NITROGEN – FREE EXTRACT DIGESTIBILITY OF COMBINATION

OF SILAGE , FORMULATED CONCENTRATE AND UREA

MOLASSES MINERAL BLOCK FED TO RAMBON CATTLE

CURRENCY DUTY

RESEARCH METHODS

PAVITHRA SANTHARA MOGAN

NIM : O61511133285

VETERINARY MEDICINE STUDY PROGRAM

FACULTY OF VETERINARY MEDICINE

AIRLANGGA UNIVERSITY

SURABAYA

2017
NITROGEN – FREE EXTRACT DIGESTIBILITY OF COMBINATION
OF SILAGE , FORMULATED CONCENTRATE AND UREA
MOLASSES MINERAL BLOCK FED TO RAMBON CATTLE

PAVITHRA SANTHARA MOGAN

NIM : O61511133285

VETERINARY MEDICINE STUDY PROGRAM

FACULTY OF VETERINARY MEDICINE

AIRLANGGA UNIVERSITY

SURABAYA

2017
 PREFACE

Praise to the presence of God who has given His grace and guidance, so that the
writer can complete the preparation of research proposals with the title Nitrogen – free extract
digestibility of combination of silage , formulated concentrate and urea molasses mineral
block fed to rambon cattle as a task of Research Methods course.

The task of preparing the proposal is the first step as a student to be able to complete
the final task in the preparation of the thesis because the proposal is a draft of activities that
contain steps that will be conducted activities by researchers and students in doing research.
If the draft of the written procedure is in accordance with the provisions of the researcher in
this case the student can conduct research activities correctly so that the results achieved in
accordance with the expected.

A researcher in this case a student in doing research activities need to get guidance
from supervisors or professionals who are experts in the field because of the ability and
knowledge of a student is still limited in addition to minimal experience. Therefore if the
research proposal that the author of this stack is approved, then the great hope of the author to
get guidance and direction from lecturers at the Faculty of Veterinary Medicine Airlangga
University Surabaya.

In preparing this research proposal, the writer realizes that there are still many
shortcomings and mistakes so the result is still far from perfect. Therefore, for the sake of
perfection of this proposal the writer expect criticism and suggestions that can broaden our
knowledge in preparing better proposals.

Finally, we would like to thank all parties and hopefully this research
proposal is useful for the readers, especially for students who take the course of Research
Method.

Surabaya, May 21, 2017

Writer

i|Page
 CONTENT

Preface………………………………………………………………………………… I
Content……………………………………………………………………………….. II
Chapter I Introduction
1.1 Background Research………………………………………………………… 1
1.2 Identification of the Problem……………………………………………….. 2
1.3 Theoretical Framework……………………………………………………... 2
1.4 The Aims of the Research………………………………………………..… 4
1.5 Outcome of Research……………………………………………………….. 4
1.6 Research Hypothesis……………………………………………………….. 4
Chapter II Literature Review
2.1 Indonesian Native Cattle Rambon…..…………………………………….. 5
2.2 Digestion in Ruminants……..……………………………………………. 6
2.3 Feed………….……………………………………………………………. 9
2.3.1Silage……..…………………………………………………………… 9
2.3.2Formulated Concentrate………………………………………………. 9
2.3.3Urea Molasses Mineral Block ………………………………………… 9
2.4 Carbohydrate Requirements……………………………………………….. 10
2.5 Digestibility of Feed……………………………………………………… 10
2.6 Nitrogen – Free Extract……………………………………………………. 11
Chapter III Methods and Materials
3.1 Research Location and Time……………..……………………………….. 14
3.2 Research Material and Equipment……………………………………….… 14
3.2.1Experimental Animals…………………………………………………. 14
3.2.2Research Materials……………………………………………………… 14
3.2.3Research Equipment…………………………………………………… 14
3.3 Methods…………………………………………………………………… 15
3.3.1Adaptation Phase and Feeding Trial…………………………………… 15
3.3.2Collection of Sample………………………………………………... 16
3.4 Experimental Design………………………………………………………. 16
3.5 Research Variable …….…………………………………………………… 16
3.5.1Independent variable……………………………………………………
16

ii | P a g e
 3.5.2Dependent variable………………………………………………………….. 16
3.5.3Controlled variable………………………………………………………… 16
3.6 Calculation of Digestibility Value……………………………………………
17
3.7 Data Analysis………………………………………………………………… 17
Reference……………………………………………………………………………….

iii | P a g e
 CHAPTER I

INTRODUCTION

1. 1 Background
The rapid development of science and technology has brought
tremendous benefits to the progress of human civilization. Types of work
that previously required considerable physical capability, now relatively can
be replaced by automatic machine tools. The robotic work system has turned
the power of human muscle with amazing acceleration. Likewise with the
discovery of new formulations of various computer capacities, as if already
able to shift the position of human brain capabilities in various fields of
science and human activity. In short, the progress of science and technology
that has been achieved now really has been recognized and felt to provide
many benefits, convenience and comfort for human life.
Science and technology for modern society is a religion. The
development of science and technology is considered as the solution of the
existing problems. While humans even worship science and technology as a
liberator who will free them from the confines of the world's fineness.
Science and technology are believed to bring human benefits to healthy
living, happiness and immortality. The contribution of science and
technology to civilization and human welfare can not be denied, but human
beings can not deny that science and technology can also bring misfortune
and misery to human life when man is wrong in using it.
The development of science and technology in the field of
information undergoes such rapid changes that any information every second
can be obtained for anyone who needs, not to mention information in the
field of food. Food fields that dikmasud is processed products and basic food
ingredients that will be processed further. The information there is a positive
but some are negative.
Positive information, such as crop cultivation, pet care and others,
while negative information include food preservation using chemicals such

1|Page
as formalin, borax and so on. All information is easily obtained by anyone
just how the use of information addressing existing information.
If we observe and follow the development of the use of chemicals to
preserve food ingredients done by food producers, then we need to remind
all interested parties from government, distributors, merchants even to
consumers that kima materials such as formalin, borax and rhodamin B are
very dangerous and deadly if consumed in a certain amount. Therefore, all
parties should not be tired to always provide counseling to all the people so
that consumers do not become parties who have to bear the consequences of
the act of rogue producers.
On the one hand, people are in need of better food products for the
present and future, ie food that is safe (does not contain ingredients that are
harmful to health), quality (has good quality in terms of color, texture and
aroma) And nutritious (containing various substances needed by the body) to
be consumed. In order for the public to meet the needs of food that is free
from damage and contamination, whether contamination of toxins, microbes,
chemical compounds and other harmful substances, food security is an
important factor to be considered and applied in food processing.
According to RI law No. 18 of 2012 on food, the third section
concerning the Regulation of Food Added Materials, article 75 states, that
any person committing food production for distribution shall be prohibited
from using food additives that exceed the set maximum threshold and / or
prohibited substances Used as food additives. However, in reality, food
traded by the community, especially processed food, is often found to
contain harmful additional substances that violate food safety criteria. Some
harmful additives that are not intended for food products are added to food
such as formaldehyde, borax, rhodamine B and methanyl yellow.
Formalin is a kind of harmful additive that is still often used freely by
irresponsible food traders or processors. This is because formalin has several
advantages, including:
1. cheaper than other preservatives
2. easy to use because in the form of solution

2|Page
 3. easy to obtain because it is traded freely
In addition, the low knowledge of traders about the dangers of formalin for
health (Widyaningsih and Murtini, 2006).
The use of hazardous chemicals such as formalin, borax, rhodamin B
or other chemicals in foodstuffs is usually done to improve the color and
texture and aroma of food and inhibit the activity of microorganisms so that
food products can be stored longer.
Traders in traditional markets and modern markets are suspected of
using hazardous materials such as formalin, borax, rhodamin B and others to
inhibit the decay process by microorganisms in foodstuffs such as shrimp,
squid, chicken, fish and some processed products such as meatballs, tofu ,
Nugget and others. All this is done so that traders do not lose when their
merchandise is not exhausted at that time, but they less realize impact will
be accepted by consumers who have repeatedly consumed foods containing
harmful substances. Various diseases have been waiting for those who
continuously consume food contaminated with harmful substances.

1.2 Problems
Foodstuffs that are traded in both modern and traditional markets are
not all safe, quality and nutritious. This is because the act of naughty traders
who do not want to lose so they use harmful materials to mengawewtkan these
foods.
Because of the act of the naughty traders like the above exposure then
the problems that appear to harm consumers. Issues arising include (1) how to
identify the physical characteristics of shrimp, squid, fish, chicken and other
beasts that are suspected to contain formaldehyde or not containing formalin;
(2) how to use laboratory qualitative analysis methods to determine shrimp,
squid, fish, chicken and other butchered or non-traded bears traded on the
market; (3) to determine the formalin content of shrimp samples, squid, fish,
chicken and other animals that showed positive results.

3|Page
1.3 Limitation Problems
Problems are so many and complex in the current study, the authors
limit with three problems, namely (1) how to identify the physical
characteristics of shrimp suspected to contain formalin and does not contain
formalin; (2) how to use laboratory qualitative methods for determining
shrimp containing formalin or not being traded on the market; (3) to determine
the formalin content of shrimp samples that showed positive results.

1.4 Problem Formulation

As has been the authors described above that the problems are so many
and complex authors have restricted to a particular object of shrimp, therefore
the author can formulate the problem becomes as follows:

1. How to identify the physical characteristics of shrimp that are suspected to

contain formalin and do not contain formalin?

2. Does the shrimp sold in the Modern Market Surabaya identified through
qualitative analysis laboratory containing formalin?

3. What is the formalin content of shrimp samples showing positive results in

qualitative analysis identified through laboratory quantitative analysis?

1.5 Research Objectives

1.5.1 General Objectives

To identify formaldehyde in shrimp are traded in the modern market in

Surabaya

1.5.2 Specific Objectives

1. To identify the physical characteristics of shrimp that are sold in

modern markets at Surabaya that is suspected to contain formalin
and does not contain formalin

2. To identify formalin on shrimp sold in the Modern Market in

Surabaya with laboratory identification through analysis qualitative.

4|Page
 3. To analyze the levels of formalin on shrimp samples that show
Positive results on qualitative analysis through laboratory
quantitative analysis.

1.6 Research Benefits

1.6.1 The Benefits of Theory

As a literature material in the development of further research

in the field of Veterinary Medicine on Veterinary Public Health.

1.6.2 Application Benefits

1. To assist government and authorized institutions in the effort to

improve the supervision of food safety, especially food of animal
origin consumed by the community.

2. Providing information for the public to understand the community

about food safety especially food of animal origin is getting better
so that society can choose food like shrimp which is safe for health
and quality life.

1.7 Research Hypothesis

Through the observation of physical characteristics, qualitative

analysis and quantitative analysis in the laboratory, the shrimp samples taken
from Modern Market in Surabaya contain formalin with varying levels of
formalin or concentration.

5|Page
 CHAPTER II

REVIEW OF RELATED LITERATURE

2.1 Hygienic Foods

Processed products and basic food ingredients such as chicken pieces,

shrimp, squid, fish and others are the food that humans need in their survival.
If humans are wrong or not careful in consuming slow food but surely
humans will be gnawed by various. Disease. Therefore food is a very
important aspect in everyday human life.

Humans in life together will form a community called society. And

this society in fulfilling all its needs depend on each other because they are
not able to meet all their needs with their own ability so that they need each
other. Because they need each other to create a variety of jobs that exist in
society, one of which is a trader.

Traders in the community sell a variety of needs required by the

community itself, one of which sells processed products and basic food
ingredients. Processed products and basic food ingredients that people need
to meet their needs vary greatly, but they need something better for the
future, which is processed products and food ingredients (food) that is safe,
more quality and nutritious for consumption.

Hygienic food is the condition and effort required to prevent food

from possible biological, chemical and other objects that may interfere with,
harm and harm human health. Safe and high quality and nutritious food is
important for the growth, maintenance and upgrading of public health and
intelligence (Alsuhendra and Ridawati, 2013; Cahyadi, 2008). Hygienic food
emerges as a dynamic problem along with the development of human
civilization and the advancement of science and technology, so that a system
is needed to monitor food since it is produced, processed, handled,
transported, stored and distributed and served to consumers. Microbiological
toxicity and chemical toxicity to foodstuffs can occur in the food handling

6|Page
 chain from starting at preprint, postharvest / processing until the time the
food product is distributed and consumed (Seto, 2001). Lack of attention to
food safety often affects health problems, such as the incidence of food
poisoning due to unhygienic processing up to the presentation and the use of
dangerous chemicals that are at risk of causing degenerative diseases, cancer
and even death (Syah, 2005).

2.2 Food Additives

The process of buying and selling in trading whether done at the level
of gorsir or retailers are not always sold out on that day and some even take a
long time for all the merchandise was sold out. For merchandise in the form of
basic ingredients food or food can not survive in the long term. Therefore
many traders use certain materials both chemical and nonchemical materials to
preserve the merchandise. This led to the emergence of the term food
additives.

Food Additives are ingredients that are deliberately added to the food
in small quantities in order to improve appearance, texture and extend the
shelf life. It also can increase the nutritional value such as protein, minerals
and vitamins (Widyaningsih and Murtini, 2006). According to Minister of
Health RI Regulation No.722 / Menkes / Per / IX / 1988, Food Additives are
ingredients that are not normally used as food and are usually not typical food
ingredients, have or have no nutritional value intentionally added to food for
(Including organoleptics) in the manufacture, processing, preparation,
treatment, packing, packaging, storage or transportation of food to produce or
be expected to produce (directly or indirectly) a component or affect the
characteristic properties of the food (Alsuhendra and Ridawati, 2013). In
general, food additives can be divided into two major parts:

1. Intentional additives, deliberate additives in food with specific intentions

and purposes, for example to improve consistency, nutritional value,
control of acidity or alkalinity and establish form and appearance and;

7|Page
2. Additive additives, the additives contained in food in very small quantities
as a result of food processing (Winarno, 1992).

The use of appropriate food additives and in accordance with the rules
will produce products with the expected quality. However, if the use of wrong
and excessive will result in the product is not safe anymore consumed. This is
caused by compounds belonging to these food additives are mostly synthetic
chemical compounds which, when used in excessive amounts or are
inconsistent with the rules, can be fatal to health (Alsuhendra and Ridawati,
2013).

Food Additives that are permitted for use on food based on Permenkes
No. 722 / Menkes / Per / IX / 1988 among others (Fardiaz, 2007):

1. Artificial sweeteners, food additives that can cause a sweet taste in foods
that have no or almost no nutritional value. Example: saccharin and
cyclamate.
2. Preservatives, food additives that can prevent fermentation, acidification or
other decomposition of foods caused by microorganisms. Commonly
added to perishable or preferred foods as bacterial or fungal growth
mediums. Examples: benzoic acid and its salts and hydroxy benzoate ester
for fruit products, soy sauce, cheese and margarine, propionic acid for
cheese and bread.
3. Dyes, food additives that can improve or colorize the food. Examples:
karmin, ponceau 4R, red erythrocine, green FCF, green S green, curcumin,
carotene, yellow kuinolin, yellow tartazin and brown caramel.
4. Flavoring and flavoring and flavor enhancers, food additives that can
provide, add or reinforce flavors and flavors. Example: monosodium
glutamate in meat products. While prohibited additives used in food
according to Regulation of the Minister of Health No. 722 / Menkes / Per /
IX / 88 include: borax, formalin, salicylic acid and salt,
diethylpirocarbonate, dulsin, potassium chlorate, chloramphenicol, and
nitrofurazon. According to the Health Minister Regulation No 1168 /

8|Page
 Menkes / Per / X / 1999 that other prohibited substances used in food are
rhodamine B, methanyl yellow and calcium bromate (Yuliarti, 2007).

2.3 Formalin

2.3.1 Definition

Formalin is a solution made from 37% formaldehyde in water.

In formalin solution is usually in add 10-15% alcohol (methanol)
which serves as a stabilizer for formalin does not have polymerization
(Mulono, 2005). Another formalin name is Formol, Morbicid, veracur
(Patnaik, 1992). Formalin was originally identified by August Wilhem
Von Hoffman in 1868. August Wilhem Von Hoffman identifies
formaldehyde when he passes methanol vapor and water over a hot
platinum spiral. The function of formalin as a disinfectant
(disinfectant) was only discovered in 1888 (Anonymous, 2006:
MOHRI R.I, and Dirjen POM, 2003). Formalin is a toxic chemical, in
which formalin toxicity has been evaluated by renowned organizations
such as IARC (International Agency For Research on Cancer), ATSR
(Agency for Toxic Substances and Disease Registry, USA) and IPC
(International Program on Chemical Safety). Formalin has been
classified by IARC into groups of compounds that are at risk of
causing cancer (Uzairu, 2009).

2.3.2 Formalin Benefits

According to Reynold (1982) for its ability to inhibit microbial
growth, formalin can be used as an effective disinfectant against
vegetative, fungal or viral bacteria, although less effective against
bacterial spores. Also according to Alsuhendra and Ridawati (2013)
the benefits of formalin are:
1. exterminator or germ killer so that it can be used for floor
cleaners, boats, warehouses and clothing and exterminator flies
and various other insects;
2. Hardener layer of gelatin and paper

9|Page
 3. Preservative of cosmetics products and nail hardener
4. As an antiseptic to sterilize medical equipment
5. As germicides and fungicides in plants and vegetables and
6. Preserving biological specimens, including corpses and skin
2.3.3 Formalin Impact For Health
Formalin is generally used as a preservative for corpses and
various types of non-food industrial materials. The use of formalin as a
food preservative is very harmful to consumers. But many irresponsible
practices are carried out by traders or food processors who add formalin
as food preservatives (Yuliarti, 2007).
According to Alsuhendra and Ridawati (2013) some negative
effects arising from formalin poisoning if entering into the human body
are:
1. Acute poisoning is a direct, visible effect short-term, and the
symptoms are loss of consciousness, anuria, vomiting, laryngeal
edema, ulceration of the gastrointestinal mucosa, diarrhea, renal
failure and ulceration of the mouth and esophagus. In high
concentrations, formalin may cause bloody diarrhea, blood urine,
vomiting of blood, gastric irritation and ultimately lead to death:
2. Chronic toxicity is a visible effect after prolonged and repeated
exposure. Symptoms include gastrointestinal irritation, vomiting,
dizziness, abdominal pain, intestinal pain and circulatory disorders.
As well as chronic formalin poisoning can also cause menstrual
disorders, infertilization, damage to the liver, brain, spleen, pancreas,
central nervous system and kidney. Possible impacts if the accumulated
levels of formaldehyde in the body exceed the limit is from the
disruption of cell function to cell death which subsequently causes
damage to the tissues and organs of the body. In the next stage can also
occur deviation from the growth
Based on previous research formalin in animal experiments
positively cause cancer so that suspected formaldehyde may cause
cancer in humans (Sihombing, 1996). Cancer can occur because

10 | P a g e
formalin that reacts with cells in the body will disrupt the arrangement
of proteins or RNA as the formation of DNA in the body. If the DNA
sequence is chaotic, then the cells will experience growth that deviates
so that there was cancer (Alsuhendra and Ridawati, 2013).

The consequences of formalin depend on the level of

formaldehyde accumulated in the body. The higher the accumulated
formaldehyde level, the worse the result. ACGIH (American
Conference of Governmental and Industrial Hygienists) set the formalin
safe threshold in the body is 0.04 ppm (Alsuhendra and Ridawati,
2013). Meanwhile, according to the International Program on Chemical
Safety (IPCS), a special body of three UN organizations, ILO, UNEP
and WHO who are concerned about the safe use of chemicals, that in
general the safe threshold of formalin in foods that can still be tolerated
in the adult body Is 1.5 mg to 14 mg per day while formalin in the form
of drinking water that can still be tolerated in the body is 0.01 ppm
(Singgih, 2013).

11 | P a g e
2.4 White shrimp
2.4.1 Description and Classification
White shrimp or also called vanname is one among nine types of
shrimp are highly valued and very popular in Indonesia because it has a
delicious taste and meat, besides the price is cheaper than tiger shrimp.

Classification of White Shrimp

Kingdom : Animalia
Sub kingdom : Metazoa
Filum : Arthropoda
Subfilum : Crustacea
Kelas : Malacostraca
Subkelas : Eumalacostraca
Superordo : Eucarida
Ordo : Decapodas
Subordo : Dendrobrachiata
Familia : Penaeidae
Sub genus : Litopenaeus
Spesies : Litopenaeus vannamei

2.4.2 Morphology of White Shrimp

Kordi (2007) explains that white shrimp has a body of

books and activity of the outer shell (eksoskeleton)

12 | P a g e
 periodically (moulting). White shrimp body parts have been
modified so that it can be used for eating, moving, and
burrowing into the mud (burrowing), and have sensory
organs, such as antenna and antenulae. Kordi (2007) also
explained that the white shrimp head consists of antennae,
antennae, and 3 pairs of maxilliped. The white shrimp head is
also equipped with 3 maxilliped pairs and 5 pairs of walking
legs (periopoda). Maxilliped has been modified and serves as
an organ to eat. At the end of the peripoda segments in the
shape of a claw (dactylus). Dactylus is on the 1st, 2nd, and
3rd legs. Abdomen consists of 6 segments. In the abdomen
there are 5 pairs (pleopods) swimming legs and a pair of
uropods (tails) that make up the fan together with the telson
(the tail).

2.5 Tiger Prawn

2.5.1 Description and Classification
Giant tiger or Penaeus monodon in Indonesia is called tiger
prawn. Tiger Prawn is one type of shrimp that many people love. This is
because of the large size and the meat is sweeter than other shrimp.

13 | P a g e
 Scientific classification of Tiger Prawn

Kerajaan : Animalia
Filum : Arthropoda
Upafilum : Crustacea
Kelas : Malacostraca
Ordo : Decapoda
Upaordo : Dendrobranchiata
Famili : Penaeidae
Genus : Penaeus
Spesies : P. Monodon

2.5.2 Morphology of Tiger Prawn

Viewed from the outside of the body of the shrimp consists of
two parts, namely the front and the back of the front is called the actual
head consisting of the head and chest that fused is called the head-chest
(cepholothorax) and the abdomen (the abdomen) there is a tail on the
back. All parts of the body and its members consist of segments
(segment) of the chest head consists of 13 segments, namely the head
itself 5 segments and chest 8 segments while the stomach consists of 6
segments. Each side of the body has a pair of limbs that are also
segmented. The whole body is covered by an outer frame called an
exoskeleton made of chitin. The framework is hardened, except on the
connections between two adjacent body parts. This makes it easier for
them to move (Mujiman and Suyanto, 2005).

2.6 Formalin Identification On Shrimp

There are several ways that can be done to identify or identify shrimp
containing formalin. The easiest way is through the observation of physical
features. The more convincing and accurate way is through laboratory testing
both qualitatively and quantitatively (Anonymous, 2010).

14 | P a g e
2.6.1 Observation of Shrimp Physical Characteristics
Based on the results of Damayanti research 2014 found
observations of physical or organoleptic characteristics of shrimp are
given formalin during cold storage as a whole can be concluded that the
cold storage of formalin shrimp until day 6 is still good and feasible to
be consumed.

2.6.2 Laboratory Testing

A. Formalin Qualitative Analysis
Qualitative analysis of formalin may be performed to indicate
the presence or absence of formaldehyde in a substance tested by
adding a certain chemical reagent in a material suspected to contain
formaldehyde resulting in a distinctive color change (Widyaningsih
and Murtini, 2006). Qualitative analysis of formalin in shrimp
samples can use several methods such as na reagent method,
chromatopic acid reagent method and Schryver reagent method.
These three methods are a fairly good method in formalin
qualitative analysis (Kuswan, 2011). But in qualitative analysis
formalin on materials suspected to contain formaldehyde such as
meat is the most commonly used method Schryver (Allport, 1951).
Suryadi et al (2008) stated that in previous studies Schryver method
has been declared as the best method for qualitative formalin
analysis because the reagent of Schryver method has a relatively
low visual detection limit of 0.2 ppm. According to Schryver (1910)
fomalin identification using Schryver reagent method can be done
by adding 2 ml of fennilhydrazine hydrochloride 1% solution, 1
ml of 1% ferrisianide (made fresh) potassium solution,
concentrated hydrochloric acid of 5 ml into the material to be tested
Where if the ingredients tested contain formalin then there will be a
change of color from colorless to red (Schryver, 1910).
B. Formalin Quantitative Analysis

15 | P a g e
 Quantitative analysis can be performed using several
methods such as volumetric titration method (Anonymous, 1994),
spectrophotometric methods (Herlich, 1990, Schryver, 1910), high
performance liquid chromatography methods (Anonymous, 1994)
and gas chromatography methods (Bianci et al., 2007) . According
to Susanti (2010) spectrophotometric method is a method of
quantitative analysis of formalin which is often used because this
method is simpler, faster, economical and has a good sensitivity in
determining levels with small concentrations in foodstuffs. The tool
used in this method of spectrophotometry is called a
spectrophotometer. As the name implies, the speculator is composed
of a spectrometer and a photometer. A spectrometer is a device that
produces rays of a spectrum of a certain wavelength and a
photometer is an absorbed light intensity measuring device.
Spectrophotometer is a device used to measure transmittance or
absorbance of a sample as a function of a particular wavelength
(Miller and Miller 2000).
Figure 3 is one of the spectrophotometer tools commonly used in
formalin quantitative tests.

According to Day (2002) spectrophotometric method is a

method of measuring the absorption of light energy by an atom or
molecule of a substance at a certain wavelength. Substances that can

16 | P a g e
be analyzed using a spectrophotometer are substances in the form of
a solution and the substance must appear colored because this
colored substance can provide uptake at visible wavelengths of
visible light. Reagents that can be used to produce a color change in
the solution for the purpose of quantitative analysis of formalin
using spectrophotometer is a reagent commonly used for qualitative
analysis of formalin one pereaksinya reagent Schryver (Kuswan,
2011). According to Schryver (1910) if Schryver reactants react
with formalin there will be a change of color from colorless to red
and can be measured uptake using a spectrophotometer at 518 nm
wavelength.
The steps to be considered in the analysis with
spectrophotometric method are: selection of maximum wavelength,
making of standard curve and determination of compound level in
sample (Gandjar and Rohman, 2007).
1. The wavelength used for quantitative analysis is the wavelength
at which maximum absorption occurs. To obtain the maximum
absorption wavelength, it is done by making a curve of the
relationship between the absorbance and the wavelength of a
standard solution at a certain concentration (Gandjar and
Rohman, 2007).
2. Standard curve making is done by measuring various known
standard concentration solution dilutions using a
spectrophotometer at maximum wavelength. The standard curve
shows the relationship between the concentration of the solution
(x axis) and the absorbance of the solution (y-axis). The function
of this standard curve is used to indicate the concentration of the
sample solution from the measurement (Underwood, 1990).
3. The determination of the concentration of the solution
compounds contained in the sample was analyzed based on the
treatment stages. The solution obtained was given treatment
according to the treatment given to the standard solution then

17 | P a g e
measured by spectrophotometry at maximum wavelength. The
result is adjusted to the comparison curve, while the sample
solution content is calculated based on the linear regression
equation: y = a + bx
Description: y = absorbance
B = slope / slope line
A = intersection point
X = concentration

18 | P a g e
 CHAPTER III

RESEARCH METHODS

3.1 Type of Research

Research of formalin content on shrimp sold in modern market in
Surabaya city is descriptive research type, that is research which give
description or description about formalin analysis on shrimp that traded in
modern market in Surabaya city. The data were obtained through observation
of the physical characteristics of shrimp and laboratory testing consisting of
two stages. The first stage of laboratory testing is formalin qualitative analysis
on shrimp samples using Schryver method and second is quantitative analysis
of formalin on shrimp sample using spectrophotometric method.
3.2 Research Time and Place
This research plan is carried out from July to October 2017. Sampling
location is in some modern markets (modern market in Darmo Permai street,
Carfour on Dukuh West Kupang road and BG Juction at Jalan Bubutan, Lotte
Mart at Pakuwon Mall Jalan Lontar 5 Surabaya, Maspion Squre road
Diponegoro and Ahmad Yani) city of Surabaya. Formal shrimp analyzes
consisting of 13 qualitative and quantitative analyzes were conducted at the
Microbiology Laboratory of the Faculty of Medicine, Airlangga University or
BPOM (Food and Drug Supervisory Agency at Jalan Jagir Wonokromo).
3.3 Research Materials
3.3.1 Sample
The number of samples used in this study is determined by the formula
of Federer (1977), namely;
(N-1) x (t-1) ≥15 Description: n = number of repetitions, t = treatment,
in this case there are 4 treatments (control group, treatment with
concentration 10%, 15%, 20%) so
(N-1) (4-1) = 15
(N-1) (3) = 15
N -1 = 15: 3
N=1+5=6

19 | P a g e
 So many repetitions are done as much as 6 times.
3.3.2 Tools and materials
The tool used in this research is 1). 10 ml measuring flask, 2).
Analytical balance sheet, 3). Destilator, 4). Glassware, 5). UV-Vis
Spectrophotometer (Cary 50 Conc.), 6). Distillation flask, 7). Test
tube, 8). Pipette scale, 9). Markers, 10). Knives, 11). Plastics, 12).
Eyedropper and 13) Ehrlenmeyer. While the material used is 1).
Shrimp sample, 2). Aquades, 3). 85% phosphoric acid, 4). Formalin
(Merck) 37%, 5). Solution of fennilhidrazine hydrochloride (Merck)
1%, 6). Solution of potassium ferrisianida (Merck) 1% and 7).
Concentrated hydrochloric acid (Merck).

3.4 Research Methods

3.4.1 Sampling
The shrimp samples are taken from 6 modern markets in
Surabaya (modern market in Darmo Permai street, Carfour at Dukuh
Kupang road and BG Juction at Bubutan Street, Lotte Mart at Pakuwon
Mall Lontar 5 Surabaya Street, Maspion Squre Diponegoro and Ahmad
Road Yani) with the number of shrimp samples taken is minimal or ±
100 grams. Shrimp samples taken from every modern market are
packaged in containers such as dry plastic. Plastic container 6 pieces
are coded with the market code for sampling. After that, the shrimp
samples were taken from the sampling site to be carried out the phase
of observation of the physical characteristics of the shrimp and the
testing stage in the laboratory.

3.4.2 Observation of Shrimp Physical Characteristics

The observation of the physical characteristics of shrimp is to observe
changes in the shape and smell of shrimp for 5 days after sampling in
some modern markets.
Parameter Value (Organoleptic)

20 | P a g e
3.5 Laboratory Testing
3.5.1 Qualitative Analysis of Formalin on Shrimp
The method used in the qualitative analysis of shrimp is the
Schryver method. The shrimp examination by the Schryver method is
used to identify formalin on shrimp in qualitative analysis. The
application of this method is used for qualitative analysis because in
previous research the Schryver method has been expressed as the best
method for formalin qualitative analysis. This is because the reagents
of the Schryver method has a relatively low visual detection limit
against formalin, which is 0.2 ppm. In this method is done sample
preparation by chopping shrimp sample then distillation. The
distillation product was then added several chemical solutions ie 2 ml
of 1% fennilhidrazine hydrochloride solution made freshly, 1 ml of 5%
ferricyanide potassium solution was made new, 5 ml of concentrated
hydrochloric acid. The presence of formalin in the sample solution
tested is shown by the formation of red color (Schryver, 1910).
3.5.2 Quantitative Analysis of Formalin on Shrimp
Formal quantitative analysis was performed to find out how
much formalin contained in shrimp samples. The method of sample
preparation on spectrophotometric quantitative analysis used the same
sample preparation method in qualitative analysis (Schryver Method).
Schryver (1910) states that the quantitative analysis of formalin in the
sample solution which showed positive results in qualitative analysis
that is by the formation of red complex compounds can be done by

21 | P a g e
using spectrophotometric method and measured uptake at 518 nm
wavelength.

22 | P a g e
 REFERENCE

Allport, Noel. L. 1951. Colorimetric Analysis. New York : Chapman & Hall, Ltd;
397-399.
Alsuhendra dan Ridawati.2013 . Bahan Toksik dalam Makanan. Rosda.Jakarta.
Anonim.1994. Methods For The Determination of Hazardous
SubstanceFormaldehyde in Air.
Anonim. 2010. Formalin Pada Makanan. http://smallcrab.com/kesehatan/650-
penggunaan-formalin-dalam-produk-pangan.
Cahyadi, W. 2008. Analisis dan Aspek Kesehatan Bahan Tambahan Pangan .
Edisi 2 Cetakan I. Bumi Aksara: Jakarta.
Damayanti, Evina. 2014. Efektivitas Kunyit Sebagai Pereduksi Formalin Pada
Udang Putih Penyimpanan Suhu Dingin. Universitas Diponegoro
Day, R. A. 2002 .Analisis Kimia Kuantitatif. Edisi Keenam. Jakarta; Penerbit
Erlangga. Hal 394, 396-404.
Fardiaz, S. 2007. Bahan Tambahan Makanan. Institut Pertanian Bogor.
Bandung.http://perpustakaan.pom.go.id/KoleksiLainnya/Buletin%20Info
%20 POM/0110.pdf.
Gandjar, I. G. dan Rohman, A. 2007.Kimia Farmasi Analisis. Cetakan II.
Yogyakarta: Pustaka pelajar. Halaman 246.
Herlich, K. 1990. Official Methods Of Analysis. 15th edition. Virginia. AOAC
Inc. Page 934.
Kuswan, A.S,. 2011. Optimasi Pereaksi Schryver dan Penerapannya Pada
analisis Formaldehid Dalam Sampel Usus dan Hati Ayam Secara
Spektrofotometri. Skripsi. Fakultas Matematika dan Ilmu Pengetahuan Alam
Program Studi Farmasi. Depok.
Kordi, K. M. Ghufran. 2007. Pengelolaan Kualitas Air dalam Budidaya Perairan.
Jakarta: PT Rineka Cipta.
Manitoba Federation of Labour. 2006. Occupational Healthcare Inc.2004.
Formaldehyde. http://www.mflohc.mb.ca. diakses pada tanggal 27 Maret
2017.

23 | P a g e
Miller JN and Miller JC. 2000. Statistic and Chemometrics for Analytical
Chemistry, 4th ed. Prentice Hall : Harlow.
Mukti, W. 2012. Analisis Spektroskopi UV-Vis “Penentuan Konsentrasi
Permanganat (KMnO4)”. Surakarta : FMIPA Universitas Sebelas Maret
Surakarta
Patnaik,P. 1992. A Comprehensive Guide to the Hazardous Properties of
Chemical Substances. New York : Van Nostrand Reinhold ; 94.
Schryver, S.B. 1910. The Photochemical Formation Of Formaldehyde in Green
Plants. Proc. Roy . Soc. London, series B 82 (554) ; 227.
Singgih, H. 2013. Uji Kandungan Formalin Pada Ikan Asin Menggunakan Sensor
Warna Dengan Bantuan FMR. Jurnal ELTEK, Vol 11 No 01.ISSN 1693-
4024.
Susanti, S. 2010. Penetepan Kadar Formaldehid Pada Tahu yang di Jual di Pasar
Ciputat Dengan Metode Spektrofotometri Uv-Vis disertai Kolorimetri
menggunakan pereaksi Nash. Skripsi. Program Studi Farmasi, Fakultas
Kedokteran dan Ilmu Kesehatan Universitas Islam Negeri Syarif
Hidayatulla. Jakarta.
Suryadi, Herman, Hayun dan Harsono, F.D. 2008. Selection Of Formalin Method
Of Analysis Based On Colour Reaction and Spectrophotometry Uv-Vis.
Proseeding Kongres Ilmiah ISFI ; 1 -10.
Suyanto, S.R dan Mujiman, A, 2001. Budidaya Udang Windu. Penebar Swadaya,
Jakarta.
Syah, D. 2005. Manfaat dan Bahaya Bahan Tambahan Pangan. Bogor:Himpunan
Alumni Fakultas Teknologi Pertanian IPB.
Underwood, AL. 1990. Analisis Kimia Kuantitatif. Jakarta : Erlangga.
Uzairu, A. 2009. Formaldehyde Levels In Some Manufactured Reguler Foods In
Makurdi, Benue State, Nigeria. (Jurnal of Applied Sciences In
Environmental Sanitation, V.N 211 -214).28
Yuliarti, N. 2007. Awas! Bahaya Di Balik Lezatnya Makanan. Andi :Yogyakarta.
Widyaningsih, T.D. dan Murtini, ES. 2006. Alternatif Pengganti Formalin
PadaProduk Pangan. Jakarta: Trubus Agrisarana.
Winarno, F.G. 1992. Kimia Pangan dan Gizi.Jakarta :Gramedia Pustaka Utama

24 | P a g e
25 | P a g e