Variation & Standardization of

Natural Products, Quality

Control & Origins of Secondary
Metabolite
REPORTERS:
Jaboli, Jun Elbert
Bernate, Jayvee
Tadeos, Denise
Nadyn
Juames, Junda
Perero, Ricky Mae
PRESENTATION OUTLINE
01 02
Phytochemical
Plant Growth Variation within
Regulators Species

03 04
Quality Control Basic Metabolic
Determination of Pathways & The
Stored Drugs Origin of Secondary
Metabolites
PLANT GROWTH
REGULATORS
 PLANT GROWTH
Plant GrowthREGULATORS
Regulators (PGR) - refers to natural
or synthetic substances that influence the growth and
development of plants.

5 RECOGNIZED GROUPS OF NATURAL

PLANT HORMONES AND GROWTH
REGULATORS • Ethylene
• Auxins • Abscisic acid
• Gibberellins
•
 PLANT GROWTH
•
REGULATORS
Auxins - function primarily in stem elongation by promoting
cell growth, the major naturally occurring auxin is indole-3-
acetic acid.
• Abscisic acid - is an ubiquitous plant hormone which play an
important role in the inhibition of seed germination and
budding. It is known as the plant stress hormone and is
involved in the response of weather stress such as tolerance
to cold and drought.
• Cytokinins - regulate immunity in plants by modulating
salicylic acid signaling and play a vital role in defense
against pathogens and insects.
 PLANT GROWTH
•
REGULATORS
Ethylene - brings about various changes to developing
plants. These includes thickening of the of the subapical
portion of the stem and reduction in the rate of elongation,
which it promotes cell growth expansion , regulates stages of
flower formation and sex expression. These are gas
responsible for fruit ripening process.
• Gibberellins - these are hormones that control and
promoting stem elongation, flowering and leaf expansion as
well as seed germination. They are also influence synthesis
of nucleic acid.
 Plant’s growth and development are under the control of
two sets of internal factors
 Nutritional factors such as the supply of carbohydrates,
proteins, fats and others constitute the raw materials
required for growth.
 Proper utilization of these raw materials is under the
control of certain “chemical messengers” which can be
classified into hormones and vitamins.
 CLASSIFICATION of PLANT
GROWTH REGULATORS
• Natural/Endogenous Hormones: Produced by some tissues
in the plant
 EXAMPLE: Indole-3-acetic acid
• Synthetic/Exogenous Hormone: Produced artificially and
similar to natural hormone in physiological activity
 EXAMPLE: Indole-3-butyric acid
 CLASSIFICATION of PLANT
GROWTH REGULATORS
ON THE BASIS OF NATURE OF FUNCTION
• Growth Promoting Hormones/Growth Promoter: Increase
the growth of plant
 EXAMPLE: Auxins, Gibberellins, Cytokinins
• Synthetic/Exogenous Hormone: Produced artificially and
similar to natural hormone in physiological activity
 EXAMPLE: Abscisic acid, Ethylene
 AUXINS
 Derived from the Greek word “auxein” (means to
grow/increase)
 Auxins may be defined as growth promoting substances which
promote growth along the vertical axis when applied in low
concentration to the shoot of the plant.
 The idea of existence of auxin was proposed by Charles
Darwin (1880) in his book “The Power of Movements in
Plants”.
 Coleoptiles of Canary grass (Phalaris canariensis) to unilateral
light and observed it to bend towards light.
 He covered the coleoptiles tip with tin foil or cut it off and
observed that coleoptiles did not bend towards unilateral light.

OCCURRENCE & DISTRIBUTION OF
AUXINS
• Occurs universally in all plants
• Where there is active growth there is auxin production.
• Growing meristem and enlarging organs produces
auxin.
• Shoot apex produces much auxin than root apex.
• Apical bud synthesizes more auxin than lateral buds.
• Developing seeds contain more auxin than matured
seeds.
• Apical bud synthesizes six times more auxin than
expanding leaves.
 STRUCTURE OF AUXINS
• Natural Auxins – are almost continuously produced by
some tissues in the plant. Also known as endogenous
growth substances
 SYNTHETIC AUXINS
 IPA (Indole Propionic Acid)
 IBA (Indole Butyric Acid)
 NAA (Naphthalene Acetic Acid)
 2,4-D (2,4 – Dichlorophenoxyacetic
acid)
 2,4,5-T (2,4,5 –
Trichlorophenoxyacetic acid)
USE OF AUXINS IN AGRICULTURE
• Rooting of Cuttings
• Seedless Fruit Production (Parthenocarpy)
• Promotion of Flowering
• Prevention of Premature Dropping of Fruits
• Germination
• Fruit Setting
• Thinning of Flower, Fruit and Leaves
• Prevention of Lodging in Cereals
• Weedicide
• Tissue Culture
 GIBBERELLINS
 Discovered by Kurosawa, a Japanese Plant Pathologist in
1928
 Rice plants infected by the fungus Gibberella fujikuroi
(syn: Fusarium moniliforme) showed excessive stem
elongation.
 Symptom is called ‘Bakane’ diseases.
 Chemical was extracted & purified and named as
Gibberellic Acid (GA)
 Now, 80 different Gibberellins are available – GA1 to
GA80.
 The most commonly occurring gibberellins is GA3.
 GIBBERELLINS
 Stimulates stem growth dramatically
 Stimulates cell division, cell elongation (for both) and
control enzyme secretions.
 Involved in overcoming dormancy in seeds and buds.
 GA translocate easily in plant (able to move freely) in
both directions.
 Used commercially in:
- increasing fruit size of seedless grapes
- stimulating seed germination and seedling growth.
 Promoting male flowers in cucumbers for seed
production
GIBBERELLINS
 CYTOKININS
 Promotes cell division
 Found in all tissues with considerable cell division
- Ex: embryos (seeds) and germinating seeds, young
developing fruits
 Roots supply cytokinins upward to the shoots.
 Interact with auxins to influence differentiation of tissues
(may be used to stimulate bud formation)
 Developing embryo shows active cell division.
 Liquid endosperm of coconut called coconut water/milk
contain cell division causing factors (Kinetin).
 Similarly, the developing endosperm of maize contain such
factors (Zeatin).
CYTOKININS
 CYTOKININS
 As roots begin to grow actively in the spring, they produce
large amounts of cytokinins that are transported to the shoot,
where they cause the dormant buds to become active and
expand.
 Tissue cultures use cytokinins to induce shoot development.
 Cytokinins may slow or prevent leaf senescence (leaf aging
or leaf fall).
 ETHYLENE (CH2=CH2)
 Growth retardant
 Ethylene promotes ripening
 Gaseous hormone  Ethylene as a gas diffuses readily
throughout the plant
 Produced in the actively growing meristems of the plant, in
senescing, ripening or ageing fruits, in senescing (ageing or
dying) flowers, in germinating seeds and in certain plant tissues
as a response to bending, wounding or bruising.
 Increase female flowers in cucumbers
 Degreening of oranges, lemons and grapefruits  Ethylene gas
breaks down chlorophyll and lets color show through.
 ABSCISSIC ACID (ABA)
 Growth retardant
 Induce stomata closing
 Inhibition of bud growth and shoot formation
 Widespread in plant body – moves readily through
plant
 ABA appears to be synthesized (made) by the leaves.
 Interacts with other hormones in the plant,
counteracting the growth – promoting the effects of
auxins & gibberellins
 ABSCISSIC ACID (ABA)
 Involved with leaf and fruit abscission (fall), onset of
dormancy in seeds and onset of dormancy (rest period)
in perennial flowers and shrubs
 ABA is effective in inducing closure of stomata in
leaves, indicating a role in the stress physiology in
plants (Ex: increases in ABA following water, heat and
high salinity stress to the plant)
 PHYTOCHEMICAL
VARIATION WITHIN
SPECIES
PHYTOCHEMICAL VARIATION W/IN
SPECIES
• Refers to the diversity of naturally occurring
chemical compounds found in different individuals
or populations of the same plant species

• These chemical compounds, known as

phytochemicals, are synthesized by plants for
various purposes.
PHYTOCHEMICAL VARIATION W/IN
 The composition SPECIES
and concentration of
phytochemicals can vary significantly within a
species due to:
• Genetic Variation
• Environmental Factors
• Developmental Stage
• Interactions with Other Organisms
PHYTOCHEMICAL VARIATION W/IN
SPECIES
PHYTOCHEMICAL VARIATION W/IN
SPECIESvariation within
 Understanding phytochemical
plant species is crucial for ensuring quality control
and optimizing medicinal purposes in several
ways:
• Consistency in Product Quality
• Identification of Bioactive Compounds
• Optimization of Cultivation Practices
• Authentication and Adulteration Detection
• Standardization of Herbal Medicines
• Pharmacological Research and Drug Discovery
QUALITY CONTROL
DETERMINATION OF
STORED DRUGS
QUALITY CONTROL DETERMINATION
OF STORED DRUGS
 Quality control of stored drugs ensures
safety, efficacy, and compliance with
regulatory standards.
 Maintaining drug quality prevents risks to
patient health and ensures effectiveness of
treatment.
OBJECTIVES OF QUALITY CONTROL
 Ensure potency and stability of active
ingredients
 Detect and prevent degradation or
contamination
 Compliance with regulatory standards (e.g.,
FDA, EMA)
FACTORS AFFECTING DRUG STABILITY
• Temperature fluctuations
• Humidity
• Light exposure
• Packaging materials
• Oxygen exposure
 QUALITY CONTROL METHODS
PHYSICAL INSPECTION
• Visual examination for changes in color, odor
& texture
• Packaging integrity check
CHEMICAL ANALYSIS
• High-performance liquid chromatography
(HPLC)
• Mass spectrometry
• Fourier-transform infrared spectroscopy (FTIR)
 QUALITY CONTROL METHODS
MICROBIOLOGICAL TESTING
• Presence of microbial contaminants
• Sterility testing
ACCELERATED STABILITY TESTING
• Exposing drugs to accelerated conditions to
predict long-term stability
QUALITY CONTROL METHODS
 REGULATORY GUIDELINES
FDA GUIDELINES: cGMP (Current Good
Manufacturing Practice)

ICH GUIDELINES: Q1A (Stability Testing

of New Drug Substances and Products)

PHARMACOPEIAL STANDARDS: USP,

BP, EP
CHALLENGES IN QUALITY CONTROL
STORAGE CONDITIONS: Maintaining
consistent temperature and humidity levels
ANALYTICAL TECHNIQUES:
Availability of sophisticated equipment and
skilled personnel
REGULATORY COMPLIANCE: Keeping
up with evolving regulations and standards
 CASE STUDIES
 EXAMPLES OF DRUG RECALLS
DUE TO QUALITY CONTROL
ISSUES

 LESSONS LEARNED AND

IMPROVEMENTS MADE
 FUTURE DIRECTIONS
 Advancements in analytical techniques
(e.g., spectroscopy, imaging)
 Integration of IoT and data analytics for
real-time monitoring
 Development of smart packaging for
enhanced stability
 CONCLUSION
 Quality control of stored drugs is essential
for ensuring patient safety and treatment
efficacy.
 Collaboration between manufacturers,
regulatory agencies, and healthcare
professionals is crucial.
 Continuous improvement and innovation are
needed to address emerging challenges.
MAJOR METABOLIC
PATHWAYS
MAJOR METABOLIC PATHWAYS
METABOLISM – the set of chemical reactions that
occur in a cell, which enable it to keep living,
growing and dividing. Metabolic processes are
usually classified as:
 Catabolism
 Anabolism
 CATABOLIC PATHWAY
 A catabolic pathway is an exergonic system that
produces chemical energy in the form of ATP, GTP,
NADH, NADPH, FADH2, from energy containing
sources such as carbohydrates, fats, and proteins.
 The end product are often carbon dioxide, water
and ammonia.
 ANABOLIC PATHWAY
 In contrast to catabolic pathways, anabolic pathways
require an energy input to construct macromolecules
such as polypeptides, nucleic acids, proteins,
polysaccharides and lipids.
 An anabolic pathway is a biosynthetic pathway
combines smaller molecules to form larger and more
complex ones.
There is a very large number of metabolic pathways. In
humans, the most important metabolic pathway are:
1.) Glycolysis - glucose oxidation in order to obtain ATP
2.) Citric acid cycle (Krebs cycle) – acetyl-CoA oxidation in
order to obtain GTP and valuable intermediates.
3.) Oxidative phosphorylation - disposal of the electrons
released by glycolysis and citric acid cycle.
4.) Pentose phosphate pathway - synthesis of pentoses and
release of the reducing power needed for anabolic reactions
5.) Gluconeogenesis - glucose synthesis from smaller
precursors, to be used by the brain.
GLYCOLYSIS or EMBDEN MEYERHOF
PATHWAY

 Glycolysis occurs in the cytosol of every cell.

 Only source of energy in erythrocytes (major
source of energy)
KREBS CYCLE or TRICARBOXYLIC ACID
 Used to generateCYCLE
energy through oxidation of
acetyl-CoA
 Used in the synthesis of NADH
 Production of amino acid
 It is located in the mitochondria of eukaryotes
 Cytosol in the prokaryotes
OXIDATIVE PHOSPHORYLATION
 Disposal of the electrons released by glycolysis and
citric acid cycle.
 enzymes relay the electrons released by substrate
oxidation to special molecules (electron acceptors)
 actually regulated in the cells.
 Takes place in mitochondria in the electron transport
chain and its sole purpose is to produce a lot of ATP.
 Another regulation of oxidative phosphorylation
undergoes common end pathway of aerobic respiration
 RESPIRATION
 Aerobic respiration happens in the inner mitochondrial
membrane which contains the relevant electron-transferring
protein complexes, each of these complexes accepts electrons
from a molecule and transfers them to a different compound,
and the full assembly is therefore termed the electron
transport chain.
PENTOSE PHOSPHATE PATHWAY
 Also known as Hexose Monophosphate Shunt
 Occurs in virtually all cell types and tissues (Liver
– 30% glucose metabolized by Pentose Phosphate
Pathway)
 Occurs in cytoplasm and produces NADPH
 Produces Trioxes, Hexoses and Pentoses
 Pentoses are necessary for nucleotide synthesis.
PENTOSE PHOSPHATE PATHWAY
In order to perform its anabolism, a cell needs not
only energy (ATP) it also needs reducing power,
under the form of NADPH.
• The pathway is very active in tissue involved in
cholesterol and fatty acid (liver, adipose tissues,
adrenal cortex, mammals glands).
• This pathway also produces ribose-5-phosphate,
the component sugar of nucleic acid.
 GLUCONEOGENESIS
 Synthesis of glucose from non-carbohydrate
precursors.
 It is important for fasting and starvation.
 It maintains blood glucose levels/concentration
 Gluconeogenesis primarily occurs in the liver
 Secondarily occurs in the kidney (occurs during
prolonged fasting as well as liver failure)
 ORIGIN OF
SECONDARY
METABOLITES
ORIGIN OF SECONDARY METABOLITES
 The metabolism can be defined as the sum of all the
biochemical reactions carried out by an organism.
 Metabolites are the intermediates and products of
metabolism and are usually restricted to small
molecules.
 Secondary metabolites are derived from primary
metabolism.
 Secondary metabolites or natural products can be
defined as: A heterogeneous group of natural metabolic
products that are not essential for vegetative growth of
CLASSIFICATION OF SECONDARY
METABOLITES
 Terpenoids and steroids
 Alkaloids
 Fatty acid-derived substances and
polyketides
 Non-ribosomal polypeptides
 Enzyme cofactors
TERPENOIDS AND STEROIDS
→ They are major group of substances derived biosynthetically
from isopentenyl diphosphate.
→ Currently, over 35,000 known terpenoid and steroid
compounds are identified.
→ Terpenoids have different variety of unrelated structures,
while steroids have a common tetra cyclic carbon skeleton and
are modified terpenoids that are biosynthesized from the
triterpene lanosterol
ALKALOIDS
→ There are over 12,000 known compounds of alkaloids.
→ Basic structures consist of basic amine group and are
derived biosynthetically from amino acids
FATTY ACID-DERIVED SUBSTANCES AND
POLYKETIDES
→ Around 10,000 compounds are identified
→ Biosynthesized from simple acyl precursors such as propionyl CoA,
acetyl CoA, and methylmalonyl CoA

NON-RIBOSOMAL POLYPEPTIDES
→ These amino acids derived compounds are biologically synthesized
by a multifunctional enzyme complex without direct RNA
transcription.

ENZYME COFACTORS
→ Enzyme cofactors are non-protein, low-molecular enzyme
SOURCES OF SECONDARY METABOLITES
→ The major sources of secondary metabolites are plants (80%
of secondary metabolite), bacteria, fungi, and many marine
organisms (sponges, tunicates, corals, and snails).
SECONDARY METABOLITES OF PLANTS
 Plant secondary metabolites represent highly economically
valuable products.
 These are used as high value chemicals such as drugs,
flavors, fragrances, insecticides, dyes, etc.
 Plants are rich in a wide variety of secondary metabolites,
such as tannins, terpenoids, alkaloids, and flavonoids, which
have been found to have antimicrobial properties.
SECONDARY METABOLITES OF PLANTS
 Plants have an almost limitless ability to synthesize aromatic
substances, most of which are phenols or their oxygen-substituted
derivatives.
 About 25,000 terpenoids are known as secondary compounds and are
derived from the five-carbon precursor isopentenyl diphosphate
(IPP).
 In total, around 12,000 known alkaloids are identified, and they
possess one or more nitrogen atoms which are biosynthesized from
amino acids.
 Many alkaloids are used in medicine, usually in the form of salts.
Some examples include vinblastine which has antitumor properties ;
quinine which has antipyretics and anti malarial properties; and
SECONDARY METABOLITES OF PLANTS
 Alkaloids are regarded as reserve materials for protein synthesis, as
protective substances discouraging animal or insect attacks, and as
plant stimulants or regulators or simply as detoxification products.
 Alkaloids currently in clinical use include the analgesics morphine
and codeine, the anticancer agent vinblastine, the gout suppressant
colchicines, the muscle relaxant tubocurarine, the anti-arrhythmic
ajmalicine, the antibiotic sanguinarine, and the sedative scopolamine.
 FUNCTIONS OF SECONDARY
METABOLITES
The major functions of the secondary metabolites
including antibiotics are:
 Competitive weapons against other living things such
as animals, plants, insects and microorganisms
 Metal transporting agents
 Agents for symbiotic relation with other organisms
 Reproductive agent
 Differentiation effectors
 Agents of communication between organisms
THANK
YOU!