Chem Practicals

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1. ELEMENTARY COMPOSITIONS OF PROTEINS
Carbon & Hydrogen
Casein was heated.
CHARRING-Carbon.
MOISTURE-Hydrogen

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1. ELEMENTARY COMPOSITIONS OF PROTEINS
Nitrogen
Soda lime + Casein -> HEAT
BLUE COLORED LITMUS PAPER

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1. ELEMENTARY COMPOSITIONS OF PROTEINS
FUSION

(2parts Na2CO3:1part KNO3)

Casein -> HEAT -> CLEAR mixture -> COOL & DISSOLVE -> FILTER

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1. ELEMENTARY COMPOSITIONS OF PROTEINS
Sulfur
Filtrate + Dilute HCl -> HEAT + BaCl2
CLOUDY WHITE LAYER
Ba2+SO4 -> BaSO4

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1. ELEMENTARY COMPOSITIONS OF PROTEINS
Phosphorous
Filtrate + Conc HNO3 + (NH4)2MoO4
YELLOW SOLUTION OF Ammonium Phosphomolybdate
3NH4++12MoO4-2+PO4-3+24H+ -> (NH4)3PO4.12MoO3 +12H2o

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2. COLOR REACTIONS OF PROTEINS
Biuret Reaction
General Test for Proteins
NaOH + CuSO4
Pink- Violet Coloration
PINK COLORATION PEPTONE

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2. COLOR REACTIONS OF PROTEINS
Biuret Reaction
General Test for Proteins
NaOH + CuSO4
Pink- Violet Coloration
VIOLET COLORATION ALBUMIN

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2. COLOR REACTIONS OF PROTEINS
Xanthoproteic Reaction
Test for the PHENYL RINGS in the TYROSINE, PHENYLALANINE & TRYPTOPHAN
HNO3+ NH4OH
YELLOW SOLUTION
SALT formation with YELLOW NITRO-substitution products and ALKALI

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2. COLOR REACTIONS OF PROTEINS
Millon;s Test
Test for PHENOL group contained in TYROSINE
Millon’s Reagent (1 Hg: 2 HNO3)
FLESH-RED PPT

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2. COLOR REACTIONS OF PROTEINS
Glyoxylic Acid/Hopkins-Cole Reaction
Test for INDOE NUCLEUS in the TRYPTOPHAN component
Hopkins Cole Rgt (Powdered Mg with cold H2o)
Glyoxylic Acid (Reduction of OXALIC acid with Mg powder)
Hopkins Cole Rgt + H2SO4
VIOLET RING @ point of contact

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2. COLOR REACTIONS OF PROTEINS
Heller’s Ring Test
Test to detect ALBUMIN
Conc. HNO3
WHITE PPT of coagulated Protein

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2. COLOR REACTIONS OF PROTEINS
Reduced Sulfur Test
Test for amino acids that contains REDUCED S GRP
NaOH + Lead Acetate
BLACK PPT (PbS)

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2. COLOR REACTIONS OF PROTEINS
Adamkiewicz Reaction
Test for INDOLE DERIVATIVES
H2SO4 & Glacial Acetic Acid (CH3COOH)
VIOLE RING/SOL’N

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3. PRECIPITATION REACTIONS OF PROTEINS
Precipitation by HEAT COAGULATION
PPT and COAGULATION occurs best near IP
1N & 5N Acetic acid (CH3COOH)
1N Acetic Acid Coagulated Best

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3. PRECIPITATION REACTIONS OF PROTEINS
Precipitation by ORGANIC SOLVENTS
NO Coagulation , protein has been completely precipitated by alcohol
Albumin + Ethyl Alcohol (CH3CH2OH)
Applied for preserving tissues

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NO PIC D:

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3. PRECIPITATION REACTIONS OF PROTEINS
Precipitation by SALTING OUT
A.AMMONIUM SULFATE
[(NH4)2SO4]
a.POSITIVE Millon’s test on PPT
b.NEGATIVE Biuret test on filtrate

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3. PRECIPITATION REACTIONS OF PROTEINS
Precipitation by SALTING OUT
B.SEPERATION OF PEPTONES & PROTEOSES
[Commercial Peptone & (NH4)2SO4]
a.POSITIVE Biuret Test on Filtrate
•Filtered: PROTEOSES
•Filtrate: PEPTONES

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3. PRECIPITATION REACTIONS OF PROTEINS
Precipitation by HEAVY METAL IONS
Salts of Heavy Metals forms INSOLUBLE METALLIC PROTEINATES
Albumin+
Pb(Ac)2 & AgNO3 & CuSO4
Pb(Ac)2 & AgNO3-> WHITE
CuSO4 -> BLUE
Albumin + NaOH PPT MORE

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3. PRECIPITATION REACTIONS OF PROTEINS

Precipitation by HEAVY METAL IONS

Salts of Heavy Metals forms INSOLUBLE METALLIC PROTEINATES

Albumin + NaOH +

Pb(Ac)2 & AgNO3 & CuSO4

Pb(Ac)2 & AgNO3-> WHITE

CuSO4 -> BLUE

Albumin + NaOH PPT MORE

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3. PRECIPITATION REACTIONS OF PROTEINS
Precipitation by ALKALOIDAL RGTS
Alkaloidal rgts- ppt proves presences of ALKALOIDS
Albumin + HCl (to acidify) + Alkaloidal Rgts
Alkaloidal Rgts:
Tannic Acid (BROWN)
Sat. Picric Acid (YELLOW)
Potassium Ferrocyanide K4[Fe(CN)6)].3H2O (WHITE)

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4. TEST FOR CARBOHYDRATES
PHYSICAL: SOLUBILITY
H20 & NaCl & Alcohol
SOLUBLE in H20 & NaCl:
Fructose, Arabinose, Sucrose, Glucose
INSOLUBLE in Alcohol: ALL

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4. TEST FOR CARBOHYDRATES
PHYSICAL: DIALYSIS
GLUCOSE-STARCH-SUCROSE
Perform MOLISCH Test

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4. TEST FOR CARBOHYDRATES
FURFURAL: Molisch Test (Alpha-Napthol Rxn)
General Test for Cabohydrates
A dehydration-condensation reaction with the formation of FURFURAL
Molisch Rgt- Alpha-Napthol in Acohol
GLUCOSE SUCROSE STARCH + Molisch Rgt + H2SO4
GLUCOSE -> VIOLET RING
SUCROSE -> DARK VIOLET RING
STARCH -> LIGHT VIOLET RING

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4. TEST FOR CARBOHYDRATES
FURFURAL: Thymol Test
Positive test for Disaccharides
GLUCOSE SUCROSE STARCH + Thymol in-alcohol + Conc. HCl
A CARMINE COLOR develops in SUCROSE

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4. TEST FOR CARBOHYDRATES
FURFURAL:
Seliwanoff’s Reaction (Resorcin –HCl Test)
Positive test for Ketoses
Seliwanoff’s Rgt- 0.5% Resorcinol + HCl
GLUCOSE FRUCTOSE ARABINOSE + Seliwanoff’s Rgt -> BOIL
LIGHT CARAMEL SOL’N developed in FRUCTOSE
Due to rxn of HYDROXYL METHYL FURFURAL w/ RESORCINOL

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4. TEST FOR CARBOHYDRATES
FURFURAL: Moore’s Test
Action of Concentration of Alkali
Glucose + conc. NaOH -> BOIL
DARK BROWN COLORED SOLUTION w/ CARAMEL ODOR
Liberation of Aldehyde w/c polymerize to form a RESINOUS substance, CARAMEL

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4. TEST FOR CARBOHYDRATES
FURFURAL: Tollen’s Phloruglucin Reaction
To differentiate PENTOSES from HEXOSES
Phloruglucin Sol’n +
GLUCOSE FRUCTOSE ARABINOSE
ARABINOSE developed into a DARK colored sol’n

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5. SPECIAL TEST FOR SACCHARIDES
Nitro-Chromic Test
Positive when: Sugar conc. is more than 1%
Presence of –CHOH group
GLUCOSE + Conc. HNO3 + 5% Potassium Chromate (K2CrO4)
BLUE SOLUTION

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5. SPECIAL TEST FOR SACCHARIDES
Mucic Acid Test
GALACTOSE + Conc. HNO3-> BOIL
OXIDATION of GALACTOSE forms the INSOLUBILITY of MUCIC ACID (an ISOMER of SACCHARIC ACID)

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5. SPECIAL TEST FOR SACCHARIDES
Reduction Test
A.Fehling’s Test
Fehling’s Rgt: Rochelle’s Salt & KOH; CuSO4
GLUCOSE FRUCTOSE SUCROSE STARCH + Fehling’s Sol’n
BRICK RED PPT positive for GLUCOSE & FRUCTOSE

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5. SPECIAL TEST FOR SACCHARIDES
B.Benedicts Test (Qualitative Test for GLUCOSE in Urine
Benedict’ Sol’n: Sodium Citrate, CuSO4, Na2CO3
0.1M/0.01M/0.02M GLUCOSE + Benedicts Sol’n
RED PPT (MOST was found in 1M)

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5. SPECIAL TEST FOR SACCHARIDES
C.Barfoed’s Test (Detects Reducing MONOSACCHARIDES)
Barfoed’s Sol’n: Copper Acetate in Acetic Acid
CARBS + Barfoed’s Sol’n
RED PPT (FIRST found in FRUCTOSE)
REDUCES if SUGAR CONC is HIGH enough & TIME of heating is LONG

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5. SPECIAL TEST FOR SACCHARIDES
Iodine Test
STARCH & DEXTRIN + dil I2 sol’n
Starch -> BLUE ->after heating-> COLORLESS
Dextrin -> VIOLET->after heating-> YELLOW
Returns to it ORINAL COLOR when COOLED

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5. SPECIAL TEST FOR SACCHARIDES
Phenylhydrazine Reaction
Test for IDENTIFYING SUGARS with FREE ALDEHDE/KETONE
Phenylhydrazine Mixture:
2 Phenylhydrazine Hydrochloride: 3 Sodium Acetate
0.5 g Phenylhydrazine + CARBS
YELLOW COLORED PPT (SUCROSE & LACTOSE PPT MORE)
Guides in Identifying: Shape of Crystals, Time Osazone Formation

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OSAZONE
Glucose -> 4-5 mins

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OSAZONE
Fructose -> 2 mins

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OSAZONE
Maltose -> Osazone INSOLUBLE in HOT H2O

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OSAZONE
Lactose -> Osazone INSOLUBLE in HOT H2O

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OSAZONE
Sucrose -> 30-35 mins (AFTER HYDROLYSIS)

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NO PIC D:

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6. GLYCOGEN
A. PREPARATION
Oyster Meat –> Mince + SAND -> grind + H2O -> BOIL -> Filter -> Wash residue -> Conc. By Boiling + Acetic Acid (CH3COOH)
OPALASCENT FILTRATE (Glycogen)

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6. GLYCOGEN
B. REACTION OF GLYCOGEN
1. To Determine helical/non- helical complexes
a. Glycogen + NaCl + I2 sol’n ---------------->RED
b. Starch + I2 sol’n --------------------> DARK BLUE

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6. GLYCOGEN
B. REACTION OF GLYCOGEN
2. To Purify Glycogen
Glycogen + 95% Alcohol (ETHANOL)
WHITE PPT (Glycogen)

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6. GLYCOGEN
B. REACTION OF GLYCOGEN
3. Reduction Properties of Glycogen
PPT (in Glycogen + Ethanol) + Benedict’s sol’n
NR Because Glycogen is a POLYSACCHARIDE and it is NOT A REDUCING SUGAR

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6. GLYCOGEN
B. REACTION OF GLYCOGEN
4. Hydrolysis of Glycogen
Glycogen + HCl -> BOIL -> COOL +NaOH + Benedicts Sol’n
YELLOW-RED PPT
Hydrolysis products of Glycogen; GLUCOSE MALTOSE

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7. LIPIDS
SOLUBILITY
Coconut Oil
SOLUBLE: Cold Alcohol, Ether CHCl3
INSOLUBLE: Hot Alcohol, NaOH, H2o, HCl

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7. LIPIDS
REACTION TOWARDS INDICATORS
Coconut Oil
•RED PPT w/ Congo Red
•NEUTRAL with Litmus Paper

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7. LIPIDS
REACTION TOWARDS INDICATORS
Rancid Oil
•DARK PURPLE PPT w/ Congo Red
•ACID with Litmus Paper

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7. LIPIDS
FORMATION OF TRANSLUCENT SPOT

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NO PIC D:

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7. LIPIDS
ACROLEIN FORMATION
Chemical Test for Fats and Glycerol
0.5 g KHSO4 + Coconut Oil -> HEAT
BURNT FAT ODOR

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NO PIC D:

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7. LIPIDS
EMULSIFICATION
A Mixture of 2 immiscible liquids with the presence of an emulsifying agent
Coconut Oil + H20 + SOAP SOL’N/ ALBUMIN SOL’N/ Na2CO3 SOL’N
PERMANENT EMULSION: SOAP & ALBUMIN SOL’N
TEMPORARY EMULSION: H2O & Na2CO3
N

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7. LIPIDS
TEST FOR UNSATURATION
Oleic Acid - Absorbs MORE I2 than Stearic
Stearic Acid
Coconut Oil
Linseed Oil - Contains MORE UNSATURATED Fatty Acids than Coconut oil

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7. LIPIDS

TEST FOR UNSATURATION

Oleic Acid + CHCl3 + Hubl’s Iodine -> ORANGE

CHCl3 + Hubl’s Iodine -> PINK

Absorbs MORE I2 than Stearic

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7. LIPIDS
TEST FOR UNSATURATION
Stearic Acid + CHCl3 + Hubl’s Iodine -> PINK
CHCl3 + Hubl’s Iodine -> PINK

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7. LIPIDS
TEST FOR UNSATURATION
Coconut Oil + CHCl3 + Hubl’s Iodine -> ORANGE
CHCl3 + Hubl’s Iodine -> PINK

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7. LIPIDS

TEST FOR UNSATURATION

Linseed Oil + CHCl3 + Hubl’s Iodine

CHCl3 + Hubl’s Iodine

Contains MORE UNSATURATED Fatty Acids than Coconut oil

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NO PIC D:

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7. LIPIDS
GLYCEROL
SOLUBILITY
SOLUBLE: H20 & Alcohol
Insoluble Ether

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7. LIPIDS
GLYCEROL
ACROLEIN TEST
KHSO4 + Glycerol
BURNT GREASE ODOR

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7. LIPIDS
GLYCEROL
NICHROME WIRE W/ POWDERED BORAX
GREEN FLAME
Due to the GLYCEROL ESTER of BORIC ACID

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NO PIC D:

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7. LIPIDS
GLYCEROL
NITRO-CHROMIC ACID TEST
Indicates presence of –CHOH groups
GLYCEROL + HNO3 + Potassium Chromate (K2Cr2O7)
BLUE SOL’N

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8. ENZYMES
OXIDASES FROM FRUITS
Darkening from exposure of APPLE, CHICO GUAVA
Due to presence of OXIDASE

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8. ENZYMES
CATALASE from POTATO
Grated potato into a fine pulp + water
•BOILED + H2O2 -> WHITE PPT
•UNBOILED + H2O2 -> BUBBLES

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8. ENZYMES
OXIDASES FROM POTATO
POTATO EXTRACT +
1% Phenol----->
1% Cathecol--->
Pyrogallol---->

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8. ENZYMES
PEROXIDASE FROM POTATO
Requires a Co-Factor, PHENOL-OXIDASE
POTATO EXTRACT + H2O2
1% Phenol----
1% Cathecol--
Pyrogallol-----

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8. ENZYMES
CATALASE FROM LIVER
Test GLOWING SPLINT
Liver + Sand -> GRIND + H2O2
GREEN FLAME
OXYGEN (gas liberated that made the flame LUMINOUS)

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9. TEST FOR NUTRIENTS IN FOODS
STARCH
F.C + I2 sol’n -> DEEP BLUE SOL’N

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Ripe Banana

Green Banana

Peanut Butter

Potato

Cheese

Cooked Rice

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9. TEST FOR NUTRIENTS IN FOODS
GLUCOSE
F.J + Benedicts sol’n -> YELLOW-RED PPT

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Ripe Banana

Green Banana

Carrots

Peanut Butter

Cheese

Fruit Juice

Cooked Rice

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9. TEST FOR NUTRIENTS IN FOODS
FATS
F. S + Ether -> TRANSLUCENT SPOT

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Ripe Banana

Green Banana

Egg Yolk

Peanut Butter

Cheese

Dilis

Uncooked Peanuts

Cooked Rice

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9. TEST FOR NUTRIENTS IN FOODS
PROTEINS
F.S + NaOH + CuSO4 -> PINK-VIOLET COLORATION

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Egg White

Egg Yolk

Peanut Butter

Cheese

Dilis

Uncooked Peanuts

Cooked Rice

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9. TEST FOR NUTRIENTS IN FOODS
MINERAL MATTER
F.S -> BURN -> FORMATION OF ASH

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Ripe Banana

Green Banana

Dilis

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9. TEST FOR NUTRIENTS IN FOODS
CARR-PRICE TEST FOR VITAMIN A
CHCl3 + F.S -> H2O bath + Antimony Trichloride [Sb(Cl3)] -> BLUE SOL’N

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Margarine

Butter

Carrots

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9. TEST FOR NUTRIENTS IN FOODS
CHOLESTEROL
ACETIC ANHYDRIDE-H2SO4 TEST
LIEBERMAN BUCHARD TEST
F.S CHCL3 + Acetic anhydride + conc. Sulphuric acid -> BLUISH GREEN SOL’N

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Egg Yolk

Peanut Butter

Cheese