Chemical Testing of Blood
MLS BIOCHEM LAB
Maria Teresa F. Mateo, RMT,MD,FPCOM,MBA
v Introduction
v Chemical test for blood
Benzidine test
Kastle-Mayer Test
Hemin test
Guaiac test
Confirmatory test
Benzidine Test
Ø a specific test used to detect the presence of
blood (RBCs or hemoglobin) in urine, stool and
other body fluids
Ø Test Principle: H2O2 with the presence of rbc
reduce benzidine solution. Result yields a blue
color complex
Ø Reagents: Benzidine Reagent (Benzidine in
Glacial Acetic Acid), 3% Hydrogen Peroxide,
Distilled Water.
Ø Method: mix 2ml of Benzidine solution with 2ml
H2O2. Take 2ml of specimen and add 2 ml of the
mixture and observe.
Ø Result: Appearance of a blue color solution
The Kastle-Meyer test
Test used to analyze evidence of blood at a crime scene.
PRINCIPLE:
Phenolphthalein reacts with hydrogen peroxide in the presence of
hemoglobin to turn from colorless to pink.
False positive: hemoglobin (meat) and certain vegetables will also
generate a positive response.
An additional benefit of this procedure is that the samples remain
intact and can be used in further testing including DNA analysis.
RESULT: Bright pink color
Hemin Test
ØAlso known as the Teichmann Test
ØPrinciple: When hemoglobin is
heated with glacial acetic acid, it
forms microscopic crystals
ØResult: Rhomboid crystals under
microscope
Method:
Ø Place a drop of blood on a glass slide.
Ø Add a drop of water, stir, and spread
uniformly by using the tip of a cover
glass.
Ø Add a drop of 0.1% NaCl solution.
Ø Dry over a low flame.
Ø Place cover glass in position and add
underneath a drop of concentrated
acetic acid.
Ø Warm as before until bubbles are
formed.
Ø Cool and examine the crystal under
the microscope.
Takayama test
Ø Also known as hemochromogen crystal
assay,
Ø Principle:
Ø Ferrous iron from hemoglobin
interacts with a pyridine solution.
Ø This interaction creates specific
crystals in the form of pyridine
ferroprotoporhyrin.
Ø Result: Red crystals when viewed
under a microscope.
Guaiac Test
Ø Detects hidden or occult blood
Ø Guaic solution (a natural resin from plants)
Ø Principle:
Guiac Test
Method:
Ø To 1 mL diluted blood, add new drops of
Guiac solution, until the resulting
solution becomes turbid.
Ø Add 1% H2O2 dropwise until a color
reaction appears.
THANK YOU.
END