Name: RAFAEL, DUNE VIENIS KAREN N.

Year &
Section: BS-Pharmacy 4A
Date Performed: November 20, 2009 Rating:
____________________

Activity No. 04
CHLOROFORM

Objectives:
1. To detect the presence of chlorofrom by performing phenylisocyanide test,
Schwartz’s test, Lustgarten’s test, cyanide test and reduction tests namely,
Fehling’s test and Tollen’s test,
2. To understand the principles behind each test, and
3. Interpret the results after performing each method of detection.

Data:
Tests
1. Phenylisocyanide Test
2. Schwartz’s Resorcinol
Test
3. Lustgarten’s Naphthol
Test
4. Reduction Tests
a. Fehling’s Test
b. Tollen’s Test
1. Cyanide Test
Data Analysis
Observation/Results
Actual: A foul odor was recognized.
Ideal (+): A very penetrating and very repulsive
odor is easily recognized(Warren, 1921).
Blank (-): No odor was recognized.
Actual: A yellowish-red color fluorescence was
observed.
Ideal (+): A yellowish-red color with a yellowish-
green fluorescence is observed (Warren, 1921).
Blank (-): A faint purple color was recognized.
Actual: It changed from blue to green and finally
turned to brown.
Ideal (+): From an evanescent blue color, it
changed to green and then to brown in contact
with air (Warren, 1921).
Blank (-): No remarkable color change was
noticed.
Actual: There was formation of red precipitate.
Ideal (+): Red precipitate appears(Warren,
1921).
Blank (-): No precipitates appeared.
Actual: Formation of silvery precipitate was
observed.
Ideal (+): A black precipitate of metallic silver
appeared (Warren, 1921).
Blank (-): No precipitation.
Actual: There was formation of blue precipitate
Ideal (+): Blue precipitation occurs (Warren,
1921).
Blank (-): No precipitation can be observed.
Phenylisocyanide Test
The penetrating and very repulsive odor of this compound is easily
recognized. A.W. Hofmann states that this test will show with certainty 1: 5,000 to
6,000 parts of alcohol. It should also be noted that chloral, chloral hydrate,
bromoform, iodoform and tetrachloromethane also give this test (Warren, 1921).

CHC3 + C6H6NH2 + 3KOH  C6H6NC + 3KCl + 3H2O

Schwartz’s Resorcinol Test

This mixture heated to boiling will develop even in very dilute solution a
yellowish red color attended by a beautiful yellowish green fluorescence. It should
be noted that chloral, bromal bromoform and iodoform also give this test (Warren,
1921).

Lustgarten’s Naphthol Test

Chloroform will produce an evanescent blue color which in contact with air
will change to green and then to brown. This color is less stable when; 8-naphthol is
used. Acidification of the blue solution will precipitate naphthol colored by a red dye
stuff. This precipitate is usually brick-red. Chloral, bromal bromoform and idoform
also give this test (Warren, 1921).

Cyanide Test
A positive test means that the distillate contained chloroform and not
because the solution is contaminated with cyanide. The following reactions take
place (Warren, 1921):
(a) CHCI3 + H 3N + 3KOH  HCN + 3KCI +
3H2O
(Chloroform) (Ammonia) (Potassium hydroxide) (Hydrocyanic acid) (Potaasium
chloride) (Water)

(b) HCN + KOH  KCN + H 2O

(Hydrocyanic acid) (Potassium hydroxide) (Potassium cyanide) (Dihydrogen
oxide)

(c) FeSO4 + 2KCN = Fe(CN)2 + K2SO4

(Ferrous sulfate) (Potassium cyanide) (Ferrous cyanide)

(d) Fe(CN)2 + 4KCN = K4Fe(CN)6

(Ferrous cyanide) (Potassium cyanide) (Potassium ferrocyanide)

(e) 3K4Fe(CN)6 + 4FeCl3 = Fe4[Fe(CN)6]3 ↓ + 12KCl

(Potassium ferrocyanide) (Ferric chloride) (Ferric ferrocyanide/Prussian blue)

Reduction Tests: Fehling’s and Tollen’s Test

These reactions are not characteristic of chloroform, because many volatile
organic substances, as formic acid and aldehydes which may occur in distillates
from animal material, reducs Fehling’s and Tollen’s reagents.

Answers to Questions:

1. What happens to chloroform when is enters the environment?

a. Air
Given that the equilibrium partitioning of chloroform is over 99%
into air, atmospheric removal processes are the most important.
Chloroform is oxidized by atmospheric hydroxyl radicals, present
naturally at an average 9.5 x 10 -3 mol cm-3 (Prinn et al., 2001). The
mechanism is typically (Euro Chlor, 2002):

(1) CHCl3 + OH∙ → CCl3∙ +H2O

(2) CCl3 + O2  CCl3OO ∙
(3) CCl3OO∙ + NO  CCl3O∙ + NO2
(4) CCl3OO∙ + HO2  CCl3O∙ + O2 + OH∙
(5) CCl3O∙  Cl∙ CCl2O
(6) CCl3O∙ + RH  CCl3OH + R∙
(7) CCl3OH  CCl2O + HCl

The first step (equation 1) is rate determining, with a rate

constant of 8.1 x 10-14 cm3 molecule-1 sec-1 at 277 oK, so that the
atmospheric lifetime of chloroform is 0.5 years [Prather, 1995]. Based
on the concentrations reported in Khalil and Rasmussen [1999], the
removal rates of chloroform by atmospheric oxidation are in the region
of 410 Gg yr-1 in the northern hemisphere and 190 Gg yr-1 in the
southern. Particularly in view of the vertical concentration profile, but
also because of uncertainties in the reaction rate constant and the
measurements themselves, these mass destruction rates have wide
ranges: from 570 to 250 Gg yr-1 (NH) and 260 to 120 Gg yr-1 (SH).
These calculations were performed using the simple two-box
hemispherical model described in McCulloch and Midgley [1996] and
so are illustrative, rather than substantive. Nevertheless, the global
total is in good agreement with the emissions that were calculated
directly in section 2 above, even if the distributions reflect a bias in the
latter towards the northern hemisphere (Euro Chlor, 2002).

a. Soil and Sediment

Several methanogens have been shown in the laboratory to
dechlorinate chloroform, particularly when promoted by metallic iron
(Fe0) [Novak et al., 1998a; 1998b]. While this has little importance in
the global mass balance of chloroform, it may be significant for
groundwater; based on the kinetic data given for carbon tetrachloride
degradation by the same organisms, the half-life for chloroform is
about 12 hours. Although the product(s) were not identified, the
mechanism of the process, driven by an unidentified enzyme, suggests
methane.

Chloroform is also degraded aerobically by Methylosinus

trichosporium. In this case, the responsible enzyme is methane
monooxygenase and the loss of chlorine follows insertion of oxygen
into the C-H bond. The product is carbon dioxide [Bartniki and Castro,
1994].

b. Water
 Although chloroform hydrolyses readily in alkaline conditions
[Hine and Dowell, 1954], the dependence of the rate on hydroxyl ion
concentration means that, under the conditions prevailing in
environmental water, its half-life is greater than 1,000 years and
hydrolysis is not an effective sink.

1. Is chloroform likely to cause cancer?

The US Environmental Protection Agency concluded that chloroform
is not likely to be carcinogenic to humans by any routes of exposure
at a dose level that does not cause cytotoxicity and cell regeneration. This
conclusion is supported by the finding that chloroform is not a strong
mutagen and is not likely to cause cancer through a genotoxic mode of
action. Hence daily exposure to 10,000 ng kg-1 body weight (600,000 ng day-
1 for an adult person) is safe from the point of view of carcinogenicity
[USEPA, 2001]. Based on a daily consumption of 1.5 kg food and 2 litres of
water based beverages per person, the chloroform levels in drinking water
(13 µgL-1 [Krasner, et al., 1989]) and foodstuff (52-71 µg.kg-1 [Crookes et al.
[1994]) are of no concern.
Conclusion:
Inhaled chloroform first passes from the air into the blood-plasma which then
transmits it to the red blood corpuscles where it may accumulate in relatively large
quantity. Air passed through blood will remove chloroform completely. Absorption of
chloroform is rapid from all parts of the body. The stimulative action of chloroform
on the mucous membranes of the respiratory passages explains such disturbances
as coughing, secretion of saliva and reflex slowing of respiration and heart-beat,
occurring at the beginning of narcosis. Dilatation of the blood-vessels of organs
living after death is due to paralysis caused by even small doses of chloroform. A
drop in blood pressure accompanies paralysis of the brain and the heart’s action is
feebler and slower. With this in mind, pharmacists play a vital role in the
management in cases of chloroform intoxication. With this activity, I understood and
acquired skills on the detection of chloroform in the laboratory aside from
management using pharmaceutical means. Chloroform is detected in the laboratory
by performing the phenylisocyanide test, Schwartz’s test, Lustgarten’s test, cyanide
test and reduction tests namely, Fehling’s test and Tollen’s test.

Bibliography

Euro Chlor. (2002). Chloroform in the Environment Occurence, Sources, Sinks and
Effects. Brussels: Euro Chlor.

Warren, W. H. (1921). Laboratory Manual for the Detection of Poisons and Powerful
Drugs. New Jersey: Read Books.