San Pedro College Biochemistry Laboratory Customize Laboratory Manual (Activity 1-4

)

Group 6 BMLS 2F Abundo, Simon Jonson Halog, John Anthony Canakan, Faraida R. Doplayna, Zipporah Paragele, Kimberly Claire Tanseco, Karen

1

ACTIVITY 1A & 1B SUBCELLULAR COMPONENTS OF THE LIVING CELL QUALITATIVE TEST FOR THE CHEMICAL COMPONENT OF THE CELL

I.

OBJECTIVES:  To determine the composition of organelles.  To understand and appreciate the different biochemical systems through separating the subcellular components of a cell.  To be able to perform different qualitative tests specific for carbohydrates, proteins, and lipids.  To compare the relative amounts of carbohydrates, proteins, and lipids.

II.

SCHEMATIC DIAGRAM (1A): Wash off blood from chicken liver using a few drops of 5% TCA (trichloroacetic acid).

Weigh the liver.

Add 5 mL of suspending medium per gram of liver.

Homogenize at low speed for 5-10 min.

Centrifuge for 10 min. and decant.

Sediment 1

Supernate 1

Add 10 mL of 5% TCA and mix. Test for carbohydrates, proteins, and lipids.

Centrifuge for 10 min. and decant.

Sediment 2

Supernate 2

Add 10 mL of 5% TCA and mix. Test for carbohydrates, proteins, and lipids. 2

Centrifuge for 10 min. and decant.

Cont. Sup 2

Sediment 3

Supernate 3

Add 10 mL of 5% TCA and mix. Test for carbohydrates, proteins, and lipids.

Add 10 mL of 5% TCA and mix. Test for carbohydrates, proteins, and lipids.

SCHEMATIC DIAGRAM (1B): Test for carbohydrates Molisch Test Benedict’s test

To 5 drops of sediment/supernate, add 5 drops Molisch reagent. Layer with 1 mL of conc. Shake sulphuric acid. Perform . control test using 1% ribose solution.

To 1 mL of Benedict’s solution, add 2 drops of the sediment/supernate.

Boil for 2 min. and cool.

Test for proteins Biuret test

Xanthoproteic test

To 5 drops of the sediment/supernate, add 5 drops of 10% NaOH and 1 drop of .5% copper sulphate.

Place 1 mL of the sediment/supernate in a test tube.

Add 1 mL of conc. Nitric acid. Perform control test using 1% albumin.

Heat and observe colour change.

Cool and add 5 drops of ammonium hydroxide.

3

Test for lipids Sudan test To 5 drops of the sediment/supernate, add 5 drops of Sudan IV Perform control test using 1% lecithin. Acrolein test Place .5 g. Of potassium bisulphate in a clean, dry test tube. Add a drop of the sediment/supernate and heat. Note the odor produced.

III. RESULTS AND OBSERVATION TESTS CARBOHYDRATES MOLISCH BENEDICT’S PROTEINS BIURET XANTHOPROTEIC LIPIDS SUDAN ACROLEIN 1% LECITHIN RED RED ACRID ODOR + + + + CONTROL 1% RIBOSE PURPLE PURPLE BRICK RED EXPECTED RESULTS SEDIMENT 1 SEDIMENT 2 SEDIMENT 3 SEDIMENT 4

1% ALBUMIN VIOLET YELLOW + + -

IV. ANALYSIS AND CONCLUSION Guide Questions: 1. Give the principle involved in each of the test used to determine the chemical components of the organelles.  Molisch test- dehydration of the carbohydrate by sulphuric acid to produce an aldehyde

4

    

Benedict’s test- reduction of copper ions by reducing sugars, which then can be detected by the formation of a brick red precipitate of cuprous oxide Biuret test- reduction of copper ions which then complexes with nitrogen atoms on peptide bonds at high pH Xanthoproteic test- nitration of benzene ring present in the structure Sudan test- ability of fat cells to selectively absorb pigments in fat dyes such as Sudan IV Acrolein test- Whenever fat is heated in the presence of a dehydrating agent, fat molecules shed glycerol in the form of the unsaturated aldehyde- acrolein.

2. Explain the principle involved in the separation scheme for the subcellular components. The principle behind the separation scheme is differential centrifugation. Tissues or cells are first homogenized to release their internal contents. Larger particles settle faster than smaller one and this provides the basis for obtaining crude organelle fractions by differential centrifugation.

V. DOCUMENTATION

Molisch test: From left: sediment 1, sediment 2, supernate 3, sediment 3, control

Benedict’s test From left: sediment 1, sediment 2, supernate 3, sediment 3

:

5

Biuret test: From left: sediment 1, sediment 2, supernate 3, sediment 3

Xanthoproteic test: From left: sediment 1, sediment 2, supernate 3, sediment 3

Sudan test: From left: sediment 1, sediment 2, supernate 3, sediment 3, control

Acrolein test: From left: sediment 1, sediment 2, supernate 3, sediment 3

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Activity 2A Qualitative Test for Proteins/ Color Reaction Test

I. OBJECTIVES    To detect proteins through the different color reaction tests. To know the significance of peptide bonds and amino acids in detecting proteins. To understand the principle behind every test.

II. SCHEMATIC DIAGRAM

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III. RESULTS AND OBSERVATIONS Qualitative test Egg Albumin Solution Expected Results (+) Not performed Brick red

Millon’s test

Not performed

Xanthoproteic test Hopkins_cole test Biuret Test Ninhydrin test Sakaguchi test

+ + + + +

yellow

yellow Violet violet Blue-violet red

IV. ANALYSIS AND CONCLUSION Guide Questions: 1. Explain the principle involved in each of the test  Millon’s Test- Millon's reagent (Hg/HNO3) gives positive results with proteins containing the phenolic amino acid “tyrosine’’, the precipitate turns flesh to red color, if proteins containing tyrosine are present. Xanthoproteic Test- determined the amount of protein soluble in a solution using conc. nitric acid. The test gives a positive result in those proteins w/ amino acids carrying aromatic groups, esp. tyrosine. The proof is neutralized w/ an alkali, turning dark yellow. Hopkin’s Cole Test- There was a violet color produced at the point of contact. Tryptophan has an indole nucleus which is responsible for the violet ring found at the junction between the two layers. Biuret Test- Reduction of Cu2+ which then complexes with the N atoms on the peptide bonds at high pH. Ninhydrin Test- Amino Acid containing a free amino group and a free carboxylic acid group that react together with ninhydrin to produce colored products. Group on the alpha-carbon and can react with ninhydrin to produce blue purple product. Sakaguchi Test- In alkaline solution, protein containing Arginine gives red color with Alpha-Napthol and sodium hypochloride.

8

V. DOCUMENTATION

Preparation of Egg Albumin solution

Test results. From left: Sakaguchi test, Ninhydrin test, Xanthoproteic test, Hopkins –cole test, Biuret test 9

ACTIVITY 2B PRECIPITATION TEST FOR PROTEINS I. OBJECTIVES    To observe the reactivity of reagents & solutions in each test To know the practical reactions in each test To Identify and observe the precipitate in each test which yield a positive result.

II. II. SCHEMATIC DIAGRAM A. Precipitation Reaction by Concentrated Mineral and Organic Acids
Place 1ml of Egg Albumin in each of the 4 test tubes

Add 5 drops of Conc. Sulfuric Acid

Add 5 drops of Conc. HCl

Add 5 drops of Nitric Acid

Add 5 drops Glacial HAc

B. Precipitation by Metallic Salts
Place 1ml of Egg Albumin in each of the 5 test tubes

Add 1ml of Dil. Mercuric Acid

Add 1ml Dil. Lead Acetate

Add 1ml Dil. Copper Sulfate

Add 1ml Dil. Ferric Chloride

Add 1ml Dil. Barium Chloride

C. Precipitation of Alkaloidal Reagents
Place 1ml of Egg Albumin in each of the 4 test tubes

-Add 1ml of Picric Acid Sol’n -Heat

- Add 1ml of TAC Acid -Heat

-Add 1ml of Tannic Acid -Heat

-Add 1ml of Phosphotungstic Acid -Heat

D. Precipitation by Alcohol
Place in each of the 3 test tubes 5ml of 95% Alcohol

-Add 2 drops of Dil. HCl -Add 3 drops of Egg Albumin

-Add 2 drops of 10% NaOH 10 -Add 3 drops of Egg Albumin

-Add 3 drops of Egg Albumin

E. Coagulation of Heat
Heat to Boiling 5ml of Albumin Solution

Add 2 Drops of Acetic Acid

F. Test for Denatured/Coagulated Proteins
Perform the following tests; a) Milllon’s Test b) Xanthoproteic Test c) Biuret Test d) Hopkins-Cole Test

Suspend each precipitate in 10 ml of Distilled Water

Place 3ml of each test tube

III. Results and Observation: C. Precipitation by Alkaloidal Reagents Chemicals Picric Acid Trichloroacetic Acid Tannic Acid Phosphotungstic Acid

Results and Observations (+) Yellowish Sol’n/Precipitate (+) Whitish Sol’n/Precipitate (+) Brownish Sol’n/Precipitate (+) Whitish Sol’n/Precipitate

D. Precipitation by Alcohol 95% Ethyl Alcohol+Dil.HCl 95% Ethyl Alcohol+10%NaOH 95% Ethyl Alcohol Results and Observations (-) Cloudy Solution/Precipitate (-) Clear Solution/Precipitate (+) Whitish, Cloudy Sol’n/Precipitate

E. Coagulation by Heat 5mL albumin sol’n + HAC Results and Observations (+) White Cloudy Precipitate

Suspended precipitate Picric acid Trichloroacetic

Millon’s Test Not performed

Xanthoproteic Test (+) (-)

Biuret Test (-) (-)

Hopkins-Cole Test (-) (-)

F. Test for Denatured/Coagulated Proteins

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acid Tannic acid Phosphotunstic acid (-) (-) (+) (-) (-) (-)

IV. ANALYSIS AND CONCLUSION
Guide Questions: 1. Give the principles involved in each of the tests. A. Precipitation reaction by concentrated mineral and organic acid Salt bridges result from the neutralization of an acid and amine on side chains. The final interaction is ionic between the positive ammonium group and the negative acid group. Any combination of the various acidic or amine amino acid side chains will have this effect. As might be expected, acids and bases disrupt salt bridges held together by ionic charges. A type of double replacement reaction occurs where the positive and negative ions in the salt change partners with the positive and negative ions in the new acid or base added. B. Precipitation by metallic salts Proteins are precipitated by salts of heavy metals, such as mercuric chloride, zinc sulfate,etc. in weak alkaline solutions, protein molecules carry negative charges and combine with positively charged metal ions to form insoluble salts which precipitate from the solution. The precipitated proteins aredenatured and this process is irreversible. C. Precipitation by alkaloidal reagents Alkaloidal reagents (e.g. tannic acid & trichloroacetatic acid) are high molecular weight anions. The negative charge of these anions counteracts the positive charge of the amino group in proteins giving a precipitate. D. Precipitation by Alcohol Alcohol denatures proteins by disrupting the side chain intramolecular hydrogen bonding. New hydrogen bonds are formed instead between the new alcohol molecule and the protein side chains. Where does the max. precipitation occur? 3rd tube. E. Coagulation by Heat The result was coagulation of the albumin solution. Egg-white is faintly alkaline. Complete precipitation takes place only in faintly acid solution. The temperature at which coagulation takes place depends to a large extent on the amount of acid and of salts present F. Test for Denatured/Coagulated Proteins The term denaturation is used more frequently than coagulation by scientific investigators at the present time to denote certain changes in proteins. Definite characteristics of the proteins are changed when they are coagulated, among which is loss of solubility in water and dilute salt solutions. In some instances and under certain conditions the coagulation process may be reversible. Manner in which denaturation may be brought about. Coagulation of proteins may be brought about by a variety of processes. But in addition to heat the action of acids, alkalies, salts, alcohol, mechanical agitation, radiation, and ultra-sonic vibrations may denature the protein and convert it from a soluble into insoluble form weight anions. The negative charge of these anions counteracts the positive charge of the amino group in proteins giving a precipitate. Therefore, even a protein is denatured it will still give a positive result from the Qualitative of proteins. 12

2-3. Using milk as test solution, how will you prove that milk is a protein? Show a schematic diagram to arrive at your answer.  Biuret solution is used to identify the presence of protein. Biuret reagent is a blue solution that, when it reacts with protein, will change color to pink-purple. Procedure:  To a test tube, add 40 drops of milk solution  Add 3 drops of Biuret reagent  Shake gently to mix  Note color change  Proteins will turn the solution into pink-purple

Schematic Diagram: To a test tube, add 40 drops of milk sol’n

Add 3 drops of Biuret reagent (sodium hydroxide 10% & copper sulfate 0.5%)

Shake gently to mix

Change in color of sol’n to pink-purple confirms presence of protein

V. DOCUMENTATION

Preparation of Egg Albumin solution

13 Precipitation by Alcohol From left: NaOh, HCL, neutral

Precipitation by Alkaloidal reagent From left: phospotungstic acid, Picric acid, Tannic acid, TCA

Hopkin’s-Cole Test From left: Tannic acid, TCA, Picric acid, Phosphotungstic acid

Biuret Test From left: Picric acid, Tannic acid, TCA, Phosphotungstic acid

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Xanthroproteic Test From left: Picric acid, Phosphotungstic acid, TCA, tannic acid

Activity 3A Enzymes I. OBJECTIVES    To be able to know the properties of an enzyme To be able to test the presence of an enzyme To be able to test the specificity of an enzyme

II. SCHEMATIC DIAGRAM Preparation of potato extract f Potato (Peeled)

Grate/blendered (100ml of distilled H2O)
Let it stand for 10 min and Stir Occasionally

Strain (through cheese cloth)

Filter (the extract through filter paper)

Biuret test

Test for catalase activity

1 ml of extract

5 ml of extract

Mix 1 ml of 3% H2O2

Add 2 ml 10% NaOH soln
Hold a splinter over the mouth of the tube

mix. add 2-3 drops of Copper sulfate 15

Add 1 ml 0.5% Benzidine

Preparation of dilute solution of salivary amylase

Rinse the collector’s mouth several times with water

Collect 1 ml of saliva

Prepare 1:8 solution of saliva by adding 10 drops (0.25 ml) saliva with 20 ml water

Use for test

Specificity of Enzyme Action

Test tube 1 2 ml 0.02 M phosphate buffer (pH 6.7) and 1 ml 0.9% Sodium chloride sol’n

Test tube 2 2 ml 0.02 M phosphate buffer (pH 6.7) and 1 ml 0.9% Sodium chloride sol’n

Add 1 ml cooked starch and 1 ml salivary amylase sol’n

Add 1 ml 1% glycogen and 1 ml salivary amylase sol’n

Stand for 15 minutes @ room temperature.

Stir the mixture

In each test tube place 1 drop of the solution in a spot plate

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Add 1 drop of Iodine sol’n. observed color produced.

REPEAT AT 5 MINUTES INTERVAL FOR 1 HOUR. III. RESULTS AND OBSERVATIONS

Biuret test Test for Catalyse activity -splinter -benzidine

Result + (violet) -

Expected Result violet Pooping sound Blue-green coloration

Test for specificity of Enzyme action Time (minutes) 5 10 Test tube 1 + + Test tube 2 -

15

+

20 25

+ +

30 35 40

+ + +

45 50 55 60

+ + + +

green Darker green w/ purple coloration Green with darker purple coloration Lesser green and purple color Presence of granule like subsatnce Lesser green and purple color Lesser green and purple color Much Lesser green and purple color green Lesser green color Almost orange Almost orange

No changes in color observed

-

-

-

-

-

IV. ANALYSIS AND CONCLUSION Guide Question: 1. Give the principle involved in Biuret test. Biuret test is based on the ability of Cu (II) ions to form a violet-colored chelate complex with peptide bonds (-CONH-groups) in alkaline conditions.

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2. How will you proved the presence of catalase? Show chemical equation. A rapid appearance of sustained gas bubbles is an indication of the presence of a catalase.

2 H2O2 → 2 H2O + O2
3. How will you prove the specific ation of salivary amylase? Enzymes are normally very specific, catalyzing at most only a few types of chemical reactions. The enzyme amylase acts on the substrate starch catalyzing its break down into simple sugars. Amylase will not catalyze the breakdown of the complex sugar sucrose into simple sugars. Indicators can be used to test for the presence of these reactions like iodine solution.in the iodine test, amylose in the starch (straight chain) forms helices where iodine molecules assemble and forming blue-black color. While amylopectin (branched portion) in glycogen forms much shorter helices and iodine are unable to assemble and only forming orange/yellow hue.

V. DOCUMENTATION

Preparation of potato extract

Testing gas evolution by splinter

Results for Buiret and catalase test

Salivary amylase solution

18

Test for specificity of enzyme TEST TUBE 1 (cooked starch)

TEST TUBE 2 (1% GLYCOGEN)

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ACTIVITY 3B FACTORS AFFECTING ENZYME ACTION

I. OBJECTIVES     To completely perform tests regarding the factors affecting enzyme action. To examine and explain the effect of temperature on enzyme. To determine the extent of digestion of enzyme through acid and base solutions with a controlled temperature. To verify the specific pH through the use of pH paper. To compare the obtained product with the standard solution.

II. SCHEMATIC DIAGRAM Effect of Temperature:
Prepare 3 test tubes each having 5ml. 1% cooked start soln +1ml saliva.

Test tube # 1: Water bath in 40ºC.

Test tube #2: Water bath in 60ºC.

Test tube #3: Water bath in 10ºC.

Take a drop after stirring and test with Iodine soln.

Take a drop after stirring and test with Iodine soln.

Take a drop after stirring and test with Iodine soln.

Perform Iodine test at 15mins interval for 1hr.

Effect of pH: Prepare 4 test tubes containing equal amts of egg white.
Measure height of coagulation for each test tube

Coagulate in hot water bath.
Test tube #3: 5ml 2% pancreatin + 10 drop 0.4% HCl. Test tube #4: 5ml 2% pancreatin + 10 drop 0.4% Na2CO3.

Test tube #1: 5ml 2% pepsin + 10 drop 0.4% HCl.

Test tube #2: 5ml 2% pepsin + 10 drop 0.4% Na2CO3.

Determine pH of each sample

Water bath test tubes @ 40ºC

Perform Biuret test of 1ml supernatant liquid.

Perform Biuret test of 1ml supernatant liquid.

Perform Biuret test Perform Biuret test Measure digestion ofof each sample of 1ml supernatant 1ml supernatant liquid. liquid.

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III. RESULTS AND OBSERVATIONS

Effect of Temperature:
Temperature 15mins Obtained Expected Color Color Powder Blue black Blue or something dark in color Dark Blue 30mins Expected Color Light Blue black Blue or something dark in color Color Dark Blue 45mins Expected Color Mossy Blue black Green or something dark in color Color Dark Blue; traces of yellow color 60mins Expected Color Mossy Blue black Green or something dark in color Color Dark Blue; traces of yellow color Orange or something light in color Result

Test Tube #1 @40 ºC

15min -

30min -

45min +/-

60min +/-

Test Tube #2 @ 60 ºC

Orange or something light in color

Orange or something light in color

Orange or something light in color

-

-

-

-

Test Tube #3 @ 10 ºC

Dark Blue

Blue black or something dark in color

Blue

Blue black or something dark in color

Green

Blue black or something dark in color

Blue

Blue black or something dark in color

+

+

+

+

Effect of pH: Test Tube Measurement (before heating) 31cm Measure ment (after heating) 30cm pH Color Expected color Result

No.1 Pepsin + HCl No. 2 Pepsin + Na2CO3 No. 3 Pancreatin + HCl No. 4 Pancreatin + Na2CO3 No. 5 Controlled

6

Brown with traces of purple color Brown with traces of purple color Purple

Purple +

31cm

29cm

9

Purple +

31cm

30cm

6

Purple +

31cm

30cm

9

Purple

Purple +

-

-

-

-

Purple

+

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IV. ANALYSIS AND CONCLUSION Guide Question: 1. Explain the mechanism of enzyme action at varying temperatures. Give the principle involved. Being a protein, an enzyme is denatured at high temperature and becomes inactive. Initially as the temperature increases, the rate of enzymatic reaction also increases but it has a limitation. When the temperature affects the molecular structure of the enzyme it gets inactivated. 2. How does different pH affect enzyme action? Explain your results using this principle. Discuss Isoelectric pH of enzyme action. Enzymes are picky with pH levels, as they are with every thing else. They have an optimal level at which they work the best, and anything above or below that level, their activity begins to slow down until they shut down all together.

V. DOCUMENTATION

After heating at controlled temperature (from Left to Right): Pancreatin (Na2CO3), Pancreatin(HCl), Pepsin (Na2CO3), Pepsin (HCl)

Biuret Test Performed (from Left to Right): Pepsin (HCl), Pepsin (Na2CO3), 0.5% peptone standard, Pancreatin (Na2CO3), Pancreatin(HCl)

Left side Spot plate (from top to bottom): 60ºC 15mins, 30mins, 45mins, 60mins. Right side Spot plate upper portion (from left to right):10ºC 15mins, 30mins, 45mins, 60mins. Right side Spot plate lower portion (from left to right): 40ºC 15mins, 30mins, 45mins, 60min. 22

Activity 4 Nucleic Acid

I. OBJECTIVES    To be able to prepare nucleic acid and perform the different tests for it. To understand the principle behind every test. To be able to chemical reactions behind every principle.

II. SCHEMATIC DIAGRAM Preparation of Nucleic Acid Liver Homogenize with 1 volume of water 1 ml homogenate + 5% TCA (stir) Centrifuge (5 minutes) Residue/sediment + alcohol-ether mixture (3:10)

Suspend with 7 ml TCA

Heat at 90°C (15 mins) Cool and centrifuge Supernatant (make up a volume to 10 ml)

Test for Purine

Orcinol test for Pentose

Orcinol test for Pentose

Test for Phosphates

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A. Test for Purine

1 ml solution + 1 ml 2N HCL

1 ml of water + 1 ml 2N HCL

Place in water bath for 20 minutes

1 ml 2N acetate buffer 10 drops 10 % copper sulfate + 5 drops sat. sodium bisulfate. Mix. Heat in a water bath for a few minutes.

Place in boiling water bath. Heat

B.

C. Diphenylalanine test for Deoxyribose

Orcinol test for Pentose

0.5 ml sample + 1 ml water 1 ml orcinol reagent

1 ml sample + 1 ml diphenylalanine reagent

Heat in water bath. Cool

Heat (5 minutes) in boiling water bath. Cool

D. Test for Phosphates

I ml sample + 5 drops NHO3

5 drops Ammonium mmolybdenate soln. Heat

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III. RESULTS AND OBSERVATIONS

Test Test for Purine

Experimental Result Test tube 1 Test tube 2 -

Expected Result White/tan flocculent ppt

Orcinol test for Pentoses Diphenylalanine test for deoxyribose

-

green blue

+ Test for phosphates yellow

IV. ANALYSIS AND CONCLUSION

Guide questions:

1. Discuss the different test used in the experiment  test for purine - The NaHSO3 reduces Cu2+ to Cu+. Interaction of purines with Cu+ results in precipitation of insoluble yellow-white complex. Orcinol test for pentoses- the basis of this test relies on the conversion of pentose sugar, ribose, present in the RNA into furfural in the presence of hot acid. Furfural reacts with orcinol in the presence of ferric chloride to give a green color. The intensity of the color produced depends on the concentrations of HCL, ferric chloride and orcinol, as well as the duration of boiling. Diphenylamine test for deoxyribose- DNA contains deoxyribose. DNA can be identified chemically with the Dische diphenylamine test. The reaction between the Dische reagent and 2-deoxypentose results in the development of a blue color. The reaction depends on the conversion of the pentose to w-hydroxylaevulinic aldehyde which then reacts with diphenylamine to give a a blue colored complex. The intensity of the blue color is proportional to the concentration of DNA. Dische reagent does not react with the ribose sugar in RNA and does not form a blue-colored complex. Test for phosphates- In the test for presence of phosphates for DNA, a yellow precipitate is obtain. The ammonium molybdate solution reacted with the sample which yields yellow crystals, phosphoammonium molybdate, which is a positive result .

2. Show chemical equations. Give the principle.

test for purine - The NaHSO3 reduces Cu2+ to Cu+. Interaction of purines with Cu+ results in precipitation of insoluble yellow-white complex.

25

orcinol test for pentoses -Deoxyribose (contained in DNA) is converted intohydroxylewulinic aldehyde when heated in concentrated H2SO4. This aldehyde reacts with diphenylamine forming blue colour complex.

Diphenylamine test for deoxyribose- this reactions depends on the conversion of the pentose to w-hydroxylaevulinic aldehyde which then reacts with diphenylalamine to give a blue colored complex.

test for phosphates- hydrolysis of pyrophosphate to phosphate forming a yellow precipitate HPO42-(aq) + 12MoO42-(aq) + 3NH4+(aq) + 23H3O+(aq)  (NH4)3[P(Mo3O10)4](yellow, s)

26

V. DOCUMENTATION

Preparation of nucleic acids (after the1st centrifugation)

The final test solutions (supernate)

Results: Test for Purine

Result: Orcinol Test

Result: Diphenylalamine Test

Result: Test for phosphates

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