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