Iopanoic acid alters thyroid hormone-related gene expression, thyroid hormone levels, swim bladder inflation, and swimming performance in Japanese medaka.

Disruption of the thyroid hormone system by synthetic chemicals is gaining attention owing to its potential negative effects on organisms. In this study, the effects of the dio-inhibitor iopanoic acid (IOP) on the levels of thyroid hormone and related gene expression, swim bladder inflation, and swimming performance were investigated in Japanese medaka. Iopanoic acid exposure suppressed thyroid-stimulating hormone ß (tshß), tshß-like, iodotyronin deiodinase 1 (dio1), and dio2 expression, and increased T4 and T3 levels. In addition, IOP exposure inhibited swim bladder inflation, reducing swimming performance. Although adverse outcome pathways of thyroid hormone disruption have been developed using zebrafish, no adverse outcome pathways have been developed using Japanese medaka. This study confirmed that IOP inhibits dio expression (a molecular initiating event), affects T3 and T4 levels (a key event), and reduces swim bladder inflation (a key event) and swimming performance (an adverse outcome) in Japanese medaka.

Effect of diisobutyl adipate on the expression of biomarker genes that respond to endocrine disruption and on gonadal sexual differentiation in Japanese medaka (Oryzias latipes).

Phthalate and non-phthalate plasticizers are used in polymer materials, such as plastic and rubber. It has recently been found that diisobutyl adipate (DIBA), which is considered an environmentally safe non-phthalate plasticizer, potentially acts as a thyroid disruptor in fish. Here, we investigated the sexual hormone effects of DIBA based on the expression levels of genes that respond to endocrine disruption and sexual hormone activity in the livers and gonads, and on gonadal sexual differentiation in Japanese medaka. Compared with the control group, the mRNA expression of chgH, vtg1, vtg2, and esr1 was significantly suppressed in the livers of DIBA exposed XX individuals. Furthermore, the mRNA expression of gsdf was significantly upregulated and downregulated in the gonads of XX and XY individuals, respectively. The mRNA expressions of esr1 and esr2b were significantly suppressed by DIBA exposure in the gonads of both XX and XY individuals. These observations suggest that DIBA has potential androgenic activity in Japanese medaka. However, normal testes and ovaries were observed in respective XY and XX medaka after DIBA exposure; therefore, these results suggest that DIBA may have weak androgenic activity.

Residues of non-phthalate plasticizers in seawater and sediments from Osaka Bay, Japan.

NPPs (Non-phthalate plasticizers) are used as alternative plasticizers to phthalate esters, but there is limited knowledge on environmental residues, and they have not been reported in Japan. A method to analyze NPPs in seawater using solid-phase extraction was developed, and the residual burden of Diisobutyl adipate (DIBA), Acetyl tributyl citrate (ATBC), Di-(2-ethylhexyl) adipate (DEHA), Di-(2-ethylhexyl) sebacate (DEHS) and Trioctyl trimellitate (TOTM) in seawater and sediment from the Osaka Bay was measured. Using an Oasis Max column and acetone as the eluting solvent, the recovery of the target substances in seawater is >68 %. In Osaka Bay, no NPPs were detected in seawater. On the other hand, ATBC and TOTM were detected in the sediment at 36-69 ng/g and 47-131 ng/g, respectively, from about half of the 14 sites, while DEHA and DEHS were detected at 83 ng/g and 181 ng/g, respectively, from only one site.

Effects of plasticizer diisobutyl adipate on the Japanese medaka (Oryzias latipes) endocrine system.

Plasticizer pollution of the water environment is one of the world's most serious environmental issues. Phthalate plasticizers can disrupt endocrine function in vertebrates. Therefore, this study analyzed thyroid-related, reproduction-related, and estrogen-responsive genes in Japanese medaka (Oryzias latipes) to determine whether non-phthalate diisobutyl adipate (DIBA) plasticizer could affect endocrine hormone activity or not. Developmental toxicity during fish embryogenesis was also evaluated. At a concentration of 11.57 mg/l, embryonic exposure to DIBA increased the mortality rate. Although abnormal development, including body curvature, edema, and lack of swim bladder inflation, was observed at 3.54 and 11.57 mg/l DIBA, growth inhibition and reduced swimming performance were also observed. In addition, DIBA exposure increased the levels of thyroid-stimulating hormone beta-subunit (tshß) and deiodinase 1 (dio1) but decreased the levels of thyroid hormone receptor alpha (trα) and beta (trß). These results suggest that DIBA has thyroid hormone-disrupting activities in fish. However, kisspeptin (kiss1 and kiss2), gonadotropin-releasing hormone (gnrh1), follicle-stimulating hormone beta (fshß), luteinizing hormone beta (lhß), choriogenin H (chgH), and vitellogenin (vtg1) expression did not change dose-dependently in response to DIBA exposure, whereas gnrh2 and vtg2 expression was elevated. These results indicate that DIBA has low estrogenic activity and does not disrupt the endocrine reproduction system in fish. Overall, this is the first report indicating that non-phthalate DIBA plasticizer is embryotoxic and disrupt thyroid hormone activity in fish.

Effects of non-phthalate plasticizer bis(2-ethylhexyl) sebacate (DEHS) on the endocrine system in Japanese medaka (Oryzias latipes).

Water pollution due to plasticizers is one of the most severe environmental problems worldwide. Phthalate plasticizers can act as endocrine disruptors in vertebrates. In this study, we investigated whether the non-phthalate bis(2-ethylhexyl) sebacate (DEHS) plasticizer can act as an endocrine disruptor by evaluating changes in the expression levels of thyroid hormone-related, reproduction-related, and estrogen-responsive genes of Japanese medaka (Oryzias latipes) exposed to the plasticizer. Following the exposure, the gene expression levels of thyroid-stimulating hormone subunit beta (tshß), deiodinase 1 (dio1), and thyroid hormone receptor alpha (trα) did not change. Meanwhile, DEHS suppressed dio2 expression, did not induce swim bladder inflation, and eventually reduced the swimming performance of Japanese medaka. These findings indicate that DEHS can potentially disrupt the thyroid hormone-related gene expression and metabolism of these fish. However, exposure to DEHS did not induce changes in the gene expression levels of kisspeptin 1 (kiss1), gonadotropin-releasing hormone (gnrh), follicle-stimulating hormone beta (fshß), luteinizing hormone beta (lhß), choriogenin H (chgH), and vitellogenin (vtg) in a dose-dependent manner. This is the first report providing evidence that DEHS can disrupt thyroid hormone-related metabolism in fish.

Thyroid hormone disruption by bis-(2-ethylhexyl) phthalate (DEHP) and bis-(2-ethylhexyl) adipate (DEHA) in Japanese medaka Oryzias latipes.

Pollution of water bodies with plasticizers is a serious environmental problem worldwide. In this study, we investigated the effects of plasticizers bis-(2-ethylhexyl) phthalate (DEHP) and bis-(2-ethylhexyl) adipate (DEHA) in Japanese medaka (Oryzias latipes). DEHP significantly increased the expression of all the genes tested: thyroid stimulating hormone beta subunit (tshß-like), tshß, deiodinase 1 (dio1), deiodinase 2 (dio2), and thyroid hormone receptor alpha (trα) and beta (trß). However, DEHA only significantly increased tshß at 7.4 µg/L but significantly decreased dio2 expression at 25.8, 111.1, and 412.6 4 µg/L, while other genes were not significantly affected. Both chemicals reduced eye size and total body length, but did not affect embryo development, hatching time and rate, and swimming performance. DEHA alone affected swim bladder inflation and not DEHP. This is the first report that not only DEHP but also DEHA disrupt thyroid hormone activity in fish. DEHP contamination (13.2 µg/L) was detected in tap water from Kobe, Japan; thus, tap water itself may disrupt thyroid hormone activity in Japanese medaka. Importantly, the effective concentration of DEHP for thyroid hormone-related gene expression and growth was close to or lower than DEHP concentrations reported in surface water elsewhere, indicating that DEHP contamination is a serious aquatic pollution.

Investigation of distribution, sources and flux of perfluorinated compounds in major southern Indian rivers and their risk assessment.

Perfluorinated compounds (PFCs) are a group of emerging contaminants still less reported in rivers, particularly southern India. Therefore, we investigated the fate of 13 PFCs in three major rivers in southern India during post-monsoon and summer seasons. Twelve PFCs were detected, with an average total PFCs of 1853 ± 1463 pg/l. However, the total PFCs recorded in ppost-monsoon and summer seasons ranged from ND (none detected) to 10,545 pg/l and ND to 4960 pg/l, respectively. Among the individual congeners, perfluoro-n-hexanoic acid (PFHxA) had the highest detection average (929 ± 710 pg/l). The higher detection of short chain PFCs signifies their increasing wide usage as an alternative to perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS). Higher levels of PFCs were observed in summer than post-monsoon season in the river Kaveri, which is mainly attributed to the decrease in river flow. A multidimensional source identification revealed domestic and commercial wastewater as the major source. A correlation analysis showed that most of the detected PFCs share the common source and undergo co-migration into rivers. The flux of PFCs into the Bay of Bengal, loaded by the Kaveri (15 kg/yr) and the Tamiraparani (2.2 kg/yr) rivers, signifies lower per capita emissions than other rivers in India and other countries. Further, the PFC levels found in the rivers can be considered safe for human consumption and aquatic organisms based on international guidelines. Being the first hand report in southern Indian rivers, the results warrant further investigation to understand the exact sources, fate and removal in detail.

Organic micropollutants in the surface riverine sediment along the lower stretch of the transboundary river Ganga: Occurrences, sources and ecological risk assessment.

The Hooghly River (HR) estuary is the first deltaic off-shoot of the perennial and transboundary river, Ganga, India. HR receives industrial and domestic waste along with storm-water run-off from Kolkata city and the adjoining districts. Organic micropollutants (OMPs) have been collectively termed for plasticizers, pharmaceuticals and personal care products, which are extensively consumed and disposed in the waste streams. Hence emerging OMPs were investigated to obtain the first baseline data from the Hooghly riverine sediment (HRS) along urban and suburban transects using gas chromatography mass spectrometry (GC-MS). The concentration range of OMPs in the HRS varied between 3 and 519 ng/g for carbamazepine, 5-407 ng/g for non-steroidal anti-inflammatory drugs (NSAIDs), 2-26 ng/g for musk ketone, 2-84 ng/g for triclosan, 2-199 ng/g for bisphenol A (BPA), 2-422 ng/g for plasticizers (phthalic acid esters (PAEs) and bis (2-ethylhexyl) adipate (DEHA)) and 87-593 ng/g for parabens. Carbamazepine concentration in sediment was an useful marker for untreated wastewater in urban waterways. High concentrations of BPA and PAEs in the suburban industrial corridor together with significant correlation between these two type of OMPs (r2 = 0.5; p < 0.01) likely reflect a common source, possibly associated with the plastic and electronic scrap recycling industries. Among all the categories of OMPs, plasticizers seems to exhibit maximum screening level ecological risk through out the study area.

Ecotoxicological assessment of cobalt using Hydra model: ROS, oxidative stress, DNA damage, cell cycle arrest, and apoptosis as mechanisms of toxicity.

The mechanisms underlying cobalt toxicity in aquatic species in general and cnidarians in particular remain poorly understood. Herein we investigated cobalt toxicity in a Hydra model from morphological, histological, developmental, and molecular biological perspectives. Hydra, exposed to cobalt (0-60 mg/L), were altered in morphology, histology, and regeneration. Exposure to standardized sublethal doses of cobalt impaired feeding by affecting nematocytes, which in turn affected reproduction. At the cellular level, excessive ROS generation, as the principal mechanism of action, primarily occurred in the lysosomes, which was accompanied by the upregulation of expression of the antioxidant genes SOD, GST, GPx, and G6PD. The number of Hsp70 and FoxO transcripts also increased. Interestingly, the upregulations were higher in the 24-h than in the 48-h time-point group, indicating that ROS overwhelmed the cellular defense mechanisms at the latter time-point. Comet assay revealed DNA damage. Cell cycle analysis indicated the induction of apoptosis accompanied or not by cell cycle arrest. Immunoblot analyses revealed that cobalt treatment triggered mitochondria-mediated apoptosis as inferred from the modulation of the key proteins Bax, Bcl-2, and caspase-3. From this data, we suggest the use of Hydra as a model organism for the risk assessment of heavy metal pollution in aquatic ecosystems.

Evaluating spatial distribution and seasonal variation of phthalates using passive air sampling in southern India.

Usage of phthalates as plasticizers has resulted in worldwide occurrence and is becoming a serious concern to human health and environment. However, studies on phthalates in Indian atmosphere are lacking. Therefore, we studied the spatio-temporal trends of six major phthalates in Tamil Nadu, southern India, using passive air samplers. Phthalates were ubiquitously detected in all the samples and the average total phthalates found in decreasing order is pre-monsoon (61 ng m-3) > summer (52 ng m-3) > monsoon (17 ng m-3). Largely used phthalates, dibutylphthalate (DBP) and diethylhexlphthalate (DEHP) were predominantly found in all the seasons with contribution of 11-31% and 59-68%, respectively. The highest total phthalates was observed in summer at an urban location (836 ng m-3). Furthermore, through principal component analysis, potential sources were identified as emissions from additives of plasticizers in the polymer industry and the productions of adhesives, building materials and vinyl flooring. Although inhalation exposure of infants was higher than other population segments (toddlers, children and adults), exposure levels were found to be safe for people belonging to all ages based on reference dose (RfD) and tolerable daily intake (TDI) values. This study first attempted to report seasonal trend based on atmospheric monitoring using passive air sampling technique and exposure risk together.

Seaweeds: A resource for marine bionanotechnology.

Marine bionanotechnology is one of the most promising areas of research in modern science and technology. Although there are multitude methods for the synthesis of nanoparticles (NPs), there is an increasing attention in developing high-yield, low-cost, non-toxic and eco-friendly procedures. The vital advantages of greener synthesis are cost-effective, reduced usage of toxic chemicals and abundant availability of resources. During the last ten years, there have been many biological entities used to elevate novel, greener and affordable methods for the metal NPs synthesis. Rate of synthesis and stability are higher for plant material mediated NPs. However, in comparison with terrestrial resources, marine resources have not been fully explored for synthesis of noble metal NPs. Our present review is designed to speculate the importance of usage of vast marine resources and its mediated NPs synthesis, in particular seaweed-mediated NPs synthesis to overcome the limitations involved in physical and chemical methods. Finally, recent advancements in greener synthesis of metal NPs, their size, distribution, morphology and applications such as antimicrobial, antifouling and anticancer potentials are briefly described along with portraying the prospective scope of research in this field without any negative impact on the environment.

Phthalate esters in water and sediments of the Kaveri River, India: environmental levels and ecotoxicological evaluations.

Phthalate esters are well known for their environmental contamination and toxicological effects as "endocrine disruptors." In this study, environmental levels of phthalate esters and ecotoxicological risk assessments were performed in one of the major rivers in India, the Kaveri. Water and sediment samples were collected during 2010-2012 representing the major stretch of the river and extracted by solid-phase and ultrasonic methods, respectively, and analyzed for six major phthalates by using a gas chromatograph-mass spectrometer. The analytical recovery for phthalates in water and sediment ranged from 79 to 121%. Results indicated that diethyl phthalate (DEP) and dimethyl phthalate were found in every sample, whereas butylbenzyl phthalate and diethylhexyl phthalate (DEHP) were detected in 92% of the water samples. Likewise, in sediment samples, DEP was found most often (94%). The total phthalates in water samples ranged from 313 to 1,640 ng/l, whereas in sediments it was 2 to 1,438 ng/g dw (dry weight) with DEHP having the highest concentration. Human health risk assessment based on drinking water consumption showed no potential risk for phthalates and also DEHP levels were safe with respect to USEPA guideline (6,000 ng/l). Further, DEHP and di-n-octyl phthalate levels in water were expected to pose little threat to sensitive organisms in the riverine ecosystem as per ECOSAR chronic values. In case of sediment, the DEHP concentration was well above the USEPA sediment guideline value. To our knowledge, this is the first study to describe the levels and ecotoxicological risks of phthalates in Kaveri River, India.

GC-MS determination of bisphenol A and alkylphenol ethoxylates in river water from India and their ecotoxicological risk assessment.

Water samples from three rivers in southern India were analyzed for octylphenol (OP), nonylphenol (NP), and bisphenol A (BPA) residues by gas chromatography and mass spectrometry. The concentrations of OP, NP and BPA ranged from ND (not detected) to 16.3 ng/L, ND to 2200 ng/L, and 2.8 to 136 ng/L with detection frequencies of 96%, 66% and 100%, respectively. All three rivers showed a similar distribution pattern of NP>>BPA>OP, however, the Kaveri river had elevated levels. The phenolic compounds in rivers are expected to cause potential toxicity to aquatic organism including crustaceans, molluscs, insects and fish. In respect to aquatic risk assessment, NP showed a greater hazard quotient (HQ) than did OP and BPA, and the highest HQ (62) was observed for fish in the Kaveri river. The backwater and estuarine NP levels may pose a risk to larvae of oysters and barnacles. Based on tolerable daily intake and reference dose, BPA and NP levels are considered safe for humans. However, NP levels in some water samples were greater than the drinking water safety limit (0.5 µg/L). This is the first report on phenolic compounds and their associated aquatic risks in Indian rivers.

Non-steroidal anti-inflammatory drugs in Indian rivers.

Pharmaceutical concentration data for Indian surface waters are currently scarce. Sewage often enters Indian rivers without prior treatment, and so previously reported environmental concentrations from regions with routinely implemented sewage treatment cannot simply be used to predict concentrations in Indian surface water. Improved knowledge of pharmaceutical concentrations in Indian waters would enable determination of potential risks posed to aquatic wildlife and human health in this region. The concentrations of five common non-steroidal anti-inflammatory drugs (NSAIDs; diclofenac, ketoprofen, naproxen, ibuprofen, and acetylsalicylic acid) were determined in surface waters from 27 locations of the Kaveri, Vellar, and Tamiraparani Rivers in southern India. The samples were extracted by solid-phase extraction and analyzed by GC-MS. The measured concentrations of four of the five drugs in this reconnaissance were relatively similar to those reported elsewhere (ND-200 ng/l); however, acetylsalicylic acid, the most readily degradable of the investigated drugs, was found at all sites and at considerably higher concentrations (up to 660 ng/l) than reported in European surface waters. This is the first report on the occurrence of NSAIDs in Indian rivers. The finding of elevated concentrations of acetylsalicylic acid is most likely a result of direct discharges of untreated sewage. Therefore, readily degradable pharmaceuticals may present larger concern in regions without consistent sewage treatment. Based on measured environmental concentrations, the risks of direct toxicity to aquatic wildlife and of humans consuming the water are discussed.

Paraben resistance in bacteria from sewage treatment plant effluents in India.

Parabens, the antimicrobial preservatives used in cosmetics, food and pharmaceuticals, are often detected in the aquatic environment. Generally, sewage treatment plants (STPs) receive community sewage containing parabens, which are ultimately released into streams/rivers. In this study, bacteria in STP effluents were evaluated for their resistance to parabens. The susceptibility was in the order of Staphylococcus aureus > Bacillus sp. >Escherichia coli > Pseudomonas aeruginosa. Gram-negative bacteria showed less susceptibility than their control and Gram-positive bacteria. Further, the bacteria were more sensitive towards butyl and ethyl parabens. Interestingly, the strains showed resistance to ≥5 mg of parabens, which is equivalent to or higher than reported environmental concentrations. The increase in paraben chain length did not enhance the susceptibility in all cases and it was understood that the activity may differ for each bacterium in the environment. This is the first profile on paraben resistance in common pathogens of Indian STPs. Paraben resistance may be developed due to continuous exposure even at sub-inhibitory and/or chronic levels in the environment and this resistance may be transferred to other pathogenic bacteria in receiving waters. Thus the study demonstrates the effectiveness of the disc diffusion method in environmental bacterial resistance assessment and addresses the risk involved in the use of parabens.

Perchlorate contamination of groundwater from fireworks manufacturing area in South India.

Perchlorate contamination was investigated in groundwater and surface water from Sivakasi and Madurai in the Tamil Nadu State of South India. Sensitive determination of perchlorate (LOQ = 0.005 µg/L) was achieved by large-volume (500 µL) injection ion chromatography coupled with tandem mass spectrometry. Concentrations of perchlorate were <0.005-7,690 µg/L in groundwater (n = 60), <0.005-30.2 µg/L in surface water (n = 11), and 0.063-0.393 µg/L in tap water (n = 3). Levels in groundwater were significantly higher in the fireworks factory area than in the other locations, indicating that the fireworks and safety match industries are principal sources of perchlorate pollution. This is the first study that reports the contamination status of perchlorate in this area and reveals firework manufacture to be the pollution source. Since perchlorate levels in 17 out of 57 groundwater samples from Sivakasi, and none from Madurai, exceeded the drinking water guideline level proposed by USEPA (15 µg/L), further investigation on human health is warranted.

Contamination and bioaccumulation of benzotriazole ultraviolet stabilizers in fish from Manila Bay, the Philippines using an ultra-fast liquid chromatography-tandem mass spectrometry.

Benzotriazole ultraviolet stabilizers (BUVSs) used in plastic products, building materials and personal hygiene products were analyzed in fishes collected from Manila Bay, the Philippines. BUVSs were detected at ng g⁻¹ level in all the fish samples, indicating their ubiquitous contamination in coastal waters. Among the targeted eight BUVSs, UV-328 was predominantly found with a mean concentration of 34.2 ng g⁻¹ lipid weight, implying large scale production and use of this compound in the Philippines. High concentrations of ∑BUVSs were found in bumpnose trevally (Carangoides hedlandensis), bluetail mullet (adult) (Valamugil buchanani), common ponyfish (Leiognathus equulus) and coral grouper (adult) (Epinephelus corallicola) indicating their active uptake and/or lower metabolic capacity to eliminate BUVSs. Among BUVSs, UV-P showed significant positive relationship (p<0.05) between concentration and fish length (r=0.29) and fish weight (r=0.31). Levels of UV-P in demersal species had positive correlation with δ¹5N, indicating that possibile sink of UV-P is bottom sediment in the bay, and ultimately accumulate through benthic food web rather than pelagic food web. To our knowledge, this is the first study on BUVSs distribution in fish from developing countries.

Determination of preservative and antimicrobial compounds in fish from Manila Bay, Philippines using ultra high performance liquid chromatography tandem mass spectrometry, and assessment of human dietary exposure.

Ultra high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) determination of four paraben preservatives (methyl, ethyl, propyl and butyl) and two antimicrobial agents (triclosan and triclocarban) belonging to personal care products (PCPs) in 20 species of fish from Manila Bay (Philippines) was performed. Detection of PCPs with greater frequency indicates the ubiquitous contamination of Manila Bay. Concentrations of total paraben were one order of magnitude higher than the antimicrobials in almost all fish, except in Stolephorus indicus and Leiognathus equulus. A positive correlation was observed between parabens concentration and fish length (r = 0.31-0.49; p<0.05 to <0.001) and fish weight (r = 0.28-0.49; p<0.05 to <0.001), but not for the antimicrobials. The estimated dietary exposure values of the four parabens in the Philippines through fish is four orders of magnitude lower than the acceptable daily intake (ADI) of 10mg/kg/day, but the values of antimicrobials are just half of the ADI of TCS. To our knowledge, this is the first report of PCPs contamination in fish from Philippines.

Multiresidue analytical method for the determination of antimicrobials, preservatives, benzotriazole UV stabilizers, flame retardants and plasticizers in fish using ultra high performance liquid chromatography coupled with tandem mass spectrometry.

A multiresidue analytical method for the determination of emerging pollutants belonging to personal care products (PCPs) (antimicrobials, preservatives), benzotriazole UV stabilizers (BUVSs) and organophosphorus compounds (OPCs) in fish has been developed using high speed solvent extraction (HSSE) followed by silica gel clean up and ultra fast liquid chromatography coupled with tandem mass spectrometry (UFLC-MS/MS) analysis. Developed extraction and clean up method yielded good recovery (> 70%) for all the four groups of emerging pollutants, i.e. antimicrobials (78.5-85.6%), preservatives (85.0-89.4%), BUVSs (70.9-112%) and OPCs (81.6-114%; except for TEP - 68.9% and TPeP - 58.1%) with RSDs ranging from 0.7 to 15.4%. Intra- and inter-day repeatabilities were less than 19.8% and 19.0%, respectively at three spiked levels. The concentrations were given in lipid weight (lw) basis, and the method detection limits were achieved in the lowest range of 0.001-0.006 ng g⁻¹ for two antimicrobials, 0.001-0.015 ng g⁻¹ for four preservatives, 0.0002-0.009 ng g⁻¹ for eight BUVSs and 0.001-0.014 ng g⁻¹ for nine OPCs. Finally, the method was successfully validated as a simple and fast extraction method for the determination of 23 compounds belonging to PCPs, BUVSs and OPCs and applied to the analysis of three species of fish from Manila Bay, the Philippines. Concentrations ranged from 27 to 278 ng g⁻¹ for antimicrobials, 6.61 to 1580 ng g⁻¹ for paraben preservatives,