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Phytochemical
Phytochemical
• phytochemicals are classes of compounds that are only found in plants that do
not also fall into the category of essential nutrients.
– phytochemicals are compounds that we ingest when we eat plants, but they are the parts
that aren’t absolutely needed by the body – in other words, they are not nutritionally
essential.
• Generally, the arils of fruit contain large amounts of organic acids, sugars, minerals,
and vitamins, but the peels contain higher amount of phenolic compounds than the
flesh.
Phytochemical classes
Class
Sub class
Classification are based on
• Biosynthetic origin
• Solubility properties
• Presence of certain key functional groups
Phenolic compound Terpenoids Nitrogen
•Recognized by their •Share lipid properties compounds
hydrophilic compounds •Biosysnthetic origin •Recognized by their
•Common origin from from isopentenyl positive responses to
the aromatic precursor phytophosphate either ninhydrin or the
shikimic acid Dragendorff reagent
Sub class
Phenolic compounds
Introduction
• Wide range of plant substances which possess in common an aromatic
ring bearing one or more hydroxyl substituent
• Phenolic substances tend to be water-soluble, since they are usually
located in the vacuole.
• The flavonoids form the largest group but simple monocyclic phenols,
phenylpropanoids and phenolic quinones all exist in considerable
numbers.
• Several important groups of polymeric materials in plants – the
lignins, melanins and tannins – are polyphenolic
• Function of some classes of phenolic compound – eg. The lignins as
structural material the cell wall; the anthocyanins as flower pigments,
• Phenolic compounds are all aromatic, so that they all show intense
absorption in the UV region of the spectrum
Phenols and phenolic acids
• The free phenols and phenolic acids are best considered
together since they are usually identified together during plant
analysis
• Acid hydrolysis of plant tissue releases a number of ether-
soluble phenolic acids
• These acids are either associated with lignin combined as ester
groups or present in the alcohol-soluble fraction of the leaf,
maybe present in the alcohol-soluble fraction bound as simple
glycosides.
• Free phenols are relatively rare in plants
• Simple phenols are included in here because their identification
is important to determining the structure of flavonoid.
Phenylpropanoids
• Naturally occurring phenolic compounds which have an
aromatic ring to which a three-carbon side-chain is attached.
• Derived biosynthetically from the aromatic protein amino
acid phenylalanine and they may contain one or more C6-C3
residue.
• The most widespread -hydroxycinnamic acids – which are
important not only as providing the building blocks of lignin
but also in relation to growth regulation and to disease
resistance.
• Four hydroxycinnamic acids are common in plants – ferulic
acid, sinapic, caffeic and p-coumaric acids.
Flavonoid pigments
• Derived from the parent substance flavone – more
general
• Classes of flavonoid
• Mainly water-soluble compounds
• Can be extracted with 70% ethanol and remain in
the aqueous layer, following partition of this
extract with petroleum ether.
Properties of different flavonoid classes
Hydrolysable tannins
- Gallotannins Esters of gallic acid 1000 – 1500
and glucose
Prototannins
- Tannin precursors Catechins (and 200 – 600
gallocatechins)
flavan-3,4-diols
Quinone pigments
• The natural quinone pigments range in colour from pale
yellow to almost to black.
• Relatively little contribution to colour in higher plants
• Frequently present in bark, heartwood or root or leaves
tissue where their colours are masked by other
pigments.
• Bacteria, fungi and lichen – pigmented by quinones
• Identification – divided into 4 groups: bezoquinines,
naphthalquinone, anthraquinones and isoprenoid
quinones
Recap - flavonoid
monoterpenes
• Chemically, terpenoids are generally lipid-
soluble
• Located in cytoplasm of the plant cell
– Essential oils sometimes occurs in special glandular
cells on the leaf surface
– Carotenoids – associated with chloroplasts in the leaf
and with chromoplasts in the petal
• Extraction – light petroleum, ether or
chloroform
• Separation – by chromatography on silica gel or
alumina using same solvent
• Difficult in detection on a microscale except carotenoids
– Because terpenoids are colourless
– No sensitive universal chromogenic reagent
• Isomerism is common: geraniol and nerol
• Mostly alicyclic compound because the cyclohexane ring usually
twisted in ‘chair’ form, different geometric conformations are
possible, depending on the substitution around the ring – often
difficult to determine
• Functions of plant terpenoids
– growth-regulating properties – abscisins (sesquiterppenoid) and
gibberellins (diterpenoid)
– Agents of communication and defense among insects
– Non-volatile terpenoids – sex hormones among fungi
– Carotenoids
• plant colour - pale yellow through bright orange to deep red
• Accessory pigments in photosynthesis
– Mono- and sesquiterpenes – distinctive smells and odours
Essential oils
• Volatile steam-distillable – characteristic scent, odour or smell
• Commercially important – natural perfumes, spices and flavoring in food
industry
• Chemically, terpene essential oils divided into 2
– Monoterpenes (C10) – boiling point 140-180 °C
– Sesquiterpenes (C15) – bp >200 ° C
• Mono – divided into three group depending on whether they are acyclic,
monocyclic, or bicyclic.
• Within each group, mono-maybe simple unsaturated hydrocarbons
(limonene) or may have functional groups and be alcohols (menthol),
aldehydes or ketones (menthone, carvone).
• Simple mono- are widespread and tend to occurs as majority of essential
oils.
• Flower and seed oils tend to have more specialized mono- present
• Sesqui- group according to the basic carbon
skeleton (same as mono-)
• Either acyclic, monocyclic or bicyclic.
• But within group – there are too many
different compounds known
• 2 special sesqui- because of the growth-
regulating properties – abscisic acid (hormone
controlling domancy in seed) and xanthinin
(auxin-antagonist)
• Isolation from plant tissues – both mono- and
sesqui- separated by extraction into ether,
petroleum or acetone.
• Classical extraction procedure – steam
distillation
• Due to the volatility – ideal for separation by
GC
Diterpenoids and gibberellins
• Comprises of a chemically heterogeneous group of
compounds
• All with C20 carbon skeleton - based on four isoprene units
• Very limited distribution compound
• Probably the only universally distributed diterpene is
acyclic parent compound of the series – phytol (present as
the ester attachment in chlorophyll)
• 3 classes of diterpenes – resin diterpenes, toxic
diterpenes and gibberellins
• Resin diterpenes
– Have protective function in nature
– Exuded from wood of trees or latex of herbaceous
plants
• Toxic diterpenes
– Poisonous
• Gibberellins
– Group of hormone – stimulate growth
– Gibberellic acid – the most popular gibberellin
Triterpenoids and steroid
• Compounds with carbon skeleton based on 6 isoprene units
• Derived biosynthetically from the acyclic C 30 hydrocarbon
• Relatively complex cyclic structure
• Most either alcohols, aldehydes or carboxylic acids.
• Colourless, crystalline, often melting, optically active
substance which generally difficult to characterize because
lack of chemical reactivity
• Widely used test – Liebermann-Burchard reaction (acetic
anhydride-conc H2SO4) – produces a blue-green colour with
most triterpenes and sterols
• Divided into at least 4 groups of compounds:
– True triterpenoid
– Steroids
– Saponin
– Cardiac glycosides occur mainly as glycosides
• Functions
– occur especially in waxy coatings of leaves and on
fruits – protective function in repelling insect and
microbial attack
– Taste properties – bitterness
• eg. Limonin – the lipid-soluble bitter principle in Citrus
fruits
Carotenoids
• C40 tetraterpenoids
• Extremely widely distributed group of lipid-
soluble pigment
• 2 principle function
– As colouring matters in flowers and fruits
– Accessory pigment in photosynthesis
• Eg in flowers – yellow colour (daffodil, marigold)
• Eg in fruits – orange or red (tomato, paprika,
palm oil)
• Over 600 known carotenoids but only a few are common in
higher plant
• Identification easy to resolve – by reference to the common
substance
• Split into two class –
– Purely hydrocarbon and contain no oxygen - based on lycopene
• Chemical structure of lycopene consists of a long chain of 8 isoprene units
joined head to tall, giving a completely conjugated system of alternate
double bond – which is the chromophore giving it colour
• Cyclization of lycopene at one end give γ-carotene
• Cyclization of lycopene at both end give β-carotene
• β-carotene isomers (α- and ε-carotene) differ in the position of the double
bond in the cyclic end units
– Oxygenated derivatives (contain oxygen) – xanthophylls
• Monohydroxycarotenes - lutein, rubixanthin
• Dihydroxy – zeaxanthin
• Dihydroxyepoxy – violaxanthin
• Glycosides – very rare in higher plant – the
best known is water-soluble crocin
• Combined forms of carotenoid
– xanthophylls esterified with fatty acid
– Eg. palmitic, oleic or linoleic acid
Nitrogen compound
Introduction
• Nitrogen compounds – nitrogen element
• Nitrogen compounds are usually basic, thus form salts with
mineral acids
• Can be extracted from plant tissue using weak acidic solvents
• Can be precipitated from extracts by addition of ammonia
• Many nitrogen compounds are charge molecules, so
electrophoresis can be used for separation
• Detection technique
– spray reagent Ninhydrin – amino acids
– Dragendorff reagent - alkaloids
Amino acid
• Plant amino acid conveniently divided into two groups (although the
division between the two groups is not entirely sharp and methods
of identifying and separating both groups are essentially the same)
– Protein amino acid
• Generally recognized to be twenty in number
• Found in plant and animal
• Glutamic, aspartic acids, glutamine and asparagine - present in larger amount and
represent a storage form of nitrogen
• Histidin, trytophan, cystein and methionine – low amount in plant tissue and cannot
be readily detected.
– Non-protein amino acid
• Only γ-aminobutyric acid regularly present in plants
• Their role in plant is not clear, although present in high concentration in seeds
• May be important as nitrogen storage material
• Amino acid are colourless ionic compounnds
• Solubility properties and high melting point –
zwitterions
• Water soluble
• Amino acid have difference charge properties,
amino acid mixture can be divided into
neutral, basic and acidic fractions by using
either electrophoresis or ion exchange
chromatograhy.
Amino acid Ninhydrin colour Charge properties
Glycine red-violet neutral
Alanine violet neutral
Serine neutral
Cysteine neutral
Threonine neutral
Valine neutral
Leucine neutral
Isoleucine neutral
Methionine neutral
Aspartic acid blue-violet acidic
Asparagines orange-brown neutral
Glutamic acid violet acidic
Glutamine neutral
Arginine basic
Lysine basic
Proline yellow neutral
Phenylalanine grey-violet neutral
Tyrosine neutral
Tryptophan neutral
Histidine basic
Amines
• Considered simply as the products of decarboxylation of amino
acid, formed by the reaction:
RCH(NH2)CO2H RCH2NH2 + CO2
• Divided into three groups
– aliphatic monoamines
• Volatile compound eg methylamine (CH3NH2), n-hexylamine (CH3(CH2)5NH2
• Have an unpleasant fish-like smell
• Function in flowers as insect attractants
– aliphatic polyamines
• Less volatile
• Still possess offensive odours
• Eg putrescine, agmatine, spermidine, spermine
• Function – growth-stimulating activity in relation to their effect on ribosomal
RNA
– aromatic amines
• Many of known aromatic amines are physiologically active and sometimes
classified as alkaloids
Alkaloids
• Largest single class of secondary plant substance
• No one definition of term alkaloid which completely
satisfactory
• Alkaloid generally includes ‘ those basic substance which
contain one or more nitrogen atom, usually in combination as
part of a cyclic system’
• Often toxic to man and many have dramatic physiological
activities; hence wide use in medicine
• Usually colourless, often optically active substance, most are
crystalline but a few liquid at room temp (eg. nicotine)
• Simple test for alkaloid – bitter taste
• Many alkaloid are terpenoid in nature (eg. solanine)
and some are best considered as modified terpenoid
• Others are mainly aromatic compound (eg. colchine) –
containing their basic group as side-chain attachment
• Specific to one family or to a few related plants – name
of alkaloid types are often derived from plant source –
eg nicotine (Nicotina tabacum), atropine ( Atropa
belladonna)
• Mostly in angiosperm, generally absent or infrequent
in gymnosperms, ferns, mosses and lower plant.
• Function of alkaloid – still obscure / not clear
• Some reported to be involved as growth
regulation, insect repellents or attractants
• Theoretically – they act as a form of nitrogen
storage in plants
• Most alkaloids are weak bases, but some, such as
theobromine and theophylline, are amphoteric
• Many alkaloids dissolve poorly in water but
readily dissolve in organic solvents, such as diethyl
ether, chloroform or 1,2-dichloroethane.
• Caffeine, cocaine, codeine and nicotine are water
soluble (with a solubility of ≥1g/L), whereas
others, including morphine and yohimbine are
highly water soluble (0.1–1 g/L).
• Alkaloids and acids form salts of various strengths.
• These salts are usually soluble in water and
ethanol and poorly soluble in most organic
solvents.
• Extraction - no single method of the extraction
from natural raw materials.
• Most methods exploit the property of most
alkaloids to be soluble in organic solvents but
not in water, and the opposite tendency of
their salts.