CARBONAID: FORMULATION OF CALCIUM CARBONATE-BASED SOLUTION

FROM ASIATIC CLAM (Corbicula fluminea) FOR POSTHARVEST MANAGEMENT

APPLICATION ON MANGO (Mangifera indica L.)

Intellectual Property Office of the Philippines (IPOPHL) Patent Code

eInvFile [ L9Y01XE07EQ2025O36 ]

A Research Project submitted to the National Level Science & Technology Fair

(Innovation Expo - Individual Category)

STEMIE-I-RO2

AINGEAL KLARYZE P. DAQUIOAG

JUVIS GEM A. DE LA CRUZ

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 RESEARCH PLAN

A.​ RATIONALE

Agriculture is an important aspect of the Philippine economy, contributing

approximately 8.6% to the Gross Domestic Product (GDP) in 2023 (Philippine Statistics

Authority). However, significant post-harvest losses pose a serious threat to the sector.

Agricultural products, particularly perishable items like mangoes, often face wastage during

distribution. Effective post-harvest management is crucial for food security, and as demand

for high-quality goods increases, the food industry must explore innovative preservation

methods.

Mango (Mangifera indica L.) is a highly valued tropical fruit, with the Philippines

being a key player in the global mango trade since the 1980s. In 2023, mango exports were

valued at P2.9 billion, making it one of the country's largest agricultural exports (Galang

2024). However, the industry is hampered by challenges such as climate change, pests,

diseases, and inadequate technology, leading to significant post-harvest losses, particularly in

regions like Pangasinan, where losses amounted to PHP 1.595 million in 2022 (S&T Media

Services 2022).

To address these issues, the researcher proposes a sustainable post-harvest

management solution that utilizes calcium carbonate derived from Asiatic clams (Corbicula

fluminea). This naturally occurring mineral has gained attention for its effectiveness in food

preservation due to its antimicrobial properties, which help maintain the quality of mangoes

during storage and transport.

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 B.​ RESEARCH OBJECTIVES

This study aims to develop a calcium carbonate-based coating solution from Asiatic

clams for the shelf-life extension of mangoes. Specifically, it seeks to answer the following:

1.​ What are the values of the following parameters for Carabao mango slices?

a.​ Weight Loss

b.​ Color values (L*, a, b, BI)

c.​ Microbial Growth (Agar Plate Count)

2.​ What are the values of the following parameters for unpeeled whole Carabao

mangoes?

a.​ Weight Loss

b.​ Color Values (L*, a, b, BI)

c.​ Titratable Acidity

d.​ Microbial Growth (Total Yeast and Molds Count)

3.​ Is there a significant difference among the four treatment solutions in terms

of the postharvest properties of sliced and whole mangoes?

4.​ Is there a significant difference among the four treatment solutions in terms

of anthracnose mitigation?

5.​ At what concentration of the coating solution is the most effective in

preserving the postharvest qualities of mangoes?​

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 C. METHODOLOGY

1.​ PROCEDURES

This section discusses the procedures that the researcher will conduct.

Figure 1. Flowchart of the Study

Figure 1 shows the general flowchart of the procedure that the researcher will

conduct. The Asiatic Clamshell (Corbicula fluminea) waste will be collected at Pinacanauan

River through the local fishermen in Peñablanca, Cagayan. After purchasing the chemicals,

the Asiatic Clamshells will then be processed into calcium carbonate and then formulated

into a coating solution. Treated and untreated mangoes will be analyzed according to their

postharvest quality parameters. A statistical analysis will then determine the coating's

effectiveness.

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Preparation of Asiatic Clams

The Asiatic clam shells (ACS) will be washed thoroughly under running water,

broken into smaller fragments, and subjected to oven drying at 60℃ for 8 hours. The dried

shells will subsequently be ground and sieved using a laboratory sieve to obtain a fine,

uniform powder.

Calcium Carbonate Extraction

a.​ Acidic Hydrolysis

The ACS powder will be immersed in 1 M HCl at a solute-to-solvent ratio of 1:10

(g/mL) for 100 minutes, maintained under constant stirring at 120 rpm and 60°C. The

mixture will then be filtered to obtain a calcium-enriched filtrate, reducing phytate content

and enhancing calcium availability for subsequent extraction. Decantation will be carried out

to isolate the precipitate.

b.​ Calcium Recovery

An alkaline treatment for calcium recovery will be carried out using a 12.5 M NaOH

solution with a solute-to-solvent ratio of 1:10 (g/mL). The calcium-enriched ACS will be

subjected to hydrolysis for 5 hours, maintained at 75°C with continuous stirring at 120 rpm.

c.​ Purification

A treatment with 0.88 M NaOH will be applied at a solute-to-solvent ratio of 1:10

(g/mL) for 180 minutes, with continuous stirring at 120 rpm and a temperature of 75°C. The

resulting precipitate will be neutralized using deionized water to achieve a pH of 7, then dried

at 90°C for 5 hours. The resulting powder will be further ground and sieved.

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Characterization of Extracted ACS Calcium Carbonate

A portion of the extracted powder will be submitted to the Department of Science and

Technology Advanced Materials Testing Laboratory (DOST-ADMATEL) for Fourier

Transform Infrared Spectroscopy (FTIR) analysis, where specific peaks associated with

calcium carbonate will be detected.

Preparation of Coating Solution

To prepare the calcium acetate solution, different amounts of calcium carbonate

powder (1%, 2%, and 3%) will be dissolved in 1000 mL of a 1% v/v aqueous acetic acid

solution, stirred at 120 rpm for 6 hours at 60°C. Separately, pectin powder will be dissolved

in water at a 5% concentration and stirred for 1 hour. The pectin solution will then be

combined with the calcium acetate solutions in a constant ratio of 3:2 v/v.

Collection of Mangoes and Application of Coating Solution

Eighteen Carabao mangoes of similar size, color, and ripeness will be collected and

washed with running water. Twelve whole mangoes (3 replicates per treatment) will be

assigned to the unpeeled group, while 20 quarter samples (5 replicates per treatment) will be

used for the sliced group. Solutions will be applied by dipping for 1 minute, followed by 10

minutes of drying in a sterile environment. Both coated and uncoated mangoes will be stored

in a controlled incubator at 25℃, with a pre-analysis of their properties conducted for

comparison.

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Postharvest Quality Analysis of Mangoes

Data for the sliced group will be gathered daily for six days, while observations for

the unpeeled group will be made at two-day intervals over a period of eight days.

a.​ Weight Loss

​ Weight loss will be determined by comparing the initial weight of the mango

samples (before treatment) with their weight at each observation point throughout the

study period.

(𝑖𝑛𝑖𝑡𝑖𝑎𝑙 𝑤𝑒𝑖𝑔ℎ𝑡 − 𝑐𝑢𝑟𝑟𝑒𝑛𝑡 𝑤𝑒𝑖𝑔ℎ𝑡)

b.​ Color Values

Browning Index

A colorimeter will be used to assess browning in mango samples by measuring

the L*, a*, and b* values.

L* value indicates lightness, ranging from 0 (black) to 100 (white).

a* value represents the green-to-red spectrum, ranging from -120 to 120.

b* value represents the blue-to-yellow spectrum, ranging from -120 to 120.

c.​ Microbial Growth (Total Yeast and Molds Count)

​ The Yeast and Mold Count (YMC) will be performed using the pour plate

method. Mango samples will be homogenized in deionized water, followed by serial

dilutions (10⁻¹ to 10⁻⁴) and plating on Dichloran Glycerol Agar (DG18). The plates

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 will be incubated at 25°C for 5 days, and the total colony-forming units (CFUs) will

be counted to evaluate microbial growth.

d.​ Titratable Acidity

The titratable acidity of the mango extract will be measured by diluting 10 mL

of the extract in 100 mL of boiled deionized water and adding four drops of pH

indicator. Titration will be carried out using 0.1 M NaOH until a persistent color

change indicates the endpoint. The volume of NaOH used will be recorded, and the

titratable acidity will be calculated based on the amount of titrant required to

neutralize the acids in the sample.

e.​ Anthracnose Mitigation

To evaluate anthracnose mitigation, infected mango peels will be cut into 2x2

cm pieces and cultured on potato dextrose agar (PDA) for 5 days. After incubation,

the fungal growth will be subcultured onto Plate Count Agar (PCA). A triangular

section on the plate will be marked, and three sterile paper discs will be placed within

this area. Then, 20 nanoliters of calcium carbonate-pectin solutions will be applied,

and the plates will be incubated at 25°C to observe the mitigation effects.

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2. RISKS AND SAFETY

In creating CARBONAID, it is essential to consider the potential risks and the safety

precautions or procedures that will protect the researcher. The most dangerous risks

associated with the research are skin irritation, chemical burns, and inhalation risks; here are

some precautions and procedures to avoid them (Occupational Safety and Health

Administration 2023; National Institute for Occupational Safety and Health 2020).

1.​ Wear proper PPE, including gloves, goggles, and lab coats, to avoid chemical

exposure.

2.​ Ensure good ventilation when working with HCl and acetic acid to avoid

inhaling fumes.

3.​ Store chemicals in clearly labeled, sealed containers and ensure proper

separation.

4.​ Be prepared for spills with neutralizers and know emergency first aid

procedures.

The Calcium Carbonate-based solution has the potential to benefit farmers and

distributors by improving mango preservation and extending shelf life. Therefore, awareness

of the risks and safety precautions associated with handling the chemicals and reagents

needed in the process is important to ensure its proper use.

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3. DATA ANALYSIS

The data gathered will be analyzed using the statistical software ‘Statistical Package

for Social Sciences’. The following tools will be utilized during the course of the study.

1.​ To compare the central tendency and variability across different treatments, the mean

and standard deviation of the weight loss will be calculated for each treatment group

using data collected from all observations over the 8-day periods.

2.​ A paired samples t-test will be employed to compare the initial and final browning

index and titratable acidity within each treatment group, determining whether

significant changes occurred over time.

3.​ One-Way Analysis of Variance (One-Way ANOVA) will be used to determine the

differences among treatments in weight loss and anthracnose mitigation of coating

solution.

4.​ Post-hoc analysis will be used to identify the specific variations between treatments.

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 D. BIBLIOGRAPHY

“Chemical Hazards and Toxic Substances - Overview | Occupational Safety and Health
Administration,” n.d.. https://www.osha.gov/chemical-hazards.

Gabriell Galang. “PH Mango, Avocado, Durian Exports Top P5 B in 2023,” Manila Bulletin.
January 25, 2024.
https://mb.com.ph/2024/1/24/ph-mango-avocado-durian-exports-top-p5-b-in-2023

"Mango – Industry Strategic Science and Technology Plans (ISPs) Platform." “Mango |
Philippine Statistics Authority | Republic of the Philippines,” April 1, 2023.
https://psa.gov.ph/major-fruit-crops/mango.

“Pocket Guide to Chemical Hazards | NIOSH | CDC,” February 8, 2020.

S&T Media Services. “Study Bares Big Postharvest Losses of PHL’s Mango, Onion, Tomato |
S&T Media Services.” BusinessMirror, March 25, 2022.
https://businessmirror.com.ph/2022/03/27/study-bares-big-postharvest-losses-of-phls-
mango-onion-tomato/.

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