1) The document describes experiments conducted to identify the presence of different phytochemicals in plant samples like apple and guava. Tests were performed to detect tannins, saponins, and flavonoids. 2) Procedures are outlined for tannins tests using ferric chloride and lead sub-acetate solutions, which detected tannins based on color changes. 3) Saponins were detected by mixing the plant extract with water and observing bubble formation. Flavonoids were detected using a lead acetate solution which produced a yellow precipitate.

1. R A J A B A L W A N T S I N G H E N G I N E E R I N G T E C H N I C A L C A M P U S
B I C H P U R I , A G R A
Submitted To Submitted by.
Dr. Ashish shukla POONAM RATHORE
Department of biotechnology 1900040540033
B.tech(bt)7th semester
HERABAL DRUG FORMULATION

2. RAPTURE BIOTECH INTERNATIONAL
(P) LTD. NOIDA

3. Table of Content
Saponins Test
Lead sub-acetate Test
Tannins Test
Introduction
Ferric chloride Test
Flavonoids Test

4. INTRODUCTION
Herbal formulations means a dosage form consisting
of one or more herbs or processed herbs in specified
quantities to provide specific nutritional, cosmetic
benefits meant for use to diagnose, treat, mitigate
diseases of human beings or animals, alter the
structure or physiology of human beings or animals.
Herbal formulations contain an active substance or
herbal substance or herbal preparation or herbal
substance in combination with one or more herbal
preparations.

5. EXPERIMENT
1. Aim;-To identify the presence of different phytochemical in a plant
sample.
2. Materials Required:- Sample powder (Apple, Guava), distilled
water, Fecl3, 10% lead acetate.
3. Theory:- Phytochemicals are naturally occurring biochemicals in
plants that help to give plants their characteristic color, flavor, smell and
texture. Apart from that, phytochemicals could prevent diseases and
inhibit pathogenic microorganisms.
The phytochemical category includes compounds recognized as
essential nutrients, which are naturally contained in plants and are
required for normal physiological functions, so must be obtained from
the diet in humans. They generally have biological activity in the plant
host and play a role in plant growth or defense against competitors,
pathogens, or predators.

6. PROCEDURE FOR EXPERIMENT
▪ Preparation of the sample extract: -
❖ Take 10 ml of water.
❖ Add 1 ml of plant extract sample (apple leaf).
❖ Pack test tube by polythin.
❖ Vortex the sample well.
❖ Leave the sample overnight.
❖ Vortex the sample again and filter.

7. Introducti
1 2 3 4 5
Take 10 ml of
water.
Pack test tube by
polythin.
Leave the sample
overnight.
Add 1 ml of plant
extract sample (apple leaf)
Vortex the sample
well.
6
Vortex the sample
again and filter.
PROCEDURE FOR EXPERIMENT

8. TANNINS TEST
Tannins are one of the major phytochemicals found in many higher
plants. Tannins have a characteristic strange smell and astringent taste
and could bind to proteins (and consequently precipitate proteins)
through the effective formation of strong complexes with proteins and
other macromolecules. Thus, they could have a major impact on animal
nutrition, including inhibition of growth rate digestive enzymes.
Types of Tannins Test
Ferric chloride Test Lead sub acetate test

9. ❖ Take 3 ml distilled water and add 1 ml plant extract
sample.
❖ Add 5% FeCl3 to the sample.
● Observations: - Blue green colour observed (apple
extract sample) and black colour (guava extract sample)
which indicates the presence of tannins.
Ferric Chloride Test

10. ❖ 1 ml of the sample extract was added to 3 drops of the lead sub-acetate
solution.
● Observations: - A cream gelatinous precipitate indicates the presence of
tannins.
Lead sub-acetate test

11. SAPONINS TEST
T Saponins are naturally occurring compounds that are
widely distributed in all cells of legume plants. Saponins,
which derive their name from their ability to form stable,
soap like foams in aqueous solutions, constitute a complex
and chemically diverse group of compounds.
Saponins are toxic chemicals that protect healthy plants
from insect, fungal, and bacterial pathogens. For this
reason, ingesting foods that contain saponins can cause
toxicity in the human body. However, severe poisoning is
rare.

12. ❖ Take 8 ml water and add 2 ml of plant extract filtrate.
❖ Vortex the sample for 15-20 minutes.
● Observation: - formations of bubbles were observerd.
PROCEDURE
Figure: - saponins test bubble formation.

13. Flavonoids Test
Flavonoids, a group of natural substances with variable
phenolic structures, are found in fruits, vegetables, grains,
bark, roots, stems, flowers, tea and wine. These natural
products are well known for their beneficial effects on health
and efforts are being made to isolate the ingredients so
called flavonoids. Phytonutrients like flavonoids have
beneficial anti-inflammatory effects and they protect our
cells from oxidative damage that can lead to disease. These
dietary antioxidants can prevent the development of
cardiovascular disease, diabetes, cancer, and cognitive
diseases like Alzheimer's and dementia.

14. ❖ Take 1 ml plant extract and add few drops Of
lead acetate to it.
● Observation: - yellow precipitate obtained.
PROCEDURE
Figure: - Flavonoids Test yellow precipitate obtained.

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1996 Jan.
• Sheehan-MP; Rustin-MH; Atherton-DJ; Buckley-C; Harris-DW;
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REFERENCES

16. Thank you!