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    Carbohydrate Post Lab Discussion

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    anon nymouse
    This document discusses carbohydrate analysis, including the structures and properties of polysaccharides like starch, cellulose, and glycogen. It describes the isolation of starch from potatoes and glycogen from chicken liver. Common tests for carbohydrates are outlined, such as the Molisch test for all carbohydrates, and iodine test for starch. The document compares acid and enzymatic hydrolysis of polysaccharides. Specific tests are defined for reducing sugars, monosaccharides, ketohexoses, pentoses, and galactose.

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    Carbohydrate Post Lab Discussion

    Uploaded by

    anon nymouse
    46 views28 pages
    This document discusses carbohydrate analysis, including the structures and properties of polysaccharides like starch, cellulose, and glycogen. It describes the isolation of starch from potatoes and glycogen from chicken liver. Common tests for carbohydrates are outlined, such as the Molisch test for all carbohydrates, and iodine test for starch. The document compares acid and enzymatic hydrolysis of polysaccharides. Specific tests are defined for reducing sugars, monosaccharides, ketohexoses, pentoses, and galactose.

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    Carbohydrate Post Lab Discussion

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    This document discusses carbohydrate analysis, including the structures and properties of polysaccharides like starch, cellulose, and glycogen. It describes the isolation of starch from potatoes and glycogen from chicken liver. Common tests for carbohydrates are outlined, such as the Molisch test for all carbohydrates, and iodine test for starch. The document compares acid and enzymatic hydrolysis of polysaccharides. Specific tests are defined for reducing sugars, monosaccharides, ketohexoses, pentoses, and galactose.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    46 views28 pages

    Carbohydrate Post Lab Discussion

    Uploaded by

    anon nymouse
    This document discusses carbohydrate analysis, including the structures and properties of polysaccharides like starch, cellulose, and glycogen. It describes the isolation of starch from potatoes and glycogen from chicken liver. Common tests for carbohydrates are outlined, such as the Molisch test for all carbohydrates, and iodine test for starch. The document compares acid and enzymatic hydrolysis of polysaccharides. Specific tests are defined for reducing sugars, monosaccharides, ketohexoses, pentoses, and galactose.

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    ANALYSIS OF CARBOHYDRATES

    POLYSACCHARIDES
    • Polymers of simple sugar units
    • Plants: Starch & Cellulose
    1. Starch: (14)
    A. Amylose (unbranched)
    B. Amilopectin (branched)
    2. Cellulose:  (14)
    • Animals: Glycogen
    PHYSICAL CHACTERISTICS OF STARCH
    • amorphous solid
    • slightly soluble in water
    • forms colloidal suspension when heated
    PHYSICAL CHACTERISTICS OF
    GLYCOGEN
    • white amorphous solid
    • Soluble in water
    • Insoluble in organic alcohol
    PHYSICAL CHACTERISTICS OF
    CELLULOSE
    • Fibrous polysaccharide
    • Insoluble in water
    POLYSACCHARIDE STRUCTURE
    ISOLATION OF STARCH
    Source: Potato/Sweet Potato/Cassava
    Isolation Procedure:

    Cell Decantation/
    Homogenization Filtration

    Basis of Separation: Difference in Solubility


    ISOLATION OF GLYCOGEN
    Source: Chicken Liver
    Isolation Procedure:
    Cell Addition of Alcohol
    Homogenization Filtration
    Acetic Acid Precipitation

    Basis of Separation:
    1. Protein Precipitation
    2. Selective Solubilization
    GENERAL TESTS FOR CARBOHYDRATES

    A. MOLISCH TEST
    – Test for all CHO
    – Reagents: conc. H2SO4, -naphthol in 95% EtOH
    – Principle:
    1) Hydrolysis of CHO to monosaccharides
    2) Dehydration of monosacharrides
    a. pentoses form furfural
    b. hexoses form hydroxymethyl furfural
    3) Condensation with -naphthol
    – Visible Positive Result: purple ring at the interphase
    MOLISCH TEST
    Formation of furfural from pentoses

    Formation hydroxymethyl furfural from hexoses


    MOLISCH TEST
    GENERAL TESTS FOR CARBOHYDRATES

    B. IODINE TEST
    – Specific Test for Starch
    – Reagent: I2 solution
    – Principle: Formation of Iodo-starch complex
    – Visible positive result:
    blue-black solution/precipitate
    HYDROLYSIS OF POLYSACCHARIDES
    ACID VS ENZYME HYDROLYSIS
    Basis for Acidic Enzymatic
    Comparisson (amylase)
    Specificity Non-specific Specific
    -all types of glycosidic (14) glycosidic bond
    linkage
    Condition -High Temperature Physiological Temperature
    Requirements
    Products Monosaccharides (glucose) Monosaccharide (glucose)
    Disaccharide (maltose)
    Oligosaccharides
    -dextrin
    -amylodextrin
    -erythrodextrin
    DIALYSIS
    • A separation technique based on ability of
    molecule to pass through a semi-permeable
    membrane until an equilibrium concentration
    is established between the outer and inner
    solution
    SPECIFIC TESTS FOR CARBOHYDRATES
    1. Benedict’s Test
    – Tests for reducing sugars (free anomeric carbon at
    the reducing end)
    – Reagents: CuSO4, Na2CO3, Na3C6H5O7
    – Principle: Reduction of Cu2+ to Cu2O (in slightly
    basic medium)
    – Visible Positive Result: brick-red ppt
    BENEDICT’S TEST
    SPECIFIC TESTS FOR CARBOHYDRATES
    2. Barfoed’s Test
    – Tests for monosaccharides
    – Reagents: CuSO4, CH3COOH
    – Principle: Reduction of Cu2+ to Cu2O (in slightly
    acidic medium)
    – Visible Positive Result: brick-red ppt
    BARFOED’S TEST
    SPECIFIC TESTS FOR CARBOHYDRATES
    3. Seliwanoff Test
    – Tests for ketohexoses
    – Reagents: resorcinol
    – Principle: condensation of resorcinol to
    hydroxymethyl furfural
    – Visible Positive Result: cherry red solution
    SELIWANOFF TEST
    SPECIFIC TESTS FOR CARBOHYDRATES
    4. Bial’s Orcinol Test
    – Tests for pentoses
    – Reagents: orcinol, FeCl3, HCl
    – Principle:
    1. dehydration of pentose to form furfural
    2. Condensation with orcinol
    3. Complexation w/ Fe3+
    – Visible Positive Result: blue green solution
    BIAL’S ORCINOL TEST
    MUCIC ACID TEST
    • Specific test for galactose and lactose
    • Reagent: conc. HNO3
    • Principle: oxidation of galactose forming
    aldaric acids (galactaric acid forms meso
    compound)
    • Visible Positive result: broken-glass like
    crystals
    MUCIC ACID TEST
    PHENYLHYDRAZONE TEST
    • Formation of ozasone crystals of CHO
    containing free carbonyl at C1 or C2
    • Reagents: phenylhydrazine, HCl, NaOAc
    • Principle: ANE, oxidation via rearrangement
    and elimination
    • Visible positive result: formation of yellow-
    orange crystals (osazone)
    PHENYLHYDRAZONE TEST
    END

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