Advantages of the method ■Screening of polycyclic aromatic hydrocarbons (PAHs) in seafood takes less than 4 minutes ■Accurate results by faster, simpler sample preparation ■Selective determination by fluorescence detection Waters Solutions <br>ACQUITY UPLC@ H-Class System with Fluorescence Detector Key words purpose INTRODUCTION <br>In the past, major oil spills, such as the 1989 Exxon Valdez oil spill and the April 2010 Gulf of Mexico oil spill, have raised concerns about the quality of seafood from these regions. Fish, crustaceans, and molluscs may come into contact with or ingest oil, posing a potential health risk to consumers. Of the many compounds found in petroleum, one important group of compounds are polycyclic aromatic hydrocarbons (PAHs). The US Environmental Protection Agency (US EPA) has identified these compounds as key pollutants1. The US Food and Drug Administration (US FDA) has identified a number of issues, including the presence of benzo(a)pyrene in fish at 3.5 x 10-2 mg/kg and the total content of phenanthrene and anthracene in oysters up to 2.0 x 103 mg/kg. 2 If the content of PAHs reaches half of the level of concern, a confirmatory experimental analysis must be performed2. In order to avoid contaminated seafood and minimize the impact on the seafood industry, a rapid screening method is needed to analyze these worrying compounds. We have proved here by using. After a simple extraction of the DisQuE Matrix Dispersion Sample Preparation Kit (QuEChERS), the ACQUITY UPLC H-Class with a fluorescence detector can complete a PAHs analysis in less than 4 minutes. test Standard: PAH certification standard, AccuStandard M 8310 Sample preparation <br> The fish meat pieces (flatfish), shelled shrimp, and shelled oysters with water were homogenized separately by a food processor according to the method described by Ramalhosa et al. 15 g of the homogenized tissue was taken into the centrifuge tube for each sample, and the certified PAH standard solution was added at three different levels. Add 5 ml of water to the fish and shrimp samples to aid in the mixing. The oysters do not require additional water. The spiked samples were thoroughly mixed and allowed to stand at room temperature for one hour. The reagent of the DisQuE tube (P/N 186004571) was added to each centrifuge tube, namely 6 g of magnesium sulfide + 1.5 g of sodium acetate and 15 mL of acetonitrile. The tube was shaken vigorously for at least 1 minute to form an emulsion of marine food tissue, buffer salts and acetonitrile. This time, it was also carried out in accordance with Ramalhosa 3's procedure because neither acetic acid was added to acetonitrile nor a secondary PSA cleaning step was performed. Initial work in our laboratory demonstrated that PSA steps are not required for liquid chromatography with fluorescence detectors (data not published). After centrifugation at 3000 rpm for 5 minutes, a portion of the acetonitrile float was transferred to an automated sampling tube for injection. The spiked samples at concentrations of 1 μg/g and 10 μg/g were diluted with 1:10 and 1:100 acetonitrile, respectively. Samples were quantified using a 6-point linear calibration curve. The standard curve is drawn using acetonitrile diluted certified standards. Results and Discussion <br> Dispersed sample preparation, also commonly referred to as QuEChERS, is an effective and rapid method of sample preparation for pesticide analysis in foods4. More recently, this method has been used to extract other contaminants from food matrices, including polycyclic aromatic hydrocarbons (3). Using the ACQUITY UPLC H-Class system, 15 fluorescent PAHs classified as key contaminants by US EPA were separated in as little as 3.5 minutes. The separation of the analytes is shown in Figure 2. The arrows point to the changes in the programmed timing wavelength. Figure 3 shows an example of a chromatogram of a shrimp, fish and oyster substrate spiked at a concentration of 10 μg/g. As shown in Figure 3D, the blank water sample, also prepared by the sample preparation procedure, showed a very clear chromatogram. Examples of unspiked marine food matrices used in this sample preparation procedure also showed no matrix interference, as shown in FIG. Each sample was quantified according to a 6-point calibration curve for each analyte. An exemplary calibration curve for benzo(a)pyrene is shown in Figure 5. The linear coefficient (R2) of all analytes was > 0.995. Polycyclic aromatic hydrocarbons were extracted from three different marine substrates by the Waters DisQuE Matrix Dispersion Sample Preparation Kit. The recovery rate and RSD percentage of shrimp, fish and oyster are shown in Tables 1 to 3. The recovery range is from 68% to 149%. Table 4 lists the recovery of a series of QC water spikes, concentrated at the listed levels, and throughout the sample preparation process described above. These results are very good for all compounds at each level of addition, except for the lowest level of addition of hydrazine in water (5 ng/g). At this low level, since the peak area is small and the baseline tilt causes the variation of the detection result of the flaw to fluctuate greatly, it can only be detected above this level. table 5 It is based on the estimated limit of 7 liters of various marine substrates at 5 ng/g. The calculation is based on US EPA 40 CFR, Appendix B to Section 136, Revision 1.15. The application note confirms the DisQuE matrix. The combination of a dispersion sample preparation kit and liquid phase fluorescence chromatography provides a rapid screening tool for PAH detection in seafood. ■Dispersed sample preparation provides a fast and efficient method for extracting polycyclic aromatic hydrocarbons from different marine substrates. Table 5 Detection limits (LOD) for spiked shrimp, fish and oysters. The standard deviation is calculated from the results of 7 single-label tests at 5 ng/g for each marine substrate, based on US EPA 40 CFR. Appendix B to Part 136, Revision 1.1. ■Given the reduced time required for sample preparation, rapid chromatographic separation is important for the analysis of samples, standards, and related QC samples by this method. references About Waters Corporation () 3 Ply Breathable Face Masks,Non-Woven Disposable face Mask,Medical Disposable Face Mask,Medical Disposable Face Mask Dongguan Keyutai Mask Co., Ltd. , https://www.maskkytai.com
DisQuETM Matrix Dispersion Sample Preparation Kit
EmpowerTM 2 software
Polycyclic aromatic hydrocarbons, PAHs, QuEChERS, fluorescence detection, food safety
The combination of the DisQuE Matrix Dispersed Sample Preparation Kit and UPLC®-FLR is demonstrated to provide a rapid screening tool for PAHs detection in seafood.
LC Condition System: ACQUIT Y UP LC with Large Capacity Flow Cell (LV FC)
H-Class
Column: PAH 4.6 x 50 mm, 3μm
Column temperature: 35 ° C
Injection volume: 10μL
Sampling rate: 20 points / sec. Detection: Program timing control fluorescence detection wavelength change software: Empower 2
Mobile phase A: Milli-Q water mobile phase B: methanol, Fisher optimal mobile phase C: acetonitrile, Fisher optimal 
Flow rate: 2.0 mL/min
Gradient program: Time flow rate %A %B %C Gradient line type (minutes) (mL/min)
0.00 2.0 30 70 0
2.25 2.0 0 70 30 6
3.50 2.0 0 0 100 6
3.60 2.0 30 70 0 6 
â– This method has proven to be more advantageous than other sample preparation techniques because accurate results can be obtained with very little sample preparation and short time. 
â– The ACQUITY H-Class system has a separation time of less than 4 minutes to meet the method requirements.
â– This solution helps labs to screen PAHs in seafood and deliver results economically and in a timely manner; so consumers can be assured of the safety of these products.
[1] USEPA Method 8310 “PolyNuclear Aromatic Hydrocarbonsâ€
Revision number 0, September 1986.
[2] Gratz et. al., “SCREEN FOR THE PRESENCE OF POLYCYCLIC
AROMATIC HYDROCARBONS IN SELECT SEAFOODS USING LCFLUORESCENCE",
USFDA Laboratory Information Bulletin,
Page: July 29, 2010
[3] Ramalhosa et al., “Journal of Separation Scienceâ€, 2009,
32, page: 3529-3538.
[4] Anastassiades et al., Journal of the AOAC Int, 2003, 86,
Page number: 412.
[5] EPA 40 CFR, Part 136, Appendix B, Revision No.: 1.1 Page 566.
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