UNEXAS Application

Environmental
Certified Quality Management DIN EN ISO 9001

Introduction

SAMPLE PREPARATION AND HPLC ANALYSIS OF PAHs IN EXTRACTED SOIL SAMPLES

Polycyclic aromatic hydrocarbons (PAH) are believed to be mutagenic and contain toxic chemicals. These chemicals contain carcinogenic properties which have led to legislative restrictions on their release in the environment. Therefore, suitable monitoring methods are extremely important. PAH pollutants are present in the air, water, and soil and have been a concern for a considerable period of time. There are several ways that they can be brought into the environment. The most predominant source is from them being readily formed during pyrolysis and incomplete combustion processes (e.g. fossil fuel burning). This application describes the automated sample extraction and analysis of PAHs in soil extract using the UNEXAS system. This system uses high performance solid phase extraction, introduces the analytes directly into the HPLC system, and is combined with a UV and fluorescence detector.

Experiment
The chromatographic separation is carried out with a binary High Pressure Gradient (HPG) system, implemented in the UNEXAS system, combined with the fluorescence HPLC monitor RF-10 AXL, and the WellChrom Spectro-Photometer K-2600. Online SPE is carried out with an UNEXAS system which controls the Autosampler Triathlon. 100 l of extracted soil samples were injected. The UNEXAS system controls the online extraction on disposable SPE cartridges as well as the direct desorption to the HPLC system.

Chromatographic Conditions Anal. Column: C18 ULTRASEP ES PAH, 250 x 4 mm ID Cartridge: HySphere Resin SH, 15-25 m, 10 x 2 mm ID Mobile Phase: Gradient acetonitrile/water (60:40 v/v) 60 100% acn in 28 min Flow Rate: 1.0 ml/min UV Detector: 254 nm FL Detector: excitation 254 nm, emission 320 nm Temperature: ambient Figure 1: Normal or split gradient cartridge elution flow scheme

Sample Preparation
The soil containing PAHs is dried and grinded with acetone in an Erlenmeyer flask before starting the extraction. Extract the soil sample for 10 minutes using a horizontal shaker. The acetone extract is mixed with water in a ratio of 4:6, evaporated to 1 ml, and placed in the autosampler vial. The Triathlon injection loop is filled with 100 l of sample mixture. After selecting the HySphere Resin SH cartridge, the UNEXAS sample preparation program is carried out as follows: 1. 2. 3. 4. 5. Activate the cartridge for 2.0 min with 1.0 ml/min methanol. Condition the cartridge for 3.0 min with 1.0 ml/min with water. Load 100 l of sample on the cartridge for 0.4 min with 1 ml/min water While loading, wash the cartridge for 0.5 min with water/methanol (v/v 1:1) at 1 ml/min and again for 2 min with water. Elute the sample to the analytical column for 5.0 min with acn (split gradient mode) at 1.0 ml/min and run the sample analysis. In the meantime, clean the SPE unit and reactivate the cartridge.

Wissenschaftliche Gertebau Dr. Ing. Herbert Knauer GmbH

UNEXAS Application
Environmental

SAMPLE PREPARATION AND HPLC ANALYSIS OF PAHs IN EXTRACTED SOIL SAMPLES

The following sample preparation program together with UNEXAS software was applied.

Figure 2: Program screen

Results Figure 3 illustrates a chromatogram of a PAH standard and an extracted soil sample. Both chromatograms are obtained after the automated online extraction and desorption to HPLC.

Figure 3: PAH standard injection on cartridge

Extract of soil sample injected on cartridge

Wissenschaftliche Gertebau Dr. Ing. Herbert Knauer GmbH

UNEXAS Application
Environmental
Certified Quality Management DIN EN ISO 9001

SAMPLE PREPARATION AND HPLC ANALYSIS OF PAHs IN EXTRACTED SOIL SAMPLES

Conclusion The recovery percentage of PAH components in soil for online SPE and HPLC with the UNEXAS system is between 95 and 105% and has a relative standard deviation (RSD) range of 1.5 4%. The unique switching diagram to concentrate and analyze PAHs allows the elution of the cartridge with 100% acn. This proves that the developed method is an excellent alternative for liquid-liquid extraction due to the higher reproducibility and recovery rate in online SPE. The overall analysis time, including the sample preparation for PAH analysis, is lower compared to the traditional liquid-liquid extraction. Additionally, online SPE HPLC for PAH analysis saves a considerable volume of solvent. Table 1: Recoveries for online SPE -HPLC Component
Naphthalene Acenaphthylene Acenaphthene Fluorine Phenanthrene Anthracene Fluoranthene Pyrene Benz(a)anthracene Chrysene Benzo(b)fluoranthene Benzo(k)fluoranthene Benzo(a)pyrene Dibenzo(a,h)anthracene Benzo(g,h,i)perylene Indeno(1,2,3-cd)pyrene

Recovery (%) RSD (%) n = 5

98 96 102 99 97 99 98 99 95 98 97 95 96 97 105 95 2.9 4.0 1.5 3.0 4.2 1.2 1.8 1.5 1.3 2.1 1.9 1.8 1.8 2.3 4.3 2.8

Wissenschaftliche Gertebau Dr. Ing. Herbert Knauer GmbH