APPROVAL SHEET

Complete report of Biochemistry Experiment, which entitled “Saliva”

arranged by :
name : Wiwiana
reg.number : 1513441007
class/group : Chemistry Education of ICP /IV
it has been checked by Assistant/ Assistant Coordinator so this report was
accepted.

Makassar, November 2017

Assistant Coordinator Assistant

Muhammad Iqbal Hidayat Uswatun Saidah A. Thalib S.Pd

ID : 1313441008

Known by,
Responsilbility Lecturer

Dra. Halimah Husain, M.Si

ID : 19641020 1990003 1 002
A. TITLE OF EXPERIMENT
Saliva
B. AIM OF EXPERIMENT
1. To test musin in saliva
2. To test tiosianat in saliva
3. To test anorganic compound in saliva
4. To test influence of temperatur at ptialin activity
5. To test Ptialin estimation
6. To determain pH that agree to saliva work
C. LITERATUR REVIEW
The hydrolysis of starch by salivary and pancreatic amylases catalyze
random hydrolysis of α(1→4) glycoside bonds, yielding dextrins, then a mixture
of glucose, maltose, and isomaltose (from the branch points in amylopectin). The
disaccharidases maltase, sucrase-isomaltase (a bifunctional enzyme catalyzing
hydrolysis of sucrose and isomaltose), lactase, and trehalase are located on the
brush border of the intestinal mucosal cells where the resultant monosaccharides
and others arising from the diet are absorbed. In most people, apart from those of
northern European genetic origin, lactase is gradually lost through adolescence,
leading to lactose intolerance. Lactose remains in the intestinal lumen, where it is
a substrate for bacterial fermentation to lactate, resulting in discomfort and
diarrhea (Murray, 2003 : 475). Saliva is speichel, Human saliva consists tp 98% of
water (Lackner, 2014 : 245).
Besides the skin, wound repair occurs in the gastronintestinal tract, in
muscle tissue following injury, and at sites of chronic inflammation, to name just
a few common examples. Primary sources of epithelial growth factors include the
submaxillary salivary gland, the duodenal Brunner’s gland, epithelial cells in the
mammary gland, and the kidneys. Growth factors are released into bodily fluids
saliva, serum, milk, and urine and travel to sites of injury and growth. Epithelial
cells at many places in the body are continuously renewed and growth factors are
released locally near these sites as well. In these situations the growth factors
operate in autocrine and paracrine manners rather than through an endocrine
mechanism (Beckerman, 2005 : 251).
Sample from multiple site presented variation in analytes results requiring
each sample to be treated as separate entity. Again requirement of skilled hand,
expensive equipment and time factor are added limitation. Collection of saliva is
less demanding and non invasive technique offers an alternative choice. Saliva is
derived from serum, gingival fluid and mucosal transudate. Whole saliva
represents a pooled sample of periodontal sites and useful for overall assessment
of Periodontitis or its risk status (Trivedi, 2012).
In most people, apart from those of northern European genetic origin,
lactase is gradually lost through adolescence, leading to lactose intolerance.
Growth factors are released into bodily fluids saliva, serum, milk, and urine and
travel to sites of injury and growth.
Atropine and scopalamine Atropine is found in the berries of the weeds
deadly nightshade and black nightshade. It is also synthesized in the leaves and
roots of Hyoscyamus muticusi. At high concentrations it is poisonous but, when
more dilute, displays a number of beneficial medical applications. Atropine
sulphate solutions (1%, w/v) are used in ophthalmic procedures to dilate the pupil.
It is also sometimes used as a pre-anaesthetic, as it inhibits secretion of saliva and
mucus in the respiratory tract, which protects the patient from
bronchoconstriction. Its ability to inhibit gastric secretion underlines its occasional
use in the treatment of some stomach ulcers. Scopalamine, found in the leaves of
the plant Hyoscyamus niger, shares some properties with atropine. Its major
medical use is to treat motion sickness (Walsh, 2003 : 29).
Saliva is responsible for the initial digestion of starch, favoring the
formation of the food bolus.(9,8) This action occurs mainly by the presence of the
digestive enzyme α-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,(20) 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.
Since several factors can influence salivary secretion and composition a precise
standard for saliva collection must be established. Such a standard would make
the test results obtained through sialometry and/or sialochemistry more helpful in
characterizing the true functional state of the salivary glands which in turn would
serve as indicators for a diagnosis when oral and/or systemic alterations are
suspected (Mrthykumar, 2013).
Glucose and galactose are absorbed by a sodium-dependent process. They
are carried by the same transport protein and compete with each other for
intestinal absorption. Other monosaccharides are absorbed by carrier-mediated
diffusion. Because they are not actively transported, fructose and sugar alcohols
are only absorbed down their concentration gradient, and after a moderately high
intake some may remain in the intestinal lumen, acting as a substrate for bacterial
fermentation. Hydrolysis of triacylglycerols is initiated by lingual and gastric
lipases that attack the sn-3 ester bond, forming 1,2-diacylglycerols and free
fatty acids, aiding emulsification. Pancreatic lipase is secreted into the
small intestine and requires a further pancreatic protein, colipase, for
activity (Murray, 2003 : 475).
Other monosaccharides are absorbed by carrier-mediated diffusion.
Because they are not actively transported, fructose and sugar alcohols are only
absorbed down their concentration gradient, and after a moderately high intake
some may remain in the intestinal lumen, acting as a substrate for bacterial
fermentation.
The proposed method does not require any laborious clean up
procedure before measurement. In addition, the method has wider linear
dynamic range with good accuracy and precision. The method shows no
interference from the common excipients and additives. This may help in
analyzing affectivity of this drug in human beings during treatment. Therefore, it
is concluded that the proposed method is simple, sensitive and rapid for the
determination of in bulk, pharmaceutical formulations and in human urine and
salivary samples (Kumar, 2011).
Salivary diagnostics is an emerging field in Proteomics. It uses salivary
proteome for the prognosis, diagnosis and management of periodontal diseases.
Saliva composed of water, electrolyte and organic molecules like amino acids,
peptides, proteins, glycoprotein and glycolipid is derived from local vasculature
originating from carotid arteries. Saliva contains biomarkers derived from serum,
gingival crevicular fluid and mucosal transudate. Many analytes associated with
periodontal diseases have been detected in this bio-fluid (Trivedi, 2012).
The results suggest that the sense of depression, as well as living
environments and lifestyle habits which induce it, increase salivary melatonin
levels for two possible reasons: anxiety increases oxidative stress in the body20
and therefore a larger volume of melatonin is produced as its antioxidant action
effectively protects the body7; and the effects of melatonin, as a ligand, on its
receptors decrease. With the aim of promoting the application of salivary
melatonin to preventive medicine, its relationships with psychiatric conditions
will be examined (Ito, 2013).
Salivary diagnostics is an emerging field in Proteomics. It uses salivary
proteome for the prognosis, diagnosis and management of periodontal diseases.
the proposed method is simple, sensitive and rapid for the determination of in
bulk, pharmaceutical formulations and in human urine and salivary samples.
Saliva. The salivary glands produce a slightly alkaline secretionwhich in
addition towater and salts contains glycoproteins (mucins) as lubricants,
antibodies, and enzymes. Amylase attacks polysaccharides, and a lipase
hydrolyzes a small proportion of the neutral fats. Amylase and lysozyme, a
murein- cleaving enzyme, probably serve to regulate the oral bacterial flora rather
than for digestion. Amylase, the most important endoglycosidase in the pancreas,
catalyzes the hydrolysis of α14 bonds in the polymeric carbohydrates starch and
glycogen. This releases maltose, maltotriose, and a mixture of other
oligosaccharides (Koolman, 2005).
 Saliva acts as a buffering system. The buffering effect of saliva is largely
due to bicarbonate/carbonate ions, and to a lesser extent to phosphate-ions and
proteins present in saliva, neutralizing acids ingested or produced by
microorganisms in the mouth. The buffer activity is usually assessed immediately
after collection of saliva sample. Saliva is an accessible fluid that can easily be
collected by the patient. Advantages of saliva testing sample are easy and non
invasive collection procedure that is neither painful nor traumatic. Saliva is
reliable for early detection of certain diseases and monitoring the disease course in
the conjunction with treatment and detection of addictive drugs (Nirmala, 2013).
Salivary amylase is a highly abundant protein in saliva. The highest
concentration of amylase was found in the saliva of the parotid gland and palatine
minor salivary glands. The most widely-known function of amylase is
endoglicosidase activity. Splitting the α-1,4-glicosidic bindings of several glycans,
such as starch (amylopectin), amylase produces oligosaccharides (dextrin)
disaccharides (maltose, isomaltose) and monosaccharide glucose. Amylase
also binds bacteria including certain bacterial piliwhich are important factors of
bacterial adhesion. Thus, amylase promotes bacterial adhesion to the
hydroxyapatite surfaces of teethwhich can lead to both advantageous
surface immune exclusion on the one hand and disadvantageous adhesion
of cariogenic or periodontopathogenic bacteria onto tooth surfaces on the other
hand (Fabian, 2012).
There has been growing interest in diagnosis based on analysis of saliva
due its simple and non-invasive collection procedure. Since saliva collection is
inherently painless, practitioners will be able to diagnose and monitor the patients’
wellbeing more frequently while approaching the sensitivity and specificity of a
blood test. Plus, the collection is also cheap and safe for both the practitioner and
patient. With that, patients especially from developing countries can be diagnosed
with minimal costs but great precision. Comprehensive analysis of human salivary
proteome through various methods is indeed necessary in order to find the most
reproducible markers for later utility in acute cardiac care. It is projected that
salivary diagnostics will be conclusive such that fewer diagnostic biomarkers can
be determined with immediate results, thus greatly improving the life quality of
patients. In the long run, more available saliva collection kits developed by
manufacturing companies will be able to aid and eventually provide solid
biomarker findings in future (Rahim, 2015).
Saliva is an accessible fluid that can easily be collected by the patient.
Advantages of saliva testing sample are easy and non invasive collection
procedure that is neither painful nor traumatic. Amylase also binds bacteria
including certain bacterial piliwhich are important factors of bacterial adhesion.
D. APPARATUS AND CHEMICALS
1. Apparatus
a. Test tube (20 units)
b. Tube rack (2 units)
c. Funnel (1 unit)
d. Measure glass 10 ml (2 units)
e. Measure glass 25 ml (2 units)
f. Beaker glass 50 ml (1 unit)
g. Beaker glass 250 ml (1 unit)
h. Beaker glass 600 ml (1 unit)
i. Spray bottle (1 unit)
j. Tripod (1 unit)
k. Gauze (1 unit)
l. Bunsen burner (1 unit)
m. Drop pipette (20 units)
n. Thermometer 100℃ (1 unit)
o. Stir bar (1 unit)
p. Stopwatch (1 unit)
q. Tube clamp (3 units)
r. Rough cloths (1 unit)
s. Soft cloth (1 unit)
2. Chemicals
a. Saliva
b. Barium chloride solution 5% (BaCl2)
c. Ammonium oxalate solution 4% ((NH4)2C2O4)
d. Acetate acid solution 2N dan 0,1 N (CH3COOH)
e. Sodium chloride solution 0,1 M (NaCl)
f. Chloride solution concentrated (HCl)
g. Mercury I chloride solution 1% (HgCl)
h. Ferri chloride solution 0,1 M (FeCl)
i. Sodium fluoride solution (NaF)
j. Iodium solution 0,01 M (I2)
k. Silver nitrate solution (AgNO3)
l. Phenol solution 2% (C6H5OH)
m. Nitrate acid solution (HNO3)
n. Ammonium Molybdate (NH4)6Mo7O24)
o. Chloroform (CHCl3)
p. Starch solution 1% ((C6H12O6)n)
q. Toluene (C6H5CH3)
r. Aquadest (H2O)
s. Buffer solution (pH 4, 5, 7, 9)
t. Millon Reagent
u. Benedict Reagent
v. Molisch Reagent
w. Tissue
x. Matches
y. Filter paper
E. WORK PROCEDURE
1. 1 Mucin Test
a. 5 mL of saliva was put into the beaker
b. Solution was added by 2 drops of asetic acid 0.1 M
c. Solution was filtrated
d. Filtrated putted into the three react tube
e. Used water as the control
f. The solution was filtered, and filtrate was test 3 drops by benedict, Molisch and
Millon reagent
g. Used water as the control
2. Thyocyanate test
a. 5 mL of saliva was put into the reaction tube
b. Solution was added by 5 drops of FeCl2 0.1 M and 1 drop of concentrated HCl.
c. Solution was added by 5 drops of HgCl 1 %
d. Used water as the control
3. Test Compilers of Saliva Inorganic Compounds
a. 15 mL saliva was added by asetic acid 2 N drops by drops until the solution
turbid
b. Mixture was heated until its boiled
c. Mixture was filtered and tested
a. To test Cl- ion, acidified the filtrate by dilute HNO3 and added 3 drops of
AgNO3 0.5 M
b. To test PO43-, acidified the filtrate by dilute HNO 3, then added by 1 mL of
Ammonium molybdate and heated
c. To test SO42- ion, acidified the filtrate by dilute HNO 3, and added 1 mL by
BaCl2 5%
d. To test Ca2+, the filtrate was added 1 mL of Ammonium oxalate 4%
4. Test the effect of Temperature on Ptyalin Activity
a. 5 mL starch solution was added into four reaction tubes,
b. First tube was put into the ice, second tube was put in room temperature, third
tube was put in water bath with temperature 38oC
c. In each tube added by 2 drops of dilute saliva (1:9) and mixed well
d. In to the fourth tube added by 2 drops of dilute saliva which has been dilute in
water bath as long as 5 minutes
e. In every 5 minutes intervals, the sample was took in every tube and test by 2
drops of I2 solution 0.01 M.
f. The breakdown speed of starch was noted
5. Test of Ptyalin Estimation
a. 10 mL of starch solution 1% added by 2 mL of NaCl 0.1 M
b. Mixture was heated in 38oC.
c. 8 tubes was prepared and filled with 3 mL of water and 3 drops of I 2 solution
0.01 M.
d. 1 mL of dilute saliva was added into the starch solution, after 30 second 2
drops of mixture was added into I2 solution
e. The time was noted when addition of mixture towards I 2 solution doesn’t occur
the color changes.
6. A pH-determination Test Suitable for salivary Work
a. 10 mL of buffer solutions (pH 4, 5, 9) was prepared, and added by 5 mL of
starch solution 1%, 2 mL of NaCl 0.1 M and 2 mL of dilute saliva (1:9).
b. Tube was heated in water bath with temperature 38oC
c. In each tube was added by 3 drops of I2 solution
7. The compound Effect face/ destroy bacteria Activities in Salivary Amylase
a. 2 mL saliva was added by 8 mL of water
b. Solution was stirred well, added 1 mL of dilute saliva into the 7 reaction tube
c. First tube was added by 5 drops of tholuene
d. Second tube was added by 5 drops of CHCl3
e. Third tube was added by 5 drops of HgCl2 1 %
f. Fourth tube was added by 5 drops phenol
g. Fifth tube was added by 0.5 mg NaF
h. Sixth tube was added by H2O
i. Let the solution in 10 minutes and shaked the solution
j. In each tube was added by 5 mL of starch solution 1%
k. Heated all the tube as long as 15 minutes
l. Every solution in each tube was divided into two parts
m. Into the first part was added by I2.
n. Every tube was observed and noted the changes that occur
F. OBSERVATION RESULT
1. Mucin Test
No Activity Result
1. 5 mL of saliva (colorless) + 2 drops Colorless
CH3COOH 0.1 M (colorless)
2. Filtered the solution Colorless
3. Saliva solution + 3 drops Benedict Blue solution
Saliva solution + 3 drops Molisch Colorless solution
Saliva solution + 3 drops Millon Turbid
1. 5 mL of H2O (colorless) + 2 drops Colorless
CH3COOH 0.1 M (colorless)
2. Filtered the solution Filter
3. Saliva solution + 3 drops Benedict Blue
Saliva solution + 3 drops Molisch Colorless solution
Saliva solution + 3 drops Millon Colorless solution

2. Thyocyanate Test
No Activity Result
1. 5 mL of saliva + 5 drops FeCl3 0.1 M Clear solution
2. + 1 drop HCl Orange
3. + 5 drops HgCl2 1 %
1. 5 mL of H2O + 5 drops FeCl3 0.1 M Colorless solution
2. + 1 drop HCl Colorless solution
3. + 5 drops HgCl2 1 % Colorless solution

3. A pH-determination Test Suitable for salivary Work

No Activity Result
1. 10 mL buffer solution (pH 4) → Colorless solution
added 2 mL NaCl 0.1 M + 5 mL
starch solution 1 % → Colorless
Heated with 38oC temperature of
water → Colorless
Purple
2. React with 3 drops I2 0.01 M
10 mL buffer solution (pH 5) → Colorless
added 2 mL NaCl 0.1 M + 5 mL
starch solution 1 % → Colorless

Heated with 38oC temperature of Colorless

water → Purple solution

3. Colorless
React with 3 drops I2 0.01 M
10 mL buffer solution (pH 7) →
added 2 mL NaCl 0.1 M + 5 mL Colorless

starch solution 1 % →
Colorless
Heated with 38oC temperature of
Purple
4. water →
Colorless
React with 3 drops I2 0.01 M
10 mL buffer solution (pH 9) →
Colorless
added 2 mL NaCl 0.1 M + 5 mL
Colorless
starch solution 1 % →
Heated with 38oC temperature of
Purple
water →
React with 3 drops I2 0.01 M

4. Test of Ptyalin Estimation

No Activity Result
.
1. 10 mL saliva + 2 mL NaCl 0.1 M → Colorless
put in hot plate 38oC
2. 3 mL H2O + 3 drops of I2 0.01 M Yellow
3. Starch solution + 1 mL saliva Turbid
4. I2 solution + saliva-starch solution
After 30 s Orange

After 60 s Orange

After 90 s Orange
 After 120 s Orange
After 150 s Orange
After 180 s Orange
After 210 s Orange
After 240 s Orange

5. The compound Effect face/ destroy bacteria Activities in Salivary Amylase

No Activity Result
1. 2 mL saliva + 8 mL H2O Colorless
2. Tube I + 5 drops Tholuen Colorless and blue
Tube II + 5 drops CHCl3 Purple formed 2 layers
Tube III + 5 drops HgCl2 1 % Turbid
Tube IV + 5 drops phenol Blue
Tube V + 0.5 mg NaF Colorless
Tube VI + 5 drops H2O Colorless
Tube VII Colorless
3.
Shaked + 5 mL starch 1 % → Hot
plate 38oC (15 minutes)
Tube Ia + I2 Turbid

Ib + benedict -

Tube IIa + I2 Turbid

IIb + benedict -

Tube IIIa + I2 Purple

IIIIb + benedict -

Tube IVa + I2 Purple

IVb + benedict -

Tube Va + I2 Purple

Vb + benedict -

Tube VIa + I2 Purple

VIb + benedict -
Purple
 Tube VIIa + I2 -
VIIb + benedict

6. Test Compilers of Saliva Inorganic Compounds

No Activity Result
1. 15 mL saliva (colorless) + 2 drops of Turbid solution
CH3COOH 2 M
2. Heat the solution Turbid solution
3. Filter the solution Colorless solution
a. To check ion Cl- Turbid solution
3 mL of filtrate (colorless) + 3 drops White precipitation
of HNO3 + 3 drops of AgNO3 0.5 M
b. To check ion PO33-
3 mL of filtrate + 3 drops of HNO3 + Yellow solution
1 mL of (NH4)6Mo7O2→ Heat solution
c. To check ion SO42-
3 mL of filtrate + 3 drops HNO3 + 1 No changes
mL BaCl2 0.1 M
d To check ion Ca2+
3 mL of filtrate + 1 mL of C2H8N2O4 No changes

7. Test the effect of Temperature on Ptyalin Activity

No Activity Result
1. 5 mL starch solution 1 %
Tube I → Put in cool water Colorless
Tube II → Room temperature Colorless
Tube III →Hot water Colorless
Tube IV →Room temperature Colorless
2.
 Tube I + 2 drops saliva Colorless
Tube II + 2 drops saliva Colorless
Tube III + 2 drops saliva Colorless
3. Tube IV + 2 drops saliva Colorless
5 minutes (first)
Tube I + 2 drops saliva + 2 drops I2 Purple
Tube I + 2 drops saliva + 2 drops I2 purple
Tube III + 2 drops saliva + 2 drops I2 purple
Tube IV + 2 drops saliva + 2 drops I2 purple
5 minutes (second)
Tube I + 2 drops saliva + 2 drops I2 Purple

Tube I + 2 drops saliva + 2 drops I2 Purple

Tube III + 2 drops saliva + 2 drops I2 Purple

Tube IV + 2 drops saliva + 2 drops I2 Purple

5 minutes (third)
Tube I + 2 drops saliva + 2 drops I2 Purple

Tube I + 2 drops saliva + 2 drops I2 Purple

Tube III + 2 drops saliva + 2 drops I2 Purple

Tube IV + 2 drops saliva + 2 drops I2 Purple

G. DISCUSSION
1. Tes Musin
Pada percobaan ini ingin diketahui kandungan musin yang terdapat dalam
saliva. Musin sebagian besar dihasilkan oleh kelenjar parotis yang merupakan
hasil dari sekresi mucus. Dan befungsi untuk membasahi makanan dan sebaagai
pelumas yang memudahkan untuk menelan makanan. Musin merupakan
kompleks dari karbohidrat atau protein dan sering disebut glikoprotein.
Pada percobaan ini saliva direaksikan dengan asam asetat dan
menghasilkan endapan putih. Penambahan asam asetat berfungsi untuk
mengendapkan musin yang terdapat dalam saliva dan CH 3COOH dapat mengubah
struktur dariprotein pada saliva, sehingga terjadi denaturasi yang menyebabkan
pengggumpalan protein. Kemudian dilakukan pemisahan endapan yang terbentuk
dengan larutan, lalu dilakukan proses penyaringan dan filtrat dibagi menjadi 2
bagianuntuk dilakukan pengujian benedict dan molish. Fungsi dari uji benedict
adalah untuk mengetahui adanya gula pereduksi dalam saliva sedangkan untuk uji
molish untuk mengetahui adanya kandungan karbohidrat dalam saliva. Pada uji
benedict menghasilkan larutan berwarna biru yang berarti saliva tidak
mengandung gula pereduksi. Pada uji molish menghasilkan larutan yang tidak
berwarna, ini berarti saliva tidak mengandung karbohidrat. Sdangkan pada uji
millons menghasilkan larutan yang juga tidak berwarna sehingga saliva
dinyatakan negative mengandung protein. Adapun reaksinya :
- Uji benedict

- Uji molish
 - Uji Millon

2. Tes tiosianat
Pada percobaan ini dilakukan pengujian terhadap ion SCN - yang terdapat
dalam saliva sebagai hasil dari pemecahan protein dengan senyawa belerang
dalam hati. Pengujian dilakukan dengan mereaksikan saliva dengan FeCl2 dan
HCl pekat sebagai katalis yang dapat mempercepat terjadinya reaksi. Reaksinya:
2SCN- + FeCl2 → Fe (SCN)2 + 2Cl-

Kemudian ditambahkan lagi dengan larutan HgCl2 yang berfungsi untuk

membentuk Hg(SCN)42- yang tidak berwarna sehingga membantu
mengidentifikasi ion SCN- pada saliva. Setelah penambahan HgCl2 diperoleh
larutan tidak berwarna yang menandakan bahwa saliva negative mengandung ion
SCN-. Reaksi yang terjadi :
4Fe(SCN)2 + 2Hg+ → 2 [Hg(SCN)4]2- + 4 Fe2+
3. Tes Penyusun Senyawa Anorganik Pada Saliva
Tujuan dari pengujian ini untuk mengetahui adanya senyawa anorganik
pada saliva antara lain ion Cl-, PO43-, SO42- dan Ca2+. Pengujian ini dilakukan
dengan cara mereaksikan saliva dengan asam asetat yang berfungsi untuk
mengendapkan glikoproptein. Dari hasil penambahan, dilakukan penyaringan
untuk memisahkan enadapan dari larutannya. Filtratnya dilakukan untuk
pengujian.
a. Ion Cl-
Filtrat yang direaksikan dengan HNO3(e) yang berfungsi sebagai katalis,
kemudian ditambahkan dengan AgNO3 menghasilkan larutan keruh dan endapan
putih yang merupakan endapan AgCl, yang menandakan bahwa saliva
mengandung ion Cl-. Reaksinya:
 Cl- + AgNO3 HNO3 ↓ AgCl + NO3-

b. Ion PO43-
Filtrat yang direaksikan dengan HNO3(e) yang berfungsi sebagai katalis,
kemudian ditambahkan ammonium molibdat yang berfungsi sebagai bahan utama
yang membentuk asam molibdat. Dari hasil penambahan diperoleh larutan kuning
yang menandakan bahwa dalam saliva tidak mengandung ion PO43-. Adapun
reaksinya:
2H3PO4 + (NH4)2 Mo7O24 → (NH4)6 (PO4)2MoO21 + 3H2O
c. Ion SO42-
Filtrat yang direaksikan dengan HNO3 (e) dan larutan BaCl2 yang berfungsi
untuk mengikat ion SO42- membentuk endapan putih yang merupakan endapan
BaSO4. Menurut teori, SO42- yang terdapat dalam saliva sangat sedikit sehingga
jika diendapkan memungkinkan tidak terjadinya endapan. Kemungkinan besar hal
inilah yang menyebakan pada hasil percobaan tidak ada endapan yang diperoleh
melainkan hanya larutan bening yang menandakan saliva tidak mengandung ion
SO42-. Reaksinya:
SO42- + BaCl2 HNO3 BaSO4 ↓ + 2Cl-
d. Ion Ca2+
Filtrat direaksikan dengan ammonium oksalat yang berfungsi untuk
mengikat Ca2+ yang membentuk endapan putih. Namun, pada percobaan tidak
diperoleh endapan putih melainkan larutan tak berwarna, yang berarti bahwa pada
saliva tidak terdapat ion Ca2+ dan tidak sesuai dengan teori uang reaksinya adalah
Ca2+ + (NH4)C2O4 → Ca C2O4 ↓ + 2NH4+
4. Tes Pengaruh Temperatur Terhadap Aktivitas Ptialin
Tujuan dari percobaan ini untuk mengetahui pengaruh temperatur terhadap
aktivitas ptialin. Ptialin merupakan hasil dari sekresi serus yang berfungsi untuk
memecah molekul amilum menjadi maltosa dengan proses hidrolisis. Saliva juga
mempunyai suhu optimal dimana dia bisa bereaksi dengan baik. Sebelum
diperoleh suhu optimal, kerja enzim meningkat dan setelah diperoleh suhu
optimal kerja enzim pada saliva menurun. Kenaikan energi kinetika molekul-
molekul yang bereaksi. Akan tetapi bila temperatur dinaikkan terus-menerus maka
energi kinetika molekul enzim menjadi besar, sehingga melampaui penghalang
energi untuk memecahkan ikatan-ikatan yang mempertahankan enzim dalam
saliva.
Pada percobaan ini larutan pati direaksikan dengan saliva encer dan
larutan iod sehingga diperoleh larutan berwarna ungu pekat. Penambahan iod
berfungsi untuk mengikat pati yang akan membentuk larutan biru. Berdasarkan
dari hasil pengamatan perlakuan pada ke-4 tabung yang berbeda-beda, pertama
didinginkan, kedua pada suhu kamar, ketiga dipanaskan pada suhu 38 0 C, dan
keempat ditambahkan saliva encer yang telah dipanaskan. Dari keempat tabung,
pada tabung ketiga warna coklat kehitaman lebih cepat hilang, ini terbukti pada
suhu 380 C karena suhu tersebut merupakan suhu optimum kerja enzim.
5. Tes Estimasi Ptialin
Tujuan dari percobaan ini untuk mengetahui waktu yang diperlukan untuk
pemecahan pati. Pada percobaan ini larutan pati direaksikan dengan larutan NaCl
dan salliva, kemudian dipanaskan pada penangas air pada suhu 38oC. Fungsi
penambahan NaCl sebagai penghambat agar pati tidak terhidrolisis. Dari hasil
pengamatan maka diketahui bahwa pada tabung nomor 7 yang lebih bening, dari
pengamatan sangat tampak pengaruh NaCl yang bertindak sebagai inhibitor
(penghambat) terjadinya pemecahan pati meskipun ditempatkan pada suhu yang
optimum enzim ptialin aktif bekerja.
6. Tes Penentuan pH yang Cocok Untuk Kerja Saliva
Pada percobaan ini digunakan larutan buffer yang direaksikan dengan
larutan pati,saliva, NaCl dan CH3COOH. Larutan buffer dengan pH yang berbeda
ditambahkan dengan pati dan NaCl kemudian dipanaskan pada suhu 38 oC setelah
penambahan saliva encer, pada buffer dengan pH 7 dan 9 ditambahkan
CH3COOH dan menghasilkan warna bening. Fungsi dari CH 3COOH adalah
mengubah struktur protein pada saliva sehingga terjadi denaturasi yang
menyebabkan penggumpalan. Dari hasil pengamatan yang dilakukan maka
diperoleh larutan yang lebih cepat bereaksi dengan iod, setelah penambahan iod
adalah larutan buffer dengan pH 7 yang ditandai dengan terjadinya perubahan
warna pada larutan menjadi warna kuning. Hal ini telah sesuai dengan teori,
dimana pH optimum saliva adalah 5,75-7,05. Fungsi dari penambahan asam
adalah untuk menginaktifkan enzim.
7. Efek Senyawa yang Menghambat / Menghancurkan Aktivitas Bakteri pada
Amilase Saliva
Pada percobaan ini dilakukan pengujian terhadap beberapa senyawa yang
dapat menghambat / menghancurkan aktivitas bakteri pada amilase saliva. Saliva
yang telah diencerkan ditambahkan toluen, kloroform, HgCl 1%, fenol 2%, NaF,
dan H2O yang masing-masing menghasilkan larutan bening. Setelah itu, masing-
masing isi tabung dibagi 2 dan diuji dengan pereaksi benedict dan Iod. Pada
toluen, kloroform, fenol 2%, NaF, dan H2O setelah direeaksikan dengan benedict
diperoleh larutan berwarna biru dan bagian yang lain setelah direaksikan dengan
iod menghasilkan larutan yang berwarna coklat. Sedangkan pada HgCl2 setelah
direaksikan dengan pereaksi benedict menghasilkan warna biru dan direaksikan
dengan iod menghasilkan larutan berwarna biru tua (keruh). Hal ini menandakan
bahwa telah terjadi penghambatan aktivitas bakteri terhadap amilase saliva.
H. CONVLUSION
Based on the experiment, we can conclude :
1. Saliva contains musin but just a little.
2. Based on the thiocyanate test, saliva isn’t contains thiosianat.
3. Saliva contains inorganic compounds such as Cl -, PO43-, SO42- and Ca2+ but in
this experiment just Cl- that positivily contained by saliva.
4. Activities Ptialin will work properly in room temperature and the maximum
temperature 38oC.
5. Estimation ptyalin that for all tube produce brown solution which is more
amylum have been hydrolysis within interval 30 seconds for all the tubes.
Based on the experiment for all tube produce blue solution which is there is
amylum not yet hydrolysis in all tubes
6. Ptialin Saliva contains enzymes that work optimally at pH 7
7. Phenol, HgCl2, NaF can inhibit the action of enzyme for only bacterial activity
is inhibited when reacted with the acid solution, the solution is corrosive, and
antibacterial agents.
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 DOCUMENTATION

Solution of Mucin test (Mollish Thiosianate

Saliva and millon) Test

compound effect Ptyaline estimation test

ory bacteria activities
alivary amylase