Development of a syringe membrane-based microextraction method based on metal-organic framework mixed-matrix membranes for preconcentration/extraction of polycyclic aromatic hydrocarbons in tea infusion.

Inevitably, the residues of polycyclic aromatic hydrocarbons (PAHs) in tea leaves will be transferred to hot tea infusion, constituting a certain drinking risk; consequently, it is imperative to develop rapid, sensitive, and robust approaches for their trace-level detection. Herein, we developed a syringe membrane-based microextraction (SMME) method for preconcentration/extraction of PAHs in tea infusions. This method utilized metal-organic framework-mixed matrix membranes (MOF-MMMs) as adsorbents, which anchored the nanoparticles of MOFs onto the surface of PVDF membrane. The UiO-66 (Zr)-based MMM possessed high Brunauer-Emmett-Teller (BET) surface area (320.5 m2 g-1) and pore volume (0.18 cm3 g-1), thus enhancing extraction/adsorption efficiency. Under optimized conditions, the limits of detection for PAHs reached as low as 0.02-0.08 µg L-1 with extraction recoveries of 85.5-102.1%, and the inter-day and intra-day precision was lower than 8.4% in tea infusions. Consequently, the SMME/HPLC-DAD method shows a great potential in conventional monitoring of PAHs in tea samples.

High crystalline magnetic covalent organic framework with three-dimensional grapevine structure for ultrasensitive extraction of nitro-polycyclic aromatic hydrocarbons in food and environmental samples.

Developing and establishing an efficient pre-treatment approach for the precise extraction of nitrated-polycyclic aromatic hydrocarbons (N-PAHs) from real-life samples is critical for ensuring their safety. In this study, a novel crystalline magnetic covalent organic framework with a grapevine structure not a single core-shell, Fe3O4@TAPT-DMTA-COF, was fabricated via chemical bonding. Unchanging the reticulated structure and high crystallinity of TAPT-DMTA-COF, the combination made this material possess not only simple operation via magnetic decantation but also remarkable chemical stability. Fe3O4@TAPT-DMTA-COF had a large surface area (1578.45 m2/g), and rich electronegative triazine-groups, which makes it become a superior magnetic enrichment material for trace N-PAHs. For N-PAHs analysis, low limits of detection (LODs) (1.43-17.24 ng/L), excellent relative standard deviations (RSDs ≤ 11.52%), and wide linearity (10-5000 ng/L) were obtained. Real-life applications based on this composite have been successfully explored by capturing the N-PAHs emitted from food and environmental samples.

A density-tunable liquid-phase microextraction system based on deep eutectic solvents for the determination of polycyclic aromatic hydrocarbons in tea, medicinal herbs and liquid foods.

A novel density-tunable liquid-phase microextraction (DT-LPME) system was developed with high-density deep eutectic solvents (DESs) as extractant and low-density organic solvents as emulsifier and density regulator. DES-rich phase was induced to form in the bottom or in the top by adjusting the emulsifier amount. This system was used to directly extract polycyclic aromatic hydrocarbons (PAHs) from liquid and solid foods, and the obtained DES-rich phase was easy to be collected for quantification. The method (LPME with HPLC-fluorescence detector) has linearity (R2 > 0.9974), detection limits of 0.6-4.2 ng L-1 for liquid foods and 0.05-0.35 ng g-1 for solid foods, recoveries of 86.2-114.9%, and intra-day/inter-day RSDs below 6.6%. The method was applied to detect PAHs in real samples, and the PAHs residue was found in honey and five solid foods. The DT-LPME method is simple, fast, green and suitable for direct extraction of analytes from both liquid and solid samples.

Air-assisted liquid-liquid microextraction based on the solidification of floating deep eutectic solvents for the simultaneous determination of bisphenols and polycyclic aromatic hydrocarbons in tea infusions via HPLC.

Deep eutectic solvents (DESs) are a new class of green "designer solvent"; its physicochemical properties can be easily tuned by adjusting DES' constituents, chemical ratio and water content. In this study, three hydrophobic DESs with low viscosity, low density, and melting points close to room temperature were designed and synthesized. Based on these DESs, an air-assisted liquid-liquid microextraction technique was developed based on the solidification of floating DESs for the simultaneous determination of bisphenols and polycyclic aromatic hydrocarbons (PAHs) via HPLC. The microextraction parameters were optimized via the Plackett-Burman design and response surface methodologies. The method shows satisfactory linearity (R2 ≥ 0.9928), a low limit of detection (0.16-0.75 µg L-1) and satisfactory precision (≤2.3%), and was successfully applied for the simultaneous determination of bisphenols and PAHs from tea infusions with satisfactory recoveries (82.0-116.6%). This method is simple, rapid, economical, environmentally compatible, dispersive solvent-frees and centrifugation-free, and has promising applications in food safety.

Determination of PAH Benzo[a]anthracene, Chrysene, Benzo[b]flouranthene and Benzo[a]pyrene in Hydro Alcoholic Herbal Extracts with Fluorescence Detector Using Solid-Phase Extraction.

An appropriate purification and quantification method has been developed for polycyclic aromatic hydrocarbons (PAH) in hydro alcoholic herbal extracts. For this, Bacopa monnieri, Camellia sinensis, Withania somnifera and Andrographis paniculata samples were extracted with modified solid-phase extraction (SPE) and the PAH were quantified using liquid chromatography coupled to fluorescence detector. Purification of herbal extract using hexane and acetone in the ratio of 1:1 followed by treatment with QuEChERS salt (6 g MgSO4 and 1.5 g sodium acetate) improved the recovery rate of PAH. Silica SPE, which accomplishes solvent exchange to hexane by cleanup method, was developed to reduce the matrix effect and quality of the result obtained was increased. The developed method can be used for regular monitoring and analysis of PAH in natural extracts so as to prevent contamination.

Biosynthesis of reusable and recyclable CuO@Magnetite@Hen Bone NCs and its antioxidant and antibacterial activities: a highly stable magnetically nanocatalyst for excellent reduction of organic dyes and adsorption of polycyclic aromatic hydrocarbons.

For the first time, through a fast, eco-friendly and economic method, the aqueous extract of the leaf of Euphorbia corollate was used to the green synthesis of the highly stable CuO@Magnetite@Hen Bone nanocomposites (NCs) as a potent antioxidant and antibacterial agent against Pseudomonas aureus, Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae pathogenic bacteria. The biosynthesised NCs were identified using the scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy, elemental mapping, X-ray diffraction (XRD), Fourier transforms infrared spectroscopy and UV-vis analytical techniques. Also, the radical scavenging activity using (2,2-diphenyl-1-picrylhydrazyl) method was used to evaluate the antioxidant activity of the NCs. The stability of nanocatalyst was monitored using the XRD and SEM analyses after 30 days from its synthesis. Furthermore, its excellent catalytic activity, recycling stability, and high substrate applicability were demonstrated to the adsorption of the polycyclic aromatic hydrocarbons of the light crude oil from Shiwashok oil fields and destruction of methylene blue and methyl orange as harmful organic dyes at ambient temperature using UV-vis spectroscopy. Moreover, the green CuO@Magnetite@Hen Bone NCs were recovered and reused several times without considerable loss of its catalytic activity.

Determination of polycyclic aromatic hydrocarbons and their nitrated and oxygenated derivatives in coffee brews using an efficient cold fiber-solid phase microextraction and gas chromatography mass spectrometry method.

Polycyclic aromatic hydrocarbons (PAHs) are food contaminants; besides, their oxygenated (oxy-PAHs) and nitrated (nitro-PAHs) derivatives have also been detected in some foods. This is worrying because these derivatives may be more toxic than PAHs. This study presents a new method for the determination of PAHs and their oxygenated and nitrated derivatives in coffee brew. The analytes were extracted by cold fiber solid phase microextraction (CF-SPME) with analysis by gas chromatography/mass spectrometry. The developed method presented good precision with intra-assay and inter-assay, ranged from 4.5 to 16.4%, and from 9.8 to 19.8%, respectively. Recovery ranged from 82.1 to96.3% and linearity showed good adjustment presenting determination coefficients (R2) from 0.980 to 0.999. The limits of quantification ranged from 0.025 to 0.224 µg L-1. The proposed method is simple, versatile, allows simultaneous extraction of PAHs, nitrated and oxygenated derivatives and was successfully applied to the analysis of commercial coffee samples. Benzo(k)fluoranthene, benzo(b)fluoranthene, pyrene, acenaphthylene and acenaphthene are the most abundant PAHs found in samples. In addition, 5,12-naphthacenequinone was the most abundant oxy-PAH and 1-nitropyrene was the most abundant nitro-PAH.

Preparation and evaluation of a reversed-phase/hydrophilic interaction/ion-exchange mixed-mode chromatographic stationary phase functionalized with dopamine-based dendrimers.

In this work, a novel dendritic stationary phase was synthesized by the repeated grafting of 1,4-butanediol diglycidyl ether (BDDE) and dopamine (DA) on the surface of silica for performing mixed-mode high-performance liquid chromatography (MHPLC). Elemental analysis (EA), thermogravimetric analysis (TGA) and Fourier transform infrared spectrometry (FT-IR) showed the successful preparation of the dendritic stationary phase. The prepared stationary phase showed the retention mechanisms of reversed-phase liquid chromatography (RPLC), hydrophilic interaction chromatography (HILIC) and ion-exchange chromatography (IEC) under different mobile phase conditions. In detail, alkylbenzenes, polycyclic aromatic hydrocarbons (PAHs) and hydrophobic positional isomers were separated successfully in the RPLC mode. The baseline separation of nucleobases, nucleosides and flavonoids was achieved under HILIC mode, respectively. Meanwhile, some acidic and basic analytes were used to evaluate the IEC mode. The effects of different chromatographic conditions, such as acetonitrile content, salt concentration and pH in the mobile phase, on the different chromatographic modes were also investigated. In addition, the application of the mixed-mode dendritic stationary phase was demonstrated by the analysis of traditional Chinese medicine (TCM), including Carthamus tinctorius L. and Abelmoschus manihot (Linn.) Medicus. Interestingly, the stationary phase also has the ability for the capture and separation of boric acids. These meaningful applications confirmed that the mixed-mode dendritic stationary phase can be potentially applied in the analysis of complex samples.

Multiwall carbon nanotube- zirconium oxide nanocomposite hollow fiber solid phase microextraction for determination of polyaromatic hydrocarbons in water, coffee and tea samples.

The purpose of this study was to evaluate the application of hollow fiber solid-phase microextraction (HF-SPME) followed by HPLC-UV to determine the ultra-trace amounts of polycyclic aromatic hydrocarbons (PAHs) as model analytes in complex coffee and tea samples. HF-SPME can be effectively used as an alternative to the direct immersion SPME (DI-SPME) method in complex matrices. The DI-SPME method suffers from serious limitation in dirty and complicated matrices with low sample clean-up, while the HF-SPME method has high clean-up and selectivity due to the high porosity of hollow fiber that can pick out analyte from complicated matrices. As a hollow fiber sorbent, a novel multiwall carbon nanotube/zirconium oxide nanocomposite (MWCNT/ZrO2) was fabricated. The excellent adsorption of PAHs on the sorbent was attributed to the dominant roles of π-π stacking interaction and hydrophobic interaction. Under the optimized extraction conditions, the wide linear range of 0.1-200 µg L-1 with coefficients of determination better than 0.998 and low detection limits of 0.033-0.16 µg L-1 with satisfactory precision (RSD < 6.6%) were obtained. The relative recoveries obtained by spiking the PAHs in water, coffee and tea samples were in the range of 92.0-106.0%. Compared to other methods, MWCNT/ZrO2 hollow fiber solid phase microextraction demonstrated a good capability for determination of PAHs in complex coffee and tea samples.

Polycyclic aromatic hydrocarbons in teas using QuEChERS and HPLC-FLD.

Polycyclic aromatic hydrocarbons (PAHs) are food-processing contaminants considered to be carcinogenic and genotoxic. Due to its drying process stage, teas may be contaminated with PAHs. The aim of the study was to validate an analytical method involving QuEChERS and HPLC-FLD for the determination of PAH4 in teas and evaluate the contamination levels in 10 different types of teas from Brazil. Recoveries varied from 54% to 99% and relative standard deviations from 1% to 21%. Limits of detection and quantification were from 0.03 to 0.3 µg/kg and 0.1 to 0.5 µg/kg, respectively. Mate tea presented the highest PAH levels, with PAH4 varying from 194 to 1795 µg/kg; followed by black (1.8-186 µg/kg), white (24-119 µg/kg), and green teas (3.1-92 µg/kg). Teas with lowest PAH4 were strawberry, lemongrass, peppermint, and boldo. Only trace levels of PAHs were detected in tea infusions, so apparently it would not affect PAH intake by Brazilian population.

Biosurfactant-assisted phytoremediation of multi-contaminated industrial soil using sunflower (Helianthus annuus L.).

This study evaluated the use of commercial rhamnolipid biosurfactant supplementation in the phytoremediation of a soil via sunflower (Helianthus annuus L.) cultivation. The soil, obtained from an industrial area, was co-contaminated with heavy metals and petroleum hydrocarbons. The remediation tests were monitored for 90 days. The best results for removal of contaminants were obtained from the tests in which the sunflower plants were cultivated in soil with 4 mg kg-1 of the rhamnolipid. Under these conditions, reductions of 58% and 48% were obtained in the total petroleum hydrocarbon (TPH) and polycyclic aromatic hydrocarbon (PAH) concentrations, respectively; reductions in the concentrations of the following metals were also achieved: Ni (41%), Cr (30%), Pb (29%), and Zn (20%). The PCR-DGGE analysis of soil samples collected before and after the treatments verified that the plant cultivation and biosurfactants supplementation had little effect on the structure of the dominant bacterial community in the soil. The results indicated that sunflower cultivation with the addition of a biosurfactant is a viable and efficient technology to treat soils co-contaminated with heavy metals and petroleum hydrocarbons.

Analysis of Recent Patenting Activities in the Field of Bioremediation of Petroleum Hydrocarbon Pollutants Present in the Environment.

BACKGROUND: In today's world one of the major environmental problems is the contamination of aquatic or terrestrial ecosystem due to spillage of hydrocarbon compounds produced due to various activities related to the petrochemical industry. In recent years, bioremediation has emerged as a promising technology for the restoration of these contaminated sites in an ecofriendly way. OBJECTIVE: The aim of present review literature is the compilation of patent documents on bioremediation of petroleum hydrocarbon pollutants to know technological advancements in this field. METHODS: This analysis was based on various criteria i.e. patenting trend over time, country-wise and assignee-wise comparisons and types of technology used in various patents. Some publicly available patent databases were used to retrieve the patent information from the year 2000 to 2016. RESULTS: Patent applications were retrieved and it was observed that different types of technological approaches were used in developing the patents. United States accounted for maximum patent publications, followed by China, Korea, Japan, Russia, Great Britain, Mexico, India and Canada in developing bioremediation technologies. US based organization DU PONT is the leading group as patent assignee followed by Biosaint Co. Ltd in Korea. CONCLUSION: Patenting activity in the field of bioremediation of petroleum hydrocarbon was not much commendable in the early 20th century. However, an increased trend was observed in the past few years. Further contribution in this aspect would help in stabilizing various global environmental as well as economic issues.

Treatment of PAH-contaminated soil using cement-activated persulfate.

In this study, a novel method for the treatment of polycyclic aromatic hydrocarbon -contaminated soil using cement-activated persulfate was developed. The removal of PAHs in soil rose with increasing initial persulfate concentration, initial Portland cement (PC) concentration, and oxidation reaction time. At an initial persulfate and PC concentration of 19.20 mmol/kg and 10% of soil weight and a reaction time of 2 h, the removal rate of PAHs reached 57.3%. Residual PAHs were mainly adsorbed within the soil granules and thus became less available. The mechanism of PC facilitating the oxidation reaction was that PC addition can increase the pH and temperature of the system. When the soil was stabilized/solidified by 10% of PC, the leaching concentration of PAHs and TOC was significantly higher than that leached from untreated soil. Persulfate oxidation decreased the leaching concentration of PAHs but increased the leaching concentration of TOC in solidification/stabilization products. The addition of activated carbon can decrease the leaching concentrations of both PAHs and TOC. Freeze-thaw durability tests revealed that the leachability of PAHs was not affected by freeze-thaw cycles. The unconfined compressive strength (UCS) of treated soil samples after 12 freeze-thaw cycles was only 49.0% of that curing for 52 days, but the UCS was still > 1 MPa. The treated soil samples can resist disintegration during the process of freeze-thaw cycles.

Novel Palm Fatty Acid Functionalized Magnetite Nanoparticles for Magnetic Solid-Phase Extraction of Trace Polycyclic Aromatic Hydrocarbons from Environmental Samples.

A novel adsorbent, palm fatty acid coated magnetic Fe3O4 nanoparticles (MNP-FA) was successfully synthesized with immobilization of the palm fatty acid onto the surface of MNPs. The successful synthesis of MNP-FA was further confirmed by X-Ray diffraction (XRD), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR) and Energy dispersive X-Ray spectroscopy (EDX) analyses and water contact angle (WCA) measurement. This newly synthesized MNP-FA was applied as magnetic solid phase extraction (MSPE) adsorbent for the enrichment of polycyclic aromatic hydrocarbons (PAHs), namely fluoranthene (FLT), pyrene (Pyr), chrysene (Cry) and benzo(a)pyrene (BaP) from environmental samples prior to High Performance Liquid Chromatography- Diode Array Detector (HPLC-DAD) analysis. The MSPE method was optimized by several parameters such as amount of sorbent, desorption solvent, volume of desorption solvent, extraction time, desorption time, pH and sample volume. Under the optimized conditions, MSPE method provided a low detection limit (LOD) for FLT, Pyr, Cry and BaP in the range of 0.01-0.05 ng mL-1. The PAHs recoveries of the spiked leachate samples ranged from 98.5% to 113.8% with the RSDs (n = 5) ranging from 3.5% to 12.2%, while for the spiked sludge samples, the recoveries ranged from 81.1% to 119.3% with the RSDs (n = 5) ranging from 3.1% to 13.6%. The recyclability study revealed that MNP-FA has excellent reusability up to five times. Chromatrographic analysis demonstrated the suitability of MNP-FA as MSPE adsorbent for the efficient extraction of PAHs from environmental samples.

Aristolic Acid Derivatives from the Bark of Antidesma ghaesembilla.

Antidesma ghaesembilla is an important medicinal and food plant in many Asian countries. Ten substances could be isolated from the dichloromethane and methanol extract: sitostenone (3), daucosterol (4), chavibetol (5), asperphenamate (6), protocatechuic acid (7), vanillic acid-4-O-ß-D-glucoside (8), 1-O-ß-D-glucopyranosyl-3-O-methyl-phloroglucinol (9), and aristolic acid II-8-O-ß-D-glucoside (10), and two new aristolic acid derivatives, 10-amino-5,7-dimethoxy-aristolic acid II (= 6-amino-9,11-dimethoxyphenanthro[3,4-d]-1,3-dioxole-5-carboxylic acid; 1) and 5,7-dimethoxy-aristolochic acid II (= 9,11-dimethoxy-6-nitrophenantro[3,4-d]-1,3-dioxole-5-carboxylic acid; 2). Exposure to humans of some of these compounds is associated with a severe disease today known as aristolochic acid nephropathy. Therefore, the traditional usage of this plant has to be reconsidered carefully.

Crude oil polycyclic aromatic hydrocarbons removal via clay-microbe-oil interactions: Effect of acid activated clay minerals.

Acid treatment of clay minerals is known to modify their properties such as increase their surface area and surface acidity, making them suitable as catalysts in many chemical processes. However, the role of these surface properties during biodegradation processes of polycyclic aromatic hydrocarbons (PAHs) is only known for mild acid (0.5 M Hydrochloric acid) treated clays. Four different clay minerals were used for this study: a montmorillonite, a saponite, a palygorskite and a kaolinite. They were treated with 3 M hydrochloric acid to produce acid activated clay minerals. The role of the acid activated montmorillonite, saponite, palygorskite and kaolinite in comparison with the unmodified clay minerals in the removal of PAHs during biodegradation was investigated in microcosm experiments. The microcosm experiments contained micro-organisms, oil, and clays in aqueous medium with a hydrocarbon degrading microorganism community predominantly composed of Alcanivorax spp. Obtained results indicated that acid activated clays and unmodified kaolinite did not enhance the biodegradation of the PAHs whereas unmodified montmorillonite, palygorskite and saponite enhanced their biodegradation. In addition, unmodified palygorskite adsorbed the PAHs significantly due to its unique channel structure.

Removal of crude oil polycyclic aromatic hydrocarbons via organoclay-microbe-oil interactions.

Clay minerals are quite vital in biogeochemical processes but the effect of organo-clays in the microbial degradation of crude oil polycyclic aromatic hydrocarbons is not well understood. The role of organo-saponite and organo-montmorillonite in comparison with the unmodified clays in crude oil polycyclic aromatic hydrocarbons (PAHs) removal via adsorption and biodegradation was studied by carrying out microcosm experiments in aqueous clay/oil systems with a hydrocarbon degrading microbial community that is predominantly alcanivorax spp. Montmorillonite and saponite samples were treated with didecyldimethylammonium bromide to produce organo-montmorillonite and organo-saponite used in this study. Obtained results indicate that clays with high cation exchange capacity (CEC) such as montmorillonite produced organo-clay (organomontmorillonite) that was not stimulatory to biodegradation of crude oil polycyclic aromatic compounds, especially the low molecular weight (LMW) ones, such as dimethylnaphthalenes. It is suggested that interaction between the organic phase of the organo-clay and the crude oil PAHs which is hydrophobic in nature must have reduced the availability of the polycyclic aromatic hydrocarbons for biodegradation. Organo-saponite did not enhance the microbial degradation of dimethylnaphthalenes but enhanced the biodegradation of some other PAHs such as phenanthrene. The unmodified montmorillonite enhanced the microbial degradation of the PAHs and is most likely to have done so as a result of its high surface area that allows the accumulation of microbes and nutrients enhancing their contact.

Three-dimensional ionic liquid functionalized magnetic graphene oxide nanocomposite for the magnetic dispersive solid phase extraction of 16 polycyclic aromatic hydrocarbons in vegetable oils.

In this paper, a novel three-dimensional ionic liquid functionalized magnetic graphene oxide nanocomposite (3D-IL@mGO) was prepared, and used as an effective adsorbent for the magnetic dispersive solid phase extraction (MSPE) of 16 polycyclic aromatic hydrocarbons (PAHs) in vegetable oil prior to gas chromatography-mass spectrometry (GC-MS). The properties of 3D-IL@mGO were characterized by scanning electron micrographs (SEM), X-ray photoelectron spectroscopy (XPS), vibrating sample magnetometer (VSM). The 3D-IL@mGO, functionalized by ionic liquid, exhibited high adsorption toward PAHs. Compared to molecularly imprinted solid phase extraction (MISPE), the MSPE method based on 3D-IL@mGO had less solvent consumption and low cost, and was more efficent to light PAHs in quantitative analysis. Furthermore, the rapid and accurate GC-MS method coupled with 3D-IL@mGO MSPE procedure was successfully applied for the analysis of 16 PAHs in eleven vegetable oil samples from supermarket in Zhejiang Province. The results showed that the concentrations of BaP in 3 out of 11 samples were higher than the legal limit (2.0µg/kg, Commission Regulation 835/2011a), the sum of 8 heavy PAHs (BaA, CHR, BbF, BkF, BaP, IcP, DaA, BgP) in 11 samples was between 3.03µg/kg and 229.5µg/kg. Validation results on linearity, specificity, accuracy, precision and stability, as well as on application to the analysis of PAHs in oil samples demonstrated the applicability to food safety risk monitoring in China.

Kingianins O-Q: Pentacyclic polyketides from Endiandra kingiana as inhibitor of Mcl-1/Bid interaction.

A phytochemical study of the EtOAc-soluble part of the methanolic extract of the bark of Endiandra kingiana led to the isolation of three new pentacyclic kingianins as racemic mixtures, kingianins O-Q (1-3), together with the known kingianins A, F, K, L, M and N (4-9), respectively. The structures of the new kingianins 1-3 were determined by 1D and 2D NMR analysis in combination with HRESIMS experiments. Kingianins A-Q were assayed for Mcl-1 binding affinity. Kingianins G and H were found to be potent inhibitors of Mcl-1/Bid interaction. A structure-activity relationship study showed that potency is very sensitive to the substitution pattern on the pentacyclic core. In addition, in contrast with the binding affinity for Bcl-xL, the levorotatory enantiomers of kingianins G, H and J exhibited similar binding affinities for Mcl-1 than their dextrorotatory counterparts, indicating that the two anti-apoptotic proteins have slightly different binding profiles.

Optimization and comparison of miniaturized extraction techniques for PAHs from crude oil exposed Atlantic cod and haddock eggs.

Two miniaturized extraction methods for a wide range of 2-6 ring polycyclic aromatic hydrocarbons (PAHs) and their alkylated homologues in small lipid-rich biota samples (≤100 mg) have been developed. Both methods utilize liquid extraction (LE) prior to a clean-up step using either normal phase solid phase extraction (SPE) or mixed-phase dispersive SPE (dSPE). Optimization of the methods was achieved by comparing the type and amount of sorbents, drying agents, and solvents used. In order to improve the limits of detection (LOD) of target PAHs under high sensitivity gas chromatography-tandem mass spectrometry analysis, specific emphasis was given to minimizing lipid co-extraction. The optimized LE-SPE method comprised extraction with dichloromethane/n-hexane (1:1, v/v) and clean-up by silica SPE, whereas the optimized LE-dSPE method comprised extraction with acetonitrile and clean-up with PSA and C18 sorbents. The methods were validated and directly compared through the analysis of Atlantic cod (Gadus morhua) and haddock (Melanogrammus aeglefinus) eggs exposed to oil. The LE-SPE method resulted in lower levels of co-extracted lipids (14.1 ± 1.7 ng/µL) than the LE-dSPE method (60 ± 14 ng/µL). Achieved PAH LODs for the LE-SPE method were typically an order of magnitude lower (<5 ng/g) than for the LE-dSPE method (<125 ng/g). The LE-SPE method offers the possibility for PAH analysis of small samples of fish eggs (~100 mg) exposed to small quantities of crude oil (~1-10 µg/L total PAHs).