A PHYTOCEMICAL ANALYSIS

OF “PHYLLANTUS NIRURI”

by:

Kathleen Joy T. Navasero

Princess Anne Maramag

Stephanie Balubal

Margarette Salas

Mharia Christina Tumamao

Approved by:

Ms. Sarah Lee G. Pagulayan,

MST
 CHAPTER I- INTRODUCTION

Plants are recognized in the pharmaceutical industry for their broad structural diversity as

well as their wide range of pharmacological activities. The biologically active compounds

present in plants are called phytochemicals. These phytochemicals are derived from various parts

of plants such as leaves, flowers, seeds, barks, roots and pulps. These phytochemicals are used as

sources of direct medicinal agents. They serve as a raw material base for elaboration of more

complex semi-synthetic chemical compounds. This paper mainly deals with the collection of

plants, the extraction of active compounds from the various parts of plants, qualitative and

quantitative analysis of the phytochemicals.

Phytochemicals are the chemicals that present naturally in plants. Now- a-days these

phytochemicals become more popular due to their countless medicinal uses. Phytochemicals play

a vital role against number of diseases such as asthma, arthritis, cancer etc. unlike

pharmaceutical chemicals these phytochemicals do not have any side effects. Since the

phytochemicals cure diseases without causing any harm to human beings these can also be

considered as “man-friendly medicines”.

The importance of medicinal plant in drug development is known to us and humans have

used them for different diseases from the beginning of human history. Traditional folk treatment

from wild plants has always guided researchers to search for novel medications to develop

healthy life for humans and animals. In addition, some medicinal plants are still obscured within

the plant which need to be scientifically evaluated.

Phyllanthus niruri may be found in profusely branched condition along with crops of

gram, wheat, pea, etc, known by the common name “gale of the wind,” “stonebreaker” (chanca
pedra), or “seed-under-leaf.” In the wild it is found growing along road sides, in street corners,

and dumps of building materials. Taxonomically, the annual herb Phyllanthus niruri belongs to

the family Phyllanthaceae of the order Malpighiales under class Magnoliopsida of the Division

Magnoliophyta. Phyllanthus niruri are erect annual herbs, 10–60 cm tall; main stem simple or

branched, terrete smooth or scabridulous in younger parts. Cataphylls, stipules 1.5–1.9 mm long,

deltoid acuminate blade 1–1.5 mm long, subulate acuminate. Leaves 3–11×1.5–6 mm elliptic

oblong obovate, oblong, or even obovate, obtuse, or minutely apiculate at apex, obtuse or

slightly inequilateral at base, petioles 0.3–0.5 mm long, stipules 0.8–1.1 mm long triangular

accuminate.

The plant is known as a small herb having wide range of medicinal properties, and it is

used widely across the world. Its taste is bitter and acts as astringent and show laxative effect.

The use of Phyllanthus niruri is gaining momentum because of its novel antiviral activity against

hepatitis B virus and for several other biological activities due to its extract such as kidney and

gallbladder stones, for cold, flu, tuberculosis, and other viral infections; liver diseases and

disorders including hepatitis, jaundice, liver cancer and jaundice, apart from these it is also

administered for diuretic, hypoglycemic and hypertension cases and it also shows anti-

inflammatory, anti-tumor, antinociceptive and anti-oxidant properties.

This phytochemical analysis covers information about medicinal uses of Phyllanthus

niruri with various pharmacological profile of the plant. The phytochemical studies were

characterized and the presence of various compounds such as lignans, phyllanthin,

hypophyllanthin, flavonoids, glycosinoids & tannins was mentioned. This study summarizes the

information about its botanical, morphological, pharmacological and biological activities of the

plant.
 CHAPTER II- REVIEW OF RELATED LITERATURE

Phyllanthus niruri is a weed found in coastal areas. It’s also known as gale of the wind or

stonebreaker. Its leaves and fruit are used as herbal medicine. Phyllanthus niruri is known for

protecting the liver. It may also combat kidney stones, hence the “stonebreaker” moniker. Keep

reading to learn more about how this plant may improve your health.

It has antioxidant properties. According to a 2014 in vitro study, an extract made from

phyllanthus niruri leaves showed strong antioxidant activity. Antioxidants fight free radicals in

the body that may cause cell damage and disease. It has antimicrobial properties. According to a

2012 studyTrusted Source, phyllanthus niruri extract has antimicrobial abilities against H. pylori

bacteria. H. pylori bacteria are common in the digestive tract and are usually harmless. But in

some cases, they can lead to peptic ulcers, abdominal pain, and nausea. Researchers also found

that the extract didn’t harm any beneficial strains of lactic acid bacteria.

It has anti-inflammatory properties. Inflammation can cause many problems throughout

the body, including skin conditions like psoriasis and chronic pain. According to a 2017 animal

studyTrusted Source, phyllanthus niruri may help reduce inflammation. For the study,

researchers injected carrageenan into the left hind paws of rats to cause inflammation. The rats

were then treated with phyllanthus niruri extract, which was found to significantly reduce the

inflammation.

It may help protect against ulcers. Findings from the same 2017 animal studyTrusted

Source also suggest that phyllanthus niruri extract can help prevent ulcers. It’s thought to reduce

gastric acid secretion and protect the stomach. It also showed potent anti-inflammatory effects

against gastric ulcers. It may help lower blood sugar. Phyllanthus niruri may also have
antidiabetic properties. According to a 2011 studyTrusted Source, the plant’s aerial parts may

help prevent glucose absorption and improve glucose storage. This may help maintain blood

sugar levels.

It may help prevent kidney stones. Phyllanthus niruri may be best known as a kidney

stone remedy. According to 2010 research, many studies have shown that it’s a powerful kidney

stone deterrent. Phyllanthus niruri can help relax the ureter after lithotripsy to help stones pass.

Lithotripsy is a procedure used to break up stones in the urinary tract. It may also prevent stones

from forming in the first place by stopping the crystals that form them from clumping together.

It may improve liver health. According to a 2017 studyTrusted Source on rats,

phyllanthus niruri may help treat nonalcoholic fatty liver disease and atherosclerosis. Both

conditions may cause insulin resistance. The study found phyllanthus niruri decreased insulin

resistance and reduced the amount of fatty acids in the liver.

A 2006 studyTrusted Source on mice also found that the antioxidants in phyllanthus

niruri may help protect the liver from acetaminophen toxicity.It may help treat acute hepatitis B.

Phyllanthus niruri may help treat acute hepatitis B infections due to its anti-viral and liver-

protecting abilities. In a 2010 studyTrusted Source, 60 people with the condition received either

a placebo or a product made of seven herbal medicines including 100 milligrams of phyllanthus

niruri. The herbal medicine group experienced faster recovery than the placebo group. However,

they reported side effects like upper abdominal pain and diarrhea. It may not help chronic

hepatitis B, though. A 2011 review of 16 randomized trials on people with chronic hepatitis B

didn’t find any significant evidence in support of the treatment. It may be anti-cancerous.

Phyllanthus niruri and other phyllanthus species may help prevent the metastasis of some lung
and breast cancers. A 2011 studyTrusted Source showed the polyphenols in the herb help prevent

the invasion, migration, and adhesion of cancer cells.

A 2012 studyTrusted Source on human colorectal and liver cancer cell lines also found

that phyllanthus niruri helped slow cancer cell growth and encouraged cancer cell death.

Phyllanthus niruri is commonly used in capsule, tea, or extract forms, and there’s no standardized

dosing recommendation. An average dose is one 500-milligram capsule per day or 1 milliliter

extract up to four times daily. For specific dosing guidelines, see your doctor or natural health

professional. Phyllanthus niruri is a popular folk remedy with powerful medicinal properties.

Some research supports its use for kidney stones, gallstones, liver health, and diabetes. Still,

much of the research is inconclusive. There’s not enough evidence to recommend the herb for

most conditions.

P. niruri has been reported to possess antidiabetic and kidney protective effects. In the

present study, the phytochemical constituents and in vitro antioxidant activity of P. niruri leaf

aqueous extract were investigated together with its effect on oxidative stress and antioxidant

enzymes levels in diabetic rat kidney. Results. Treatment of diabetic male rats with P. niruri leaf

aqueous extract (200 and 400 mg/kg) for 28 consecutive days prevents the increase in the amount

of lipid peroxidation (LPO) product, malondialdehyde (MDA), and the diminution of superoxide

dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activity levels in the kidney

of diabetic rats. The amount of LPO showed strong negative correlation with SOD, CAT, and

GPx activity levels. P. niruri leaf aqueous extract exhibits in vitro antioxidant activity with IC50

slightly lower than ascorbic acid. Phytochemical screening of plant extract indicates the presence

of polyphenols. Conclusion. P. niruri leaf extract protects the kidney from oxidative stress

induced by diabetes.
 A preclinical study was performed to determine if the extract from Phyllanthus niruri

(PN) plays a protective role against liver cirrhosis induced by thioacetamide (TAA) in rats.

Initially, acute toxicity was tested and the results showed that the extract was benign when

applied to healthy rats. Next, the therapeutic effect of the extract was investigated using five

groups of rats: control, TAA, silymarin, and PN high dose and low dose groups. Significant

differences were observed between the TAA group and the other groups regarding body and liver

weights, liver biochemical parameters, total antioxidant capacity, lipid peroxidation, and

oxidative stress enzyme levels. Gross visualization indicated coarse granules on the surface of

the hepatotoxic rats’ livers, in contrast to the smoother surface in the livers of the silymarin and

PN-treated rats. Histopathological analysis revealed necrosis, lymphocytes infiltration in the

centrilobular region, and fibrous connective tissue proliferation in the livers of the hepatotoxic

rats. But, the livers of the treated rats had comparatively minimal inflammation and normal

lobular architecture. Silymarin and PN treatments effectively restored these measurements closer

to their normal levels. Progression of liver cirrhosis induced by TAA in rats can be intervened

using the PN extract and these effects are comparable to those of silymarin.

A herb of the EUPHORBIACEAE family that grows up to 60 cm. The plant is bitter in

taste, the leaves are small, green, and short-petioled with a thin and glaucous under surface. The

flowers are unisexual, monoecious, minute, greenish and inconspicuous, short-stalked and borne

in pairs in the axils of the leaves. The fruit is a capsule, globose, slightly depressed at the top

with 6 enervations. In the roots, the secondary growth starts very early and is well pronounced.

There is a distinct cambium. No starch grains, mineral crystals or latex vessels are seen in either

the root or stem.

 CHAPTER III- METHODOLOGY

3.1. Collection of Plants

Plants under consideration may be collected either from wild forests or from herbariums.

When plants are collected from wild, there is a risk that they have been incorrectly identified.

The major advantage of wildlife plants is that they will not contain any pesticides. After the

plants are collected from wild or from herbarium, they have to be processed for cleaning in order

to prevent the deterioration of phytochemicals present in plants.

3.2. Cleaning of Plants

After plants collection they have to be cleaned properly. The cleaning process may

involve the following steps. Cleaning, washing, peeling or stripping leaves from stems. Cleaning

has to be done by hands in order to get better results.

3.3. Drying

The main purpose of drying is to remove the water content from plants so that the plants

can be stored. Plants have to be dried immediately as soon as the plants collection or this will

lead to spoilage of plant materials. The drying consists of two methods. Drying can be done

either by natural process or by artificial process.

3.3.1. Natural Process

Natural process includes sun-drying. Sometimes plants are placed on drying frames or on

stands, to be air-dried in barns or sheds. But this may take few weeks for complete drying. The

time depends on temperature and humidity.

3.3.2. Artificial Drying

Artificial drying can be done with the help of artificial driers. This process will reduce the

drying time to several hours or minutes. The common method that is followed in drying

medicinal plants is warm-air drying. In this process plants are placed in the plates of drier on

which warm air is blown. This method is mainly applicable to fragile flower and leaves and this

requires large number of workers since loading and unloading of plants has to be done manually.

3.4. Powdering

After complete drying of plants, they have to be powdered well for further analysis

3.5. Extraction

Plants may undergo further analysis and examination such as extraction to identify its

various components and benefits.

3.5.1 Methods of Extraction

•Plant Tissue Homogenization

Plant tissue homogenization in solvent has been widely used by researchers. Dried or

wet, fresh plant parts are grinded in a blender to fine particles, put in a certain quantity of solvent

and shaken vigorously for 5 - 10 min or left for 24 h after which the extract is filtered. The

filtrate then may be dried under reduced pressure and re-dissolved in the solvent to determine the

concentration. Some researchers however centrifuged the filtrate for clarification of the extract
• Serial Exhaustive Extraction

It is another common method of extraction which involves successive extraction with

solvents of increasing polarity from a non-polar (hexane) to a more polar solvent (methanol) to

ensure that a wide polarity range of compounds could be extracted. Some researchers employ

Soxhlet extraction of dried plant material using organic solvent. This method cannot be used for

thermolabile compounds as prolonged heating may lead to degradation of compounds.

• Soxhlet Extraction

Soxhlet extraction is only required where the desired compound has a limited solubility

in a solvent, and the impurity is insoluble in that solvent. If the desired compound has a high

solubility in a solvent then a simple filtration can be used to separate the compound from the

insoluble substance. The advantage of this system is that instead of many portions of warm

solvent being passed through the sample, just one batch of solvent is recycled. This method

cannot be used for thermolabile compounds as prolonged heating may lead to degradation of

compounds.

• Maceration

In maceration (for fluid extract), whole or coarsely powdered plant- drug is kept in

contact with the solvent in a stoppered container for a defined period with frequent agitation until

soluble matter is dissolved. This method is best suitable for use in case of the thermolabile drugs.

• Decoction
 This method is used for the extraction of the water soluble and heat stable constituents

from crude drug by boiling it in water for 15 minutes, cooling, straining and passing sufficient

cold water through the drug to produce the required volume.

• Infusion

It is a dilute solution of the readily soluble components of the crude drugs. Fresh

infusions are prepared by macerating the solids for a short period of time with either cold or

boiling water.

• Digestion

This is a kind of maceration in which gentle heat is applied during the maceration

extraction process. It is used when moderately elevated temperature is not objectionable and the

solvent efficiency of the menstrum is increased thereby.

•Percolation

This is the procedure used most frequently to extract active ingredients in the preparation

of tinctures and fluid extracts. A percolator (a narrow, cone-shaped vessel open at both ends) is

generally used. The solid ingredients are moistened with an appropriate amount of the specified

menstruum and allowed to stand for approximately 4 h in a well11 l closed container, after which

the mass is packed and the top of the percolator is closed. Additional menstruum is added to form

a shallow layer above the mass, and the mixture is allowed to macerate in the closed percolator

for 24 h. The outlet of the percolator then is opened and the liquid contained therein is allowed to

drip slowly. Additional menstruum is added as required, until the percolate measures about three
quarters of the required volume of the finished product. The marc is then pressed and the

expressed liquid is added to the percolate. Sufficient menstruum is added to produce the required

volume, and the mixed liquid is clarified by filtration or by standing followed by decanting.

• Sonication

The procedure involves the use of ultrasound with frequencies ranging from 20 kHz to

2000 kHz; this increases the permeability of cell walls and produces cavitation. Although the

process is useful in some cases, like extraction of rauwolfi a root, its large-scale application is

limited due to the higher costs. One disadvantage of the procedure is the occasional but known

deleterious effect of ultrasound energy (more than 20 kHz) on the active constituents of

medicinal plants through formation of free radicals and consequently undesirable changes in the

drug molecules.