Angeles University Foundation

Angeles City, Pampanga

College of Allied Medical Professions

Department of Medical Technology

Routine Stool Examination:

Chemical Phase

Submitted by:
Group3 BSMT-3C
Bautista, Joseph Ryan C.
Gamboa, Lois Danielle L.
Gundran, Angelika Joy

Introduction

Gross or macroscopic examination of stool may not always reveal the

presence of blood, which is why chemical examination for occult blood is
employed. Occult blood, generally, is not present. If there is occult blood detected
in the stool specimen, it indicates either infection or some disorder of the digestive
system.
Other chemical tests for fecal specimen include detection of reducing
substances and determination of pH. Strongly acidic stool (pH below 5.5) indicates
an excess of carbohydrates in the diet. It is nonpathologic. But, if fermentation is
present, it may be due to lactose intolerance. Then, it is pathologic. Strongly
alkaline stool (pH above 7.5) indicates an excess of protein in the diet. It is non
pathologic. Reducing substances are generally found in stools of infants suffering
from diarrhea.

Flowchart/Brief Summary of the Procedure

Guaiac Test
It is performed using a slide that contains paper squares coated with guaiac,
a chemical derived from resin of a tree of the genus Guaiacum. A small portion of
fecal/stool specimen is applied to the paper squares in the special slide. Then, a
developer solution containing hydrogen peroxide is added to the paper. If blood is
present in the specimen, the iron (Fe) in the hemoglobin catalyzes the reaction
between the guaiac and the hydrogen peroxide, forming a blue color. Fecal occult
blood determination is based on the detection of the pseudoperoxidase activity of
hemoglobin.

Results and Interpretation

Negative for Occult

Blood

Positive for Occult

Blood

Results Obtained: Negative for Occult Blood Test

Manner of Reporting
o Trace very faint blue
o + - faint blue
o ++ - distinct blue
o +++ - dark blue
o ++++ - prussian blue

Answer to Research Questions

1. State the principle of fecal occult blood determination.
o Fecal occult blood determination is based on the pseudoperoxidase
activity of hemoglobin in which liberates oxygen from the reaction
of stool and hydrogen peroxide to oxidize the guaiac reagent.
2. What are the different indicators that are used in the detection of fecal
occult blood?
o Guaiac resin - When the hydrogen peroxide is dripped on to the
guaiac paper, it oxidizes the alpha-guaiaconic acid to a blue colored
quinone.
o Benzidine - the suspected fluid is treated with glacial acetic acid and
ether, and the latter is then decanted and treated with hydrogen
peroxide and a solution of benzidine in acetic acid; the presence of
blood is indicated by a bluish colour turning to purple.
o Imipramine hydrochloride (IPH) and desipramine hydrochloride
(DPH)- proposed as new reagents for detection of fecal occult blood.
The usefulness of IPH and DPH in occult blood detection has been

examined and compared with benzidine and stanoccult methods.

The results show that the proposed reagents are selective and
sensitive and gives reproducible results. The proposed methodology
is much less subject to vegetable peroxidase, iron and vitamin C
interference and can be performed on patients who are on a normal
diet.

Updates/Related Journal
Digestive Diseases and Sciences
October 1997, Volume 42, Issue 10, pp 2064-2071
Performance
Characteristics
and
Comparison
of
Two
Immunochemical and Two Guaiac Fecal Occult Blood Screening Tests
for Colorectal Neoplasia
A new immunochemical test for stool Hb, FlexSureOBT, was compared with the
immunochemical HemeSelect and guaiac Hemoccult II and Hemoccult SENSA
tests. Blinded development of test cards smeared with stools having added human
blood showed better analytical sensitivity of FlexSure OBT (0.2 ml blood/100g
feces), than Hemoccult SENSA (0.5 ml) or Hemoccult II (1.0 ml). All four stool
tests were prepared by 403 subjects having endoscopic examinations. The guaiac
tests and FlexSure OBT were easy to prepare and develop. The positivity rate of
Hemoccult SENSA was 8.7%, Hemoccult II 6%, FlexSure OBT 4.2%, and
HemeSelect 3.4%. In this population, guaiac tests were as sensitive as
immunochemical tests for clinically significant colorectal neoplasia, but with
significantly lower predictive positive values. A combination of a sensitive guaiac
test (Hemoccult SENSA) and a specific confirmatory test for human Hb (FlexSure
OBT) provided high specificity, comparable to HemeSelect.

References:
o B. Estridge, Basic Clinical Laboratory Techniques, 2011, pp. 680-681
o S. Strasinger, Urinalysis and Body Fluids, 5th Edition, pp. 245-251
o http://www.aarogya.com/conditions-and-diseases/diagnostic-tests/physical-achemical-examination.html