1

INTRODUCTION

Background of the Study

The Naso species also known as Surahan is characterized with a strong fishy odor

and off-flavor primarily because it fed on algae. It is relative to the microalgae-based

foods on the market which often have a ‘grassy’ taste, intense green color and fishy odor

that many consumers consider unpleasant (Coleman, Van Poucke, Dewitte, Ruttens,

Moerdijk-Poortvliet, Latsos & Robbens (2022). This characteristic affects its market

value and demand wherein market price is relatively low in the Mercedes Fish Port.

Additionally, it is also the reason why most unicorn fish from by-catch were not landed in

the local fish port because fishermen tend to keep them for personal consumption while

some portions of their catch were sold to local fish vendors.

The researchers sought to eliminate fishy odor and off-flavor to add to its value

and raise its demand to the local market. Gonzales, Baleta & Bolaños, (2017), evaluated

the acceptability of calamansi leaves, guava leaves and lemon grass aqueous extracts as

off-odor and off-flavor remover and flavor enhancer to dried Parrot fish (Scarus

rivulatus) in terms of color, aroma, taste and texture and general acceptability. The

present study only used the calamansi leaves and guava leaves extract as neutralizing

agent to remove the fishy odor and off-flavor using Surahan. Parrotfish and Surahan are

characterized of strong fishy flavor and off odor primarily due to the same habitat and

same food being consumed throughout their lives. Gonzales et. al (2017) used dried

parrot fish and the researchers of the present study steamed the fish and both of them

steamed the leaves to facilitate extraction.

 2

Several studies were conducted on guava (Psidium guajava) and calamansi

(Citrus microcarpa) and it revealed that both possess antibacterial and antioxidant

property that may be applicable to neutralize or remove the fishy odor and off flavor of

unicorn fish (Naso spp.). Because of these, the researchers made use of the guava and

calamansi leaves extracts as a neutralizing agent where the subject fish will be marinated

under different marinating period to make sure that the extract will sooth thoroughly

removing the fishy odor and off flavor of short-nose unicorn fish.

The researcher used the extracts from guava and calamansi leaves as a

neutralizing agent to remove the fishy odor and off flavor of short-nose unicorn fish

(Naso spp.) which was evaluated by the respondents through organoleptic test.

Additionally, the fish was steamed to make sure that there are no factors that will affect

its quality attributes other than the Guava (Psidium guajava) and Calamansi (Citrus

microcarpa) leaves extract. Furthermore, in order to make the leaves soft for blending,

the researchers subjected the leaves to steaming and made use of it as neutralizing agent

to the unicorn fish to be prepared for the respondents.

Importance of the Study

The result of this study provided information on the removal of off-flavor and

fishy odor of short-nose unicorn fish (Naso spp.) using guava (Psidium guajava) and

calamansi (Citrus microcarpa) leaves extracts. It would be beneficial to Camarines Norte

State College, Government/agency, fish vendor, consumer, future researcher, and local

community. The result of this study would benefit the school for it will encourage the
 3

school administration through their extension services to provide the community

knowledge on how to neutralized the fishy odor and off flavor of Surahan. The

Government and other concerned agency may promulgate programs on the value addition

of Surahan with the help of this study. It would also increase the commercial value of

short-nose unicorn fish because with the help of this study the fish can be consumed

without fishy odor and off-flavor. Moreover, it would provide the consumer the

information on how to remove off-flavor and fishy odor using guava and calamansi

extract for them to enjoy fish consumption. Future researchers who wish to conduct

experimental research related to the topic may use the study which will serve as their

references. Lastly, the local community would generate the idea that the guava and

calamansi leaves which are abundant within their locality can be used to neutralize or

remove fishy odor and off-flavor of the Naso spp.

Objectives of the study

The general objective of this study is to utilize guava (Psidium guajava) and

calamansi (Citrus microcarpa) leaves extracts as a neutralizing agent to remove the fishy

odor and off flavor of short-nose unicorn fish (Naso spp). Specifically, it aimed to: 1)

evaluate the sensory acceptability of unicorn fish as to appearance, odor, taste, and

texture and the general acceptability in terms of marinating time, and 2) determine the

microbial count of the most acceptable treatment such as Escherichia coli, coliform

bacteria, aerobic bacterial count, yeast and molds.

Scope and Limitation

 4

This study focused on assessing the acceptability of short-nose unicorn (Naso

spp.) in terms of appearance, odor, taste and texture. The researchers utilized Guava

(Psidium guajava) and Calamansi (Citrus microcarpa) leaves extract as a neutralizing

agent to remove the fishy odor and off flavor of the subject fish. The organoleptic

characteristics of short-nose unicorn fish with Guava (Psidium guajava) and Calamansi

(Citrus microcarpa) leaves extract were determined in terms of appearance, taste, odor

and texture with the use of 7-point Hedonic scale wherein the product was evaluated by

thirty (30) respondents. Data gathered was analyzed using Analysis of Variance

(ANOVA).

This study was limited on the use extract from guava and calamansi leaves and

the extract from other parts of guava and calamansi were not included, excluding also the

use of other plants or herbs as a neutralizing agent to remove the fishy odor and off-flavor

of the fish. Other species of fish aside from short-nose unicorn (Naso spp.) was also not

included in this study. The extraction processed that were applied by the researchers were

manually and without the use of mechanical means. The phenolic compound extraction

was not also done by the researchers. The researchers conducted a preliminary study

before the final formulation of the four different treatments and identified the proportion

of the extract to water where the fish was marinated. The study was conducted at the wet

laboratory of Camarines Norte State College- Institute of Fisheries and Marine Sciences,

Barangay San Roque, Mercedes Camarines Norte on March 30, 2023.

 5

REVIEW OF RELATED LITERATURE

Contents and property of Guava and Calamansi leaves

In the study of Pongsak & Parichat (2010), it was found out that guava leaves

have four antibacterial flavonoids (morin-3-O-lyxoside, morin-3-O-arabinoside,

quercetin, and quercetin-3-Oarabinosi de) that were isolated from fresh and dried

Psidium guajava leaves and their concentrations were determined. Moreover, guava

leaves’ flavonoids were also shown to have bacteriostatic effects on fish pathogenic

bacteria including Aeromonas hydrophila, Aeromonas salmonicida subsp. salmonicida,

Flavobacterium columnare, Lactococcus garvieae, Streptococcus agalactiae and Vibrio

salmonicida (Rattanachaikunsopon and Phumkhachorn, 2007). The findings showed that

the flavonoid isolated from guava leaves (morin-3-O-lyxoside, morin-3-O-arabinoside,

quercetin and quercetin-3-Oarabinoside) might be potential biologically active

compounds for use as food preservatives to improve the shelf-life and the safety of foods.

Moreover, Olatunde, Della Tan, Shiekh, Benjakul, & Nirmal (2021) examined the

ethanol guava leaf extract with different chlorophyll removal process using sedimentation

process treating the pacific white shrimp. Antibacterial and anti-melanosis properties of

all extracts were examined. The quality changes were monitored compared to those

treated with 1.25% sodium metabisulphite and the control (without any treatment) during

12 days of storage at 4 °C. Changes in microbial and chemical qualities were lower in

shrimps treated with GLE-S solution as compared to others.

 6

Furthermore, guava (Psidium guajava) leaves have been traditionally used to

manage several diseases such as rheumatism, diarrhea, diabetes mellitus, and cough. In

this present investigation, antifungal and antibacterial property of guava leaves were

estimated using Bacillus subtilis (Gram positive bacterial strain), Escherichia coli (Gram

negative bacterial strain), Saccharomyces cerevisiae (Yeast, fungal strain) and

Aspergillus niger (Mould, fungal strain) strains. It was found that guava leaves are rich in

phenols, flavonoids and tannins whereas components like alkaloids, flavonoids, saponins

and triterpenes are present in comparatively lesser amounts. As polyphenols have strong

antimicrobial property, it can be concluded that rich source of phenols, flavonoids and

tannins are the probable cause of anti-microbial property of guava leaves (Das &

Goswami, 2019).

Based on the study of Gonzales, Baleta & Bolaños, (2017), the acceptability of

calamansi leaves, guava leaves and lemon grass aqueous extracts as off-odor and off-

flavor remover and flavor enhancer to dried Parrot fish (Scarus rivulatus) was evaluated

in terms of color, aroma, taste and texture and general acceptability. Dried parrot fish

treated with guava leaf extract is the most preferred and highly acceptable by the

panelists. Analysis of the sensory and organoleptic evaluation also showed that dried

parrot fish treated with either guava leaf, lemon grass and calamansi leaf extract is highly

acceptable based on the different criteria of evaluation. Similarly, the guava leaf extract

also masked the off-odor of dried Parrot fish.

Moreover, Ostonal, Arugay, Bassig, Iñigo, Mahmudah, Dichoson, & Cabajes,

(2021) investigated the effects of brine solution pre-treatment and addition of calamansi

juice extract on oyster mushroom kroepoek sensory and physicochemical (pH, color,
 7

moisture, water activity) properties. Results revealed that the pre-treatment and addition

of calamansi juice extract significantly affected. However, treatment 1 (5% brine solution

and 10 ml calamansi juice extract) was chosen as the most preferred since it had the

lowest rank among samples, hence was subjected to laboratory tests along with the

control. Physicochemical tests revealed a significant decrease in pH of both raw and

cooked indicating longer shelf stability.

Effects of other extracts to process fish

Guiñares, Agbon, Campeon & Belinario (2014) tested the effect of garlic extract

to smoked sardines. Their study revealed that smoked sardines treated with different

concentrations of garlic extracts (5 g, 10 g, and 15 g) generally had lower peroxide, free

fatty and acid values than the control or untreated smoked sardine samples. On the other

hand, smoked sardines treated with garlic extracts showed marked reductions in aerobic

plate count (cfu/g) during the 30 days of frozen storage (-18 0C). The result agreed with

the study conducted by Kumolu-Johnson and Ndimele (2011) when organoleptic

parameters was evaluated on the hot-smoked catfish which showed that the control

samples with no ginger extract received a lower panel score than the ginger paste-treated

samples.

Moreover, according to Bou, Claret, Stamatakis, Martínez, & Guerrero (2017),

the effect of adding citric acid directly or encapsulated on the quality and shelf-life of

ready-to-eat sea bass patties were observed. Microbial growth and total basic volatile

nitrogen were maintained at relatively low levels up to 8 weeks of storage with respect to

oxidative stability, the addition of encapsulated citric acid minimized secondary

 8

oxidation values more efficiently than its direct addition, regardless of the concentration.

This is in agreement with the decreased fishy odor observed in those patties containing

encapsulated citric acid. Accordingly, sensory analysis showed that the addition of

encapsulated citric acid at resulted in lower scores in fish aroma compared to that of the

control.

Neutralization process to reduce off flavor and off-odor of fish

According to Chakraborty, & Joseph (2015), crude Sardinella longiceps oil was

refined in different stages such as degumming, neutralization, bleaching, and

deodorization. Phosphoric acid (1%) was found to be an effective degumming reagent. It

demonstrated that unfavorable odor-causing components, particularly low molecular

weight carbonyl compounds, could successfully be removed by the refining process.

Moreover, Nilsuwan, Chantakun, Chotphruethipong, & Benjakul, (2021), uses hydrolysis

to produce fat-free Hydrolyze Collagen powder from salmon skin via optimization of

one-step hydrolysis using mixed proteases (papain and Alcalase) at different levels which

reveals that no fat content with lowest fishy odor intensity. The combined processes

including hydrolysis and defatting could therefore render hydrolyze collagen powder free

of fat and negligible fishy odor. While De Oliveira, Minozzo, Licodiedoff &

Waszczynskyj, (2016), deodorized tuna (Thunnus albacares) by-product oil obtained by

enzymatic hydrolysis.

Synthesis State-of-the Art

Pongsak & Parichat (2010) determined that guava leaves have four antibacterial

flavonoids while the present study uses guava leaves as an off-odor and off-flavor
 9

remover. Secondly, Olatunde et al. (2021) examined the effect of the ethanol guava leaf

extract in treating the Pacific white shrimp while the present study used guava leaf extract

under varying marinating period in unicorn fish (Naso sp.). Moreover, Gonzales et al.

(2017) tested the acceptability of calamansi leaves, guava leaves and lemon grass

aqueous extracts as off-odor and off-flavor remover and flavor enhancer to dried Parrot

fish (Scarus rivulatus) wherein they evaluated the subject in terms of color, aroma, taste

and texture and general acceptability which is similar with the present study.

In addition, Ostonal et al. (2021) investigated the effects of brine solution pre-

treatment and addition of calamansi juice extract on oyster mushroom kroepoek sensory

and physicochemical (pH, color, moisture, water activity) properties while the present

study used calamansi leaves extract to short-nose unicorn fish (Naso spp.) to determine

its acceptability. Furthermore, Guiñares et al. tested the effect of garlic extract to smoked

sardines and Kumolu-Johnson and Ndimele (2011) evaluated the organoleptic parameters

on the hot-smoked catfish using ginger extract which is the same with the present study

which determined the acceptability of the subject using the guava and calamansi leaves

extract as a neutralizing agent. Lastly, Chakraborty et. al (2015) used neutralizing process

such as degumming, neutralization, bleaching, and deodorization to remove the

unfavorable odor from crude oil derive from Sardinella longiceps. On the other hand,

Nilsuwan et. al (2021) and de Oliveira et. al (2016) used hydrolysis to remove fishy odor

while the present study used marination with the aid of leaf extracts to neutralize the off-

flavor and fishy odor

All of the abovementioned studies dealt with the content and property of guava

leaves and calamansi juice extracts as an additive to food. It also features the effects of
 10

other extract to smoked sardines and catfish and the different neutralizing process to

remove fishy odor respectively.

MATERIALS AND METHODS

Research Design

The researchers used the factorial design to assess the organoleptic characteristics

and microbial of Short-nose unicorn fish (Naso spp.) wherein researchers experimented

on the guava and calamansi leaves extract as a neutralizing agent to remove the fish odor

and off flavor under different marinating period. Four (4) different treatments including

the control treatment with three replicates each were applied in the study. This study used

panelists stationary style wherein the panelists where stationary in their designated post

and the sample were presented to them directly which were coded, in order to diminished

or avoid any mistake by the respondents in tasting proper.

Subject of the Study

The subject of the study is the short-nose unicorn fish (Naso spp.) marinated using

guava (Psidium guajava) and calamansi (Citrus microcarpa) leaves extracts in different

marination period. The acceptability of the subject fish in terms of appearance, odor,

taste, texture and its general acceptability was evaluated. The most acceptable treatment

was subjected to microbial analysis of aerobic plate count (APC), coliform count, yeast

and mold and Escherichia coli.

 11

Experimental Design

Four different treatments were used in this study namely, Treatment 0 ( T 0) which

is the unmarinated sample as the control treatment, Treatment 1 ( T 1) was marinated in

500 g of guava leaves extract with 500 ml of water, while Treatment 2 ( T 2) was

marinated in 500 g of calamansi leaves with 500 ml of water and Treatment 3 ( T 3) in 250

g of guava leaves and 250 g calamansi leaves with 500 ml of water. Each treatment was

applied with 500 ml of water except for the controlled treatment (T 0) (Gonzales et al.,

2017) which is 1:1 ratio. with the leaves for the purpose of boiling for 15 minutes to

soften the leaves for easy extraction. Each treatment has three replicates wherein

Replicate 1 represents one hour marinating period, two (2) hours marinating period for

Replicate 2, three (3) hours for Replicate 3 respectively.

Table 1. Composition of Samples per Treatment

Treatments Components Replicates (Marinating period)

1 hour 2 hours 3 hours

Treatment 0 Control (not marinated) T 0 R1 T 0 R2 T 0 R3

Treatment 1 500 g guava leaves extract T 1 R1 T 1 R2 T 1 R3

with 500 ml of water

Treatment 2 500 ml calamansi leaves T 2 R1 T 2 R2 T 2 R3

extract with 500 ml of

water

Treatment 3 250 ml guava leaves extract T 3 R1 T 3 R2 T 3 R3

and 250 ml calamansi

 12

leaves extract with 500 ml

of water

Procedure

Procurement of raw materials and other ingredients. The short-nose unicorn fish

(Naso spp.) was purchased at Mercedes Fish Port while some was purchased directly to

fishermen within the vicinity of the Municipality of Mercedes, Camarines Norte which

weighs on average from 300 g- 500g. On the other hand, the guava and calamansi leaves

was gathered within the locality.

Preparation of Materials. All materials were prepared such as basin used in

washing of the short-nose unicorn fish and blender for refining the leaves. The measuring

cup were utilized to measure the quantity of water used in the different treatment, plastic

container for marinating, knife for degutting the fish and steamer for cooking (Daniel et

al., 2019). Thermometer was also used in monitoring the fish temperature while

preparing different treatments and the aluminum foil for wrapping the fish in order to

minimized the damage to the fish skin while steaming.

Extracting. The leaves were collected within the backyard of the researcher and

the weight of each plant material was pre- determined according to the experimental

design. The leaves were steamed for 15 minutes, to soften the leaves and then the plant

materials were grinded using blender and were squeezed using a clean fine mesh net cloth

to remove the extract. Each plant extracts were prepared separately. Each leaves was

determined by preparing an equal proportion of plant material to ratio 500 grams plant

material: 500 ml of water, 1:1 by (Gonzales et al., 2017). The filtered solution was stored
 13

in the refrigerator until used. After the entire process, 923 ml of mixture of water and

leaves extract were produced.

Marinating of short-nose unicorn fish. The fish in Treatments 1, 2, and 3 were

marinated for 1 hour, 2 hours, and 3 hours respectively as its replicates using the prepared

proportion under room temperature to preserve the fish and maintain its freshness while

the Treatment 0 was not marinated.

Steaming. The fish was wrapped in an aluminum foil and steamed for 30 minutes

to avoid other factors that will affect the fish other than the guava and calamansi extract

and in order to prevent unwanted damages to the fish while steaming.

Data Collection

The data was collected through an organoleptic test which evaluated the four (4)

different treatments using 7-Point Hedonic Scale and determined the general acceptability

of the samples. According to Singh-Ackbarali & Maharaj (2014), the preferred number of

respondents in a sensory evaluation should be from twenty (20) to fifty (50) respondents.

In this study, the researchers selected thirty (30) respondents to evaluate the product.

Population and samples. Stratified convenient sampling was utilized in the study

to determine the respondents during the sensory evaluation. This sampling technique

helps in making smaller strata and has an equal representation in accordance to the

population of each category. The respondents or panelists were composed of IFMS

students. The number of panelists in each section was determined based on their overall

population. The formula below was followed.

Research Instrument and its Administration. The researchers used sensory

evaluation sheet with the used of 7-point hedonic Scale to test the acceptability of short-
 14

nose unicorn fish (Naso spp.) considering the different attributes such as appearance,

odor, taste, and texture. The steamed fish was evaluated by thirty (30) respondents in

panel stationary style wherein panelists were stationary on their designated post and the

samples were given to them directly. Prior to the evaluation, the respondents signed a

consent form for food allergies consideration. The respondents evaluated all of the

samples which were randomly placed in an individual container/plate. In addition, they

were required to gargle water after tasting every sample in order to remove the after taste

that may affect their taste perception of the preceding samples. The panelists chosen were

students of 3rd and 4th year students of IFMS because they have already backgrounds on

post-harvest practices of fish.

Hedonic Scale. A 7-point scale patterned after the hedonic scale was used as a

tool in making a descriptive evaluation of the quantitative data obtained from the survey

n−1
results with range of 0. 86 which was obtain using the formula; R= , to make the
n

range equally distributed in a 7-point Hedonic scale, determining the average response.

Parameters and Measuring Procedure. Hedonic testing gives an idea to

producers about the likes of the end user. The panelist will be composed of thirty (30)

respondents to test the general acceptability of the product in terms of appearance, odor,

taste and texture. This study used sensory evaluation coded samples which were assessed

by thirty (30) respondents (Singh-Ackbarali & Maharaj, 2014).

Arrangement of samples and coding. The samples evaluated by the respondents

were arranged in accordance with the experimental layout design (Figure 1). The samples

were given the following codes such as T 0 R1 = 371, T 0 R 2 = 412,T 0 R3 = 491 T 1 R1 = 512,

T 1 R2 = 606,T 1 R3 = 667, T 2 R1 = 753, T 2 R2 = 808, T 2 R3 = 837, T 3 R1 = 891, T 3 R2 = 948,

 15

T 3 R3 = 356 (Kortei, Odamtten, Obodai, Akonor, Wiafe-Kwagyan, Buckman, & Mills,

2020). The respondents were unaware where the samples fell unto the random codes

labeled in each sample. The samples are coded with arbitrary selected 3-digit numbers to

avoid bias and order randomly, it was not arranged by treatments but rather according to

the arrangement provided above. (Sharif, Butt, Sharif & Nasir, 2017).

Tools for Data Analysis

The data gathered was evaluated using the Two-Way Analysis of Variance

(ANOVA) without replication to determine whether there is a significant difference

among the treatments with the aid of Microsoft Excel. Average Weighted Mean (AWM)

was used to determine the best lot among the sample.

∑ f (w)
AWM=
N

Where:

X – Computed weighted mean

– Summation

f – Frequency

w – Average of response

N – Total number of respondents

 16

RESULTS AND DISCUSSION

The results of the organoleptic test on the acceptability of short-nose unicorn fish

(Naso spp.) in terms of appearance, odor, taste, texture and general acceptability with the

use of guava and calamansi leaves extract as a neutralizing agent for removing fish odor

and off flavor was presented in this chapter. This also discussed the result of the

microbial analysis conducted which determinined the microbial count of all of the

treatment such as E. coli, Coliform bacteria, Aerobic Plate count (APC), yeast and mold.

Appearance
6
5.46 5.38 5.36 5.37

0
0
Treatment 0 Treatment 1 Treatment 2 treatment 3

Figure 1. Graph of appearance of different treatments

 17

In order to determine the acceptability of quality attributes of all samples, the

researchers analyzed and gathered the average weighted mean (AWM) of each treatment

using Microsoft Excel (2010). In terms of appearance, appearance with the control having

the highest average weighted mean (AWM) of 5.46 while Treatment 1 has an AWM of

5.38 the rest of the treatments garnered an AWM of 5.37 which are all interpreted as like

moderately and Treatment 2 has AWM 5.26 which is interpreted as like slightly. The

extracts of calamansi and guava leaves affected the appearance of short-nose unicorn fish

is also in accordance with the result of the study of Gonzales, Baleta & Bolaños, (2017)

wherein the fish marinated in guava leaves and calamansi extract obtain the lowest

acceptability in terms of appearance because of dark discoloration it imparted. Unlike the

control treatment which was not marinated in guava and calamansi leaves extracts, the

samples from the three (3) treatments manifested discoloration due to the natural pigment

present in the leaves particularly the chlorophyll.

Odor
6

5.01 5 4.92
5

4.07
4

0
Treatment 0 Treatment 1 Treatment 2 Treatment 3
 18

Figure 2. Graph of odor of different treatments

In terms of odor, T0 and T1 had an AWM of 5.01 and 5.0 respectively which

were interpreted as like slightly. Treatment 2 that has been marinated in calamansi leaves

extract has the highest AWM of 4.92 with the same interpretation. Lastly, Treatment 3

which is marinated in the mixture of guava and calamansi leaves extract with an AWM of

4.07 was interpreted as neither like nor dislike. The result was supported by the study of

Gonzales, Baleta & Bolaños, (2017) wherein the fish that was marinated in calamansi

extract obtained the lowest acceptability in terms of odor while the fish that was

marinated with guava obtained the highest AWM next to the fish marinated in the lemon

grass extract. The treatment which was marinated in the mixture of calamansi leaves

extract obtained the lowest acceptability because the odor of the two leaves extract is not

compatible with each other which is characterized of indescribable odor while treatment

marinated in the calamansi obtain an acceptability score of 4.9 out of 5.0 due to its

citrucy and tangy smell. On the other hand, guava leaves extract is characterized with

fruity and sweet attributes and a distinct grapefruit-like aroma and the control rely only

on the natural fishy odor of the subjected fish which is highly acceptable for the

respondents.
 19

Taste
6
5.29 5.27
5.04 5.07
5

0
0
Treatment 0 Treatment 1 Treatment 2 Treatment 3

Figure 3. Graph of taste of different treatments

The organoleptic test in terms of taste showed that the control treatment has the

lowest AWM of 5.04, followed by Treatment 2 with 5.07 then by Treatment 3 with 5.27

which were all interpreted as like slightly and Treatment 1 with 5.29 was interpreted as

like moderately. The table revealed that the short-nose unicorn fish (Naso spp.) marinated

in guava leaves and calamansi leaves extract obtained the highest result compared to

other treatments. The result is in accordance to the study of Opeña, Baleta & Bolaños

(2017) wherein they used brine solution, calamansi extract, guava extract and lemon

grass extract as their treatments in which it revealed that the smoked surgeon fish

(Acanthurus sp.) marinated in guava leaves extract obtained the highest acceptability

weighted mean of 4.17 out of 5.0 as the highest score this is due to the tea-like taste of

the guava leaves extract (Linnea Covington, 2021).

 20

Texture
6
5.46 5.38 5.5

5
4.36

0
0
Treatment 0 Treatment 1 Treatment 2 Treatment 3

Figure 4. Graph of texture of different treatments

Results showed that in terms of texture, Treatment 0 or the unmarinated samples

had the lowest AWM of 4.36 which is interpreted as neither like or dislike; among other

treatments which gained 5.46, 5.38 and 5.5 respectively which are all interpreted as like

moderately. The study of Opeña, Baleta & Bolaños (2017) revealed that the smoked

surgeon fish (Acanthurus sp.) marinated in guava leaves extract obtained the highest

acceptability weighted mean of 4.05 out of 5.0 in terms of texture. Moreover, according

to Forde & De Graaf (2022), the effect of texture on food intake is mainly operational

through eating rate where harder, chunkier and more viscous texture result in lower

eating rate which refers to the amount of kcal consumed per minutes where harder,

chunkier and viscous food decrease appetite of the consumer resulting to lower eating

rate. The result of this study reveals that fish which were not marinated were juicy and
 21

those fish that were marinated using the guava and calamansi leaves extract were dry and

not flaky resulting to lower average weighted mean.

General Acceptability
6

0
Appearance Odor Taste Texture

Treatment 0 Treatment 1 Treatment 2 Treatment 3

Figure 5. Graph of different treatments in terms of appearance, odor, taste and

texture

The sensory evaluation determined the general acceptability of all treatments. As

shown in Figure 5, the most acceptable treatment is Treatment 1 which is the samples

marinated in guava leaves extract with an AWM of 5.28 followed by Treatment 2

samples marinated in calamansi leaves extract with an AWM of 5.19, Treatment 3

marinated in the mixture of guava and calamansi leaves extract with 5.11 AWM and

Treatment 0 as the control treatment with an AWM of 4.97. The result revealed that

short-nose unicorn fish marinated in guava leaves extract is the most acceptable in terms

of appearance, odor, taste and texture followed by the fish marinated in the mixture of

guava and calamansi leaves extract and then the treatment marinated in calamansi leaves

extract while the unmarinated fish was least acceptable based on the preference of the
 22

respondents. In the study of Opeña, Baleta & Bolaños 2017); Gonzales, Baleta &

Bolaños, 2017), the fish that were marinated in guava leaves gathered the highest general

acceptability compared to the other treatments.

Table 2. General Acceptability of Short-nose Unicorn Fish in terms of Marinating

Time

TREATMENTS WEIGHTED MEAN INTERPRETATION

Control 5.0 Like slightly

R1 5.3 Like slightly

R2 5.3 Like slightly

R3 5.2 Like Slightly

In terms of the general acceptability with regards to the marinating period of 1, 2

and 3 hours, all of the replicates including the control are interpreted as like slightly with

the weighted mean of 5.0 (control), 5.2 (R3) and 5.3 for R1 and R2. The results revealed

that the samples of short-nose unicorn fish marinated under 1 hour and 2 hours are the

most acceptable for the panelists.

Table 3. Data Analysis of different treatment using ANOVA single factor

Summary Count Sum Average Variance

T0 3 15.65 5.216667 0.018533

T1 3 15.92 5.306667 0.005233

T2 3 15.49 5.163333 0.003333

 23

T3 3 15.9 5.3 0.0073

1hr 4 21.02 5.255 0.024367

2hrs 4 20.89 5.2225 0.001692

3hrs 4 21.05 5.2625 0.009958

Source of SS Df MS F P-value F crit

Variatio

0.01428
Rows 0.042867 3 9 1.315265 0.353473 4.757063
0.00180
Columns 0.003617 2 8 0.166454 0.850441 5.143253
0.01086
Error
0.065183 6 4
Total 0.111667 11

The data analysis revealed that there is no significant difference among the

treatments marinated in different proportions of marinating ingredients and marinating

periods. This is signified by having a higher P-value which is 0.353473 and 0.850441

against the alpha of 0.05. Two-way ANOVA is used when there is a single independent

variable or factor, and the goal is to investigate if variations or different levels of that

factor have a measurable effect on a dependent variable. A higher P value than the alpha

means that no effect that was observed.

 24

Microbial Analysis of the Most Acceptable Treatment

Table 4. Microbial Composition of Treatment 1

Test parameter/s Results Regulatory limit

E. coli <3.0 *MPN/g 11

Yeast and Mold < 10 CFU/g <1000 CFU/g

Aerobic Plate Count < 10 CFU/g 5 x 105 CFU/g

Coliforms < 10 CFU/g 10 CFU/g

SOURCE: PNS/FDA 26:2010 and FDA circular no. 2013-010

The microbial composition of the most accepted treatment (Treatment 1) which

was marinated in guava leaves extract was presented in Table 9. Based on the result

obtained from the microbial analysis conducted by the Bureau of Fisheries and Aquatic

Resources Regional Office No. V (BFAR), the E. coli level of the sample was <3.0

*MPN/g against the regulatory limit of 11 MPN/g. If the result exceeded within the

prescribed regulatory limits it would result to food poisoning or other health risk.

Moreover, higher E. coli in the body would result to severe stomach pain, bloody

diarrhea and kidney failure (nhsinform.scot).

Table 4 showed results of the microbial analysis conducted by the Central Bicol

State University of Agriculture – Food Laboratory (CBSUA) for yeast, mold, Aerobic

Plate Count (APC) and coliforms of sample wherein all tests’ results were within the

acceptable limit for human consumption based from the regulatory limit set by the

Philippines National Standard (PNS) and Food and Drug Agency (FDA). On the other

hand, higher number of coliforms can cause upset stomach, vomiting, fever, or diarrhea
 25

(Michigan.gov). Counts of yeast and mold above the regulatory limits can trigger rhinitis

and asthma. In addition, exposure to fungi is associated with a number of other illnesses

including allergic bronchopulmonary mycoses, allergic fungal sinusitis and

hypersensitivity pneumonitis (ncbi.nlm.nih.gov). The results implied that the fish

subjected to microbial tests was fresh and the handling practices the researchers applied

was in accordance with the Good Manufacturing Practices to ensure food safety.

SUMMARY, CONCLUSION AND RECOMMENDATION

Summary

In terms of the organoleptic attributes and general acceptability, Treatment 1

marinated in 500 ml of guava leaves extracts was found to be the most acceptable with an

average weighted mean of 5.28 compared to Treatment 2 marinated in 500 ml of

calamansi leaves extract with 5. 16 AVW and Treatment 3 in a mixture of 250 ml guava

and 250 ml calamansi leaves had a 5.05 AVW. The least accepted treatment was the

unmarinated samples which gained an AVW of 4.97. Additionally, based on the

 26

evaluators’ responses, the samples marinated in one (1) and two (2) hours were the most

accepted. The ANOVA also showed no significant difference among the samples with p-

value of 0.614967. The organoleptic tests’ results were determined through the sensory

evaluation conducted using 7-Point Hedonic Scale where a total of thirty (30)

respondents evaluated the samples, thus interpreted as like slightly.

Furthermore, the result of the microbial analysis of the most accepted treatment

(Treatment 1) revealed that the microbial composition of the sample was below the

regulatory limits for E. coli, coliform count, APC and yeast and mold.

Conclusion

Obtained results suggests that guava leaves and calamansi extracts as neutralizing

agents helps in the elimination of off-flavor and fish odor in the steamed short-nosed

unicorn fish (Naso spp.) thus improved the sensory attributes of the product which were

more accepted by the panel of evaluators. The marination period also plays an important

role in neutralizing the odor and flavor of the fish wherein based on the respondents, the

fish marinated for about 1-2 hours was suggested to get the most preferred appearance,

aroma, taste and texture. Sensory evaluation on the quality attributes as well as

considering the marination period showed no significant difference at all. However,

considering the highest mean score of short-nosed unicorn fish marinated in different

amount of leaves extracts under different marinating time, hence, revealed that

marinating the fish with guava leaves extracts alone under 1-2 hours was the most

promising technique to neutralize its odor and flavor.

 27

Low levels of E. coli, coliform, APC and yeast and mold were determined based

form the microbial analyses conducted in which the study inferred that the subject’s

microbial composition is lower than the regulatory limits therefore ensuring the safety of

the consumers.

Recommendation

Although there was no significant difference between the samples, the researchers

still recommend the formulation for the most acceptable treatment in neutralizing the

fishy odor and off flavor of the short-nose unicorn fish. However, the formulation can be

also considered on other fish species that share the same qualities as the subject.

Additionally, the researchers advise marinating the fish for just an hour to give it the most

desired sensory qualities and increase its acceptability in consumer’s preferences.

LITERATURE CITED

Bou, R., Claret, A., Stamatakis, A., Martínez, B., & Guerrero, L. (2017). Quality changes
and shelf‐life extension of ready‐to‐eat fish patties by adding encapsulated citric
acid. Journal of the Science of Food and Agriculture, 97(15), 5352-5360.

Daniel, Jarabejo & Macaro (2019). Acceptability of shellfish as empanada filling.

Unpublished thesis. CNSC-IFMS.

Chakraborty, K., & Joseph, D. (2015). Production and characterization of refined oils
obtained from Indian oil sardine (Sardinella longiceps). Journal of Agricultural
and Food Chemistry, 63(3), 998-1009.
 28

Das, M., & Goswami, S. (2019). Antifungal and antibacterial property of guava (Psidium
guajava) leaf extract: Role of phytochemicals. Int. J. Health Sci. Res, 9(2), 39-45.

de Oliveira, D. A., Minozzo, M. G., Licodiedoff, S., & Waszczynskyj, N. (2016).

Physicochemical and sensory characterization of refined and deodorized tuna
(Thunnus albacares) by-product oil obtained by enzymatic hydrolysis. Food
Chemistry, 207, 187-194.

Gonzales, R. O., Baleta, F. N., & Bolaños, J. M., (2017). Acceptability of selected herbs
as off- odor remover and flavor enhancer for dried parrot fish Scarus rivulatus.

Guiñares, R. C., Agbon, M. C. E. B., Campeon, A. S. B., & Belinario, M. F. M. T.

(2014). Effects of Garlic Allium sativum Extract on Lipid Oxidation,
Microbiological, and Organoleptic Qualities in Hot-smoked Sardines, Sardinella
longiceps During Frozen Storage. International Journal of Fisheries and Aquatic
Studies, 1(6), 87-94.

Kortei, N. K., Odamtten, G. T., Obodai, M., Akonor, P. T., Wiafe-Kwagyan, M.,
Buckman, S., & Mills, S. W. N. O. (2020). Sensory evaluation, descriptive
textural analysis, and consumer acceptance profile of steamed gamma-irradiated
Pleurotus ostreatus (Ex. Fr.) Kummer kept in two different storage
packs. Scientific African, 8, e00328.

Kumolu-Johnson, C. A., & Ndimele, P. E. (2011). Anti-oxidative and anti-fungal effects

of fresh ginger (Zingiber officinale) treatment on the shelf life of hot-smoked
catfish (Clarias gariepinus, Burchell, 1822). Asian journal of Biological
sciences, 4(7), 532-539.

Lytou, A. E., Panagou, E. Z., & Nychas, G. J. E. (2017). Effect of different marinating
conditions on the evolution of spoilage microbiota and metabolomic profile of
chicken breast fillets. Food microbiology, 66, 141-149.

Nilsuwan, K., Chantakun, K., Chotphruethipong, L., & Benjakul, S. (2021). Development
of hydrolysis and defatting processes for production of lowered fishy odor
hydrolyzed collagen from fatty skin of sockeye salmon (Oncorhynchus nerka).
Foods, 10(10), 2257.

Olatunde, O. O., Benjakul, S., & Vongkamjan, K. (2018). Antioxidant and antibacterial
properties of guava leaf extracts as affected by solvents used for prior
dechlorophyllization. Journal of food biochemistry, 42(5), e12600.

Olatunde, O. O., Della Tan, S. L., Shiekh, K. A., Benjakul, S., & Nirmal, N. P. (2021).
Ethanolic guava leaf extracts with different chlorophyll removal processes:
 29

Anti-melanosis, antibacterial properties and the impact on qualities of

Pacific white shrimp during refrigerated storage. Food Chemistry, 341, 128251.

Omoba, O. S., & Azeez, H. I. (2016). Quality characteristics and overall acceptability of
steamed and boiled blue whiting fish (Micromesistius poutassou)
cracker. Nutrition & Food Science, 46(6), 857-870.

Opeña, A. M., Baleta, F. N., & Bolaños, J. M. (2017). Sensory, organoleptic, and
proximate composition of smoked surgeon fish Acanthurus sp. using selected
herbs as flavor enhancer. Int. J. Food Sci. Nutr, 2(1), 174-181.

Ostonal, J. M., Arugay, M. B., Bassig, B. T., Iñigo, H. B. R., Mahmudah, N. A.,
Dichoson, H. K. E., ... & Cabajes, A. T. S., (2021). Effects of Pre-treatment and
Addition of Calamansi Juice Extract on Oyster Mushroom Kropek Sensory and
Physicochemical Properties.

Pongsak, R., & Parichat, P. (2010). Contents and antibacterial activity of flavonoids
extracted from leaves of Psidium guajava. Journal of Medicinal Plants
Research, 4(5), 393-396.

Rattanachaikunsopon, P., & Phumkhachorn, P. (2007). Bacteriostatic effect of flavonoids

isolated from leaves of Psidium guajava on fish pathogens. Fitoterapia, 78(6),
434-436.

Sharif, M. K., Butt, M. S., Sharif, H. R., & Nasir, M. (2017). Sensory evaluation and
consumer acceptability. Handbook of food science and technology, 361-386.

Singh-Ackbarali, D., & Maharaj, R. (2014). Sensory evaluation as a tool in determining

acceptability of innovative products developed by undergraduate students in food
science and technology at the University of Trinidad and Tobago. Journal of
Curriculum and Teaching, 3(1), 10-27.

Tabassum, P., Khan, S. A. K. U., Siddiqua, M., & Sultana, S. (2018). Effect of guava leaf
and lemon extracts on postharvest quality and shelf life of banana cv. Sabri (Musa
sapientum L.): Effect of plant extract on banana shelf life. Journal of the
Bangladesh Agricultural University, 16(3), 337-342.

Topuz, O. K. (2016). Effects of marinating time, acetic acid and salt concentrations on
the quality of little tunny fish (Euthynnus alletteratus) fillet. Journal of Food
Processing and Preservation, 40(6), 1154-1163.
30
 31

APPENDICES

APPENDIX A

Republic of the Philippines

Camarines Norte State College,
Institute of Fisheries and Marine Sciences
San Roque, Mercedes, Camarines Norte

APPLICATION FOR ORAL DEFENSE

Name: Vianne M. Buenaobra & Mark M. Cereno

Degree: BS Fisheries BS Fisheries

 32

Title: GUAVA LEAVES AND CALAMANSI LEAVES EXTRACT AS

NEUTRALIZING AGENT FOR REMOVING FISH ODOR AND OFF FLAVOR
OF SHORT-NOSED UNICORN FISH (Naso spp).

Date of Final Defense: May 2023

Time:

Member of the Evaluation Advisory Committee

Chairperson: JOHN CHRISTOPHER C. NOLIAL _________ __________

Printed Name Signature Date

Member 1. ANNE MARIE L. CARREON _________ __________

Printed Name Signature Date

Member2. ALIN D. QUIBRAL _________ __________

Printed Name Signature Date

APPENDIX B
Republic of the Philippines
CAMARINES NORTE STATE COLLEGE
Institute of Fisheries and Marine Sciences
San Roque, Mercedes, Camarines Norte

APPLICATION FOR ORAL DEFENSE

Date:

MANUEL B. ALBERTO, PH.D.

Dean/Campus Director

Sir:
 33

We respectfully request for paper presentation of our thesis manuscript titled:

“GUAVA LEAVES AND CALAMANSI LEAVES EXTRACT AS
NEUTRALIZING AGENT FOR REMOVING FISH ODOR AND OFF FLAVOR
OF SHORT-NOSED UNICORN FISH (Naso spp)” in Camarines Norte State College
Institute of Fisheries and Marine Sciences. Hoping for your consideration and approval
on this matter.
Thank you very much.

VIANNE M. BUENAOBRA MARK M. CERENO

BS Fisheries BS Fisheries

Favorably Endorsed

JOHN CHRISTOPHER C. NOLIAL

Chairperson

ALIN D. QUIBRAL
Researcher Coordinator
For Dean’s Office Use
Submitted the thesis manuscript
Seminar date applied for is the prescribed time to allow the review of Manuscript
by Examining Committee/Panel members.

MANUEL B. ALBERTO, Ph.D.

Campus Director

APPENDIX C

Republic of the Philippines

Camarines Norte State College,
Institute of Fisheries and Marine Sciences
San Roque, Mercedes, Camarines Norte

PERMISSION TO CONDUCT STUDY AND TO USE COLLEGE FACILITIES

FOR THE CONDUCT OF UNDERGRADUATE THESIS

March 29, 2023

MANUEL B. ALBERTO, Ph.D.

Dean/Campus Director
Sir:
 34

We have the honor to request permission from your office to conduct study titled
“GUAVA LEAVES AND CALAMANSI LEAVES EXTRACT AS
NEUTRALIZING AGENT FOR REMOVING FISHY ODOR AND OFF FLAVOR
OF SHORT-NOSE UNICORN FISH (Naso spp.)”.
Laboratory facilities and materials to be used/borrowed/rented for the experiment
are blender, measuring cup and thermometer.
Duration of the Study: March 30, 2023.
Thank you very much.
Very respectfully yours,

VIANNE M. BUENAOBRA
Student Applicant

MARK M. CERENO
Student Applicant
Recommending Approval:

MONICALYN V. BORLAGDAN
Adviser

Approved:
MANUEL B. ALBERTO, Ph.D.
Dean/Campus Director
APPENDIX D

Republic of the Philippines

Camarines Norte State College,
Institute of Fisheries and Marine Sciences
San Roque, Mercedes, Camarines Norte

CERTIFICATE OF THE SECRETARY

This is to certify that all suggestions given by the panel of Examiners in

connection with the Oral Examination Vianne M. Buenaobra and Mark M. Cereno in the
 35

defense of their thesis “GUAVA LEAVES AND CALAMANSI LEAVES EXTRACT

AS NEUTRALIZING AGENT FOR REMOVING FISHY ODOR AND OFF

FLAVOR OF SHORT-NOSE UNICORN FISH (Naso spp.)”.

were complied with.

SHIRLEY A. ALBERTO
Secretary
Oral Examination Committee

APPENDIX E

Republic of the Philippines

Camarines Norte State College,
Institute of Fisheries and Marine Sciences
San Roque, Mercedes, Camarines Norte

CERTIFICATION OF THE EDITOR

This is to certify that the thesis of Vianne M. Buenaobra and Mark M. Cereno

titled “GUAVA LEAVES AND CALAMANSI LEAVES EXTRACT AS

 36

NEUTRALIZING AGENT FOR REMOVING FISHY ODOR AND OFF FLAVOR

OF SHORT-NOSE UNICORN FISH (Naso spp.)”was edited by the undersigned.

RONALD O. DE LEMIOS, MAEd

Editor

APPENDIX F

Republic of the Philippines

Camarines Norte State College,
Institute of Fisheries and Marine Sciences
San Roque, Mercedes, Camarines Norte

CERTIFICATION OF THE AUTHOR

This is to certify that the researcher study by the undersigned titled “GUAVA

LEAVES AND CALAMANSI LEAVES EXTRACT AS NEUTRALIZING AGENT

 37

FOR REMOVING FISHY ODOR AND OFF FLAVOR OF SHORT-NOSE

UNICORN FISH (Naso spp.)”is original and not yet submitted to any publishers for

consideration. This is to certify that works, pictures, figures, and texts used from other

sources are properly acknowledged. Given this 2023 in Camarines Norte State College

Institute of Fisheries and Marine Sciences, San Roque Mercedes, Camarines Norte.

VIANNE M. BUENAOBRA MARK M. CERENO

Researchers

APPENDIX G

Republic of the Philippines

Camarines Norte State College,
Institute of Fisheries and Marine Sciences
San Roque, Mercedes, Camarines Norte

REQUEST FOR A THESIS ADVISER

March 29, 2023

MONICALYN V. BORLAGDAN
Instructor I

Madam:
 38

We, the undersigned BS Fisheries IV students, are the proponents of the thesis titled
“GUAVA LEAVES AND CALAMANSI LEAVES EXTRACT AS
NEUTRALIZING AGENT FOR REMOVING FISH ODOR AND OFF FLAVOR
OF SHORT-NOSE UNICORN FISH (Naso spp.)”. We would like to request your
assistance as adviser of our research study.

We are confident that your experience and expertise will help us complete our study on
time. We look forward to your favorable consideration and approval. Thank you and God
bless!

Very respectfully yours,

MARK M. CERENO

VIANNE M. BUENAOBRA

Noted:

SHIRLEY A. ALBERTO, MAEd

Thesis Writing Professor

Conforme:

MONICALYN V. BORLAGDAN
Instructor I
APPENDIX H

CURRICULUM VITAE

Personal Data

Name: Vianne M. Buenaobra

Nickname: “Amaya”

Age: 23

Date of Birth: November 6.1999

Place of Birth Purok-8, Barangay 8, Alawihao, Daet, Camarines Norte

Civil status: Single

Address: Aurora Street, Purok-5, Barangay 5, Daet, Camarines Norte

 39

Father’s Name: Victor O. Buenaobra

Mother’s Name: Marry Ann M. Buenaobra

Educational Background

Elementary: Urban Poor Alawihao Resettlement Elementary School (UPARES)

Barangay Alawihao, Daet, Camarines Norte
2010-2011
Secondary: Our Lady of Lourdes College Foundation (OLLCF)
Vinzons Ave, Daet, 4600 Camarines Norte
2018-2019
Tertiary: Camarines Norte State College (CNSC)
Institute of Fisheries and Marines Sciences
Mercedes, Camarines Norte
2022-2023
Course: Bachelor of Sciences in Fisheries
2022-2023

CURRICULUM VITAE
Personal Data

Name: Mark M. Cereno

Nickname: “Mark”

Age: 23

Date of Birth: April 4, 2000

Place of Birth Barangay Mancruz, Daet, Camarines Norte

Civil status: Single

Address: Purok-1, Barangay San Roque, Mercedes, Camarines Norte

Father’s Name: Rodel P. Cereno

Mother’s Name: Maricel M. Cereno

 40

Educational Background

Elementary: San Roque Elementary School

Barangay San Roque, Mercedes, Camarines Norte
2011-2012

Secondary: San Roque National High School

Barangay San Roque, Mercedes, Camarines Norte
2016-2017

Tertiary: Camarines Norte State College (CNSC)

Institute of Fisheries and Marines Sciences
Mercedes, Camarines Norte
2022-2023

Course: Bachelor of Sciences in Fisheries

2022-2023

APPENDIX I

Interpretation of average response

Value Average Weighted Mean Interpretation

7 6.3-7.0 Like very much

6 5.4-6.2 Like moderately

5 4.5-5.3 Like slightly

4 3.6-4.4 Neither like nor dislike

3 2.7-3.5 Dislike slightly

2 1.8-2.6 Dislike moderately

1 1.0-1.8 Dislike very much

 41

DESCRIPTIVE PROFILING
Score Organoleptic Quality
Appearance Odor Taste Texture General
(color) (odor) (Flavor) (flakiness) Acceptability
7 The fish flesh There is no fishy There is no The fish meat is The attributes
color is bright odor and smell of fishy flavor and very flaky and are very
white guava and taste of guava moist acceptable
calamansi extract and calamansi
extract

6 The fish flesh There’s a slightly There’s a The fish meat is The attributes
color is smell of guava slightly taste of moderately are moderately
moderately bright and calamansi guava and flaky and moist acceptable
white extract calamansi
extract
5 The fish flesh There’s a There’s a The fish meat is The attributes
color is slightly moderately smell moderately slightly flaky are slightly
bright white of fishy odor and taste of guava and moist acceptable
smell of guava and calamansi
and calamansi extract
extract
 42

4 The fish flesh is The fishy odor The fishy flavor The fish meat is The attributes
no longer bright and smell of and smell of flaky but are either
white but has a bit guava and guava and slightly dry acceptable/not
of freshness calamansi extract calamansi acceptable
is slightly strong extract is
moderately
strong

3 The fish flesh The fishy odor The fishy flavor The fish meat is The attributes
color is slightly and smell of and smell of flaky but are slightly not
fresh white calamansi extract guava and moderately acceptable
is moderately calamansi
strong extract is strong

2 The fish flesh The fishy odor The fishy flavor The fish meat is The attributes
color is and smell of and smell of flaky and dry are moderately
moderately fresh calamansi extract guava and not acceptable
white is strong calamansi
extract is very
strong

1 The fish flesh The fishy odor The fishy flavor The fish meat is The attributes
color is fresh and smell of and smell of not flaky and are very not
white guava and guava and very dry acceptable
calamansi is very calamansi
strong extract is
extremely
strong
Source: Nurhamidah, N., & Dara, W. (2022). Analysis of the Nutritional value of Puyu fish (Anabas testudineus) and
Acceptability of the Processing. Sainstek: Jurnal Sains dan Teknologi, 14(1), 46-50.

APPENDIX J

Coding System

T0 Replicate Product code

1 371

2 412

3 491

T1

1 512
 43

2 606

3 667

T2

1 753

2 808

3 837

T3

1 891

2 948

3 356

APPENDIX K

RESEARCH INSTRUMENT
SENSORY EVALUATION SHEET
Research Title: GUAVA LEAVES AND CALAMANSI LEAVES EXTRACT AS NEUTRALIZING
AGENT FOR REMOVING FISHY ODOR AND OFF FLAVOR OF SHORT-NOSE UNICORN FISH
(Naso spp.)

Proponents: Mark M. Cereno & Vianne M. Buenaobra

Name of evaluator:_____________________________Age:_______Gender:____________

Direction: Check (/) the number of your rating in which 7 is the highest and 1 is the lowest in terms of
Appearance, odor, taste and texture. Drink water in every sample testing to remove the after taste of every
sample. Each score has its interpretation as follows; 7 - Like very much, 6 - Like moderately 5 - Like
slightly, 4 - Neither like nor dislike, 3 - Dislike slightly, 2 - Dislike moderately and 1 - Dislike very much.
 44

Sam Apperance Odor Taste Texture

ple
1 2 3 4 5 6 7 1 2 3 4 5 6 7 1 2 3 4 5 6 7 1 2 3 4 5 6 7
371

412

491

512

606

667

753

808

837

891

948

356

Note: the general acceptability is being determined by the average of the responses in terms of appearance, odor, taste and texture

Comments and Recommendation:

______________________________________________________________________________________

_______________________
Signature of the respondent

CONSENT FORM

Research Title: GUAVA LEAVES AND CALAMANSI LEAVES EXTRACT AS

NEUTRALIZING AGENT FOR REMOVING FISHY ODOR AND OFF FLAVOR OF
SHORT-NOSE UNICORN FISH (Naso spp.)

Proponents: Mark M. Cereno & Vianne M. Benaobra

Name of
evaluator:_____________________________Age:_______Gender:____________

Address:________________________________________________________________

Check the box below:

QUESTION YES NO
 45

1. I confirm that I have read and understand the consent form

date_____________ for the above study and have the
opportunity to ask question.
2. I understand that my participation is voluntary that I am
free to withdraw at any time for any reason.
3. I am aware that the researchers have received the proper
training on food safety and have prepared the food using
hygienic practices.
4. I confirm that I don’t have any food allergies relating to
short-nose unicorn fish or any seafood or sensitive or under
medication which restrict free food choice and I am fit to
participate in this study.
5. I agree to make to take part in the above study and my
participation will provide important information to the
researcher of Camarines State College-Institute of Fisheries
and Marine Sciences on Guava and calamansi leaves
extract as neutralizing agent for removing fishy odor and
off flavor of short-nose unicorn fish (naso spp.)

_______________________
Signature of the respondents

APPENDIX L

Statistical Analysis
 46

CODE Appearance Total AWM

Score 1 2 3 4 5 6 7
371 1 4 10 8 7 156 5.2
412 1 5 10 7 7 164 5.47
491 1 3 3 7 9 7 171 5.7
total 1 5 12 27 24 21
PRODUC
T 2 15 48 135 144 147 491 5.46
512 2 4 10 6 8 165 5.5
606 1 6 7 9 7 155 5.17
667 3 7 8 6 6 164 5.47
total 6 17 25 21 21
PRODUC
T 18 68 125 126 147 484 5.38
753 1 1 9 5 6 8 158
808 1 2 7 5 6 9 160
837 5 6 6 4 9 156
total 2 8 22 16 16 26
PRODUC
T 4 24 88 80 96 182 474 5.26
891 1 2 2 7 10 8 167
948 2 0 8 6 7 7 157
356 2 2 3 11 2 10 159
total 5 4 13 24 19 25
PRODUC
T 10 12 52 120 114 175 483 5.37
APPENDIX L

Statistical Analysis

CODE Odor Total AWM

Score 1 2 3 4 5 6 7
371 1 4 6 9 5 5 148
412 5 4 10 6 4 145
491 1 2 5 9 6 7 158
total 2 11 15 28 17 16
PRODUCT 4 33 60 140 102 112 451 5.01
512 5 4 9 8 4 152
606 1 6 4 8 5 6 148
667 3 2 5 8 6 6 150
total 4 13 13 25 19 16
PRODUCT 8 39 52 125 114 112 450 5
753 1 5 6 8 5 5 146
 47

808 2 3 3 9 7 6 154
837 2 4 6 10 3 5 143
total 5 12 15 27 15 16
PRODUCT 10 36 60 135 90 112 443 4.92
891 1 6 2 8 6 7 153
948 5 6 6 7 6 153
356 1 4 4 10 7 4 150
total 2 15 12 24 20 17
PRODUCT 4 45 48 120 120 119 456 5.07

APPENDIX L

Statistical Analysis

CODE Taste total AWM

Score 1 2 3 4 5 6 7
371 1 0 7 3 7 7 5 146
412 1 2 8 5 9 5 154
491 1 1 3 4 7 7 7 154
total 2 2 12 15 19 23 17
PRODUCT 2 4 36 60 95 138 119 454 5.04
512 1 4 4 3 10 8 161
606 1 5 4 5 5 10 158
667 1 5 4 5 6 9 157
total 3 14 12 13 21 27
PRODUCT 6 42 48 65 126 189 476 5.29
 48

753 3 3 2 9 7 6 152
808 1 0 5 5 4 9 6 152
837 2 4 5 6 5 8 152
total 1 5 12 12 19 21 20
PRODUCT 1 10 36 48 95 126 140 456 5.07
891 1 1 2 6 5 7 8 156
948 4 8 4 6 8 156
356 5 3 7 5 10 162
total 1 1 11 17 16 18 26
PRODUCT 1 2 33 68 80 108 182 474 5.27

APPENDIX L

Statistical Analysis

CODE Texture Total AWM

Score 1 2 3 4 5 6 7
371 0 0 2 5 8 8 7 163
412 3 5 9 8 5 157
491 1 6 7 12 4 162
total 6 16 24 28 16
PRODUC
T 18 64 120 168 112 482 4.36
512 2 3 9 6 10 169
606 3 3 9 9 6 162
667 1 1 4 11 7 6 160
total 1 6 10 29 22 22
PRODUC 2 18 40 145 132 154 491 5.46
 49

T
753 2 6 10 5 7 159
808 3 4 8 8 7 162
837 4 3 8 5 10 164
total 9 13 26 18 24
PRODUC
T 27 52 130 108 168 485 5.38
891 2 5 4 8 11 171
948 5 6 2 8 9 160
356 2 3 10 9 6 164
total 9 14 16 25 26
PRODUC
T 27 56 80 150 182 495 5.5

APPENDIX M

RESULT OF MICROBIAL ANALYSIS

 50

APPENDIX M

RESULT OF MICROBIAL ANALYSIS

 51

APPENDIX N

DOCUMENTATION
 52

Raw Materials

Treatments with their replicates

 53

Blending of leaves and straining

Marinating of short-nose unicorn fish

 54

Steamed fish for sensory evaluation

Sensory Evaluation
55