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DSpace at VNU: Asia–Pacific mussel watch for emerging pollutants: Distribution of synthetic musks and benzotriazole UV stabilizers in Asian and US coastal waters

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High concentra-tions of musks and BUVSs were detected in mussels from Japan and Korea, where the levels were com-parable or greater than those of PCBs, DDTs and PBDEs.. HHCB and AHTN hav

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Asia–Pacific mussel watch for emerging pollutants: Distribution of synthetic

musks and benzotriazole UV stabilizers in Asian and US coastal waters

Haruhiko Nakataa,⇑, Ryu-Ichi Shinoharaa, Yusuke Nakazawaa, Tomohiko Isobeb, Agus Sudaryantob, Annamalai Subramanianb, Shinsuke Tanabeb, Mohamad Pauzi Zakariac, Gene J Zhengd, Paul K.S Lamd, Eun Young Kime, Byung-Yoon Minf, Sung-Ug Wef, Pham Hung Vietg, Touch Seang Tanah,

Maricar Prudentei, Donnell Frankj, Gunnar Lauensteink, Kurunthachalam Kannanl,m

a Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555, Japan

b

Center for Marine Environmental Studies (CMES), Ehime University, Bunkyo-Cho 2-5, Matsuyama 790-8577, Japan

c

Faculty of Science and Environmental Studies, Universiti Putra, 43400 Serdang, Malaysia

d

Department of Biology and Chemistry, City University of Hong Kong, 83, Tat Chee Avenue, Kowloon, Hong Kong, China

e

Department of Biology, Kyung Hee University, Hoegi-Dong Dongdaemun-Gu, Seoul 130-701, South Korea

f

Environmental Engineering, Kyungnam University, 449 Wolyoung-Dong, Kyungnam 631-701, South Korea

g Hanoi National University, 19 Le Thanh Tong Street, Hanoi, Vietnam

h Member of the Economics, Social and Culture Observation Unit (OBSES) of the Council of Minister, Cambodia

i

Science Education Department, De La Salle University, 2401 Taft Avenue, 1004 Manila, Philippines

j

TDI-Brooks International, 1902 Pinon, College Station, TX 77845, USA

k

1305 East West Highway National Oceanic and Atmospheric Administration, National Centers for Coastal Ocean Science, Silver Spring, MD 20910, USA

l

Wadsworth Center, New York State Department of Health, P.O Box 509, Albany, NY 12201-0509, USA

m

Department of Environmental Health Sciences, School of Public Health, State University of New York at Albany, Empire State Plaza, P.O Box 509, Albany, NY 12201-0509, USA

a r t i c l e i n f o

Keywords:

Asia–Pacific mussel watch

Emerging pollutant

Synthetic musk

Benzotriazole UV stabilizers

Concentrations and distribution

a b s t r a c t

We analyzed 68 green and blue mussels collected from Cambodia, China, Hong Kong, India, Indonesia, Japan, Korea, Malaysia, Philippines, Vietnam and the USA during 2003 and 2007, to elucidate the occur-rence and widespread distributions of emerging pollutants, synthetic musks and benzotriazole UV stabi-lizers (BUVSs) in Asia–Pacific coastal waters Synthetic musks and BUVSs were detected in mussels from all countries, suggesting their ubiquitous contamination and widespread distribution High concentra-tions of musks and BUVSs were detected in mussels from Japan and Korea, where the levels were com-parable or greater than those of PCBs, DDTs and PBDEs Significant correlations were found between the concentrations of HHCB and AHTN, and also between the concentrations of UV-327 and UV-328, which suggest similar sources and compositions of these compounds in commercial and industrial products To our knowledge, this is the first study of large-scale monitoring of synthetic musks and BUVSs in Asia– Pacific coastal waters

Ó 2012 Elsevier Ltd All rights reserved

1 Introduction

In recent years, there is an increasing public and scientific

con-cern on the occurrence and contamination by emerging pollutants,

such as synthetic musks and benzotriazole UV stabilizers (BUVSs)

in the environment The two polycyclic musks, HHCB (CAS#:

1222-05-5) and AHTN (CAS#: 1506-02-1), are important products

of the fragrance industry, which occupy 90–95% of the total

pro-duction of polycyclic musks (Worldwide propro-duction: 5600 tons in

lists HHCB as a high-production-volume chemical (HPV), which

suggests more than 450 tons of the compound is annually pro-duced in or imported into the US (USEPA, 2003) Although poten-tial effects and ecotoxicological concerns of the musks have been indicated (HERA, 2004), these compounds are used in personal care products, such as perfumes, body creams, lotions, and deodorants

as ingredients, at the maximum concentrations of several thou-sands oflg/g levels (Reiner and Kannan, 2006)

BUVSs are a class of plastic additives that are used in polypro-pylene and ABS (Acrylonitril, Butadiene and Styrene) copolymer products, which in turn are employed in building, automobile and consumer materials Among several BUVSs, UV-P (CAS#: 2440-22-4), UV-234 (CAS#: 70321-86-7), UV-326 (CAS#: 3896-11-5), UV-327 (CAS#: 3864-99-1), UV-328 (CAS#: 25973-55-1), and UV-329 (CAS#:3147-75-9) are commonly used in Japan The domestic production and import of UV-327 were 2436 tons

0025-326X/$ - see front matter Ó 2012 Elsevier Ltd All rights reserved.

⇑ Corresponding author Tel./fax: +81 96 342 3380.

E-mail address: nakata@sci.kumamoto-u.ac.jp (H Nakata).

Contents lists available atSciVerse ScienceDirect

Marine Pollution Bulletin

j o u r n a l h o m e p a g e : w w w e l s e v i e r c o m / l o c a t e / m a r p o l b u l

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between 2004 and 2009 (Nakata et al., 2010), but it decreased

dra-matically in 2010, only 3 tons (METI, 2012), probably due to the

availability of an alternative in the Japanese market UV-320

(CAS#: 3846-71-7) is prohibited from production, usage and

im-port in Japan since 2007, because of its potential for

bioaccumula-tion (BCF: up to 10,000; NITE, 2009) and toxicities to liver and

other tissues in laboratory mammals (Ema et al., 2008) The

no-observed adverse effect levels (NOAEL) of UV-320 in male and

fe-male rats were 0.1 mg/kg/day and 2.5 mg/kg/day, respectively

(Hirata-Koizumi et al., 2008) The significant gender difference in

NOAELs of UV-320 implies its endocrine disrupting potentials

The occurrence and concentrations of polycyclic musks and

BUVSs has been reported in the environment HHCB and AHTN

have been detected in air (Karrenborn and Gatermann, 2004; Peck

com-pounds were also found in high trophic species in coastal waters,

such as marine mammals and seabirds, suggesting their

concentrations of HHCB and AHTN are detected in wastewater

USA (Simonich et al., 2000), and several Asian countries (Guo

et al., 2010; Shek et al., 2008a,b; Zheng et al., 2007; Nakata and

Shinohara, 2010) This indicates that a major source of musks into

the environment may be WWTPs Further, HHCB were detected in

air and seawater from remote areas of the Northern Atlantic Ocean

(Xie et al., 2007) and in seals from the Antarctica (Schiavone et al.,

2009) These observations suggest that polycyclic musks,

espe-cially HHCB, have a potential of long-range transport, similar to

several persistent organic pollutants (POPs), but studies on

large-scale environmental monitoring of musks have been limited so far

BUVSs have been detected in marine organisms, such as

lug-worms, bivalves, fish, and birds from Japanese coastal waters

mammal tissues (Nakata et al., 2010), due to their lipophilic

prop-erties (log Kow: 6.95 for 327, 7.25 for 328) The BCF of

UV-327 between seawater and marine mammals was estimated to

be 33,300 (Nakata et al., 2010), which was similar to that reported

for a legacy POP, HCHs (BCF: 37,000;Tanabe et al., 1984) in a

wes-tern North Pacific food-chain Recently,Kim et al (2011)analyzed

fish samples from Manila Bay, Philippines, and indicated the

ubiq-uitous contamination and bioaccumulation of UV-P in the coastal

ecosystem High concentrations of BUVSs were detected in

influ-ents, effluent, and sewage sludge in Japan (Nakata et al., 2010;

Kameda et al., 2011) and China (Zhang et al., 2011), implying that

WWTP is a source of BUVSs into the environment However,

simi-lar to synthetic musks, studies on occurrence and environmental

concentrations of BUVSs are chiefly local in nature, there is a lack

of data on the widespread contamination and distribution of

BUVSs in the environment

The Mussel Watch Programs (MWPs) have reported POPs

con-tamination in coastal environments The National Oceanographic

and Atmospheric Administration (NOAA) launched MWP in 1986,

and more than 140 chemicals have been measured in bivalves

collected from nearly 300 monitoring sites along the US coast

concen-trations in 2009 (Kimbrough et al., 2009) As bivalves, such as

mus-sels and oysters, are sessile organisms that filter and accumulate

particles from water, the contaminant levels in their tissue are a

good indicator for understanding local pollution In Asia,Monirith

such as PCBs, DDTs, chlordanes (CHLs), HCHs, hexachlorobenzene

(HCB) in mussels collected from 12 countries during 1994 and

2001.Isobe et al (2007)reported spatial distribution of polycyclic aromatic hydrocarbons (PAHs) and phenolic compounds in mus-sels from Southeast Asian countries.Ramu et al (2007)analyzed persistent organochlorines and polybrominated diphenylethers (PBDEs) in mussels from Asian coastal waters during 2003 and

2005 However, till to-date, large-scale monitoring for emerging pollutants, synthetic musks and BUVSs, in Asia–Pacific coastal waters is not available

In this study, we analyzed two polycyclic musks (HHCB and AHTN) and four BUVSs (UV-320, UV-326, UV-327, UV-328) in blue and green mussels collected from Asian and US Pacific coasts to understand the status of contamination and geographical distribu-tion of these chemicals The correladistribu-tions between compounds were examined to elucidate the production/usage and discharge profiles

of these compounds Further, concentrations and accumulation patterns of musks and BUVSs were compared with those of POPs which were reported previously

2 Experimental section 2.1 Reagents

HHCB and AHTN standards were obtained from Dr Enrenstorfer GmbH (Augsburg, Germany) Four benzotriazole standards,

UV-320, UV-326, UV-327, and UV-328 were purchased from Wako Pure Chemicals, Co Ltd., Tokyo, Japan A deuterated PAH standard (d10-phenanthrene) obtained from Cambridge Isotope Laboratory (Andover, USA) was used as the surrogate standard

2.2 Samples Fifty-three green mussels (Perna viridis) and blue mussels (Myti-lus edulis) were collected from coastal waters of Asian countries, such as Cambodia (n = 2), China (n = 5), Hong Kong (n = 8), India (n = 3), Indonesia (n = 2), Japan (n = 7), Korea (n = 17), Malaysia (n = 4), Philippines (n = 2), and Vietnam (n = 3) during 2003 and

2007 Blue mussels were also obtained from the Pacific coast of the USA (n = 15) during 2004 and 2005 Detailed information on mussel samples is shown in Supporting Information (SI-1) Soft tis-sues of mussels collected from each location were pooled and homogenized, and kept at 20 °C until chemical analysis 2.3 Analytical method

The musks and BUVSs were analyzed by the method described previously (Nakata et al., 2007, 2009) with slight modifications Briefly, approximately 5 g of mussels were dried with anhydrous sodium sulfate and extracted with a mixture of dichloromethane and hexane (8:1) using a Soxhlet apparatus An aliquot of the ex-tract was used for lipid measurement by gravimetric analysis

d10-phenenthrene was spiked into the extract as a surrogate stan-dard For clean-up of the extract, a gel permeation chromatography column packed with Biobeads-S-X3 (Bio-Rad Laboratories, Hercu-les, CA, USA) and silica-gel column containing 5% deactivated Wako-gel C-200 (Wako Pure Chemical Co Ltd, Japan) were used for lipid removal, as well as removal of pigment and other extrane-ous components A mixture of 50% hexane in dichloromethane and 5% diethyl ether in hexane was used as mobile phase for GPC and silica-gel columns, respectively The eluate from the columns was

interfaced with a mass spectrometer (GC–MS, Agilent 6890 and 5973N Series)

Determination and quantification of polycyclic musks and BUVSs were performed using a GC–MS in SIM mode Ions were monitored at m/z 243, 258, 213 for HHCB; m/z 243, 258, 159 for

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AHTN; m/z 308, 323, 252 for 320; m/z 300, 315, 272 for

326; m/z 342, 344, 357 for 327; m/z 322, 351, 336 for

used was a HP-5MS fused silica capillary column (30 m  0.25 mm

i.d., 0.25lm film thickness; Agilent Technologies, USA) The oven

temperature was programmed from 80 °C to 160 °C at a rate of

10 °C/min and held for 10 min, and the temperature was increased

to 300 °C at a rate of 3 °C/min, with a final hold time of 15 min The

temperatures of injector and detector of GC–MS were set at 270 °C

and 300 °C, respectively Helium was used as a carrier gas

2.4 Quality control

A standard mixture containing all musks and BUVS analyzed in

this study was used to determine recovery rates of the compounds

through the analytical procedure Salad oil (0.5 g), that did not

con-tain the target compounds, was spiked with 50 ng of the standard

mixture Three replicate analyses were performed, and the average

recoveries of HHCB, AHTN, UV-320, UV-326, UV-327, and UV-328

were 114 ± 0.4%, 92 ± 1.6%, 114 ± 12%, 122 ± 11%, 114 ± 14%, and

110 ± 8.8%, respectively The recoveries of the surrogate standard,

d-phenanthrene, spiked into all mussel samples ranged from 72%

to 125% A procedural blank was analyzed with every set of 6

sam-ples a check for laboratory contamination and to correct sample

values, if necessary The concentrations were reported as below

the limit of detection, if the peak height was not greater than a

sig-nal to noise ratio of 3 times the blank values The limit of

detec-tions (LODs) of musks and BUVSs were 0.8, 0.4, 0.05, 0.10, 0.12,

and 0.15 ng/g (wet wt.) for HHCB, AHTN, 320, 326,

UV-327, and UV-328, respectively

2.5 Statistical analysis

Arithmetic mean (AM) and geometric mean (GM) were used to

describe the results Concentrations below the LOD were

substi-tuted with a value equal to the LOD for the calculation of AM and GM Data analysis by Speaman’s rank test was conducted using Excel Statistics (Esumi Co Ltd, Tokyo, Japan)

3 Results and discussion 3.1 Concentrations and distribution-synthetic musks Synthetic musks and BUVSs were detected in most of the sam-ples analyzed in this study The highest concentrations were found for HHCB (mean: 1300 ng/g lipid wt., Geometric mean [GM]:

430 ng/g), followed by AHTN (mean: 230 ng/g, GM: 90 ng/g),

UV-326 (mean: 150 ng/g, GM: 46 ng/g), UV-328 (mean: 130 ng/g, GM: 60 ng/g), UV-327 (mean: 68 ng/g, GM: 33 ng/g), and UV-320 (mean: 7.6 ng/g, GM: 4.7 ng/g) (Table 1) The detection frequencies

of HHCB were 81%, which were greater than those of AHTN (60%), UV-328 (65%), UV-326 (57%), and UV-327 (56%), and UV-320 (5.9%) This suggests extensive production and usage of HHCB, resulting in ubiquitous contamination in Asia–Pacific coastal waters

High concentrations of HHCB were found in mussels from Korea (mean: 2300 ng/g lipid wt., GM: 830 ng/g), Japan (mean: 2300 ng/

g, GM: 670 ng/g), Philippines (mean: 3300 ng/g, GM: 2800 ng/g), Malaysia (mean: 2200 ng/g, GM: 1500 ng/g) and Indonesia (mean:

1500 ng/g, GM: 1500 ng/g) (Table 1,Fig 1) The highest HHCB con-centrations in mussels were found in Kohyongsong Bay, Geoje, Korea, at 14,000 ng/g lipid wt., (SI-2) Elevated concentrations of HHCB were also found in mussels from Busan Bay (4500 ng/g), and Ulsan Bay (3400 ng/g), but HHCB was not detected in mussels from three stations, Wonmumpo, Gwangyang Bay and Chunsoo Bay in Korea Large variations in HHCB concentrations were also found in samples from the USA and Japan In San Francisco Bay, HHCB concentrations in mussels were between 1600 and

2200 ng/g lipid wt., whereas the concentrations in other locations

in the West Coast of the US were below 200 ng/g In Japan, HHCB

Table 1

Concentrations (ng/g lipid wt.) of polycyclic musks and benzotriazole UV stabilizers in mussels from Asia–Pacific coastal waters.

Country/region N of sample Sampling year Lipid (%) Polycyclic musks Benzotriazole UV stablizers

HHCB AHTN UV-320 UV-326 UV-327 UV-328 Cambodia 2 2004 1.6 280 (260) * 70 (57) ND ND ND 120 (110)

[100] ** [50] [0] [0] [0] [100] China 5 2004 0.64 270 (220) 190 (150) ND 60 (33) 84 (65) 96 (52)

[80] [100] 0 [40] [80] [60] Hong Kong 8 2004 1.4 710 (300) 110 (76) ND 91 (18) 93 (48) 200 (75)

[63] [63] [0] [25] [75] [75]

Indonesia 2 2003 1.2 1500 (1500) 180 (170) ND 33 (22) 58 (45) 120 (110)

[100] [100] [0] [50] [100] [100] Japan 7 2007 2.8 2300 (670) 860 (200) 33 (13) 450 (260) 38 (15) 120 (93)

[71] [57] [57] [100] [43] [100] Korea 17 2005 1.5 2300 (830) 220 (100) ND 210 (90) 100 (56) 220 (150)

[82] [47] [0] [76] [65] [94] Malaysia 4 2004 1.8 2200 (1500) 370 (290) ND 42 (12) ND 24 (14)

[100] [100] [0] [25] [0] [25] Philippines 2 2004 1.1 3300 (2800) 490 (460) ND 120 (50) 150 (150) 170 (140)

[100] [100] [0] [50] [100] [100] USA 15 2004–2005 1.1 430 (210) NA ND 130 (70) 61 (45) 69 (33)

[93] [0] [0] [80] [73] [20]

Total 68 2003–2007 1.5 1300 (430) 230 (90) 7.6 (4.7) 150 (46) 68 (33) 130 (60)

[81] [60] [6] [57] [56] [65] ND: Less than detection limit (HHCB: 0.80, AHTN: 0.40, UV-320: 0.05, UV-326: 0.10, UV-327: 0.12, UV-328: 0.15 ng/g wet wt.).

NA: Data not available.

* Parenthesis: Geometric mean (GM) concentration.

**

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concentrations in Osaka Bay mussels were more than 10-fold

greater than those of Ariake Sea, western Japan This may be due

to the distance between sampling stations and point source of

pol-lution, such as WWTP Elevated concentrations of HHCB were

de-tected in influent, effluent, and sewage sludge samples in several

countries (Eschke, 2004; Simonich et al., 2000; Guo et al., 2010;

Shek et al., 2008a,b; Zheng et al., 2007; Nakata et al., 2010) The

highest concentrations of HHCB were found in mussels from Tsim

Sha Tsui in Hong Kong (SI-2), where WWTP effluent outfall from

four plants are located (Shek et al., 2008a,b) These results are in

accordance with a previous study which determined polycyclic

musks in mussels from Hong Kong (Shek et al., 2008a,b)

Another synthetic musk fragrance, AHTN, was detected in

mus-sels from all countries except for Vietnam and the USA AHTN was

not identified in the US mussels because of the existence of

inter-ference peaks in the GC–MS chromatograms Similar to HHCB, high

concentrations of AHTN were found in mussels from Japan (mean:

860 ng/g lipid wt., GM: 200 ng/g), Malaysia (mean: 370 ng/g, GM:

290 ng/g), Philippines (mean: 490 ng/g, GM: 460 ng/g), and Korea

(mean: 220 ng/g; GM: 100 ng/g) (Table 1) The lower

concentra-tions of AHTN compared to HHCB may be explained by different

amounts of production/usage between these two compounds It

was reported that the use volumes of HHCB and AHTN were

1427 and 358 tons in Europe in 2000 (HERA, 2004), respectively

AHTN concentrations in WWTP samples were approximately 5–

2004; Simonich et al., 2000; Guo et al., 2010; Shek et al.,

2008a,b; Zheng et al., 2007; Nakata et al., 2010), reflecting lower

levels of discharge of AHTN into the aquatic environment AHTN

and HHCB concentrations were generally low in mussels from

India and Vietnam While further research is needed because of the limited number of samples analyzed, this provides some basic information on the extent of distribution of these compounds in coastal areas

3.2 Concentrations and distribution-benzotriazole UV stabilizers Among the four BUVSs analyzed, UV-326 and UV-328 were the predominant compounds in mussels, and they were detected in samples from China, Hong Kong, Indonesia, Japan, Korea, Malaysia, Philippines, and the USA (Table 1,Fig 1) The detection frequency

of UV-326 was the highest in Japan (100%), followed by the US West Coast (80%) and Korea (76%) High concentrations of

UV-326 were found in Japanese mussels (mean: 450 ng/g lipid wt., GM: 260 ng/g), especially in Osaka Bay (maximum: 1500 ng/g; SI-1) In Korea, UV-326 concentrations in mussels from Ulsan Bay were between 850 and 1200 ng/g, which were approximately one order of magnitude greater than those of other sampling sites in that country Elevated concentrations of UV-326 were found in two stations at Hong Kong, Tsim Sha Tsui (450 ng/g) and Sai Wan

Ho (230 ng/g) These levels were similar to UV-326 concentrations

in mussels from Tokyo Bay, Japan (120–390 ng/g) On the other hand, UV-326 concentrations in mussels from Indonesia, Malaysia and Philippines were generally low, although the concentrations of musks in mussels in these countries were relatively high These observations may suggest the existence of point sources of

UV-326 in Eastern Asian countries

The distribution of UV-326 in the US West Coast samples was less variable, and mean concentration was 130 ng/g (GM:

70 ng/g) This is inconsistent with the results of synthetic musk

3 µg/g (lipid wt )

Cambodia

USA

Korea Japan

China

Hong Kong

Philippines

Indonesia

India

Vietnam Malaysia

ND ND

ND ND

HHCB

1 µg/g (lipid wt )

ND ND

ND ND

Cambodia

USA

Korea Japan

China

Hong Kong

Philippines

Indonesia

India

Vietnam Malaysia

ND ND ND ND ND

UV-328

Fig 1 Geographical distribution of HHCB and UV-328 in blue and green mussels collected from Asia–Pacific coastal waters.

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concentrations; elevated levels of HHCB were found in mussels

from San Francisco Bay (Table SI-2) Previous studies reported

the occurrence of BUVSs including UV-326 in wastewater and

sludge samples25,31, suggesting that WWTP is one of the sources

of environmental discharge of BUVSs However, spatial similarity

in the distribution of UV-326 concentrations in US mussels implies

the presence of another source of this compound into the

environ-ment Recent investigations showed the occurrence of BUVSs in

road dust samples originated from automobiles (Nishidome et al.,

2011), which may be a potential source of BUVSs into the aquatic

environment UV-326 was not detected in mussels from Cambodia,

India, and Vietnam, probably due to the small amounts of

produc-tion and usage of this compound

UV-327 was detected in mussels from China, Hong Kong,

Indo-nesia, Japan, Korea, Philippines and the USA High concentrations

of UV-327 were found in samples from Philippines (mean:

150 ng/g, GM: 150 ng/g), followed by Korea (mean: 100 ng/g,

GM: 56 ng/g), Hong Kong (mean: 93 ng/g, GM: 48 ng/g), and Japan

(mean: 38 ng/g, GM: 15 ng/g), but the concentrations showed large

variations among stations Similar to the spatial distribution of

UV-326, higher residue levels of UV-327 were found in samples from

Tsi Sha Tsui in Hong Kong, Ulsan Bay in Korea, and Baccor cavire

in Philippines However, UV-327 was not found in Osaka Bay

sam-ples, due to an interference peak in the GC–MS analysis The mean

concentration of UV-327 was generally low in Chinese mussels

(mean: 84 ng/g; GM: 65 ng/g), but high detection frequencies were

observed (80%) A similar profile was found for the US samples, at

mean concentrations and detection frequencies of 61 ng/g (GM:

45 ng/g) and 73%, respectively These results may indicate

wide-spread release of UV-327 from non-point source into the aquatic

environment UV-327 was not detected in mussels from Cambodia,

India, Malaysia, and Vietnam, which was similar to what found for

UV-326

UV-328 was detected in mussels from all countries, except for

India and Vietnam (Fig 1,Table 1) The highest concentrations of

UV-328 were found in mussels from Tsim Sha Tsui, Hong Kong

(830 ng/g lipid wt.), followed by Ulsan Bay (620 ng/g lipid wt.),

On-san Bay (590 ng/g) in Korea, Sai Wan Ho, Hong Kong (430 ng/g) and

Tokyo Bay, Japan (370 ng/g) (SI-2) The detection frequency of

UV-328 in mussels was high in Japan (100%) and Korea (94%), but the

concentrations showed a large spatial variation, approximately 5–

10-fold differences among various locations This profile is similar

to that for UV-326 concentrations, suggesting the existence of a

point source of BUVSs in the vicinity of Ulsan Bay and Tokyo Bay

Similar to Korean and Japanese samples, UV-328 concentration

varied considerably in the US mussels High concentrations of

UV-328 was detected in mussel from San Francisco Bay, at level

of 310 ng/g lipid wt., but samples from other locations showed

lower concentration or less than detection limit This is in

accor-dance with the spatial distribution of HHCB in mussels from the

West Coast of the US As described earlier, BUVSs were detected

in wastewater samples as well as in road dust, but no mass-balance

analysis is available so far A quantitative investigation is needed

to determine potential sources of BUVSs into the aquatic

environment

While the numbers of samples analyzed for each country was

limited, UV-328 was detected in all samples collected from

Cambo-dia, Indonesia, and Philippines (Table 1) Among four BUVSs

ana-lyzed, UV-328 was only detected in Cambodian mussels, and the

levels were comparable with those of Hong Kong and Japan This

implies that the usage of UV-328 in Cambodia may be extensive

compared to other BUVSs UV-328 was detected in Philippines

samples at a concentration of 270 ng/g lipid wt.Kim et al (2011)

reported the occurrence of BUVSs in fish samples collected from

Manila Bay, Philippines The high concentrations and frequent

detection of UV-328 in fish suggest widespread use of this

compound in urbanized areas of Philippines UV-328 was not identified in mussels from India and Vietnam, which was similar

to that found for other BUVSs

UV-320 was only detected in mussels from Japan at low concen-trations (mean: 33 ng/g lipid wt., GM: 13 ng/g) It has been re-ported that UV-320 is bioaccumualtive and highly toxic, the Japanese government prohibited the production, use, and import

of UV-320 since 2007 (NITE, 2009) While UV-320 is detected in road dusts in recent years (Nishidome et al., 2011), the exposure and the concentrations in environmental matrices is expected to decrease in the future Lack of detection of UV-320 in mussel sam-ples from all countries, except for Japan, indicates less usage of this compound in the Asia–Pacific region

3.3 Correlation between compounds The relationships between concentrations of musks and BUVSs

in mussels were examined using Spearman’s rank correlation analysis A significant correlation between HHCB and AHTN concentrations was found, except for one mussel collected from Kohyongsong Bay Korea (Fig 2) This is consistent with the results

of previous studies investigating marine organisms and sediment (Nakata et al., 2007); suggesting that sources and profiles of envi-ronmental exposure to HHCB and AHTN are similar However, there were two different regression lines, with high and low slopes,

with high slope were mainly composed of mussels from Osaka Bay and Tokyo Bay, Japan The slope value of the regression line was 0.32, which was approximately 2.5-fold greater than that of another group (slope: 0.13) These results may imply that large amounts of AHTN have been discharged into coastal waters of Ja-pan Significant correlations between HHCB and AHTN concentra-tions were also found in wastewater samples from Japan, Korea, China and Hong Kong A slope value of the regression line between HHCB and AHTN concentrations in wastewater samples was 0.42

in Japan (Nakata et al., 2010), which was apparently greater than those of China (0.09;Zheng et al., 2007), Hong Kong (0.18;Shek

et al., 2008a,b) and Korea (0.34;Guo et al., 2010) While temporal and seasonal variations in slope may be present in WWTPs, large amount of production and usage of AHTN should be considered

in Japan

A significant positive correlation was found between UV-328 and UV-327 concentrations in mussels (r2= 0.64, p < 0.01;Fig 3) This correlation was also found in surface water, and sediments

in Japan (Nakata et al., 2009; Kameda et al., 2011) and fish from Manila Bay, Philippines (Kim et al., 2011), which indicates the sim-ilar sources of these compounds, such as plastic materials How-ever, mussel samples could be classified into two groups, Group

A and B, based on UV-328 concentrations (Fig 3) Group A includes mussels containing low concentrations of UV-328 (<250 ng/g li-pid wt.), which showed a significant correlation and a high slope value of the regression line (0.71) In contrast, mussels of Group

B contained high concentrations of UV-328 (>250 ng/g), but the slope of correlation (0.27) was approximately one-third of that of Group A This group mainly consists of mussels from polluted sites, such as Ulsan Bay, Tsu Sha Tsui, and Tokyo Bay in Korea, Hong Kong and Japan, respectively These observations may suggest the occurrence of highly contaminated sites with UV-328 in East Asia, although their potential sources are not well known

A significant correlation was also observed between UV-328 and UV-326 concentrations, when two samples from Osaka Bay were removed from the calculation (Fig 4) As described earlier, high concentrations of UV-326 were detected in mussels from

Osa-ka Bay, probably due to the presence of a point source in this re-gion Furthermore, a significant correlation was found in the relationship between UV-327 and UV-326 (r2= 0.32, p < 0.01)

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These results were consistent with those reported in the previous

studies which analyzed BUVSs in sediment (Nakata et al., 2009)

and fish (Kim et al., 2011), suggesting that exposure profiles of

BUVSs into the aquatic environment may be similar in the

Asia–Pa-cific regions, except for some locations which are proximal to point sources It is interesting to note that there was a lack of significant relationship between concentrations of synthetic musks and BUVSs in mussels, as has been reported in earlier studies

0 500 1,000 1,500 2,000 2,500 3,000

0 2,000 4,000 6,000 8,000 10,000 12,000 14,000 16,000

Osaka-1 (Japan)

Osaka-2 (Japan)

Tokyo-1 (Japan) Tokyo-2 (Japan) Dalian (China)

Kohyonsong bay (Korea)

y=0.13x+44

(r 2=0.94, p<0.01)

y=0.32x+240

(r 2=0.98, p<0.01)

HHCB concentration (ng/g lipid wt.)

Fig 2 Relationship between HHCB and AHTN concentrations in mussels collected from Asian coastal waters.

0 50 100 150 200 250 300 350

UV-328 concentration (ng/g lipid wt.)

250 ng/g

Group A y=0.71x+20

(r2=0.51, p<0.01)

(HK)

(HK)

(KOR)

(KOR)

(KOR) (JPN)

(US)

(JPN) (CHN)

(KOR)

Group B y=0.27x+54

(r2=0.67, p<0.01)

Fig 3 Relationship between UV-328 and UV-327 concentrations in mussels from Asia–Pacific coastal waters.

UV-328 concentration (ng/g lipid wt.) 0

400 800

y=0.76x+45 (r 2=0.41, p<0.01)

excluding Osaka samples Osaka-1, JPN

Osaka-2, JPN

1,600

1,200

Fig 4 Relationship between UV-328 and UV-326 concentrations in mussels from Asia–Pacific coastal waters.

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3.4 Comparison to legacy POPs

In order to understand the status of contamination and

geo-graphical distribution by emerging pollutants in Asian coastal

waters, concentrations of synthetic musks and BUVSs in mussels

obtained in this study were compared with those reported for

leg-acy POPs previously (Fig 5) Interestingly, GM concentrations of

musks (sum of HHCB and AHTN concentrations) were the highest

among the compounds analyzed in mussels from Cambodia,

Indonesia, Japan, Korea, Malaysia and Philippines The GM

concen-trations of musks were 1800 ng/g lipid wt., in mussels from

Malay-sia, which were more than 20-fold greater than those of legacy

2007) In Philippines and Indonesia, musks concentrations were

3300 ng/g and 1700 ng/g lipid wt., respectively, followed by DDTs

(1400 ng/g in Philippines, 730 ng/g in Indonesia) While the

num-bers of samples analyzed were limited for each countries,

exten-sive monitoring of musks contamination is necessary as their

concentrations are high in mussels BUVSs were also detected in

mussels from Philippines (GM: 340 ng/g), which were comparable

with those in Japan and Korea BUVSs were also found in almost all

fish samples from Manila Bay, Philippines, implying

bioaccumula-tion potential and ubiquitous contaminabioaccumula-tion by BUVSs in this

country (Kim et al., 2011)

The GM concentrations of musks in Japanese and Korean

mus-sels were higher than those of PCBs and DDTs Further, BUVSs in

mussels showed the second highest concentrations, at levels of

370 ng/g in Japan and 300 ng/g in Korea While the details of

pro-duction and usage of musks and BUVSs are not available, large

amount of these compounds, especially HHCB, UV-326 and

UV-328, may be produced and used in both countries However,

comparable concentrations of HHCB and PCBs were observed in

bi-valves collected from the Ariake Sea, western Japan The

concentra-tions of HHCB in oyster and clams were 760 ng/g and 700 ng/g

(Nakata et al., 2007), respectively, which were similar to those of

PCBs at the concentrations of 250–600 ng/g in mussels, oysters

elevated concentrations of PCBs in mussels were reported in

urbanized areas, such as Osaka Bay and Tokyo Bay at the GM

con-centrations of 2600 ng/g lipid wt., (Ueno et al., 2010) and 1600 ng/

g lipid wt., respectively Additional monitoring studies on musk

concentrations in Japanese coastal waters may be necessary to

compare concentrations between musks and PCBs and to delineate

temporal trends in concentrations The GM concentrations of musks and BUVS in mussels were similar for Japanese and Korean samples This is in accordance with HHCB concentrations in marine mammal tissues; average concentrations of HHCB in blubber tis-sues of finless porpoises were 29 ng/g lipid wt., in Japan (Nakata

et al., 2007) and 37 ng/g lipid wt., in Korea (Moon et al., 2011) The musks concentrations in Japanese and Korean mussels were one order of magnitude greater than those of PBDEs, reflecting the different exposure amounts between compounds

In China and Hong Kong, DDT concentrations were high in mus-sels, and musks showed the second highest concentrations in this study The GM concentration of musks was 370 ng/g lipid wt., in mussels from China, which was approximately 8-fold greater than that of PCBs (45 ng/g lipid wt.).Shek et al (2008a,b)reported that mean concentrations of HHCB in mussels from Hong Kong were

1200 ng/g lipid wt., which was also higher than PCBs concentra-tions in Hong Kong (400 ng/g;Ramu et al., 2007) The residue lev-els of musks and BUVSs were generally low in musslev-els from India and Vietnam

This study revealed the occurrence and widespread contami-nation by emerging pollutants, synthetic musks and BUVSs, in Asia–Pacific coastal waters However, little information on ecotoxicological implications of musks and BUVSs are available Further monitoring studies are needed to assess their trends and risks associated with accumulation and distributions in the aquatic environment

Acknowledgements The US samples were collected as a part of the NOAA’s Status and Trends Mussel Watch program We thank Ms Amanda Brew-ster and Dr Juan Ramirez (TDI Brooks International College Station, Texas) for the provision of mussels samples from the US Pacific coast for analysis Thanks are also due to Prof Hideshige Takada (Tokyo University of Agriculture and Technology, Japan) for provid-ing mussel samples analyzed This study was partly supported by the Grant-in-aid for Scientific Research (C) (No 20510067) and Scientific Research (S) (No 20221003) from Japan Society for the Promotion of Science (JSPS), Global Environmental Research Fund (RF-094) of the Ministry of the Environment, Japan, and Global COE Program and program under the Special Coordination Funds for Promoting Science and Technology from the Japanese Ministry

of Education, Culture, Sports, Science and Technology (MEXT) Appendix A Supplementary data

Supplementary data associated with this article can be found, in

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