Analysis_of_Saliva.docx

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Analysis of Saliva John Emmanwel Aljhu C. Del Rosario Department of Biology, College of Science, PUP Sta. Mesa, Manila 1

ABSTRACT

Saliva which contains mucins, amylase and other proteins is a viscous fluid with a pH around 6.8. In this experiment collection of three samples of saliva was done to test each of its components such as amylase, mucins, calcium and inorganic phosphate and to determine if these are present in the samples. Results (in the form of colored precipitate and changes in color of the solution) that were obtained were all positive for each component of the saliva which indicates that these components are present in the samples.

Key words: Saliva, mucins, amylase, calcium, inorganic phosphate Introduction

Methodology

Saliva contributes to the digestion of food because it lubricates and allows the swallowing of food more convenient. It contains enzyme such as amylase which is responsible for  breaking down of starch into simpler sugars that can be later absorbed by the small intestine. Mucin is a glycoprotein which can be  precipitated  precipitated by acetic acid at around pH 4.5 has a function in saliva. It provides protection in oral activity. Calcium and inorganic phosphate are also an essential component of the saliva that can also be precipitated using acetic acid (for calcium) and concentrated nitric acid (for inorganic phosphate).

Collection of Saliva

Objective

The aim of this experiment is to test if there is a presence of amylase, mucins, calcium and inorganic phosphate in the saliva which can  be seen as as colored precipitates. precipitates.

The mouth was rinsed to get rid of food  particles. About 10 ml of warm water was taken into mouth and moved it about by the tongue for nearly a minute. Fluid was collected in a clean  beaker. The tests were performed with this diluted saliva. Test for Amylase To 2.5 ml of starch solution in a test tube, 1 ml of 0.1M phosphate buffer, pH 7.0 was added followed by 1 ml of 0.1 M NaCl solution. 1 ml of salivary fluid was also added to the mixture. Mixed. At intervals of one minute, one drop of the mixture was transfer to depressions in a spot plate, containing a drop of iodine solution. Test for Mucins A 1% acetic acid was added from a 2mL of salivary fluid drop by drop.

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Test for Calcium To 2 ml of salivary fluids, 5 drops of 1% acetic acid and 5 ml of 2% potassium oxalate solution were added. Test for Inorganic Phosphate To 2 ml of salivary fluids, few drops of concentrated nitric acid were added followed by a pinch of ammonium molybdate. The solution was then warmed. Results and Discussion

Saliva was rich in the enzyme amylase which is the main enzyme to breakdown starch. Amylase hydrolyses α  1,4 glycosidic bonds in starch at random. The break-down products do not have capacity to bind iodine. In this test using iodine there is no large enzymatic reaction happens in sample A and B because it results a  black blue color. But there is a brown in C which can determined small amount of enzymatic reaction. Test for Mucins

Test for Amylase

A

B  Figure 2.Test for mucins shows a thread-like  precipitate in the three samples of salivary  solution.

C

 Figure 1.Test for amylase with different saliva  sample.

A thread-like precipitate was formed which indicates the presence of mucins. Test tube C has the longest thread-like precipitate which indicates a greater amount of mucin is  present on it. Test tube A and B are the same. Mucin is a glycoprotein constituent of mucus. The precipitate was formed because of the acetic acid which is around pH 4.5. Test of Mucins is  performed to know if there are any symptoms of abnormalities that are happening to your body. Increased mucin production occurs in many adenocarcinomas, including cancers of the  pancreas, lung, breast, ovary, colon and other tissues.

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Test for Calcium

of inorganic phosphate present in the saliva. Inorganic phosphate combined with ammonium molybdate to form a yellow colored ammonium  phosphomolybdate. The precipitate ammonium  phosphomolybdate is the inorganic salt of phosphomolybdic acid. It contains the remarkable phosphomolybdate ion complex. Conclusion

 Figure 3.Test for calcium shows a white  precipitate formed as calcium oxalate. After the addition of acetic acid, reaction is evident. The previously slightly clear solution turns white. Then after the potassium oxalate was added small granules of precipitate was formed. Only, small amount of precipitate was collected. A faint white precipitate was formed. Calcium ions are precipitated as calcium oxalate under neutral or slightly acidic conditions. Test for Inorganic Phosphate

These particular tests for the different components of the saliva which gave a positive in all test generally indicates that these components are always present in the saliva like the amylase which in the experiment gives a  black blue color, without the presence of it starch in the food we eat will never be break down because starch is necessary to break down into glucose monomer by amylase through hydrolyzing the α 1,4 glycosidic bonds of  starch to form glucose monomer. Positive results are in the form of colored precipitates like canary yellow colored ammonium phosphomolybdate in the test for inorganic phosphate while a white thread-like precipitate was formed in the test for mucins which indicates the presence of mucins . All in all, analysis of saliva is necessary to determine what substances are present in saliva. References

Hayman, J.M Jr., and Johnston, Sara M.: The Excretion of Inorganic Phosphate, J. Clin investigation 11:607 (May) 1932 Power, M. H., and Wakefield, E. G.: A, Proc. Staff Meet, Msyo Clin. 6: 401 (July 8) 1931  Figure 4.Test for inorganic phosphate shows a canary yellow (ammonium phosphomolybdate)  precipitate. In this experiment, test tube A and C has the lesser precipitate formed while test tube B has the greater which indicates a greater amount

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