CHAPTER I

THE PROBLEM AND ITS BACKGROUND

INTRODUCTION

Lactic acid (2-hydroxypropanoic acid, C3H6O3) is an organic compound discovered

by Carl Wilhelm Scheele in 1780 where he successfully isolated it from the sour milk

(Castillo Martinez et al., 2013). The production can either be naturally or synthetically.

This organic acid is viscous, nonvolatile liquid at room temperature and hygroscopic

because of its miscibility in water and ethanol above melting point (Organic Materials

Review Institute (OMRI), 2015).

Nowadays, the demand for lactic acid is increasing continuously. Around 370,000

metric tons of lactic acid was produced in the year 2017 and a rapid increase of demand

every year at the rate of 5-8%. It is of earnest importance because of its use as a raw material

for the production of various products by fermentation and different industries like food,

pharmaceutical, cosmetics and textile industries. The hydroxyl and carboxyl groups that

are present on the acid’s structure can be converted into useful chemicals such as esters

and bio solvents. The most important application of lactic acid is its use as feedstock for

the production of the bio polymer polylactic acid (PLA) (Battula, et.al., 2018). PLA is

biodegradable and environmentally friendly that is used as an alternative to plastics made

from petrochemicals. Its use would significantly alleviate waste disposal problems. Other

application is on medical devices (Abdel-Rahman, et.al., 2011). Because of the wide range

of lactic acid’s use there is a need for a greater production of it.

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Potentiality of Lactic Acid Production from Chapter I. The Problem and Its Background
Indian Almond (Terminalia catappa) Mesocarp
Aycardo, M.C.A., Celis, C.M.L., Montoya, E.C.A.

Synthetic production of lactic acid involves chemical synthesis. Natural production

involves raw materials containing high percentage of carbohydrates. Examples are the

lignocellulosic biomass to undergo treatment and fermentation. Compare to the synthetic

production, natural production of lactic acid is more economical and favorable due to

environmental concerns and limited nature of petrochemical feedstocks (Wee, et.al., 2006).

Through this, range of research and development programs are being put through to

ascertain other raw materials for the production. Indian Almond (Terminalia catappa),

commonly known in the Philippines as Talisay, is a raw material that can be considered in

the production of lactic acid.

Indian Almond (Terminalia catappa) is a tree found throughout the Philippines

along seashore. The world’s production of this fruit stands at 700,000 tons annually.

Talisay tree bears abundant fruits and usually falls on the ground when ripened. The seed

of the fruits can be eaten and are very nutritious containing significant amount of high-

quality proteins and chemicals (Bolaji, et.al., 2013). Once the seed of the fruit was eaten,

the mesocarp part is left and is already considered as waste. Physicochemical analysis of

dried Indian Almond mesocarp shows that it contains 12.65% of ash, 84.93% of

carbohydrates, 0.37% of oil, 0.1% of protein, 1.35% moisture, 316 mg/g of glucose, and

1.30 mg/g of tannin with 3434.5 kcal/kg calorific value (Nwosu, et.al, 2008). It can be

noticed that 84.93% of Indian Almond mesocarp is carbohydrates which makes it a

potential raw material for lactic acid production since carbohydrate is the precursor for its

production. Even though studies also show that there is a significant amount of

carbohydrates in the seed, it cannot be considered due to the fact that the seeds can be eaten.

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Potentiality of Lactic Acid Production from Chapter I. The Problem and Its Background
Indian Almond (Terminalia catappa) Mesocarp
Aycardo, M.C.A., Celis, C.M.L., Montoya, E.C.A.

STATEMENT OF THE PROBLEM

The study aimed to know the potentiality of lactic acid production from Indian Almond

(Terminalia catappa) mesocarp. Specifically, this study aimed to answer the following:

1. Is there a significant difference on the effect of varying molar concentration of sulfuric

acid (0.3, 0.4, 0.5 M) in the physicochemical properties in the extraction of

carbohydrates from Indian Almond mesocarp in terms of the following parameters?

1.1.Moisture

1.2.Ash

1.3.Crude protein

1.4.Crude fiber

1.5.Crude fat

1.6.Nitrogen free extract/ carbohydrates

2. How do the following varying conditions affect the yield of lactic acid?

2.1.pH (7, 8, 9)

2.2.Lactic Acid Bacteria (1/4, 1/3, ½ LAB culture)

NULL HYPOTHESES

1. There is no significant difference on the effect of varying molar concentration of

sulfuric acid (0.3, 0.4, 0.5 M) in the physicochemical properties in the extraction of

carbohydrates from Indian Almond mesocarp in terms of the following parameters:

1.1.Moisture

1.2.Ash

1.3.Crude protein

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Potentiality of Lactic Acid Production from Chapter I. The Problem and Its Background
Indian Almond (Terminalia catappa) Mesocarp
Aycardo, M.C.A., Celis, C.M.L., Montoya, E.C.A.

1.4.Crude fiber

1.5.Crude fat

1.6.Nitrogen free extract/ carbohydrates

2. There is no significant difference on the effect in the yield of lactic acid upon

varying the following conditions:

2.1.pH (7, 8, 9)

2.2.Lactic acid bacteria (1/4, 1/3, ½ LAB culture)

SIGNIFICANCE OF THE STUDY

This research aimed to make use of carbohydrates that was extracted from the

mesocarp of Indian Almond that undergone fermentation and know if there is a potential

that lactic acid can be produced. The wide range of the use of lactic acid made this study

significant because it will contribute on the researches on finding renewable sources for

the production of lactic acid.

This study provided data on the yield of carbohydrate that can be extracted from

the Indian Almond mesocarp and the yield of lactic acid after fermentation making it

significant for the future studies that will further optimize the process for the production of

lactic acid. This study also encompasses the methods used upon obtaining the data.

The data obtained can also be used as a basis for future studies regarding the

application of the other component of Indian Almond mesocarp.

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Potentiality of Lactic Acid Production from Chapter I. The Problem and Its Background
Indian Almond (Terminalia catappa) Mesocarp
Aycardo, M.C.A., Celis, C.M.L., Montoya, E.C.A.

SCOPE AND DELIMITATION OF THE STUDY

The study conducted is only limited to the potentiality of lactic acid production

from Indian Almond (Terminalia catappa) mesocarp. Indian Almond was chosen as the

raw material of the study because of its abundance in the locality to which the use of the

high carbohydrate content of the fruit’s mesocarp remains unexplored.

The processes involved in the study are carbohydrates extraction by acid hydrolysis

and fermentation using lactic acid bacteria to convert the extracted carbohydrates to lactic

acid. Varying molar concentration of sulfuric acid for acid hydrolysis to extract

carbohydrates was employed and varying substrate’s pH and lactic acid bacteria dosage for

the fermentation of carbohydrates to form lactic acid.

The milling process of the mesocarp was performed at the university’s Unit

Operations Laboratory wherein Wiley Mill was used. The fermentation process was done

in the Chemical Engineering Laboratory 1. Analytical testing of the substrate from the acid

hydrolysis of milled mesocarp was done by Lipa Quality Control Center (LQCC) and the

testing for the potentiality of the lactic acid production after fermentation was done by SGS

Philippines, Inc. The chosen laboratory testing centers offer quality inspection, verification,

testing, and certification services.

CONCEPTUAL FRAMEWORK OF THE STUDY

The study aimed to know the potentiality of lactic acid production from the

mesocarp of Indian Almond (Terminalia catappa). The figure below shows the conceptual

framework used in the study.

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Potentiality of Lactic Acid Production from Chapter I. The Problem and Its Background
Indian Almond (Terminalia catappa) Mesocarp
Aycardo, M.C.A., Celis, C.M.L., Montoya, E.C.A.

INPUT PROCESS OUTPUT

• Indian Almond • Indian Almond • Carbohydrates
Mesocarp Mesocarp • Lactic Acid
• Lactic Acid Pretreatment
Bacteria (LAB) • Acid Extraction of
• Acid Carbohydrates
• Characterization of
Carbohydrates
• Lactic Acid Bacteria
Preparation
• Fermentation
• Characterization of
Lactic Acid

Figure 1.1. Conceptual Framework of Potentiality of Lactic Acid Production from Indian

Almond (Terminalia catappa) Mesocarp

DEFINITION OF TERMS

Acid Hydrolysis. It is a process in which a protic acid is used to catalyze the cleavage of

a chemical bond via a nucleophilic substitution reaction, with the addition of the elements

of water (H2O). For example, in the conversion of cellulose or starch to glucose (Rennie &

Law, 2016). In this study, acid hydrolysis is the method use to extract carbohydrates from

the milles Indian Almond mesocarp.

Anaerobic Fermentation. a method cells use to extract energy from carbohydrates when

oxygen or other electron acceptors are not available in the surrounding environment

(Cammack et al., 2008).

Carbohydrate. It is an abundant and diverse group of compounds that have the general

formula Cn(H2O)n. The smallest are monosaccharides like glucose, the largest are complex

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Potentiality of Lactic Acid Production from Chapter I. The Problem and Its Background
Indian Almond (Terminalia catappa) Mesocarp
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polymers such as starch, cellulose, and glycogen that are indeterminate in length (Lackie,

2010). In this study, carbohydrate is the intermediate extracted after the acid hydrolysis of

milled Indian Almond mesocarp.

Enzymatic Hydrolysis. It is a process in which enzymes facilitate the cleavage of bonds

in molecules with the addition of the elements of water (Rennie & Law, 2016).

Fermentation. the chemical breakdown of a substance by bacteria, yeasts, or other

microorganisms, typically involving effervescence and the giving off of heat (Rennie &

Law, 2016). In this study, fermentation is the process used for the conversion of

carbohydrates to lactic acid.

Heterolactic Fermentation. A type of lactic acid fermentation by which small yields of

lactic acid are produced and much of the sugar is converted to carbon dioxide and other

products (Cammack et al., 2008).

Indian Almond. An Asiatic tree "(Terminalia catappa") widely cultivated and naturalized

in the tropics and having fruit with edible seeds (Davidson & Jaine, 2014). In this study,

Indian Almond fruit is the raw material used.

Lactic Acid. A hygroscopic organic acid C3H6O3 present normally especially in muscle

tissue as a by-product of anaerobic glycolysis, produced in carbohydrate matter usually

by bacterial fermentation, and used especially in food and medicine and in industry

(Allaby, 2019). In this study, lactic acid is the product of carbohydrate fermentation using

lactic acid bacteria.

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Potentiality of Lactic Acid Production from Chapter I. The Problem and Its Background
Indian Almond (Terminalia catappa) Mesocarp
Aycardo, M.C.A., Celis, C.M.L., Montoya, E.C.A.

Lactic Acid Bacteria (LAB). Any of various bacteria chiefly of the

genera Lactobacillus and Streptococcus that produce predominantly lactic fermentation

on suitable media and some of which are used in the commercial production of lactic

acid and as cheese and butter starters (Bender, 2014). In this study, LAB is the type of

bacteria used for the fermentation of carbohydrates to produce lactic acid.

Mesocarp. The middle layer of the pericarp of a fruit, between the endocarp and the

exocarp. It is usually the fleshy layer of a fruit wall (Bender, 2014). In this study, Indian

Almond mesocarp was used as its physicochemical analysis shows that it contains a large

amount of carbohydrate.

Nutrient Agar. Nutrient agar is used as a general-purpose medium for the cultivation of

less fastidious microorganisms, can be enriched with blood or other biological fluids

(HiMedia Laboratories Pvt. Ltd., 2018). In this study, nutrient agar is the media used to

culture the lactic acid bacteria.

pH. It is a measure of the concentration of hydrogen ions in a solution, and therefore of its

acidity or alkalinity (Schaschke, 2014). In this study, pH is one of the conditions that varies

to know its effect on the yield of lactic acid.

Substrate. It is the base on which an organism lives, grow, or obtains its nourishment

(Schaschke, 2014). In this study, substrate is the extract from the acid hydrolysis that will

undergo fermentation.