CHM 3123L-2 Analytical Chemistry 3 Laboratory

First Semester, SY 2022-2023 1 JANUARY 2023

Experiment No. 3
Analysis of Trihalomethanes in Drinking Water
De Guzman, A.J., Duave, J.E., Fudalan, X.R., Sison, S.T.R
College of Science, Pamantasan ng Lungsod ng Maynila

Abstract

Chlorination is one of the techniques used for water purification and disinfection. However, due to the reaction
of chlorine with the naturally organic compounds such as humic acid and fulvic acid, trihalomethanes (THMs)
are produced, namely: chloroform (CHCl3), dichlorobromomethane (CHCl2Br), bromomethane (CHClBr2), and
bromoform (CHBr3). Gas chromatography technique was applied for the analysis of THMs in water. A two (2)
meter long glass column with an internal diameter of two (2) millimeters (mm) was used with a 10% squalene
stationary phase on an 80/100 mesh Chromosorb, W-AW packing material with a 1-5μL aliquot of pentane
injected onto the chromatograph while being operated at 65°C with a flow rate of 25 mL/min were the optimal
experimental conditions for the chromatographical analysis were the experimental parameters of the study. The
results of THMs analysis from gas chromatography showed that the drinking water sample are well within the
drinking standard (80 – 100 ppb).

Keywords: Trihalomethane, gas chromatography, drinking water

volatile organic compounds (Dewulf & Van

1. INTRODUCTION
Drinking water must be ensured to be safe
for consumption, especially to its consumers.
Hence, water purification and disinfection
techniques are developed to meet the standards
established. One of the main techniques used is
chlorination, wherein chlorine is applied to
drinking water to deactivate microorganisms as
well as protect the water from the repropagation of
microorganisms in its totality (Cho, Kong, Oh,
2002).
However, one of the aftermaths of
chlorination is the production of trihalomethanes,
or THMs. This is due to the reaction of chlorine
with the naturally occurring organic compounds,
i.e., humic acid [C9H9NO6] and fulvic acid
[C14H12O8] (Rook, 1976; Bellar, Lichtenberg, &
Kroner, 1974). The THMs, namely: chloroform
[CHCl3], dichlorobromomethane [CHCl2Br],
bromomethane [CHClBr2], and bromoform
[CHBr3], are considered to be carcinogens, thus,
human exposure must be avoided, if not,
minimized (Craun, 1985; Simpson & Hayes, 1998).
Analytical methods have been developed
and utilized for the analysis of THM content; all of
which are related to gas chromatography. Methods
such as: (1) static headspace technique, (2) liquid-
liquid extraction, (3) purge and trap, and (4) solid-
phase microextraction technique are used in order
to analyze samples with possible high contents of
Langenhove, 1999; Kurán & Soják, 1996).
The current study aims to develop rapid and
efficient method for the analysis of THMs in
drinking water through gas chromatography (GC).
This further provides a thorough investigation
regarding the concentration of THMs in drinking
water.
2. METHODOLOGY
The sample was collected in a 40 mL glass
vial with a TFE-faced septum. The vial was to the
top to avoid the formation of air bubbles. A
reducing agent, 25 mg of ascorbic acid, was added
to quench the production of trihalomethanes. The
vial was then sealed and stored at 4 °C for less than
14 days. A standard stock solution for each
trihalomethane was then prepared by adding 9.8
mL of methanol to a 10 mL volumetric flask. The
flask was allowed to stand for a few minutes until it
was completely dry before it was weighed to the
nearest ±0.1 mg. With a 100 μL syringe, 2 drops of
trihalomethane were added directly to the methanol
in the flask. The flask was then reweighed before
the solution was diluted to 10 mL. The prepared
solutions were then transferred to a 40 mL glass
vial with a TFE-lined screw top and stored away
from light exposure at -10° to -20°C. The solutions
were diluted even further with methanol in
preparing multicomponent working solutions.

Page 1
CHM 3123L-2 Analytical Chemistry 3 Laboratory
First Semester, SY 2022-2023 1 JANUARY 2023

In another volumetric flask, an internal analyzed under 67°C in a Chromosorb W-AW

standard was prepared by dissolving 1,2- packing material with a flow rate of 25 mL/min.
dibromopentane in hexane. A few milliliters of the
prepared internal standard solution were added to 3.2. Reaction Mechanism
pentane until the concentration of 30 μg 1,2- Chlorine is one of the chemicals used to
dibromopentane/L was achieved. In the preparation disinfect drinking water before consumption
of the samples for analysis, 5 mL of the solution throughout most cases. Trihalomethanes (THM)
was withdrawn from each vial before the vials were are produced when organic substances, for aquatic
capped and weighed to the nearest ±0.1 mg. humic and fulvic acids, which are often present in
Following that, 2 mL of pentane and internal water, react with chlorine during the chlorination of
standard solution were added to each vial while surface water (Masoud et al., 2019). A study found
vigorously shaking for 1 minute to thoroughly mix that the relationship between THM concentration
the components. The solution was then allowed to and humic substance levels is convex, indicating
separate into two phases for 2 minutes. Using a that there are multiple steps in the reaction. The
glass pipet, approximately 1 mL of the pentane, or production of organochlorine intermediates and its
the upper phase, was transferred to a 1.8 mL screw- conversion to THM take place in the first and
top vial with a TFE septum and stored at 4°C. The second steps of the reaction, respectively.
original sample vials were then rinsed, dried, and Additionally, the kinetic equations for THM
weighed to the nearest ±0.1 mg to calculate the generation as a function of the precursor and
weight of the sample. chlorine concentrations under regulated pH and
In the analysis of the samples through gas temperature conditions are derived using a
chromatography, a 2-meter-long glass column with mechanistic model that is based on the reactivity of
an internal diameter of 2 mm was used with a 10% chlorine with model compounds (Adin et al.,
squalene stationary phase on an 80/100 mesh 1991).
Chromosorb W-AW packing material. A 1-5μL
aliquot of the pentane solution was injected into the 3.3. Analysis of Trihalomethanes in Drinking
gas chromatograph, which operated at 67°C with a Water
flow rate of 25 mL/min.
The data on the concentration of the Compound Peak Area Concentration
standard and peak area were used to calculate the (sample) (in ppb)
concentration of each trihalomethane present in the a. CHCl3 1.56 ×10 4 1.25 x 10−4
sample. It is also important to note that proper b. CHCl2Br 4 −5
safety measures, protective equipment, and 5.13 ×10 1.32 x 10
techniques were observed throughout the c. CHClBr2 1.49 ×10 4 9.36 x 10−4
experiment to minimize hazards and errors. a. CHBr3 1.76 ×10
4
9.47 x 10
−5

Table 1. Concentration of Trihalomethanes present in drinking water.

3. RESULTS AND DISCUSSION

When chlorine, which is commonly used
to disinfect water, interact with other naturally
3.1. Gas Chromatography
occurring chemicals in the water, a group of
Analytical chemistry uses a technique
disinfection byproducts known as trihalomethanes
called gas chromatography to separate volatile
(THM) are produced. These harmful
organic chemicals based on their physical
byproducts have no color and evaporate from the
characteristics and identify organic substances. The
water into the atmosphere (Hood, 2005). A study
stationary phase of the column in a gas
conducted by US.EPA recommends that the
chromatograph, which is typically used in gas
maximum level of at least four THM species
chromatography, is made up of a long column
present in drinking water sources should fall under
filled with tiny particles and coated with a thin
or must not exceed 80 μg/L. Additionally, the
coating of adsorbent. The carrier gas on the other
World Health Organization has established a limit
hand, which carries the sample, is known as the
of 100 μg/L for the total THM concentration.
mobile phase.
(Sadeghi et al., 2019).
Injected samples exist in the gas phase;
hence, samples must be heated to be vaporized

( )(
before passing through a column (State, 2016). In concentration of standard peak area ( sample )
this experiment, a glass column that is 2 meters ppb= x
peak area ( sample ) peak area ( standard
long and 2 millimeters in diameter with a 10%
squalene stationary phase was utilized for the concentration of standard
analysis of the sample. The injected aliquot of Equation 1.
pentane solution which measures 1-5 μL was Through mathematical interpolation (Eq.
1), among the four trihalomethanes analyzed,

Page 2
CHM 3123L-2
First Semester, SY 2022-2023
CHCl3 yielded the highest concentration (M =
−4
1.25 x 10 ), while CHBr3, on the other hand had
the lowest (M = 9.47 x 10−5) (Table 1).
The concentration of the samples reported
in ppb are relatively low, hence, the drinking water
analyzed is permissible for safe drinking, since
high concentrations of THM in drinking water can
cause risks and dangers that may be fatal.
Compound Retention time (min)
a. CHCl3 20.2
b. CHCl2Br 30.1
c. CHClBr2 38.3
a. CHBr3 45.8
Table 2. Retention time of each compound
Figure 1. Graphical representation of retention time per compound
Moreover, from Table 1, it can be
determined that CHBr3 has the higher retention
time. Hence, it can be deduced that CHBr3 has the
highest retention time, therefore is the most soluble
of all the compounds.
With the small increments of the
concentration of each THM compound, all of
which are within the range of 80 – 100 parts per
billion (ppb), it can be concluded that the analyte is
safe for consumption.
3.4. Possible Experiment Errors and
Improvements.
Despite being an analytical method, gas
chromatography is nevertheless subject to some
errors. This is a result of analytical errors that occur
while the experiment is being conducted; these
sources may result from contamination,
measurement mistakes, and mechanical or
instrument flaws, to name a few (Marius Miricioiu
et al., 2016).
A relatively low flow rate, suitable for the
sample utilized in this experiment, of 25 mL/min
was used. An overly high column temperature can
be a cause of short retention time and poor
separation since all components largely remain in
the gas phase. Since, if the compound does not
make contact with the stationary phase, the
retention time will decrease because interaction
between the components and the stationary phase is
Analytical Chemistry 3 Laboratory
1 JANUARY 2023
necessary for separation to take place. (Gas
Chromatography Theory, 2016).
The preparation of the gas chromatograph
prior to the actual experiment plays an important
role in the result, hence there are measures used to
improve the results; resolution in gas
chromatography. During analysis, three factors
should be considered for a better resolution.
Temperature should be relatively low to allow the
interaction of analytes in the column with the
stationary phase since the interaction allows the
separation. Furthermore, the size of the column
used should be appropriate in dimensions for the
conducted analysis and should be accompanied by
proper flow rate operation (Axion Labs, 2022).
4. CONCLUSION
Gas chromatography was applied to
determine trihalomethane (THM) content in
drinking water. Concentrations per volatile
compound as well as its peak area were analyzed.
A two (2) meter long glass column with an internal
diameter of two (2) millimeters (mm) was used
with a 10% squalene stationary phase on an 80/100
mesh Chromosorb, W-AW packing material with a
1-5μL aliquot of pentane injected onto the
chromatograph while being operated at 65°C with a
flow rate of 25 mL/min were the optimal
experimental conditions for the chromatographical
analysis.
Ppb from the sample data that has been
acquired, were correlated to the retention time that
has been sufficed. Where, the most soluble of all
the compounds, CHBr3, has the lowest
concentration, hence, its concentration in the
drinking water sample is little. Moreover, the
concentration of each THM compound is little in
comparison to the standard of 80 – 100 parts per
billion (ppb), therefore, it is safe for consumption.
Lastly, from the data acquired, it can be
concluded that gas chromatography is gravely
feasible for the analysis of drinking water.
5. REFERENCES
Adin, A., Katzhendler, J., Alkaslassy, D., & Rav-
Acha, Ch. (1991). Trihalomethane
formation in chlorinated drinking water: A
kinetic model. Water Research, 25(7), 797–
Page 3
CHM 3123L-2 Analytical Chemistry 3 Laboratory
First Semester, SY 2022-2023 1 JANUARY 2023

805. https://doi.org/10.1016/0043- Marius Miricioiu, Niculescu, V., Oana Romina

1354(91)90159-n Dinca, & Mihaela Emanuela Craciun.
Axion Labs (2022). How do you improve (2016, March). Analytical Errors in
resolution in gas chromatography? Routine Gas Chromatography Analysis.
https://axionlabs.com/chromatography- ResearchGate; Revista de Chimie SRL.
training/how-do-you-improve-resolution-in- https://www.researchgate.net/publication/29
gas-chromatography/ 9536353_Analytical_Errors_in_Routine_G
‌Bellar, T. A., Lichtenberg, J. J., & Kroner, R. C. as_Chromatography_Analysis#:~:text=Thes
(1974). The Occurrence of Organohalides e%20errors%20may%20include
in Chlorinated Drinking Waters. Journal %20contamination,gas%20caused%20by
(American Water Works %20pressure%20reducers.
Association), 66(12), 703–706. Masoud, M. S., Ismail, A. M., & El-Hoshy, M. M.
https://www.jstor.org/stable/41267646 (2019). Kinetics and thermodynamics of the
Cho, D.-H., Kong, S.-H., & Oh, S.-G. (2003). formation of trihalomethanes. Applied
Analysis of trihalomethanes in drinking Water Science, 9(4).
water using headspace-SPME technique https://doi.org/10.1007/s13201-019-0981-1
with gas chromatography. Water R
‌ ook, J. J. (1976). Haloforms in Drinking
Research, 37(2), 402–408. Water. Journal (American Water Works
https://doi.org/10.1016/s0043- Association), 68(3), 168–172.
1354(02)00285-3 https://www.jstor.org/stable/41268395
Craun, G. F. (1985). Epidemiologic studies of Sadeghi, H., Nasseri, S., Yunesian, M., Mahvi, A.
organic micropollutants in drinking H., Nabizadeh, R., & Alimohammadi, M.
water. Science of the Total (2019). Trihalomethanes in urban drinking
Environment, 47, 461–472. water: measuring exposures and assessing
https://doi.org/10.1016/0048- carcinogenic risk. Journal of
9697(85)90350-x Environmental Health Science and
‌Dewulf, J., & Van Langenhove, H. (1999). Engineering, 17(2), 619–632.
Anthropogenic volatile organic https://doi.org/10.1007/s40201-019-00374-
compounds in ambient air and natural x
waters: a review on recent developments S
‌ impson, K. L., & Hayes, K. P. (1998). Drinking
of analytical methodology, performance water disinfection by-products: an
and interpretation of field Australian perspective. Water
measurements. Journal of Research, 32(5), 1522–1528.
Chromatography A, 843(1-2), 163–177. https://doi.org/10.1016/s0043-
https://doi.org/10.1016/s0021- 1354(97)00341-2
9673(99)00225-3 State, N. (2016). Gas Chromatography [YouTube
Gas Chromatography Theory. (2016). Ucla.edu. Video]. In YouTube.
https://www.chem.ucla.edu/~bacher/Genera https://www.youtube.com/watch?
l/30BL/gc/theory.html v=X8hQUQHOYlA
Hood, E. (2005). Tap Water and Trihalomethanes:
Flow of Concerns
Continues. Environmental Health
Perspectives, 113(7), A474.
https://www.ncbi.nlm.nih.gov/pmc/articles/
PMC1257669/
‌Kurán, P., & Soják, L. (1996). Environmental
analysis of volatile organic compounds in
water and sediment by gas
chromatography. Journal of
Chromatography A, 733(1-2), 119–141.
https://doi.org/10.1016/0021-
9673(95)01121-8

Page 4
CHM 3123L-2 Analytical Chemistry 3 Laboratory
First Semester, SY 2022-2023 1 JANUARY 2023

Annex A
Calculation for the ppb

Page 5