Introduction

Polycyclic aromatic hydrocarbons (PAHs) are a class of lipophilic and

ubiquitous environmental pollutants, consisting of multiple fused rings. They can enter the environment through natural sources (e.g. forest fires and volcanoes) and anthropogenic activities (e.g. incomplete combustion of fossils fuels and organic matter). They are also present in smoked/barbecued foods.1,2

Many PAHs exhibit toxic, mutagenic and carcinogenic behaviors, and due to their lipophilic nature they are readily absorbed from the gastrointestinal tracts of mammals.3,4 While hundreds of PAHs exist, sixteen are on the U.S. Environmental Protection Agency’s (EPA) priority pollutant list, and are routinely monitored via EPA Methods 550.1 (drinking water), 610 (waste water), and 8310 (ground water and solid waste).5,6,7 These regulated methods employ routine analysis by HPLC, using UV and fluorescence detection. Depending on the matrix, additional PAHs are often also monitored as part of the analysis, including alkyl substituted PAHs such as 1-methylnapthalene and 2-methylnapthalene.

Fast Analysis of 18 PAHs Using a Quasar SPP PAH Column

A P P L I C A T I O N B R I E F

Author:Kathryn Lawson-Wood

PerkinElmer Ltd. Seer Green, UK

Liquid Chromatography

2

Superficially Porous Particle (SPP) columns are packed with particles made of a solid, non-porous core surrounded by a shell of a porous silica material, which have properties similar with those of the fully porous silica materials conventionally used in HPLC. SPP columns provide significant improvements in efficiency, resulting in improved resolution between critical peak pairs, as well as increased sensitivity. Additionally, due to the shorter diffusion path within the SPP particles and their reduced surface area, reductions in run times can be observed. This increases sample throughput and generates savings in solvent consumption.8

This application brief describes the use of a Quasar™ SPP PAH column for the analysis of 18 PAH compounds (including 16 of those outlined in the EPA methods, and two which are commonly added).

Experimental Conditions

Method Parameters

HPLC method parameters are shown in Table 1. It is important to note that EPA methods 550.1, 610 and 8310 require detection via UV and fluorescence. The data generated here, however, simply uses detection via photodiode array (PDA) to demonstrate the excellent resolving power of the Quasar SPP PAH column.

Solvents and Samples

An EPA Method 8310 Quality Control Check solution, containing 18 PAH compounds in acetonitrile (5 – 100 µg/mL), was used for this analysis. All solvents were HPLC grade and standards were filtered using a 0.45 μm PTFE filter (P/N 02542909).

Results and Discussion

The analysis of the EPA Method 8310 Quality Control Check solution (18 PAH compounds) using the Quasar SPP PAH (100 x 3.0 mm, 2.6 µm) is shown in Figure 2 at 280 and 254 nm. Diode array detection allowed spectral peak identification (Table 2).

All 18 compounds were baseline resolved (Rs ≥ 1.5) in just over 12 minutes. The minimum resolution value occurred between peaks 4 and 5 (2-methylnapthalene and acenaphthene), with an Rs of 2.28. Excellent peak shape was observed, with tailing factors (5 % peak height) for all peaks between 1.01 – 1.09.

With the excellent resolution provided by the Quasar SPP PAH column, analysis time could be reduced by operating at a higher flow rate, or by using a shorter SPP column. Due to the shorter diffusion distance into the particles, SPP columns allow operation at higher optimum linear velocities in comparison with fully porous columns.

Figure 1: Chemical structures of the 16 PAH compounds specified by EPA Methods 550.1, 8310 and 610, and two additional commonly added compounds (1-methylnapthalene and 2-methylnaphthalene).

Figure 2. EPA Method 8310 Quality Control Check solution chromatograms at 280 and 254 nm.

Instrument PerkinElmer Flexar™ with PDA Plus™ Detector

Column 100 mm 3.0 mm 2.6 µmQuasar SPP PAH

(N9307264)

Mobile Phase A: Water B: Acetonitrile

Gradient ProfileStep 1: Linear gradient from 50 – 100 % B in 8 minStep 2: Hold for 5 min at 100 % B

Flow Rate 0.77 mL/min

Temperature 30° C

PDA Detector 280 and 254 nm

Injection Volume 2 µL

Table 1. HPLC method parameters for the analysis of PAHs.

3

4. D. Lerda, Polycyclic Aromatic Hydrocarbons (PAHs) Factsheet, European Commission Joint Research Centre, European Union, 2011, 4th Ed.

5. United States Environmental Protection Agency, “Determination of Polycyclic Aromatic Hydrocarbons in Drinking Water by Liquid Solid Extraction and HPLC with Coupled Ultraviolet and Fluorescence Detection,” Method 550.1, 1990, EPA Environmental Monitoring Systems Laboratory, Office of Research and Development, Cincinnati, Ohio 45268.

6. United States Environmental Protection Agency, “Determination of Polycyclic Aromatic Hydrocarbons in Municipal and Industrial Wastewater,” Method 610, 1982, EPA Environmental Monitoring Systems Laboratory, Office of Research and Development, Cincinnati, Ohio 45268.

7. United States Environmental Protection Agency, “Determination of Polycyclic Aromatic Hydrocarbons in Ground Water and Wastes,” Method 8310, 1986, EPA Environmental Monitoring Systems Laboratory, Office of Research and Development, Cincinnati, Ohio 45268.

8) PerkinElmer Brownlee Superficially Porous Particle (SPP) HPLC Columns, https://www.perkinelmer.com/CMSResources/Images/44-151014APP_Performance_Brief.pdf Date accessed: Jan 2021.

Consumables Used

Conclusions

• The Quasar SPP PAH column offers a highly selective separation of PAH compounds, with excellent peak shape and baseline resolution.

• In this application, baseline resolution of 18 PAHs (including 16 of those specified in EPA methods) has been achieved in just over 12 minutes.

• With a solid core and outer porous silica coating of the SPP phase, up to 50 % improvement in cost and efficiency can be realized in comparison with traditional porous silica columns.

References

1. J.E. Balmer, H. Hung, Y. Yu, R.J. Letcher, and D.C.G. Muir, Sources and environmental fate of pyrogenic polycyclic aromatic hydrocarbons (PAHs) in the Arctic, Emerging Contaminants, 2019, 5, 128-142.

2. F. Malhat, N. Loutfy, M.Allah I. El Menyawi, and M. Tawfic Ahmed, Review of Contamination by Polycyclic Aromatic Hydrocarbons (PAHs) in Egyptian Aquatic Environment, Polycyclic Aromatic Compounds, 2019, DOI: 10.1080/10406638.2019.1684325.

3. H. I. Abdel-Shafy, and M. S.M. Mansour, A review on polycyclic aromatic hydrocarbons: Source, environmental impact, effect on human health and remediation, Egyptian Journal of Petroleum, 2016, 25, 107-123.

Component Description Part Number

Columns Quasar SPP PAH (100 x 3.0 mm, 2.6 μm) N9307264

HPLC Vials 2 mL Amber 9 mm Screw Top Vial with Write-on Patch and Fill Lines (100/Pack)

N9307802

HPLC Vial Caps 9 mm Screw Top Blue (Polypropylene) Cap with PTFE/Silicone Pre-Slit Septa (100/Pack)

N9306203

PEEK Fittings Finger Tight for 1/16’’ OD PEEK Tubing 09920513

Stainless SteelFittings Ti Hybrid w/Flat Wrench Ferrule/Nut N9306301

Syringes Syringe 1 mL BD Luer-Lok Disposable, (100/Pack)

02542890

Syringe Filters 0.45 μm PTFE (Hydrophobic) Syringe Filter, 13 mm

02542909

Peak Compound Concentration (µg/mL)

1 Napthalene 100

2 Acenaphthylene 100

3 1-methylnapthalene 100

4 2-methylnapthalene 100

5 Acenaphthene 100

6 Fluorene 100

7 Phenanthrene 100

8 Anthracene 100

9 Fluoranthene 10

10 Pyrene 10

11 Benzo(a)anthracene 10

12 Chrysene 10

13 Benzo(b)fluoranthene 10

14 Benzo(k)fluoranthene 5

15 Benzo(a)pyrene 10

16 Dibenzo(a,h)anthracene 10

17 Benzo(g,h,i)perylene 10

18 Indeno(1,2,3-cd)pyrene 10

Table 2. PAH peak identification and concentrations.

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Quasar SPP PAH Columns, Guards and Guard Holder Part Numbers

P/N Description Length (mm)

ID (mm)

Particle size (µm)

N9307260 Quasar SPP PAH 50 2.1 2.6

N9307261 Quasar SPP PAH 100 2.1 2.6

N9307262 Quasar SPP PAH 150 2.1 2.6

N9307263 Quasar SPP PAH 50 3.0 2.6

N9307264 Quasar SPP PAH 100 3.0 2.6

N9307265 Quasar SPP PAH 150 3.0 2.6

N9307266 Quasar SPP PAH 50 4.6 2.6

N9307267 Quasar SPP PAH 100 4.6 2.6

N9307268 Quasar SPP PAH 150 4.6 2.6

N9307269Quasar SPP PAH Guard Cartridge (3/Pack)

5 3.0 2.6

N9307270Quasar SPP PAH Guard Cartridge Holder

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