Effectivity of Orange Lirio (Hippeastrum puniceum) Bulb Extract

In Treating Fungal Infections

Ryan Mark O. Diosay

Craig Jaden B. Bordas
Mat Makisig J. Villanueva
Ray Aldrine B. Crujedo
Chaira Mae B. Agbot
Nicole C. Sillacay

Submitted in partial fulfillment of the requirements in Science 10

Immaculate Heart of Mary Academy

Mr. Guerrero T. Nucos

Research Adviser

February, 2020
 ABSTRACT

The plant Hippeastrum puniceum is a widely seen ornamental plant. The fresh bulbs of the

plant were used traditionally for healing wounds, curing tumors and piles. As till date no relevant

scientific studies had been conducted on this plant. The present study is conducted to scientifically

validate the pharmacognostic and physicochemical parameters. The microscopic evaluation of the

bulb shows the presence of starch grains, mucilage cells and xylem fibers. Physicochemical

parameters like ash values, extractive values, crude fiber content and fluorescence analysis were

also established. Preliminary phytochemical screening of various extracts showed the presence of

alkaloids, carbohydrates, flavonoids, tannins, saponins, terpenoids, proteins and amino acids. Total

phenolic and flavonoid content of various bulb extracts were determined by Folin Cio-Calteau and

Aluminum chloride colorimetric method respectively. The choloform extract showed highest

amount of phenolics and the highest amount of flavonoid was present in ethyl acetate extract.
 Chapter I

INTRODUCTION

A. Background of the Study

People, researchers and scientists these days tend to study more on herbal plants as

alternatives for some products especially those that are common in the environment. One of the

examples is the Orange Lirio (Hippeastrum puniceum) bulb extract which is said to be good in

curing fungal infections.

Orange lirio (Hippeastrum puniceum) is an ornamental plant widely known in the country.

The fresh bulb of the plant is traditionally used to treat wounds, tumors and stomachaches done by

the locals. However, there is still no known relevant scientific research about its medical use. (Dr.

Farrah, 2018)

In this study, the researchers try to prove the effectivity of the plant’s bulb extract against

fungal infections which is a problem of all age groups. The researchers want to prove that there

are plants that are more effective against other diseases especially in skin. The researchers also

want to scientifically validate the pharmacogenetic and physicochemical parameters of the plant.

According to a recent study, the bulb of the plant is found out to contain chemical compounds

especially alkaloids that is an emerging therapeutic aid against fungal diseases. Aside from that, it

also protects the plant from insect attack and can be an antibiotic agent. (Stuart, 2015)

By understanding the concept of this research, one can acknowledge that the plant’s bulb

is not just as a part of the plant’s structure but also as an alternative for curing fungal infections.

Some people are using organic exfoliant seen in advertisements to exterminate skin infections such

as alipunga (Tinea pedis), ringworm (Tinea corporis), and an-an (Tinea versicolor). Due to the
side effects that are observed in the usage of these, the researchers conducted this to help their

fellowmen look for solutions and alternatives that are not harmful to one’s health.

B. Statement of the Problem

Orange lirio is known for curing infections/aches internally and externally. It helps heal

infections easily for it has chemical substances that rejuvenates or renew dead cells that can be

found in the skin. It kills bacteria without using harsh ingredients; it helps promote hygiene without

irritating the skin from within. (Stuart, 2015)

In this study, it is proposed to answer the question: “How effective is the orange lirio

(Hippeastrum puniceum) bulb extract in treating fungal infections?”

1. What properties of the extract of the Orange lirio bulb make fungal infections curable?

2. What kind of fungal infections can the bulb extract treat?

3. What is the most effective way to get the extract of the orange lirio bulb?

4. Is using this as an alternative safe?

5. Does it have side effects on one’s health?

6. Does this cost more expensive or cheaper than ordinary anti-fungal products?

C. Hypothesis

Hypothesis – The orange lirio bulb extract is very effective in healing fungal infections.

Null hypothesis – The orange lirio bulb extract is not (so) effective in healing fungal infections.

D. Significance of the Study

 Orange lirio is very common in the environment. Aside from its availability, it is also very

easy to cultivate. Using the bulb of orange lirio as an alternative for anti-fungal products deepens

the possibility of making treatments out of these due to the therapeutic properties of its extract.

Researchers – This study would help the student researchers to be aware and

knowledgeable of the process involved in extracting an orange lirio bulb for a treatment.

Customers – This study would help the customers especially housewives, students, teens,

etc. that uses orange lirio bulb as part of their everyday life. It would help to maximize the quality

of the product and at the same time meet the demands of their market.

Institution – This study would enable the institution to provide an environment of learning

and encouraging students to conduct researches that has a valid impact in the society.

The study aims to produce an exfoliant out of Orange lirio bulb extract. The product to be

produced will be effective because of the phytonutrients from the bulb of Hippeastrum puniceum

that will surely help renew and maintain the glowing, smooth and beautiful skin. Moreover, it is

also eco-friendly since it doesn’t have harmful elements on it that can cause severe damage to your

skin.

E. Scope and Delimitation of the Study

The study talks about how the bulb extract of the Orange Lirio (Hippeastrum Puniceum)

can be possibly used as an alternative treatment for fungal skin infections like alipunga (Tinea

pedis), ringworm (Tinea corporis), an-an (Tinea versicolor), and athlete’s foot. The bulb of the

ornamental plant used for this study contains “Alkaloids” which is widely used by various

synthetic agents in its clinical practices by studying the effectiveness and its anti-fungal potential.
 The study limits the use of the exfoliant in the face because it may irritate the skin

especially those who have sensitive ones. Despite this, the study of orange lirio bulb and getting

its extract is often useful since it has the ability to treat, cure, and disinfect fungal infections.

F. Definition of Terms

Potency – the strength or effectiveness of something

Bulb – a rounded part of some plants that is under the ground and that grows into a new plant

Pharmacognostic – natural molecules of a plant that can be used to make medicines, food and

other agricultural and consumer products

Physicochemical – being physical and chemical

Alkaloids – any of a class of nitrogenous organic compounds of a plant which have physiological

actions on humans

Therapeutic – producing good effects on your body or mind

Exfoliant – a mechanical or chemical agent that is applied to the skin to remove dead cells from

the surface

Cultivate – to grow or raise (something) under conditions that you can control

Phytonutrients – a substance found in certain plants which is believed to be beneficial to human

health and help prevent various diseases

 Chapter II

REVIEW OF RELATED LITERATURE

Orange Lirio (Hippeastrum Puniceum)

Hippeastrum puniceum, known as Fire Lily, Easter Lily, Lily of the Palace, Common

Amaryllis and Ridderstjerne is a perennial bulbous plant which is native to some parts of tropical

regions of South America and it has become naturalized in some places. It produces 3 to 6 bright

green, glossy strap-shaped (lorate) leaves, 12-24 inches long and 1”- 1 ¼ inches wide that taper to

a point. The flowers are shaped like a trumpet and borne in an umbel on a stem. The petals, or

more precisely tepals, are orange-red with paler bases. The bulb consists of medicinal components

and it is used against asthma, constipation, bronchitis and other respiratory diseases. It also

contains the phytochemicals, namely Galanthamine and Lycorine (Witherill, 2012).

Tribal and Herbal Medicine Uses

The orange lirio (Hippeastrum puniceum) is used by many for treating stomachaches.

There are different countries who use this flower for medicinal purposes. In India, the fresh bulbs

are used to heal wounds, piles and tumors, while in Jamaica, it is used to produce plaster with

bread or called as Eryngium foetidum which is used on swelling, sores, abscesses and ulcers. For

the people in French Guiana, they locally used orange lirio on treating whooping cough. Flower

infusion in their country is considered as an antispasmodic. In NW Guyuna, the roots of the flower

are used to cure asthma, biliousness to induce vomiting and as laxative. It is said that Hippeastrum

puniceum may have therapeutic compositions because it is locally practiced in medicine as a

remedy for swellings and wounds (Mitchell, Ahmad, 2006).

Fungal Infections

Fungi stays in certain parts of the body where the skin surfaces touch: under the breasts,

between the toes, and in the genital area. Generally, most fungal skin infections are mostly caused

by yeasts (like Malassezia furfur or Candida) or called as dermatophytes, such as Epidermophyton,

Microsporum, and Trichophyton. Most fungi can be seen in the epidermis’ topmost layer (stratum

corneum) and it does not go deeper. Obese people are most likely to obtain these infections for

they have excessive skinfolds, specifically if the skin is broken down and it becomes irritated.

Individuals with diabetes are likely to be more vulnerable to fungal skin infections as well (Aaron,

2018).

Strangely, fungal infections on one part of the body can cause rashes on parts of the body

that are not infected. For example, a fungal infection on the foot may cause an itchy, bumpy rash

on the fingers. These eruptions (dermatophytids, or identity or id reactions) are allergic reactions

to the fungus. They do not result from touching the infected area (Aaron, 2018).

Plant Chemicals

The Hippeastrum puniceum bulbs were collected in the Catimbau National Park, PE. The

fresh plant material was triturated and macerated with methanol for 48h at room temperature. With

a methanolic extract the selective extraction for alkaloids was carried out and other fractions were

obtained: Hp01 (Hexane), Hp02 (Ethyl Acetate) and Hp03 (Ethyl Acetate: Methanol 3:1). Later,

these fractions were submitted to GC / MS analysis and to classical fractionation and purification

techniques. The examination of extracts and fractions by GC-MS showed the presence of 11

alkaloids, such as 9-O-demethyllycoramine, lycoramine, galantamine, assoanine, kirkine, 8-

demethylmaritidine, pancratinine, 11-hydroxyvittatine, pseudolicorine, 2α-hydroxyhomolycorine

and lycorine. Of these, four were isolated and identified from the class of isoquinoline alkaloids,
9-O-demethyllycoramine, 9-demethyl-2α-hydroxyhomolycorine, lycorine and tazettine (Jamal,

Soprani, Andrade, Bastida, Borges, 2018).

Microscopic examination of bulbs showed starch grains, mucilage cells and xylem fibers.

Phytochemical screening of various extracts yielded alkaloids, carbohydrates, flavonoids, tannins,

saponins, terpenoids, proteins and amino acids. A chloroform extract showed the highest phenolic

content (Deepa, Kuriakose, 2014).

Study evaluated an aqueous extract of bulbs of Hippeastrum puniceum for anti-

inflammatory activity using protein denaturation and proteinase inhibition methods and

antioxidant activity by iron chelating and total antioxidant assay. Results showed that antioxidant

and anti-inflammatory is possible to be attributed to flavonoids and phenolics (Stuart, 2015).

Antifungal Agents

Flavonoids are phenolic compounds widely distributed in the plant kingdom. These natural

compounds have been known to be active against a variety of microorganisms. An overview of

the recent papers on the antifungal activity of flavonoids which represent a potential alternative to

conventional fungicides is presented (Lourenço, 2013).

Terpenoid phenols, including carvacrol, are components of oregano and other plant

essential oils that exhibit potent antifungal activity against a wide range of pathogens, including

Candida albicans, Staphylococcus aureus, and Pseudomonas aeruginosa (Rae et. al. 2010).

EtOAc fraction showed a broad spectrum of antimicrobial activity and gallic acid had better

antifungal activity, while tannin was active against all microorganisms tested (Carvalho et. al.

2018).

Saponins exhibit antimicrobial properties, guarding your body against fungi, bacteria and

viruses. At the same time, they improve immune function by stimulating the production of T-cells.
Additionally, they act as antioxidants and scavenge oxidative stress. That's why these compounds

are used in some vaccines (Picincu, 2018).

 Chapter III

METHODOLOGY

We must know the respective uses and it is also necessary to know the alternatives, if some

materials are not available or accessible. With these guidelines, the flow of conducting this

experiment will have little to no interruptions or hindrances that could possibly disturb the process

if making this experiment.

Materials and Equipment:

The materials needed are the following: orange lirio bulbs, vodka, egg yolks, olive oil,

water, wire whisk, bowl, empty jars, and kitchen knife.

Procedure:

A. Extraction

Slice the orange lirio bulb and triturate into small mince. Then, macerate it with vodka

and leave the solution in room temperature for 48 hours. Vodka is used because it has 40%

ethanol, and ethanol is used in the preparation of essences and flavorings in medicines and

drugs like anti-fungal and anti-inflammatory products. After 2 days, the orange lirio bulb

extract will be obtained.

B. Emulsification

Beat the egg yolks in a bowl with a wire whisk. Many proteins in egg yolks can act

as emulsifiers because they have some amino acids that repel water and some amino acids

that attract water. Lecithin is another important emulsifier in egg yolk. Known as a

phospholipid, it’s a fatlike molecule with a water-loving “head” and a long, water-fearing

“tail.” The tail gets buried in the fat droplet and its head sticks out of the droplet surface into

the surrounding water. This establishes a barrier that prevents the surface of the fat droplet

from coming into contact with the surface of another fat droplet.

Afterwards, gently pour 1 cup of olive oil and ¼ cup of water into the egg yolks and

mix. Olive oil acts as an anti-oxidant, which is a substance that prevents oxidation. When

applied to the skin, anti-oxidants may prevent premature aging. Also, some research suggests

that putting olive oil on the skin after sun exposure may fight off cancer-causing cells. After

mixing, pour the orange lirio bulb extract and mix again. Finally, pour the finished solution

into 3 empty jars.

Experimental Design:

In order to determine the quantity of the orange lirio bulb extract needed in order for this

experiment to be effective, the researches prepared 3 empty jars that will be filled with varying

amounts of the extract.

Table 1: Jar Extract Samples

Number of teaspoons
Test Subjects
with the extract added

Jar A 1 teaspoon of lirio bulb extract

Jar B 2 teaspoons of lirio bulb extract

Jar C 3 teaspoons of lirio bulb extract

Testing:

According to some studies, the orange lirio bulbs have not been found to be applicable to

humans especially because of the presence of alkaloids. Without further evidence, the extract must

not be used because of its possible harmful effects from its properties.
 Chapter IV

RESULTS AND DISCUSSION

Table 2: Chemical Constituents of Orange Lirio Bulb Extract

Constituent Uses

Lycoramine Anti-inflammatory

Galantamine Anti-inflammatory

Assoanine Antioxidant

Tazettine Antioxidant

Kirkine Antioxidant

Pancratinine Antioxidant

Lycorine Antioxidant

Flavonoids Antifungal

Terpenoids Antifungal

Tannins Antifungal

Saponins Antifungal
 Carbohydrates Provide and Store Energy

Important in the biosynthesis of hormones,

Proteins
enzymes, and membrane channels

As shown in the table above, it shows the contents of the orange lirio bulb extract and its

uses. Out of all the contents in the table, there are four of these that are antifungal. These are the

flavonoids, saponins, terpenoids, and tannins. These three are chemical substances found in the

orange lirio bulb that contain antifungal properties that helps eradicate fungi and bacteria.
 Chapter V

SUMMARY, CONCLUSIONS, AND RECOMMENDATIONS

Summary:

This section presented an overview of our study on the effectivity of orange lirio

(Hippeastrum puniceum) bulb extract in treating fungal infections. The researchers made an

investigatory project that aims to know and to produce an antifungal cream by using the extraction

of the orange lirio's bulb. With the simple materials and ingredients gathered and used, we are able

to conduct our study and prove if it is effective or not.

Based on the observations in the methodology, the bulb of the plant is harmful to apply on

the skin of the humans because of its harmful composition. It contains alkaloids with reported

toxicity, especially to cats. Bulbs considered the most poisonous part of the plant. Toxicity is

attributed to phenanthridine alkaloid derivatives i.e., lycorine, crinidine, clivacetine clivorine,

cliviasine and clividine. Manifestations of poisoning may include gastrointestinal (vomiting,

salivation, diarrhea, abdominal pain), convulsions, cardiac arrhythmias, low blood pressure and

respiratory depression.

Conclusion:

The constituents of the orange lirio bulb extract contained flavonoids, tannins, saponins,

and terpenoids, which have antifungal properties. Therefore, there is a slight possibility that the

orange lirio bulb extract cream is very effective in healing fungal infections.

Recommendations:
 The researchers have found out that the extract from the orange lirio bulb is said to have

both antifungal properties and toxicity. Further studies are recommended to prove the antifungal

properties like the flavonoids, tannins, saponins and terpenoids to be suitable for the human skin

besides the presence of alkaloids like lycorine, crinidine, clivacetine clivorine, cliviasine and

clividine that cause its toxicity. Without enough evidence, the researchers suggest that the orange

lirio bulb should not be used in the human skin.

Moreover, this study will serve as a precursor for other researchers who conducted similar

study or to those who will have interest in investigating this scientific problem. This science

investigatory project can be used to back up their research. To sum it up, the researchers emphasize

the need for careful test and analysis in conducting this study.
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