APPROVAL SHEET

Complete report of Biochemistry Experiment with the title Saliva, which

made and arranged by:
Name : Jumriana Jufri
Class : ICP Chemistry A
ID : 1413440011
Group : II (Two)
After Checked and consulted by Assistant and Coordination Assistant, so this
report is accepted.

th
Makassar, November 2016
Assistant Coordinator Assistant

Ida Masita Marwa Karim

Known By,
Responsibility Lecturer,

Hardin, S.si.,S.pd.,M.pd
 CHAPTER 1
INTRODUCTION

A. Background
Many of the complaints that may arise in the oral cavity. One complaint is a
complaint of dry mouth or xerostomia. This situation is generally associated with
reduced salivary flow, but sometimes the amount or salivary flow remained normal
but the person complaining of dry mouth.
Dry mouth symptoms can be acute or chronic, temporary or permanent, and less
or somewhat perfect. In what form of dry mouth symptoms arise, depending on
penyebabmya. Many factors can cause dry mouth, such as radiation in the neck and
head, Sjogren's syndrome, systemic diseases, side effects of drugs, stress and age.
Reduced saliva production is kept up accompanied by changes in the composition of
saliva which resulted largely salivary function can not run smoothly. This has resulted
in some complaints in patients with dry mouth, such as difficulty in chewing and
swallowing food, difficulty in speaking, reduced sensitivity to taste, difficulty in
wearing dentures, mouth tasted like burnt and so on.
Given the importance of saliva and the consequences caused by due to reduced
salivary flow, then it is necessary countermeasures against patients with dry mouth
symptoms. Treatment is given depends on the cause and severity of dry mouth. In this
paper will discuss the factors that cause and treatment of dry mouth symptoms.
B. Formulation Of Experiment
1. What the organic compound that contain in saliva ?
2. What the inorganic compound that contain in saliva ?
3. How the influence temperature againt phtialin activities ?
4. What kind of pH that correct to saliva?
5. What the effect of compound the activities of bactery on analyze saliva
C. Objectives Of Experiment
1. To know the organic compound that contain in saliva
2. To know the inorganic compound that contain in saliva
3. To know the influence temperature againt phtialin activities.
4. To know the pH that correct to saliva.
5. To know the effect of compound the activities of bactery on analyze saliva
D. Benefit Of Experiment
1. Apprentice can find out that organic compound that contain in saliva
2. Apprentice can find out that inorganic compound that contain in saliva
3. Apprentice can find out the influence temperature againt phtialin activities.
4. Apprentice can find out the pH that correct to saliva.
5. Apprentice can find out the effect of compound the activities of bactery on
analyze saliva

CHAPTER II
PREVIEW OF LITERATURE

1. Tests mucin
Saliva is produced by three pairs of major glands, the parotids, the
submandibulars, and the sublinguals, located outside the mouth, and hundreds of
minor glandseach the size of a pinhead and located just below the oral
epithelium . As judged by magnetic resonance imaging, the volume of the parotid
gland is about 2.5 times that of the sub-mandibular gland and eight times that of
the sublingual gland . Similar relationships are obtained when the comparisons
are based on gland weights, the parotid gland weighing 1530 g (Gray1988). The
saliva from the parotid and submandibular glands reaches the oral cavity via long
excretory ducts (7 and 5 cm, respectively), the parotid duct (also called Stensens
duct) opening at the level of the second upper molar, and the submandibular duct
(Whartons duct) opening on the sublingual papilla. In about 20% of the
population, the parotid duct is surrounded by a small accessory gland.Sublingual
saliva empties into the submandibular duct via the major sublingual duct
(Bartholins duct) or directly into the mouth via a number of small excretory ducts
opening on the sublingual folder. Likewise, the saliva of minor glands, such as of
the buccal, palatine (located just in the soft palate), labial, lingual, and molar
glands, empties into the mouth directly via small, separate ducts just traversing
the epithelium (Tandler and Riva1986). Unless saliva is collected directly from
the cannulated duct, the saliva in the mouth will be contaminated by the gingival
crevicular uid, blood cells, microbes, antimicrobes, cell and food debris, and
nasopharyngeal secretion. Consequently, mixed saliva (whole saliva) collected
by spitting or drooling is not pure saliva, although the term salivais usually
used.
 (Ekstrom, 2012:3-4)
According to the opinion of Rosen,2002:2-7 is :
a. parotid gland represents the largest salivary gland, averaging 5.8 cm in the
craniocaudal dimension, and 3.4 cm in the ventral-dorsal dimension. The
average weight of a Parotid gland is 14.28 g. It is irregular, wedge shaped,
and unilobular. The Parotid has been described as having 5 processes (3
superficial and 2 deep), thus making it very difficult to surgically removal all
parotid tissue. It lies in the parotid compartment, a triangular space which
also contains CN VII and its branches, sensory and autonomic nerves, the
External Carotid artery and its branches, the Retromandibular (Posterior
Facial) vein, and Parotid lymphatics.
b. Submandibular gland weighs the weight of the Parotid. It is often
referred to as the Submaxillary gland because of the tendency of British
anatomists to refer to the mandible as the submaxilla. This gland lies in the
submandibular triangle formed by the anterior and posterior bellies of the
Digastric muscle and the inferior margin of the mandible. The gland is
positioned medial and inferior to the mandibular ramus partly superior and
partly inferior to the base of the posterior half of the mandible. The gland
forms a C around the anterior margin of the Mylohyoid muscle, which
 divides the Submandibular gland into a superficial and deep lobe. The deep
lobe comprises the majority of the gland. The Marginal Mandibular branch of
CN VII courses superficial to the Submandibular gland and deep to the
Platysma. As is the case with the Parotid gland, the Submandibular gland is
invested in its own capsule, which is also continuous with the superficial layer
of deep cervical fascia.
c. Sublingual Gland This is the smallest of the major salivary glands. The
almond shaped gland lies just deep to the floor of mouth mucosa between the
mandible and Genioglossus muscle. It is bounded inferiorly by the
Mylohyoid muscle. Whartons duct and the Lingual nerve pass between the
Sublingual gland and Genioglossus muscle. Unlike the Parotid and
Submandibular glands, the Sublingual gland has no true fascial capsule. Also
unlike the Parotid and Submandibular glands, the Sublingual gland lacks a
single dominant duct. Instead, it is drained by approximately 10 small ducts
(the Ducts of Rivinus), which exit the superior aspect of the gland and open
along the Sublingual fold on the floor of mouth. Occasionally, several of the
more anterior ducts may join to form a common duct (Bartholins duct),
which typically empties into Whartons duct. Of note, the ducts of the
sublingual glands are too small for the injection of contrast, making a
sialogram of this gland impossible
According to the opinion of Rosen,2002:2-7 is :
a. Parotid gland is the majority of the parotid gland can be easily assessed by
ultrasound however the deep portion of the gland may be seen with difficulty
and the portion of the parotid gland which lies medial to the mandible cannot
be identified with consistency. The masseter muscle is located deep to the
anterior part of the superficial parotid, lateral to the ramus of the mandible.
The inferior portion of the parotid gland may be referred to as the cervical
lobe. Within the parenchyma the retromandibular vein is often identified,
lying lateral to the external carotid artery. The plane of the retromandibular
 vein through the parotid can be used to differentiate between the superficial
and deep part of the parotid gland.
b. The submandibular gland is triangular shaped, with a homogeneous
echogenic structure, identified at the posterior border of the mylohyoid
muscle. The facial artery and vein are located posterior to or within the gland,
the facial artery passing superiorly behind the submandibular gland, over the
inferior body of the mandible. Non dilated intraglandular ducts are usually not
identified, but may be seen as faintly visible narrow, confluent tubules.
Wharton's duct (main duct) is identified between the mylohyoid and
hyoglossus muscles, colour flow imaging may help in differentiating it from
the adjacent lingual vessels.
c. The sublingual gland is distinguished from the genioglossus muscles as a
echogenic mass lying lateral to the genioglossus, deep to the mylohyoid.
There may be a direct communication with the submandibular gland situated
dorsally. The ducts lead to the sublingual caruncle, in the anterior part of floor
of the mouth, they cannot be identified with ultrasound ( Gritzman,2011:3-4).
So my opinion is The Parotid gland is a purely serous salivary gland. Of note, the
Parotid gland is unique in that it contains many fat cells; in fact, the adipocyte to
acinar cell ratio in the Parotid is 1:1. The Submandibular gland is mixed, but
predominantly serous. Approximately 10% of its acini are mucinous. The Sublingual
gland is mixed, but predominantly mucous. Of the major salivary glands, the
Sublingual gland utilizes a simple system of transport, whereas the Parotid and
Submandibular glands involve elaborate networks. Salivary gland stroma is rich in
lymphocytes and plasma cells, which are responsible for the production of IgA. IgA
fuses with the secretory piece on the basal membrane, is then transported across the
epithelial cell, and released into the ductal lumen as secretory IgA.
2. Test thiocyanate ( SCN- )
The SCN" concentrations in whole saliva were in the same rnge and higher
in smokers than in nonsmokers, s reported by others (l,17,18).In stimulated
parotid saliva the concentrations found (1.27 2.40mmol/1) are higher than those
reported by Pruitt et al. of0.76 0.55 (SD) (19).The SCN- concentrations in
parotid and whole saliva are about 30 times higher than in plasma. Thus SCN"
must accumulate in the parotid gland and be secreted througha process which is
still unexplained. This is also the case for hypocyante (OSCN"),a degradation
product of SCN"(19), which functions s an antimicrobial agent (20). Extremely
high flow rates (u to 3 ml/min) could lower the SCN" concentration by dilution.
However, it is improbable that it could account solely for these large
concentration differences. The levels of SCN" in saliva are also influenced by diet
and smoking habits and show a larger scatter than the plasma values
( Degiempietro,1987:6).
Measured saliva SCN is compared to self-reports of average cigarette
smoking. Mean levels and the percentage of students above selected points (2
85 ,ug/ml and 2 100 ,ug/ml) increase with the reported level of smok- ing. In self-
declared "quit" smokers, the levels were similar to light and nonsmokers. Mean
SCN levels were not significantly different between the nonsmoker, once or twice
experimenter, few per month, and quitter categories. Paired comparisons of mean
SCN levels indicated that SCN levels were significantly higher for those in
categories 4, 5 and 6 (smoking more than a few/month) compared to lower level
smoking categories. Within these three heavier smoking categories, mean SCN
was significantly higher as cigarette consumption increased(Luepker,1981:3).
SCN concentration in body fluids appeared to be important in classifying
patient as smokers or non smokers, in determining some clinical condition, and in
specimen validity testing in firencic drug testing. The human saliva samples were
diluted and the anions were separated by an extractive alkylatin technique.
Thiocyanate is usually present in saliva,serum and urine in low concentration. It is
principal metabolic product of cyanide metabolism. High concentration may be
due to HCN ingestion from tobacco smoke and metabolic convention to
SCN(Paul,2006:1).
 So my opinion is Thiocyanate (also known as rhodanide) is the anion [SCN] -.
Thiocyanate is the conjugate base of thiocyanic acid. A commonly found derivatives
including salts of potassium thiocyanate (KSCN) and sodium thiocyanate (NaSCN).
Organic compounds containing the functional group SCN are also called
thiocyanate. For example, mercury (II) thiocyanate is widely used in the manufacture
of fireworks. Thiocyanate is also known to be an important part in the biosynthesis
hypothiocyanite by lactoperoxidase. Therefore, the presence of thiocyanate or
thiocyanate reduction compounds damage the human immune system. Thiocyanate
itself is a metabolite of sodium nitroprusside sodium, assisted catalysis rodanese.
Thiocyanate (SCN-) is a derivative of cyanide (HCN), which has the properties
goitrogenik, in the body inhibits decision-iodine by the thyroid gland which is useful
for the formation of thyroid hormones. Impaired formation of thyroid hormone causes
enlargement of the thyroid gland is often called a goiter. On a deeper level by weight,
impaired formation of thyroid hormone can cause stunted physical growth, disrupt
the central nervous system, mental retardation, and even can make a person blind
and deaf.

3. Test inorganic compounds

Comprised of both inorganic and organic compounds, saliva is distinguished
by it high volume compared to salivary gland weight, high potassium
concentration, and low osmolarity. The large relative volume of saliva production
is due to its higt secretion rate, which can go up to 1 ml per gram of
salivary gland perminute . Saliva is mostly hypotonic to plasma, but its
osmolarity increases with increasing rate of secretion, and at its highest rate
saliva approaches isotonicity . The concentration of electrolytes in
saliva also changes with varying secretion rates. Within the salivary gland,
potassium(K+) concentration is always high while sodium(Na+) concentration
is low compared to that found in plasma .With increasing flow rates, however,
Na+ concentration increases,while K+ concentration initially decreases slightly
and then levels off to a constant level . Chloride (Cl-) concentrations follow the
same general pattern as Na+ concentrations . In other words,Na+ andCl- are
generally secreted and then slowly reabsorbed along the course of the salivary
system, from acinus to duct . The salivary concentration of bicar-bonate(HCO3-)
is hypertonic compared to in plasma ex-cept at lower rates of secretion
(Chrishtopher,2012:11).
Saliva is composed of water , organic and inorganic molecules, but a large
intra- and inter-subject variability in composition is reported. (10) salivary
calcium and phosphate concentrations increase with age showing peak values
around menopause. Therefore we suggest that menopause is reflected in saliva as
elevated levels of calcium and phosphate (11) Saliva also plays an important role
in maintaining the integrity of dental tissues due to the presence of calcium,
phosphorous and other inorganic ions as this environment is known to facilitate
remineralization of incipient lesions or demineralized zones of enamel. Thus
calcium and phosphorous in saliva forms a natural defence mechanism against
dissolution of teeth.(12) Positive correlations have been shown between high
salivary calcium content and periodontitis and between high salivary calcium
content and number of intact teeth. It was also found that subjects with
periodontitis have more intact teeth and more intact molars than subjects who are
free of the disease.Therefore, the present concept is that periodontitis affected
subjects have higher intraoral mineralization potential(Prashaanti,2016:2).
The concentration of salivary calcium varies with the SF and is not affected
by diet. However, diseases such as cystic fibrosis and some medications such as
pilocarpine cause an increase in calcium levels. Depending on the pH, salivary
calcium can be ionized or linked. Ionized calcium is important for establishing
equilibrium between the calcium phosphates of enamel and its adjacent liquid.
Non-ionized calcium can be linked to inorganic ions (inorganic phosphate,
bicarbonate, fluoride), to small organic ions (citrate), and to macromolecules
 (statherin, histidine-rich peptides, and proline-rich proteins). A special case of the
combination of calcium is its strong link with amilase, where it acts as a co-factor
necessary for the enzyme function. Inorganic orthophosphate found in saliva
consists of phosphoric acid (H3PO4) and primary (H),secondary (HPO4), and
tertiary (PO4inorganic phosphate ions. The concentrations of these ions depend
on salivary pH and vary in accordance with the SF. As the flow increases, the total
concentration of inorganic phosphate diminishes.The most important biological
function of this ion is to maintain the dental structure. Another function, discussed
previously, is its buffer capacity, relevant only in unstimulated
SF(Almeida,2005:4).
So my opinion is Saliva is the colorless and viscous fluid which is secreted by
the salivary glands (parotid gland, submandibular gland and sublingual gland) and
many small glands scattered in the mucosa of the oral cavity. Salivary glands
produce approximately 1-1,5 dm3 of saliva per day. The density of saliva is
about1,002-1,012 g/cm3. Human saliva consists of 99% water and the rest are
organic and inorganic compounds and gases.

4. Temperature Enzim ptyalin

 Saliva is responsible for the initial digestion of starch, favoring the formation
of the food bolus. This action occurs mainly by the presence of the digestive
enzyme -amylase (ptyalin) in the composition of the saliva. Its biological
function is to divide the starch into maltose, maltotriose, and dextrins. This
enzyme is considered to be a good indicator of properly functioning salivary
glands, contributing 40% to 50% of the total salivary protein produced by the
glands. The greater part of this enzyme (80%) is synthesized in the parotids and
the remainder in the submandibular glands. Its action is inactivated in the acid
portions of the gastrointestinal tract and is consequently limited to the
mouth(Almeida.2005:4).
Produced a bell-shaped curve with the highest peak indicating the optimum
temperature for enzymatic activity. At 4C, enzymatic reaction of salivary amylase
occurs slowly or not at all due to lack of energy and heat. As the temperature
increases, its enzymatic also increases up until the optimum temperature. Figure 1
shows that the optimum temperature of salivary amylase ranges from 32C to
37C.This applies to the human body since salivary amylase is suitable to function
within these temperatures. After 37C, the graph then steeply declines as a result
of loss of activity. At 50C and 70C, salivary amylase is denatured. The molecular
conformation of the enzyme becomes altered as the hydrogen bonds responsible
for
its secondary, tertiary and quaternary structures are broken(Rodillas,2005:6).
5. pH
Most enzymes are active only over a narrow pH range and have an optimal
pH, at which reaction is the fastest. An increase or decrease in pH also causes
denaturation in enzymes, thereby affecting their activity. Table 2 shows the results
obtained on how enzyme activity of salivary amylase is affected by pH. At pH 4,
the salivary amylase is in a too acidic environment to function. As pH decreases,
certain amino acids like aspartate and glutamate are protonated, causing them to
lose their net negative charge which consequently denatures the enzyme. The
 optimum pH for the action of salivary amylase ranges from 5.6 to 6.9 (Talwar &
Srivastava, 2006). This is consistent with the peak found between pH 4 and 6 in
Figure 2. However, the curved peaked highest at pH 10. Inconsistencies with the
results obtained can be attributed to human error such as inaccuracies in
measurement and timing during the experiment. Ideally at pH 10, salivary
amylase is denatured due to high alkalinity. As pH increases, certain amino acids
such as lysine and arginine are deprotonated, causing them to lose their net
positive charge which also results to enzyme denaturation. The activity of
enzymes may be markedly changed by any alteration in pH, which in turn, alters
electrical charges on the enzyme. Changes in charge affect the ionic bonds that
contribute to the enzymes tertiary and quaternary structure, thereby changing the
proteins conformation and activity. Thus, pH-activity relationship of enzymes is
dependent on the amino acid side chains present in the enzyme(Rodillas,2005:6).

6. Estimation ptialin
Salivary amylase is the enzyme produced by the salivary glands. Formerly
known as ptyalin, it breaks down starch into maltose and isomaltose. Amylase,
like other enzymes, works as a catalyst. All catalysts are enzymes, but not all
enzymes are catalysts. A catalyst is a substance that hastens a chemical reaction
but does not become part of the end product. Amylase digests starch by
catalyzing hydrolysis, which is splitting by the addition of a water molecule. The
presence and absence of starch can be confirmed by several tests such as the
iodine test, Benedicts and Fehlings test. In general, a blue-black color indicates
the presence of starch (Rodillas,2005:4).
7. The effects of compounds that inhibit/destroy bacterial activity in salivary
amylase.
The enzyme inhibitors are low molecular weight chemical compounds. They
can reduce or completely inhibit the enzyme catalytic activity either reversibly or
permanently (irreversibly). Inhibitor can modify one amino acid, or several side
chain(s) required in enzyme catalytic activity. To protect enzyme catalytic site
from any change, ligand binds with critical side chain in enzyme. Safely, chemical
modification can be done to test inhibitor for any drug value. In drug discovery,
several drug analogues are chosen and/or designed to inhibit specific enzymes.
However, detoxification or reduced toxic effect of many antitoxins is also
accomplished mainly due to their enzyme inhibitory action. Therefore, studying
the aforementioned enzyme kinetics and structure-function relationship is vital to
understand the kinetics of enzyme inhibition that in turn is fundamental to the
modern design of pharmaceuticals in industries [Sami et al. 2011]. Enzyme
inhibition kinetics behavior and inhibitor structure-function relationship with
enzyme active site clarify the mechanisms of enzyme inhibition action and
physiological regulation of metabolic enzymes as evidenced in following chapters
in this book. Some notable classic examples are: drug and toxin action and/or
drug design for therapeutic uses e.g., iodoacetamide deactivates cys amino acid in
enzyme side chain; methotrexate in cancer chemotherapy through semi-
selectively inhibit DNA synthesis of malignant cells; aspirin inhibits the synthesis
of the proinflammatory prostaglandins; sulfa drugs inhibit the folic acid synthesis
essential for growth of pathogenic bacteria and so many other drugs. Many life-
threatening poisons, e.g., cyanide, carbon monoxide and polychlorinated
biphenols are all enzyme inhibitors(Sharma,2012:1-2).
 CHAPTER III
METHODE OF EXPERIMENT

A. Apparatus and Chemicals

a. Apparatus
b. Small Test Tubes 12 pieces
c. Big Test Tubes 6 pieces
d. Neraca Analytic 1 piece
e. Small tube rack 2 pieces
f. Big test tube rack 1 piece
g. Volumetric glass 10 ml 2 pieces
h. Volumetric glass 25 ml 1 piece
i. Dropped pipette 10 pieces
j. Baker glass 1000 ml 1 piece
k. Baker glass 500 ml 1 piece
l. Baker glass 250 ml 1 piece
m. Baker glass 250 ml 2 pieces
n. Funnel 2 pieces
o. Spray bottle 1 piece
p. Asbestos gauze 2 piece
q. Spritus 2 pieces
r. Tripod 2 pieces
s. Stopwatch 2 pieces
t. Stir bar 1 piece
u. Wood clamp 1 piece
2. Chemicals
a. Saliva
b. Acetic acid 0.1 M and 2 N (CH3COOH)
c. Millon reagent
d. Benedict reagent
e. Molisch reagent
f. Iron (III) chloride 0.1 M (FeCl3)
g. Hydrochloric acid concentrated (HCl(p))
h. Mercurium (I)chloride (HgCl)
i. Amylum
j. Sodium chloride 0.1 M (NaCl)
k. Iod solution (I2)
l. Buffer solution
m. Aquadest (H2O)
n. Phenol (C6H5OH)
o. Nitrate acid diluted (HNO3)
p. Silver nitrate (AgNO3)
q. Filter paper
r. Tissue
s. Matches
t. Ice
u. Label
v. Toluene (C6H5CH3)
B. Work Procedure
1. Mucin test
5 ml of saliva in reaction tube added 2 drops of acetic acid 0.1 M. Used air as
control, separate the precipitate and do Millon test, benedict est and millon.
2. Thiocianate test
Into 5 ml of saliva in tube reaction added 5 drops of FeCl 0.1 M solution and 1
drops of concentrated HCl, then added 5 drops of HgCl 1% that can formed
Hg(II) thiocianate that mot have color.
3. Inorganic test
Into 15 ml of saliva added acetic acid 2 N drop by drop until the mixture was
turbid, then heated until boiled, then filter divided into three part:
a. To test Cl- the filtrate was add to HNO 3 diluted and add some drops of
AgNO3 0.5M
b. PO43- test, add HNO3 diluted then add 1 ml of BaCl2 5%
c. For Ca2+ ion, add into filtrate 1 ml of NH4 oxalate 4%
4. Test the effect of temperature in ptyalin activity
Add each of 5 ml starch 1% into 4 of tube reaction that cleaned. First tube
reaction was cooled in ice cube, the second tube in room temperature, the third
tube was hate until the temperature was 380C. And the fourth tube was boiled,
into the four tube add 2 ml of diluted saliva with ratio (1:9), and mix it well into
tube number 4, add 2 drops of I2 0,01 M
5. Ptialin test estimation
Into 100 ml of starch solution 1% added 2 ml of NaCl 0,1 M solution and placed
in boiled water that have 380C of temperature. After that add 1 ml of saliva
diluted (1:9) into the starch of solution. Add prepared 8 tube reaction that is
 contain 3 ml of water and 3 drops I 2 0,01 M. In interval 30 second add again into
tube 2 until 4 minutes.
6. Test determine suitable pH for saliva work
Prepared 10 ml of buffer solution tha have pH 8; 7,4; 6,,8; 6 and 5,2 into that
bufferof solution. After that add 5 ml of starch solution 1%. 2 ml NaCl 0,1M and
2 ml diluted saliva (1:9). Placed the tube into boiled water with 38% of
temperature. Add to each tube I2 solution int tube with pH between 8-7,4 must
added acid before add I2 solution.
7. The effect of compound that inhibitor/breakdown the activity bacteria in a
mylase of saliva
Diluted 2 ml of saliva with 8 ml of water and mixed it well add 1 ml of this
diluted of saliva in 7 tube reaction. Next into tube reaction each add toluene 5
drops in I2, 5 tube and 5 drops of chloroform in tube 2.5 drops of phenol 2% in
tube 4 and 0.5 mg NaF. In tube 5 then 5 drops of water i tube 6, let it during 10
minutes and shaked it. After that add into each tube 5 ml of starch solution 1%.
Placed all tube in warmbath with the 380C of temperatute during 5 minutes. In
the end divided the contain of each tube reaction into 2 part, in first part add I 2,
and other part add benedict.
 CHAPTER IV
RESULT AND DISCUSSION

A. Observation Result

1. Test of Musin
No Activities Result
1. Saliva (5 ml) + CH3COOH (2 ml) Colorless solution
filtered Filtrate Colorless solution
a. Filtrate + reagent of mollisch Turbid solution
b. Filtrate + reagent of benedict Blue solution
c. Filtrate + reagent of millon Colorless solution

2. Test of Thiosianat
No Activities Result
1. 5 ml saliva + 5 drops FeCl2 0,1 M Red precipitate
 Mixture + 1 drop HCl concentrate Turbid solution
. Turbid solution + H2O (2 ml) + 10 drops Colorless solution
HgCl
3. Test of decomposer of Inorganic Compounds in Saliva

No Activities Result
1. 15 ml saliva + 10 drops CH3COOH Turbid solution
2. Turbid solution was filtered Colorless solution
3. Colorless solution was divided into 4 Colorless solution
tube
Test Cl- ion
Form precipitate
Tube I: filtrate + 3 ml HNO3 + AgNO3
Test Ca2+ ion
Form green solution
Tube II: 3 ml HNO3 + Amonium molibdat
5% Colorless solution
Tube III: 3 ml HNO3 + BaCl2 0,1 M Turbid solution
Tube IV: 3 ml HNO3 + (NH4)2C2O4

4. Test of the effect temperature for activity of enzyme

No Activities Result
 1. Put starch solution 1% (5 ml) into 4 Colorless solution
tube
2. Tube I (at room temperature)
5 ml starch 1% + 2 ml saliva solution Turbid solution
At 5 minutes added iod 0.01 M Yellow solution

Tube II (at ice water)

5 ml starch 1% + 2 ml saliva solution Turbid solution

At 5 minutes added iod 0.01 M Blue solution

Tube III (at 38 )

Turbid solution
5 ml starch 1% + 2 ml saliva solution Blue solution
At 5 minutes added iod 0.01 M
Tube IV (with heated saliva) Turbid solution
5 ml starch 1% + 2 ml saliva solution Blue solution
At 5 minutes added iod 0.01 M

5. Test of estimation ptialin

No Activities Result
1. 10 ml starch 1% + 2 ml NaCl 0.1 M and Turbid solution

heated at 38
2. Yellow solution

Put 3 ml H2O + 3 Drops I2 0,01 M in 8

3. Brown solution with time is 30
tube + 1 ml saliva
. second for 8 tube
Put 2 drops mixture (starch 1 % and NaCl
0.1 M into 8 tube
6. Test of determine suitable pH for activities saliva
No Activities Result
1. 10 ml buffer pH 9 + 2 ml NaCl 0,1 M Colorless solution
Colorless solution + 2 ml saliva than Yellow solution
 heated (38 )+ CH3COOH 5 drops +

iod 5 drops
2. Colorless solution
10 ml buffer pH 7 + 2 ml NaCl 0,1 M
Yellow solution
Colorless solution + 2 ml saliva than

3. heated (38 )+ iod 5 drops Colorless solution

10 ml buffer pH 5 + 2 ml NaCl 0,1 M Purple solution
colorless solution + 2 ml saliva than
4. Colorless solution
heated (38 )+ iod 5 drops
Blue solution
10 ml buffer pH 4 + 2 ml NaCl 0,1 M
Colorless solution + 2 ml saliva than

heated (38 )+ iod 5 drops

7. The effect of compounds which can destroying of bacteria in amylase saliva

No Activities Result
1. 2 ml saliva + 8 ml H2O Colorless solution
2. Tube 1
1 ml saliva + 5 drops toluene (let Colorless solution
stand 10 minutes)
Colorless solution + 5 ml starch Colorless solution and there are
3. Tube 2 bubble
1 ml saliva + 5 drops chloroform Colorless solution
(let stand 10 minutes)
Colorless solution + 5 ml starch Colorless solution and there are
bubble
Tube 3
4.
1 ml saliva + 5 drops HgCl2 1% (let stand
Colorless solution
10 minutes)
Colorless solution + 5 ml starch
 Tube 4 Colorless solution
5. 1 ml saliva + 5 drops phenol (let stand
10 minutes) Colorless solution
Colorless solution + 5 ml starch
Tube 5 Colorless solution
6. 1 ml saliva + 5 drops NaF (let stand 10
minutes) Colorless solution
Colorless solution + 5 ml starch
Tube 6 Colorless solution
7. 1 ml saliva + 5 drops water (let stand 10
minutes) Colorless solution
Colorless solution + 5 ml starch
Solution in each tube divided into two Colorless solution
8. part
Part 1 (tested with I2)
Tube 1 ( Toluene)
Tube 2 (chloroform) Yellow solution
Tube 3 (HgCl2)
Yellow solution
Tube 4 (Phenol)
Tube 5 (NaF) Brown solution
Tube 6 (water) Yellow solution
Part 2 (tested with reagent benedict) Yellow solution
Tube 1 ( Toluene) Yellow solution
Tube 2 (chloroform)
Tube 3 (HgCl2)
Tube 4 (Phenol) Blue solution
Tube 5 (NaF) Blue solution
Tube 6 (water) Blue solution
Blue solution
Blue solution
 Blue solution
B. Discussion
1. Tes Musin
Percobaan ini bertujuan untuk membuktikan adanya Musin dalam saliva.
Musin adalah suatu glikoprotein atau karbohidrat protein yang dikeluarkan ale klenjar
sublingual dan kelenjar submandibular. Percobaan ini saliva ditambahkan dengan
asam asetat, penambahan asam asetat berfungsi untuk mengendapkan musin dan juga
akan mendenaturasi protein dalam musin sehingga strukturnya menjadi tidak larut
dan mengendap dan endapannya berwarna putih. Kemudian untuk membuktikan
bahwa itu benar-benar musin. Maka endapan dipisahkan dengan filtrate. Kemudian
diuji dengan millon, pereaksi molisch dan pereaksi benedict (disaring jika ada
endapan ) tapi pada percobaan ini tidak disaring karena tidak terdapat endapan setelah
penambahan asam asetat.
Pengujian dengan pereaksi millon, pereaksi ini terbuat dari larutan merkuro
dan merkuri dalam HNO3. Tujuan dari pengujian ini adalah untuk mengetahui adanya
kandungan protein dalam saliva. Dari hasil percobaan diperoleh endapan putih. Hasil
ini menunjukkan uji positif adanya protein dalam saliva dan telah sesuai dengan teori
apabila pereaksi ini ditambahkan dengan protein akan menghasilkan endapan putih.
Adapun reaksi tang terjadi :

NH2 NH2
HO CH2 CH COOH + HgNO3 HgO CH2 CH COOH + HNO3
endapan Putih

Pengujian dengan pereaksi molisch, pereaksi ini terbuat dari alfa naftol dan
H2SO4. Tujuan dari pengujian ini adalah untuk mengetahui adanya karbohidrat dalam
saliva. Hasil yang diperoleh dari percobaan ini yaitu larutan keruh. Hasil yang
didapat tidak sesuai dengan teori dimana saliva mengandung amilase yang dapat
mengubah amilum menjadi glukosa atau suatu karbohidrat yang jika ditambah
dengan pereaksi molisch akan membentuk cicncin ungu. Terbentuknya cincin akibat
terjadinya reaksi kondensasi anara Furfural dan alfa naftol. Adapun reaksinya
adalah :

CH2OH OH O
O O
OH + H2SO4 C +
HO OH HOCH2 OH
O H
OH
hidroksi alfa naftol C C
metil O CH2OH
glukosa H
fulfural

cincin ungu

Pengujian dengan pereaksi benedict, pereaksi ini mengandung larutan

kuprisulfat, Na2CO3 dan natrium sulfat. Tujuan pengujian ini adalah untuk
mengetahui keberadaan gula pereduksi dalam saliva misalnya glukosa. Dari hasil
percobaan diperoleh larutan berwarna biru yang menandakan bahwa tidak adanya
gula pereduksi dalam saliva. Hasil yang diperoleh tidak sesuai dengan teori uji positif
menghasilkan endapan merah bata. Terbentuknya endapan disebabkan karena gula
pereduksi dapat mereduksi Cu2+ menjadi Cu+ yang kemudian mengendap manjadi
Cu2O. Adapun reaksinya adalah :

CH2OH CH2OH
O O O
OH + 2Cu2+
+ 5OH-
OH
C + Cu2O + 3H O
2
HO OH H endapan
HO
OH merah
OH bata
glukosa

2. Tes Tiosianat
Percobaan ini bertujuan untuk mengetahui adanya ion tiosianat (SCN -) dalam
saliva. Penambahan FeCl2 berfungsi untuk mengikat SCN- sedangkan HCl pekat
berfungsi sebagai katalis untuk mempercepat reaksi pengikatan SCN- oleh FeCl2.
Adapun reaksinya :
 2 SCN- + FeCl2 [Fe(SCN-)2] + 2 Cl-
Campuran kemudian ditambahkan dengan HgCl 0,1 N menghasilkan larutan
berwarna merah . HgCl berfungsi untuk membentuk senyawa kompleks [Hg(SCN) 4]2-
yang merupakan senyawa tak berwarna. Hasil yang diperoleh sesuai dengan teori
dimana menurut teori pada uji positif ion tiosianat menghasilkan larutan larutan
merah karena mengandung sianida yang bersifat meracun. Adapun persamaan
reaksinya adalah:
4 Fe (SCN)2 + 2 Hg2+ 2 [Hg(SCN)4]2- + 4 Fe2+
3. Tes Penyusun Senyawa Anorganik Saliva
Percobaan ini bertujuan untuk mengetahui adanya senyawa-senyawa
anorganik salam saliva, seperti Cl-, PO43-,SO42-, dan Ca2+. Penambahan asam asetat
bertujuan untuk mengendapkan musin, setelah penambahan asam asetat, larutan
dipanaskan dan terbentuk endapan putih. selanjutnya dilakukan penyaringan untuk
memisahkan endapan dan filtratnya. Setelah itu dilakukan pengujian terhadap
filtratnya. Filtrat dibagi 4 dan dilakukan uji terhadap ion Cl-, PO43-,SO42-, dan Ca2+.
a. Ion Cl-
Penambahan HNO3 encer pada percobaan ini adalah sebagai katalis sedangkan
sedangkan AgNO3 berfungsi mengikat ion Cl- membentuk endapan putih. Dari hasil
percobaan diperoleh larutan keruh dan terdapat endapan putih . Hasil yang diperoleh
sesuai dengan teori, seharusnya yang di peroleh yaitu terapat endapan putih. Adapun
reaksinya :
Cl- + AgNO3 AgCl + NO3
Endapan putih
b. Ion PO43-
Penambahan HNO3 berfungsi sebagai katalis. Amonium molibdat
[(NH4)2MoO4] berfungsi mengikat ion PO43- membentuk senyawa berwarna hijau,
senyawa berwarna hijau yang menandakan mengikat PO 43. Dari hasil percobaan
diperoleh larutan berwarna hijau hasil yang diperoleh telah sesuai dengan teori.
Adapun reaksinya yaitu:
PO43- + 3(NH4)2MoO4 + 23H+ (NH4)2P(MoO10)3 +12 H2O
c. Ion SO42-
Penambahan HNO3 berfungsi sebagai katalis sedangkan BaCl2 berfungsi
untuk mengikat ion SO42- membentuk endapan putih. Dari hasil percobaan diperoleh
larutan tak berwarna hasil yang diperoleh tidak sesuai dengan teori, seharusnya
terbentuk endapan putih. Adapun reaksinya yaitu:
SO42- + BaCl2 + HNO3 BaSO4 + HNO3 + 2Cl-
endapan putih
d. Ion Ca2+
Penambahan NH4C2O4 berfungsi untuk mengikat Ca2+ menjadi CaC2O4 yang akan
mengendap membentuk endapan putih yang menandakan dalam larutan terdapat ion
Ca2+. Hasil yang diperoleh larutan keruh dan terdapat endapan putih, hasil yang
diperoleh sesuai teori. Adapu reaksinya yaitu :
Ca2+ + NH4C2O4 CaC2O4 + NH4
Endapan putih
4. Tes Pengaruh temperature terhadap aktivitas ptyalin
Percobaan ini bertujuan untuk mengetahui suhu optimal sehingga enzim
ptyalin (amilase) dapat bekerja maksimal ada 4 perlakuan pada percobaan ini yaitu
pada suhu kamar , pada air es, pada suhu 38 oC dan didihkan saliva akan bereaksi
dengan pati. Saliva akan menghidrolisis pati menjadi maltose maupun glukosa.
Reaksi hidrolisis tersebut akan berlangsung secara optimal pada suhu tertentu untuk
mengidentifikasi ada tidaknya pati yang terhidrolisi oleh enzim ptyalin maka
ditambahkan I2 yang akan bereaksi dengan amilum membentuk kompleks berwarna
biru tua. I2 berperan sebagai pemberi warna yang akan merubah barna biru menjadi
bening yang menandakan bahwa amilum telah terhidrolisis.
Namun pada percobaan ini semua larutan berwarna biru dan tidak terjadi
perubahan setelah penambahan I2 Hasil yang diperoleh tidak sesuai dengan teori
dimana pada suhu kamar enzim ptyalin bekerja dengan normal, pada suhu 38 oC
enzim ptyalin bekerja dengan baik sehingga dapat menghidrolisis pati menjadi
glukosa dan maltose. Pada suhu rendah ptyalin bekerja kurang baik karena enzin
ptyalin blum aktiv sehingga membutuhkan waktu yang lebih lama untuk
menghidrolisis pati. Sedangkan ada suhu tinggi enzim ptyalin tidak dapat mengubah
ataupun dapat menghidrolisis amilum menjadi glukosa dan maltose karena pada suhu
tersebut (suhu tinggi) enzim ptyalin akan rusak.
5. Tes Estimasi Ptialin
Percobaan ini bertujuan untuk mengetahui kemampuan enzim ptyalin
mengubah amilum menjadi maltose. Saliva yang digunakan ditempatkan pada suhu
38oC terlebih dulu untuk mencapai tingkat optimum kerja enzim tersebut. Selanjutnya
dilakukan penambahan NaCl 0,1 M untuk menghambat proses pemecahan pati
sehingga menghambat reaksi pati dengan enzim ptyalin (amilase). NaCl merupakan
inhibitor nonkompetitif terhadap pati. Pengukuran waktu aktivasinya dilakukan
dengan menambahkan iod pada samper dengan interval waktu 30 detik. Hal ini untuk
mengetahui aktivasi enzim melalui hasil hidrolisisnya dimana warnanya akan berubah
dari biru tua menjadi cokelat yang menandakan semakin banyak pati terhidrolisis .
Berdasarkan hasil pengamatan didapat larutan keruh dan urutan tabung yang cepat
berubah warnanya yaitu mulai dari warna biru kurang pekat sampai pekat sekali
antara lain : Tabung 4,2,3,6,7,1,5, dan 8. Hal ini berarti semakin banyak waktu yang
digunakan maka kerja enzim semakin lambat. Ini tidak sesuai dengan teori yang
menyatakan bahwa titik akromatik berada pada 60 detik. Dimana warna biru hilang
dan hanya cokelat yang merupakan iod bebas.
6. Tes penentuan pH yang cocok untuk kerja saliva
Percobaan ini bertujuan untuk mengetahui pH yang sesuai sehingga amilase
dapat bekerja optimal. Pada percobaan ini digunakan larutan buffer 4, 5, 7, dan 9.
Keempat larutan buffer ditambah larutan pati 1%, NaCl, dan saliva encer. Pati
meripakan bahan atau pereaksi, sedangkan NaCl berfungsi menghambat hidrolisis
pati. Kemudian larutan dengah pH 7 dan 9 diasamkan dengan CH3COOH, harus
diasamkan agar reaksi I2 dapat berlangsung karena I2 bekerja pada pH asam. Pada
larutan pH 4 menghasilkan warna biru, larutan buffer 5 menghasilkan warna ungu,
larutan buffer 7 menghasilkan warna hijau (bening), larutan buffer 9 menghasilkan
larutan tak berwarna. Hasil yang didapat ini tidak sesuai dengan teori, menurut teori
pada pH 7 dan suhu 380C larutan menjadi bening karena pada pH dan suhu tersebut,
enzim amilase bekerja optimal mampu menghidrolisis amilum menjadi maltosa,
penambahan iod setelah dipanaskan pada suhu 380C berfungsi sebagai indikator
7. Tes Efek Senyawa yang menghambat/ menghancurkan aktivitas bakteri pada
amilase saliva
Percobaan ini bertujuan untuk mengetahi senyawa atau zat yang dapat
menghambat aktivitas bakteri pada amilase saliva. saliva ditambahkan toluen, fenol
2%, kloroform, HgCl2 1%, NaF dan air pada tabung yang berbeda. Selanjutnya saliva
ditambahkan pati sehingga amilase dapat berkerja menghidrolisis pati. Mempercepat
reaksinya maka tiap tabung dipanaskan. kemudian setiap tabung di bagi 2 bagian.
Bagian pertama dengan pereaksi benedict dan bagian kedua diuji dengan iod. dilihat
dari tingkat keasamannya, semakin asam suatu larutan menyebabkan amilase tidak
bekerja. Kemudian untuk uji dengan iod hasil yang diperoleh untuk toluene,
kloroform, dan air menghasilkan warna merah cokelat dan HgCl 3, fenol, dan NaF
larutan berwarna biru. Hal ini berarti HgCl3, fenol dan NaF diperoleh warna biru
untuk uji iod yang berarti pati tidak dapat terhidrolisis dan warna biru pada uji
benedict yang menandakan tidak ada gula pereduksi. Sehingga disimpulkan HgCl 3,
Fenol yang dapat menghambat kerja enzim amilase.

CHAPTER V
CONCLUSSION AND SUGGESTION

A. Conclussion
Based on experiment the conclusion is:
a. Based on test effect temperature toward enzym activity that optimum
temperature of saliva is 380C.
b. Based on mucin test there is mucin in saliva, and there is no carbohydrate, and
reducing sugar.
c. Based on experiment there is SO42- and Cl- that is there is white precipitate and
 negative result for Ca2+ and PO43- that produce colorless solution.
d. Based on the thiocyanate test there is no thiocyanate in saliva that test
e. pH that suitable for saliva activity is 5,7 and 9
f. Based on experiment the compound that iterference of bacteria activities is
HgCl2 and phenol
B. Suggestion
a. Hopefully apprentice must check the reagent condition in lab
b. Must be carefull in mixturing the solution.

DAFTAR PUSTAKA

Almeida,Patricia Del Vigna. 2015, Saliva Composition And Function: A

Comprehensive Review. Journal of Contemporary Dental Practice.Vol.9.
No.3

Chrishtopher. 2012, Anatomy And Function Saliva. Boston: Spinger

Degiempietro,P and E.Pehein. 1987, Determination of Thicyanate and Saliva Without

Deproteinisation and Its Validation as Smoking Parameter. Journal of
Determination of Thicyanate.Vol.25.No.10
Extrom, Jorgen. 2012, Saliva and The Control of its Secretion. Italy: Pharmacologi
Gritzman,Noberts S. 2001, Sonography of the Salivary Glands and Soft
Tissue of the Neck. Journal of Uropean Course Book. Vol.2No.5

Luepker, Russellv. 1981, Saliva Thicyanate: A Chemical Indicator of agarette

Smoking in adolescent. Journal of Scientific Product. Vol7.No.12

Paul, Buddhap. 2006, Cyanide and Thiocyanate in Human Saliva by Gas

Chromatography mass Spectrometry. Journal of Analuytical Toxicology.
Vol.3

Prashaanti,N. 2016, A Study on Association of Salivary Calcium and Phosphate in

Oval heat. Journal of Pharmaceutical Science and Research. Vol.8.No.7

Rodillas, Gae Khail. 2005, Effect Temperature and pH on Enzymatic Activity of

Salivary Amylase. Journal of Scientific. Vol.3.No.1

Rosen, Federick.S. 2001, Anatomy and Physology of the Salivary Glands. Journal of
Anatomy.Vol.6.No.2

Sharma,Rakers. 2012. Enzyme Inhibition and Bioaplication.China: Intechopen

JOURNAL OF EXPERIMENT
Tittle of Experiment : Saliva
th
Day, Date of Experiment : Monday , November 2016
Name : Jumriana Jufri
Register Number : 1413440011
Class/ Group : ICP A of Chemistry/ II
Members : 1. Andi tenri Tayu
2. Khairul Azhar
3. Yuli Ratnadilla
 Assistant : Marwah Karim

A. PURPOSE OF EXPERIMENT
6. To know the organic compound that contain in saliva
7. To know the inorganic compound that contain in saliva
8. To know the influence temperature againt phtialin activities.
9. To know the pH that correct to saliva.
10. To know the effect of compound the activities of bactery on analyze saliva
B. APPARATUS AND CHEMICALS
1. Apparatus
a. Small Test Tubes 15 pieces
b. Small tube rack 2 pieces
c. Test tube rack 2 pieces
d. Volumetric glass 10 ml, 25 ml @ 2 pieces
e. Dropped pipette 5 pieces
f. Baker glass 250 ml 1 piece
g. Funnel 2 pieces
h. Big test tube 6 pieces
i. Spray bottle 1 piece
j. Asbestos gauze 1 piece
k. Spritus 1 piece
l. Tripod 1 piece
m. Kasa asbes 1 piece
n. Stopwatch 1 piece
o. Stir bar 1 piece
p. Wood clamp 1 piece
3. Chemicals
a. Saliva
b. Acetic acid 0.1 M and 2 N (CH3COOH)
c. Millon reagent
 d. Benedict reagent
e. Molisch reagent
f. Iron (III) chloride 0.1 M (FeCl3)
g. Hydrochloric acid concentrated (HCl(p))
h. Mercurium (I)chloride (HgCl)
i. Amylum
j. Sodium chloride 0.1 M (NaCl)
k. Iod solution (I2)
l. Buffer solution
m. Aquadest (H2O)
n. Phenol (C6H5OH)
o. Nitrate acid diluted (HNO3)
p. Silver nitrate (AgNO3)
q. Filter paper
r. Tissue
s. Matches
t. Ice
u. Label
v. Toluene (C6H5CH3)
C. WORK PROCEDURE

1. Mucin test

acetate acid
0,1 M
separate
the
precipitate Separate the precipitate and test
H2O as a Millon, Benedict and Mollish
control
5 mL saliva
 2. Thiocianate test

5 mL saliva
5 drops FeCl 1 drop HCl 5 drops HgCl
0,1 M concentrated 1%

used H2O as
control
5 mL saliva

3. Inorganic test

acetate acid 2
N until the
solution heated the filtered
turbid solutinon
15 mL saliva

a. To test Cl- ion

Acidity with
HNO3 solution
some drops
AgNO3 0,5
M

3 mL filtrate

b. To test PO43- ion

 Acidity with 1 mL
HNO3 solution amonium
molidbat

Heated

3 mL filtrat

c. To test SO42- ion

Acidity with
1 mL of
HNO3 solution
BaCl2 5%

3 mL filtrate

d. To test Ca++ ion

1 mL of NH4-
oxalate 4%

After an hour and then take a good look

because of the possibility of sediment
formed little

3 mL filtrate
 4. Test the effect of temperature in ptyalin activity

5 mL pati 5 mL pati 5 mL pati 5 mL pati

solution 1% solution 1% solution 1% solution 1%

1 2 3 4

first second thirt

2
1 3
with temperature
380C

at the temperature
at ice water 250C
 2 drops saliva solution 2 drops saliva solution

1
2 3
with temperature 380C

at ice water
at the temperature
250C

2 drops saliva test with 2 drops I2

have a boil
solution 0,01 M

every 5 minute grab Record the speed of the

4 sample from each tube starch solution into each
tube

5. Ptialin test estimation

2 mL NaCl 0,1
M

with temperature
380C

10 mL starch
solution 1%
 3 mL of water

1 2 3 4 5 6 7 8

3 drops I2 0,01M

1 2 3 4 5 6 7 8

2 drops of starch mix saliva

1 mL saliva
solution

1 2 3 4 5 6 7 8

10 mL strach
solution 1%
 record the time when the addition of a mixture
starch saliva does not cause discolaration

6. Test determine suitable pH for saliva work

Larutan-larutan buffer

5 mL larutan pati 1% 10 mL 10 mL 10 mL 10 mL 10 mL

1 2 3 4 5

1 2 3 4 5 pH 8 pH 7,4 pH 6,8 pH 6 pH 5,2

2 mL NaCl 0,1 M
2 mL saliva encer

1 2 3 4 5 1 2 3 4 5
 Temperature
380C I2 solution

Determine the tube where the

color disappears reached
24 24

7. The effect of compound that inhibitor/breakdown the activity bacteria in

amylase of saliva

2 mL saliva 8 mL water mixture

solutio
1 mL saliva cair
n

1 2 3 4 5 6 7
 5 drops 5 drops 0,5 mg
5 drops 5 drops 5 drops
1% fenol NaF
toluene CCl4 HgCl2 H2O
2%

1 2 3 4 5 6

for each tube is 5 mL starch solution1%

let stand for 10 minutes occasionally shaken

1 2 3 4 6
5

1 2 3 4 5 6

each tube is devide into two parts

Temperatur
380C

Selama 1 2 3 4 6
15 menit 5
24
 each tube in part 1 added I2solution

1 2 3 4 6
5

each tube in part 2 added Benedict

2 3 4 5 6
record the observation result
foe aech tube

Makassar, 19 November 2016

Assistant Apprentice

Marwah Karim Jumriana Jufri

JOURNAL OF EXPERIMENT
 Tittle of Experiment : Saliva
th
Day, Date of Experiment : Monday , November 2016
Name : Jumriana Jufri
Register Number : 1413440011
Class/ Group : ICP A of Chemistry/ II
Members : 1. Andi tenri Tayu
2. Khairul Azhar
3. Yuli Ratnadilla
Assistant : Marwah Karim

D. PURPOSE OF EXPERIMENT
11. To know the organic compound that contain in saliva
12. To know the inorganic compound that contain in saliva
13. To know the influence temperature againt phtialin activities.
14. To know the pH that correct to saliva.
15. To know the effect of compound the activities of bactery on analyze saliva
E. APPARATUS AND CHEMICALS
2. Apparatus
q. Small Test Tubes 15 pieces
r. Small tube rack 2 pieces
s. Test tube rack 2 pieces
t. Volumetric glass 10 ml, 25 ml @ 2 pieces
u. Dropped pipette 5 pieces
v. Baker glass 250 ml 1 piece
w. Funnel 2 pieces
x. Big test tube 6 pieces
y. Spray bottle 1 piece
z. Asbestos gauze 1 piece
aa. Spritus 1 piece
ab. Tripod 1 piece
 ac. Kasa asbes 1 piece
ad. Stopwatch 1 piece
ae. Stir bar 1 piece
af. Wood clamp 1 piece
4. Chemicals
w. Saliva
x. Acetic acid 0.1 M and 2 N (CH3COOH)
y. Millon reagent
z. Benedict reagent
aa. Molisch reagent
ab. Iron (III) chloride 0.1 M (FeCl3)
ac. Hydrochloric acid concentrated (HCl(p))
ad. Mercurium (I)chloride (HgCl)
ae. Amylum
af. Sodium chloride 0.1 M (NaCl)
ag. Iod solution (I2)
ah. Buffer solution
ai. Aquadest (H2O)
aj. Phenol (C6H5OH)
ak. Nitrate acid diluted (HNO3)
al. Silver nitrate (AgNO3)
am. Filter paper
an. Tissue
ao. Matches
ap. Ice
aq. Label
ar. Toluene (C6H5CH3)
F. WORK PROCEDURE
 2. Mucin test

acetate acid
0,1 M
separate
the
precipitate Separate the precipitate and test
H2O as a Millon, Benedict and Mollish
control
5 mL saliva

3. Thiocianate test

5 mL saliva
5 drops FeCl 1 drop HCl 5 drops HgCl
0,1 M concentrated 1%

used H2O as
control
5 mL saliva

4. Inorganic test

acetate acid 2
N until the
solution heated the filtered
turbid solutinon
15 mL saliva

e. To test Cl- ion

Acidity with
HNO3 solution
some drops
AgNO3 0,5
M

3 mL filtrate
f. To test PO43- ion

Acidity with 1 mL
HNO3 solution amonium
molidbat

Heated

3 mL filtrat

g. To test SO42- ion

Acidity with
1 mL of
HNO3 solution
BaCl2 5%

3 mL filtrate

h. To test Ca++ ion

1 mL of NH4-
oxalate 4%

After an hour and then take a good look

because of the possibility of sediment
formed little

3 mL filtrate
 5. Test the effect of temperature in ptyalin activity

5 mL pati 5 mL pati 5 mL pati 5 mL pati

solution 1% solution 1% solution 1% solution 1%

1 2 3 4

first second thirt

2
1 3
with temperature
380C

at the temperature
at ice water 250C
 2 drops saliva solution 2 drops saliva solution

1
2 3
with temperature 380C

at ice water
at the temperature
250C

2 drops saliva test with 2 drops I2

have a boil
solution 0,01 M

every 5 minute grab Record the speed of the

4 sample from each tube starch solution into each
tube

6. Ptialin test estimation

2 mL NaCl 0,1
M

with temperature
380C

10 mL starch
solution 1%
 3 mL of water

1 2 3 4 5 6 7 8

3 drops I2 0,01M

1 2 3 4 5 6 7 8

2 drops of starch mix saliva

1 mL saliva
solution

1 2 3 4 5 6 7 8

10 mL strach
solution 1%
 record the time when the addition of a mixture
starch saliva does not cause discolaration

7. Test determine suitable pH for saliva work

Larutan-larutan buffer

5 mL larutan pati 1% 10 mL 10 mL 10 mL 10 mL 10 mL

1 2 3 4 5

1 2 3 4 5 pH 8 pH 7,4 pH 6,8 pH 6 pH 5,2

2 mL NaCl 0,1 M
2 mL saliva encer

1 2 3 4 5 1 2 3 4 5
 Temperature
380C I2 solution

Determine the tube where the

color disappears reached
24 24

8. The effect of compound that inhibitor/breakdown the activity bacteria in

amylase of saliva

2 mL saliva 8 mL water mixture

solutio
1 mL saliva cair
n

1 2 3 4 5 6 7
 5 drops 5 drops 0,5 mg
5 drops 5 drops 5 drops
1% fenol NaF
toluene CCl4 HgCl2 H2O
2%

1 2 3 4 5 6

for each tube is 5 mL starch solution1%

let stand for 10 minutes occasionally shaken

1 2 3 4 6
5

1 2 3 4 5 6

each tube is devide into two parts

Temperatur
380C

Selama 1 2 3 4 6
15 menit 5
24
 each tube in part 1 added I2solution

1 2 3 4 6
5

each tube in part 2 added Benedict

2 3 4 5 6
record the observation result
foe aech tube

Makassar, 19 November 2016

Assistant Apprentice

Marwah Karim Jumriana Jufri

 DOCUMENTATION

1. Test mucin 2. Test thiocyanate

3.Test inorganic compounds in saliva 4. Effect temperature for enzyme activity

5. Test estimation ptialyn

6. Test of pH determination that proper to saliva activity

7. effect compound that advoid the activity of bactery amylase saliva