STONE ANALYSIS
Common biological calculi
. Renal calculi
. Gall stone analysis
. Salivary duct stone ( lesser extent )
RENAL CALCULI
any part of kidney
Chemical substances
1. Phosphate
2. Salts of Calcium
3. Uric Acid
4. Organic substances –
Cholesterol,Bilirubin,Xanthin
Conditions lead to CALCULOGENESIS
a. Hypercalciuria
b. Hyperoxaluria
c. both
Inhibition of Calculogenesis
@ Citric acid
@ Pyrophosphate
@ Glycosaminoglycans
@ Tamm Horsefall protein
ANALYSIS OF RENAL STONE
Physical Appearance
1. Colour
2. surface
3. consistency
4. Size
5. large stone ,cut into layers to see the
Prior layers
to Test
1. Separated
2. Wash
3. Dry
4. Crushed
TEST OBSERVATION CHEMICAL NATURE
1. APPEARANCE a. Coloured a. URIC ACID & URATE
b. Small size b. Uric Acid
2. SOLUBILITY
a. WATER a. Insoluble In cold a. URIC ACID & URATE
Soluble on warming,
Ppt on coooling
b. Dil Hcl b. 1. soluble b.1. OXALATE , PHOSPHATE
b.2. Insoluble b.2. URIC acid
if soluble, add b.a. Amorphous ppt b.a. PHOSPHATE
AMMONIUM HYDROXIDE
b.b. Crystaline Ppt b.b. Triple PHOSPHATE
c. Dil ACETIC ACID c. soluble c. Phosphate
1. TEST FOR CALCIUM
Stone powder in test tube
Add Diluted Hcl
Heat
Add 1 % glacial acetic acid
Add ammonium powder
White precipitate
PRINCIPLE
Formation of Calcium Oxalate
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PRINCIPLE
3. TEST FOR URIC ACID
Stone powder
10 % KOH solution
14 % NaCO3 solution
Benedict ‘ s uric acid reagent
BLUE COLOUR
PRINCIPLE
Uric acid in ALKALINE medium, REDUCES
Phosphotungstic acid to
PHSOSPHOTUNGSTEN BLUE
4. TEST FOR PHOSPHATE
Stone powder
Add Conc HNO3
Add Ammonium Molybdate
Mix & Heat
Canary Yellow colour
PRINCIPLE
Formation of Ammonium PhosphoMolybdate
5. TEST FOR OXALATES
Stone powder
Add conc. Sulphuric acid
boil
Add dilute solution of Potassium Permanganate
Disappearance of VIOLET colour
GALL STONE
Usually forms in gall bladder
Colour depends on the pigment content,
it varies from white/ brown/ dark green / near black
It may be
1. pure cholesterol
2. cholesterol and calcium
3. cholesterol / calcium / bile pigment
GALL STONE ANALYSIS
STEP 1 •Extraction with ETHER
•LIBERMANN- BURCHARD
STEP 2 REACTION
•TEST FOR CALCIUM &
STEP 3 •PHOSPHORUS
• TEST FOR BILE PIGMENTS
STEP 4
STEP -1
EXTRACTION WITH ETHER
Stone Powder With ETHER I In A Test Tube In Warm Water
filter
Evaporation of ether
Residue dissolve with 95 % Ethanol
Crystallisation
Observe crystal under microscope
Notched CRYSTAL
Step 2
LIBERMANN- BURCHARD REACTION
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For cholesterol
PRINCIPLE
In the presence of Acetic Anhydride ,
cholesterol and Sulphuric acid gives green
colour.
Step 3
test for CALCIUM & PHOSPHORUS
Add Residue
and Diluted Hcl Filter
Filtrate
separated
Test for TEST FOR
CALCIUM PHOSPHORUS
TEST FOR TEST FOR
CALCIUM PHOSPHORUS
•Filtrate , add with •To the Filtrate add
Glacial Acetic acid Conc Nitric acid and
Ammonium Molybdate
and Ammonium •Heat
Oxalate • canary Yellow
•WHITE PRECIPITATE precipitate
Step 4
Test for BILE PIGMENTS
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Appearance of GREEN colour - BILIRUBIN
PRINCIPLE
Bile pigments present in urine – adsorbed to the
precipitated BARIUM SULPHATE,FeCl3 in FOUCHET’S reagent,
oxidises BILIRUBIN to GREEN coloured BILIVERDIN, BLUE
Coloured BILICYANIN, BROWN coloured BILIFUSCHIN.
THANK YOU
