Purpose: Colorimetric spot test for alkaloids, especially morphine and codeine.
Procedure:
Slowly add 1 ml concentrated sulfuric acid to 1 ml of cold fuming nitric acid in a chilled container.
Mix well and use immediately.
⚠️ Use extreme caution due to strong acids and fumes.
Dragendorff’s Reagent
Purpose: Detection of alkaloids in plant extracts or formulations.
Procedure (stock solution method):
Stock A: Dissolve 1.7 g bismuth subnitrate in 20 ml glacial acetic acid.
Stock B: Dissolve 40 g potassium iodide in 100 ml distilled water.
Mix 10 ml of A and 20 ml of B.
Dilute with distilled water to make 100 ml.
Store in a dark bottle; stable for weeks.
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DETERMINATION OF DISSOLVED OXYGEN IN WATER (WINKLER METHOD)
AIM:
To determine the dissolved oxygen (DO) content in water.
APPARATUS:
BOD bottles, burette, pipette.
REAGENT:
Manganese sulfate, alkaline iodide, starch, sodium thiosulphate.
PRINCIPLE:
DO oxidizes Mn²⁺ to Mn⁴⁺, which liberates iodine that is titrated.
PROCEDURE:
Add MnSO₄ and alkaline iodide to water sample.
Allow precipitate to form and settle.
Add H₂SO₄ to dissolve.
Titrate liberated iodine with sodium thiosulphate using starch as indicator.
CALCULATION:
DO (mg/L) = Volume of thiosulfate used × Normality × 8
DETERMINATION OF CITRIC ACID IN FRUIT JUICE
AIM:
To estimate citric acid content in fruit juice.
APPARATUS:
Burette, conical flask, pipette.
REAGENT:
0.1M NaOH, phenolphthalein.
PRINCIPLE:
Citric acid neutralized by NaOH; endpoint indicated by pink color.
PROCEDURE:
Pipette 10ml of juice into a flask.
Add 2–3 drops phenolphthalein.
Titrate with NaOH to pink endpoint.
CALCULATION:
% Citric Acid = Titre × 0.064 × 100 / Volume of sample
DETERMINATION OF CHLOROPHYLL IN LEAF EXTRACT
AIM:
To estimate chlorophyll content in a leaf sample.
APPARATUS:
Spectrophotometer, centrifuge, test tubes.
REAGENT:
80% acetone.
PRINCIPLE:
Chlorophyll is extracted and measured at specific absorbance values.
PROCEDURE:
Grind 1g of leaf in 10ml acetone.
Centrifuge and collect supernatant.
Measure absorbance at 645 nm and 663 nm.
CALCULATION:
Use Arnon's formula:
Chlorophyll a = 12.7(A663) – 2.69(A645)
Chlorophyll b = 22.9(A645) – 4.68(A663)
