Phenols in Effluent using High-Performance Liquid Chromatography

Aims
This study seeks to use high-performance liquid chromatography(HPLC) to quantitatively
and qualitatively analyse three phenols.

Introduction
HPLC performs on the basis of the difference in vapour pressures of molecules in a mixture.
These molecules are separated into a mobile phase(solvent mixture) and stationary
phase(the column) which are pumped through and vapourised. A UV spectrometer is
utilised as a detector for retention times from which concentration can be calculated.

Experimental
Three solutions of varying concentrations of mixed phenols are prepared into volumetric
flasks and made up with 50% methanol:water. These three solutions were each injected into
the HPLC which displayed concentration and peak areas that was used to construct a
calibration curve. In the following order phenols unknown, A B & C, repeat of phenol
unknowns A B & C and blank were injected into the HPLC and their chromatograms with
table of retention times were printed out.
For this particular experiment, the HPLC auto sampler was not working thus the injections
were performed manually.

Figure 1. Sketch of HPLC [1]

Results

Figure 2. Calibration curve computed through mixed phenols of varying concentrations.

Unknown A – Concentrations of phenols

Name Retention time Area Height Concentration(ppm)

Phenol 5.408 238282 22177 27.246
o-Chlorophenol 8.125 314065 23095 26.346
m-Cresol 6.988 518552 41756 55.173

Unknown B – Concentrations of phenols

Name Retention time Area Height Concentration(ppm)

Phenol 5.414 330519 30364 37.793
o-Chlorophenol 8.127 597957 43221 50.160
m-Cresol 6.992 241378 19292 25.682

Unknown C – Concentrations of phenols

Name Retention time Area Height Concentration(ppm)

Phenol 5.303 443255 41352 50.684
o-Chlorophenol 7.952 442536 32676 37.122
m-Cresol 6.842 240378 19412 25.576
Analytical Results with Uncertainties
Three unknowns were tested for phenol content and are listed below. Uncertainty from the
HPLC is deemed to be negligible since combined with the software package its very
accurate.
Unknown A
Phenol 27 ± 0.27 ppm
o-Chlorophenol 26 ± 0.26 ppm
m-Cresol 55 ± 0.55 ppm
Unknown B
Phenol 38 ± 0.38 ppm
o-Chlorophenol 50 ± 0.50 ppm
m-Cresol 27 ± 0.27 ppm
Unknown C
Phenol 51 ± 0.51 ppm
o-Chlorophenol 37 ± 0.37 ppm
m-Cresol 26 ± 0.26 ppm

Discussion and Conclusions

All 3 unknowns contained phenol concentration levels that were well above safe Australia
water marine & aquatic life standards. Which states estimated threshold level in water for
m-cresol 0.2 ppm, Phenol 1-10 ppm and o-chlorophenol 0.0001-0.015.[3] Fish and other
aquatic organism would cease to exist in these polluted waters and they definitely would be
unsuitable for human consumption at these toxic levels. Furthermore according to
Australian water drinking standards the levels are also too high and would be unsafe to
drink as it would cause adverse health effects. The toxicity of phenols is over 100x too high
as such home owners should not let children play in waters polluted from which these
samples were collected.

References
[1] Clark. J (2016) High performance liquid chromatography Available at:
http://www.chemguide.co.uk/analysis/chromatography/hplc.html Accessed: 11/9/2017
[2] Ball. G (2017), CHEM3901 Lab Manual, The University of New South Wales, Kensington,
NSW, Australia
[3] Australian Water Association (2017) Australian and New Zealand Guidelines for Fresh
and Marine Water Quality. Page 4.4-16 Table 4.4.5 Available from :
https://www.environment.gov.au/system/files/resources/e10f8ee3-54b4-4e90-8694-
50b6a3194b9d/files/nwqms-guidelines-4-vol2.pdf Accessed: 11/9/2017