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BIO LAB

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Allyssa Bianca Granada
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4 views

BIO LAB

Uploaded by

Allyssa Bianca Granada
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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BIOCHEMISTRY LABORATORY NOTES

CARBOHYDRATES

● Isolation of Liver glycogen: extraction of polysaccharides from animal tissue


using TCA to denature proteins and solubilize glycogen.
○ Chemicals used:
■ TCA (10% for grinding and 5% for rinsing)- denature proteins and
extract glycogen from the liver tissue.
■ 95% ethanol- precipitate glycogen from the aqueous extract.
■ Sodium Chloride (NaCl)- help in promoting flocculation of glycogen.
● Isolation of Plant starch: involve grinding the plant material to release starch
granules.

GENERAL TESTS FOR CARBOHYDRATES


● Molisch Test: reacts with molisch reagent in the presence of concentrated sulfuric
acid to form a purple complex.
○ Detect general carbohydrates (mono and poly)
○ Positive: Purple Ring
○ Chemicals used: 2-3 drops molisch reagent, 1-2 mL concentrated sulfuric
acid (H2SO4)
● Anthrone Test: reacts with anthrone reagent under acidic conditions to produce
a blue-green color.
○ Detect polysaccharides
○ Positive: Blue-green color
○ Chemicals used: 2 mL anthrone reagent
● Iodine Test: Iodine forms a blue complex due to the helical structure of amylose.
○ Detects starch
○ Positive: Blue-black color
○ Chemicals used: few drops of 0.01 M Iodine solution

SPECIFIC TESTS FOR CARBOHYDRATES


● Seliwannof’s Test: Ketoses react with seliwanoff’s reagent and concentrated HCl
to produce a cherry-red color.
○ Detect ketoses
○ Positive: Cherry-red color
○ Chemicals used: 5 mL Seliwanoff reagent, 1 mL sugar sol.
● Bial Orcinol: pentoses react with orcinol reagent and HCl to produce a
blue-green color.
○ Detect pentoses
○ Positive: Blue-green color
○ Chemical used: 4.5 mL Orcinol Reagent, 0.5 mL sugar sol.
BIOCHEMISTRY LABORATORY NOTES

● Mucic Acid test: Hexoses produce mucic acid upon oxidation with nitric acid.
○ Detects galactose (from lactose)
○ Positive: White precipitate
○ Chemicals used: 1 mL Nitric Acid (HNO3), 50 mg sugar
● Osazone Formation Test: Reducing sugars reacts with phenylhydrazine to form
osazones, which are yellow crystals.
○ Detect reducing sugars(can be identified)
○ Positive: Yellow crystalline precipitate
○ Chemicals used: 2 g Phenylhydrazine, 4 g sodium acetate, 3-10 drops
glacial acetic acid, 5 ml sugar sol.
● Fehling’s Test: reducing sugars reduces copper (II) ions to copper (I) oxide in
alkaline solution.
○ Detect reducing sugars (presence only, no specific identification)
○ Positive: Red Precipitate
○ Chemicals used: 1 ml each of Fehling’s A and B diluted w 4 ml water, 8
drops sugar sol.
● Barfoed’s Test: Monosaccharides reduce copper (II) ions in an acidic solution to
form red precipitate, while disaccharides do not.
○ Sample Detected: Monosaccharides
○ Positive: Red precipitate
○ Chemicals used: 3 mL Barfoed’s reagent, 10 drops sugar sol.
● Benedict’s Test: Reducing Sugars reduce copper (II) ions in alkaline solution to
form colored precipitates.
○ Detect reducing sugars (presence and amount only, no specific
identification)
○ Positive: Green/ Yellow/ Red precipitate depending on conc.
○ Chemicals used: 2 mL Benedict’s reagent, 5 drops sugar sol.

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