Republic of the Philippines

Sorsogon State College

Sorsogon City Campus
Sorsogon City
________________________________________________________________________
JHON DAVE SURBANO

BSED-SCIENCE 1A

ANALYTICAL CHEMISTRY

The incident began when a park ranger found a dead white-tailed deer near a pond in the Land
Between the Lakes National Recreation Area in western Kentucky. The ranger and the chemist were able
to find the cause of death so that further deer kills might be prevented. But they badly found the
decomposed carcass. Thus, there’s no fresh organ tissue could be gathered because of the advanced
state of decomposition. However, a few days after several investigations, the ranger discovered two
more dead deer near the same location. The investigators noticed that grass surrounding nearby power
line poles was wilted and discolored, whereas the investigator speculated that a herbicide might have
been used on the grass. A common ingredient in herbicides is arsenic which when expose to animals and
plants can cause chemical reactions and later on death. which the compound is the disodium salt of
methane arsenic acid (CH 3AsO(OH)2), which is very soluble in water. and therefore, the case study
focuses on confirming the presence of arsenic in both the grass and deer organs to determine its
concentration in the sample.

Speculation on the presence of arsenic was made after the proper observing and investigation
on the area. However, in determining the quantity of arsenic in the acquired biological samples of both
grass and deer organs. The investigations based or used the published methods of the Association of
Official Analytical Chemists (AOAC). In this method, the arsenic distilled as arsine, AsH 3, and then
determined by colorimetric measurement.

The biological sample used for testing was acquired through collecting samples of the discolored
dead grass along with the samples from the organs of the deer that were both found in the area of the
event.

After acquiring the sample, it was then transferred to the laboratory. The deer were dissected,
and the kidneys were removed for analysis, it is because the suspected pathogen (arsenic) is rapidly
eliminated from an animal through its urinary tract. The kidney was then cut into pieces and
homogenized in a high-speed blender. Three 10-gram samples of homogenized kidney tissue from each
deer were placed in porcelain crucibles which it is served as replicates for the analysis. Then, the
procedure called dry ashing was made to obtain an aqueous solution for analysis. Dry ashing served to
free the analyte from organic material and convert it to arsenic pentoxide. The dry solid in each sample
credible was then dissolved in dilute HCI, which converted the As 2O5 into water-soluble arsenic acid
(H3AsO4).
 In order to estimate the substances that can interfere on the analysis. Arsenic was separated
from other substances by converting it into arsine AsH 3 which the solutions resulting from the deer and
grass were combined with Sn2+, and a small amount of iodide ion was added to catalyze the reduction of
H3AsO4 to H3AsO3, the H3AsO3 was then converted to AsH3 by the addition of Zinc metal). The entire
reaction was carried out in flasks equipped with a stopper and delivery tube and the arsine was been
collected in the absorber solution. This arrangement ensured that interferences were left in the reaction
flask and that only arsine was collected in the absorber in special transparent containers which is called
cuvettes.

The reduction of H3AsO4 to H3AsO3 according to the following equation:

H3AsO4 + SnCl2 + 2 HCl  H3AsO3 + SnCl4 + H2O

The conversion of H3AsO3 into AsH3 by the addition of zinc metal as follows reaction:

H3AsO3 + 3 Zn + 6 HCl  AsH3(g) + 3 ZnCl2 + 3 H2O

To determine the amount of arsenic in each sample the chemist measured the intensity of the
red color formed in the cuvettes with an instrument called UV-Vis spectrophotometer. A calibration
curve was generated by measuring the absorbance of several solutions that contain known
concentrations of analyze. The result shows that the color becomes more intense the arsenic content of
the standards increases from 0 to 25 parts per million (ppm).

Arsenic Concentration (ppm) Absorbance

0.0 (blank) 0.00

5.0 0.13

10.0 0.28
15.0 0.42

20.0 0.59
25.0 0.71

Deer #1 (unknown concentration) 0.47

Deer #2 (unknown concentration) 0.63

Calibration curve is used to determine the concentration of arsenic. The absorbances of the
solutions in the cuvettes are measured using a spectrophotometer. The color intensity of each solution
is represented by its absorbance in which increases from 0 to about 0.72 as the concentration of arsenic
increases from 0 to 25 parts per million. The absorbance values are then plotted in the curve and then
read the corresponding concentrations. The lines leading from the cuvettes to the calibration curve
shows that the concentrations of arsenic in the two deer samples were 16 ppm and 22 ppm,
respectively. Arsenic in kidney tissue of an animal is toxic at levels about 10 ppm, so it was probable that
the deer were killed by 600 ppm arsenic. Thus, the investigators concluded that the deer had probably
died as a result of eating the poisoned grass.

Statistical methods were used to analyze the data in the experiment. For each of the standard
arsenic solutions and the deer samples, the average of the three absorbance measurements was
calculated. The average absorbance for replicates is a more reliable measure of the concentration of
arsenic then a single measurement. To find the best straight line among the points and to calculate the
concentrations of unknown samples along with their statistical uncertainties and confidence limit, least
squares analysis of the standard data was been used. Theses methods are reliable and therefore the
result of the analysis is indeed true.

The overall process illustrates how analytical chemistry is used in solving a problem or in this
paper Case Study: Toxicology Investigation (Based on the case study Deer Kill)