution) for reducing sugars• Potassium iodobismuthate (precipitate alkaloid)• Potassium tetraiodomercurate (Mayer’s reagent) precipitate alkaloid• Ruthenium red (gums and mucilages)• Sodium carbonate (disintegrating fibres)• Sodium hypochlorite (clearing, defatting)• Sudan III (oils or suberized walls)• Sulphuric acid (charring, dissolve all but little action on suberin)
Reagents
• AFLP: Amplified Fragment Length Polymorphism
• SSR : Simple Sequence Repeat (microsatellite)
• Sequencing rDNA, ITS: Internal Transcribed Spacer and chloroplast genes
• RAPD: Random Amplified DNA Polymorphism
• RFLP : Restrictive Fragment Length Polymorphism
DNA Profiling
Method of Choice • Level of polymorphism
• Reliability
• Robustness
• RAPD– Easy to start but difficult to get reliable results
• RFLP– Obsolete, low resolution and high cost
• AFLP– Highly polymorphic, easier to start but difficult to data base
• SSR– Highly polymorphic, expensive to develop, used for intra-specific
analysis or closely related species.• SCAR
– Suitable for differentiation of targeted herbs.• Sequencing
– Good for differentiating plants from different families and genera, not polymorphic enough for intervening sequences.
Comparison among Methods
-- get more detail information --
Plant Nomenclature Taxonomy, In: Trease and Evans Pharmacognosy, 15th ed., p.10.
--read and discuss--
addendum
LABORATORY ACTIVITIES
PREPARATION OF STARCH- ISOLATION OF AMYLUM -
Read again and discuss related topics from course material of FA2205 General Pharmacognosy
A procedure for starch production was given in some detail in a Roman treatise by Cato in 184 BCE. Grain was steeped in water for ten days and then pressed. Fresh water was added. Mixing and filtration through linen cloth gave a slurry from which the starch was allowed to settle. It was washed with water and finally dried in the sun.
Raven et al., 2009, Starch – Chemistry and Technology, p.2.
STARCHHandbook of Pharmaceutical Excipients, 5th ed., 2006.
STARCHHandbook of Pharmaceutical Excipients, 5th ed., 2006.
Read and discuss:
Handbook of Pharmaceutical
Excipients, 5th ed., 2006,
p.725-726.
Handbook of Pharmaceutical Excipients, 5th ed., 2006, p. 730.
A procedure for starch production was given in some detail in a Roman treatise by Cato in 184 BCE. Grain was steeped in water for ten days and then pressed. Fresh water was added. Mixing and filtration through linen cloth gave a slurry from which the starch was allowed to settle. It was washed with water and finally dried in the sun.
Raven et al., 2009, Starch – Chemistry and Technology, p.2.
STARCH, PREGELATINIZEDHandbook of Pharmaceutical Excipients, 5th ed., 2006.
Handbook of Pharmaceutical Excipients, 5th ed., 2006.
Read and discuss:
TAPIOCA/CASSAVA STARCH PRODUCTION
Raven et al., 2009, Starch – Chemistry and Technology, p.548.
TAPIOCA/CASSAVA STARCH PRODUCTION
Raven et al., 2009, Starch – Chemistry and Technology, p.548.
TAPIOCA/CASSAVA STARCH PRODUCTION
rice starch
corn starch
Handbook of Pharmaceutical Excipients, 5th ed., 2006.
Compare:
Pregelatinized Starch
Handbook of Pharmaceutical Excipients, 5th ed., 2006.