Name : Siti Maryam binti Mohd Ramzi

Proposed Title : Eco-friendly Protein Extraction from Moringa Oleifera

INTRODUCTION

Moringa oleifera is the most widely cultivated species of the genus Moringa, which is the

only genus in the family Moringaceae. Moringa is a fast-growing, drought-resistant tree

which could be widely found in north-western India, and widely cultivated in tropical and

subtropical areas of Asia, South Africa, South America, the islands of Caribbean and Pacific.

The tree is also known as the ‘magical tree’ by Western researchers for its outstanding and

abundance of benefits that it could offer. Even so, it is not commonly known by the locals

due to its non-local origin.

Moringa is known to be a vitamin source or the cheapest supplementary food because it

could provide nutrients to promote ones’ health. It contains vitamin A more than carrot, iron

more than spinach, vitamin C more than orange, calcium more than milk, potassium more

than banana and its protein quality rivals that of eggs and milk and could be easily be

digested.

Traditionally, the tree is used to treat diarrhea, gut inflammation, parasites, arthritis, joint

pain, gout, sore throat, urinary tract infections and anaemia. This is because moringa has

antibacterial property, including helicobacter pylori bacteria which causes ulcer and gastritis,

and anti-inflammatory. However, the scope of this study is focusing on the extraction of

protein from its leaves and seeds using an eco-friendly method which the quality of the yield

is as good as of eggs and milk and easily digested. The justification of why protein is to be

extracted from moringa is because of its high protein content, of about 2g of protein from 12g

of dried moringa seeds and leaves, respectively. The grout of its seeds and leaves produced

during the extraction could be used as livestock feed.

PROBLEM STATEMENT

There are a few ways that have been used to extract protein done by researchers (Martínez-

Maqueda, Hernández-Ledesma, Amigo, Miralles, & Gómez-Ruiz, 2013). It starts with the

technic of breaking down the cell wall of the material to be used. There are five categories

for this purpose which are mechanical homogenization, ultrasound homogenization,

pressurised homogenization, temperature treatment and lastly, chemical and osmotic

breakdown. Based on previous researches, two methods which were considered suitable for

post-extraction from moringa are ultrasound homogenization for its leaves and temperature

treatment for its seeds.

The dilution of protein is also important as it is the key point in preparing the protein

samples. It is used to isolate protein from other substances present in the sample. Organic

solvent, reported by other researches, could be used to extract protein from food sources.

Commonly used method in extracting protein from plant sources is the coagulation of

trichloroacetic acid (TCA) or also known as acetone. The rising awareness of the

environmental issue causes the usage of organic solvent to be avoided.

Later, extraction using water as solvent started to gain the attention of the researchers. The

protein yield is not as much as the extraction using organic solvent which makes the

researchers to come up with another method using alkali. However, high pH level would

cause the protein obtained to disintegrate which could lower the percentage of the yield. The

quality of the protein yield is also low due to the side reaction of the product with alkali.

Moreover, a huge amount of water is needed to flush out the remaining alkali in the sample

which will produce a lot of waste water.

Latest research reported the usage of sodium salt and calcium to extract protein from plant

sources is the simplest method proposed due to the availability of the reagent. In this

research, a type of solvent called ionic liquid which is known for its eco-friendly property is

proposed to be used to extract protein from moringa. Ionic liquid is a type of salt and chosen

for its liquidised state, also the use of ionic liquid has not been commercialised by others

researchers as to date. Theoretically, it could extract protein exactly like of the extraction

using sodium salt and calcium. In this research, the ionic liquid to be used is the eutectic

mixture of ethyltrimethylammonium (choline) chloride and urea which is also known as deep

eutectic solvent (DES). DES is chosen for its affordable price and non-toxic property (Singh,

Lobo, & Shankarling, 2012).

OBJECTIVES

The objectives of this research are:

1. To extract protein from moringa seeds and leaves.

2. To determine the presence of protein by analysing the presence of amino acids.

SCOPE OF STUDY

It should be noted that only moringa seeds and leaves would be used as the raw material for

this research. The solvent to be used in this research is DES.

METHODOLOGY

Preparation of leaf powder

Preparation of seed powder

Note : it is highly preferable to execute steps 3 to 6 right before the extraction process is

conducted to prevent any aging effects.

Strip leaves

Wash with clean water and drain

Wash with 1% saline (3-5 mins) and drain

Wash with clean water and drain

Dry using drier till moisture content less

than 10% at 50°C-55°C

Mill with blender till fine powder form

Sieve powder to remove stalks and large contaminants

Dry using drier at 50°C for 30 mins till

moisture content below 7.5%

Powder store in air tight container and

ready to be used

Wash pods with clean water

Sun-dry pods till could be opended

easily

Retrieve seeds Dehusk and deshell seeds

Mill using blender till fine powder form

Powder store in air tight container and

ready to be used

Preparing crude extract

Extracting protein from crude extract

Lipid removal using 95% ethanol or petroleum ether

Filter mixture and dry at room temperature

Mix filtered solid with water

Stir mixture for 30 mins

Filter the seperated solution

Filtrate ready to be used

Mix crude extract with DES

Filter or centrifudge misture to obtain the white precipitate formed

Suspend in water and filter to remove insoluble materials

Freeze dry product

White protein powder is obtained

ANALYSIS METHODOLOGY

1. HPLC

2. FTIR

ReferencesMartínez-Maqueda, D., Hernández-Ledesma, B., Amigo, L., Miralles, B., & Gómez-Ruiz, J. Á. (2013).

Extraction/Fractionation Techniques for Proteins and Peptides and Protein Digestion. In F. Toldrá, L. Nollet, & (eds.), Food Microbiology and Food Safety 2 (pp. 21-50). New York: Springer Science+Business Media.

Singh, B. S., Lobo, H. R., & Shankarling, G. S. (2012). Choline chloride based eutectic solvents: Magical catalytic system for carbon–carbon bond formation in the rapid synthesis of β-hydroxy functionalized derivatives. Catalysis Communications(24), 70-74. Retrieved from www.elsevier.com/locate/catcom