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Concentrations of perfluorinated acids in livers of birds from Japan and Korea
Chemosphere 49 (2002) 225–231 www.elsevier.com/locate/chemosphere
Concentrations of perfluorinated acids in livers of birds from Japan and Korea Kurunthachalam Kannan a,*, Jae-Won Choi b, Naomasa Iseki c, Kurunthachalam Senthilkumar c, Dong Hoon Kim a, Shigeki Masunaga c, John P. Giesy a a
c
National Food Safety and Toxicology Center, Department of Zoology, Institute for Environmental Toxicology, Michigan State University, East Lansing, MI 48824, USA b National Institute for Environmental Studies, Onogawa 16-1, Tsukuba, Ibaraki 305-8506, Japan Graduate School of Environment and Information Sciences, Yokohama National University, 79-7 Tokiwadai, Hodogaya-ku, Yokohama 240-8501, Japan Received 1 March 2002; received in revised form 23 May 2002; accepted 30 May 2002
Abstract Livers of birds collected from Japan and Korea (n ¼ 83) were analyzed to determine the concentrations of perfluorooctanesulfonate (PFOS), perfluorooctanesulfonamide (FOSA), perfluorooctanoic acid (PFOA) and perfluorohexanesulfonate (PFHS). PFOS was found in the livers of 95% of the birds analyzed at concentrations greater than the limit of quantitation (LOQ) of 10 ng/g, wet weight. The greatest concentration of PFOS of 650 ng/g, wet weight, was found in the liver of a common cormorant from the Sagami River in Kanagawa Prefecture. Concentrations of PFOS in bird livers from Japan and Korea were within the ranges of values reported for those from the United States and certain European countries. PFOA and PFHS were found in 5–10% of the samples analyzed. The greatest concentrations of PFOA and PFHS in bird livers were 21 and 34 ng/g, wet weight, respectively. FOSA was found in all the samples (n ¼ 10) of cormorants collected from the Sagami River in Japan. The greatest concentration of FOSA in cormorant liver was 215 ng/g, wet weight. There was no significant correlation between the concentrations of PFOS and FOSA in cormorants collected from the Sagami River. These results suggested that the distribution of FOSA is localized. No ageor gender-specific differences in fluorochemical concentrations could be discerned in birds. Ó 2002 Elsevier Science Ltd. All rights reserved. Keywords: PFOS; Perfluorinated compounds; Birds; Asia
1. Introduction Concern about fluorinated organic compounds, particularly perfluorinated (fully fluorinated) compounds, is increasing. Perfluorooctanesulfonate (PFOS) and related perfluorinated acids are shown to be globally
*
Corresponding author. Tel.: +1-517-432-6321; fax: +1-517432-2310. E-mail address: [email protected] (K. Kannan).
distributed, environmentally persistent and bioaccumulative (Giesy and Kannan, 2001, 2002). PFOS, perfluorooctanesulfonamide (FOSA), perfluorooctanoic acid (PFOA) and perfluorohexanesulfonate (PFHS) have been reported to occur in blood sera of humans (Hansen et al., 2001). Similarly, PFOS, FOSA, PFOA and PFHS have also been identified in tissues of wildlife from various parts of the world (Giesy and Kannan, 2001; Kannan et al., 2001a,b, 2002a,b,c). Earlier studies have reported the occurrence of these compounds in wildlife collected from North America and certain European
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K. Kannan et al. / Chemosphere 49 (2002) 225–231
countries. In this study, concentrations PFOS, FOSA, PFOA and PFHS in livers of birds collected from Japan and Korea are reported to provide information on the extent of contamination in these industrialized Asian countries.
2. Materials and methods Liver samples from forty individuals belonging to six species of birds were collected from Japan during June 1998 to February 1999 except gray herons, which were collected during 1997–1998. Samples of Japanese birds were from Rishiri Island (Hokkaido), Haneda airport (Tokyo), Atsugi city and the Sagami River in Kanagawa Prefecture and Gyotoku wild bird observatory in Chiba, Tokyo. Kanagawa Prefecture is located just south of highly urbanized areas such as Tokyo and Yokohama is one of the major industrial cities located in Kanagawa Prefecture (Fig. 1). Sea gulls were collected from Rishiri Island and common cormorants were collected from the Sagami River. Gyotoku wild bird observatory is a rehabilitation center for birds. Forty three individuals belonging to 10 species of birds were collected from the Nakdong River estuary in Korea from 1993 to 1994, except two black-tailed gulls, which were collected in 1997. The Nakdong River estuary is located along the east coast of Korea near Pusan (Fig. 1). Several industrial complexes including chemical manufacturers, oil refineries and heavy industries are located near this estuary. The estuary has been designated as a natural conservation area to protect wildlife. Among several species of birds analyzed from Korea,
the black-tailed gull is a resident, piscivorous bird. Black-tailed gulls were also collected in Nan Do, which is located on the west coast of Korea and Hong Do is located south of the Nakdong estuary. Most other species studied are migratory birds that winter in the Nakdong River estuary. Bird species analyzed in this study are listed in Tables 1 and 2. Prior to dissection, body weight, length and sex of birds were assessed. Livers were stored at )20 °C until analysis. Concentrations of PFOS, FOSA, PFOA and PFHS in liver were measured using high performance liquid chromatography (HPLC) with electrospray tandem mass spectrometry (Hansen et al., 2001). A liver homogenate of 1 g of liver in 5 ml of high purity Milli-Q water was prepared. One ml of the homogenate, 1 ml of 0.5 M tetrabutyl ammonium hydrogen sulfate solution (adjusted to pH 10), and 2 ml of 0.25 M sodium carbonate buffer were added to a 15-ml polypropylene tube for extraction. After thorough mixing, 5 ml of methyl-tertbutyl ether (MTBE) was added, and the mixture was shaken for 20 min. The organic and aqueous layers were separated by centrifugation, and an exact volume of MTBE (4 ml) was removed from the solution. The aqueous mixture was rinsed with MTBE and separated twice. The solvent was allowed to evaporate under nitrogen before being reconstituted in 0.5 or 1 ml of methanol. The sample was vortexed for 30 s and passed through a 0.2 lm nylon mesh filter into an autosampler vial. Analyte separation was performed using a HewlettPackard HP1100 liquid chromatograph modified with low dead-volume internal tubing. Ten ll of extract was injected onto a 50 2 mm (5 lm) Keystone Betasilâ C18
Fig. 1. Map of Japan and Korea showing sampling locations.
Table 1 Concentrations (ng/g, wet weight) of perfluorinated acids in livers of birds from Japan Date of collection
Location
Sex
Body weight (g)
Length (cm)
PFOS
FOSA
PFOA
PFHS
Sea gull (n ¼ 14) Larus crassirostris
Jun-98
Rishiri Island, Hokkaido
M
658 (586–706)
487 (468–521)
53 (23–89)
<75
<19
<7.5–34a
Sea gull (n ¼ 7) L. crassirostris
Jun-98
Rishiri Island, Hokkaido
F
504 (470–570)
468 (435–510)
40 (<19–53)
<75
<19
<7.5
Sea gull (n ¼ 1) L. crassirostris
Sep-98
Haneda Airport, Tokyo
NA
789
535
230
<75
<19
<7.5
Spot-billed duck (n ¼ 1) Anas poecilorhyncha
Jun-98
Gyotoku bird observatory
F
722
500
160
<75
<19
<7.5
Black-headed gull (n ¼ 1) Larus ridibundus
Jun-98
Gyotoku bird observatory
M
196
401
<19
<75
21
<7.5
Black-eared kite (n ¼ 1) Milvus lineatus
Jul-99
Atsugi city, Kanagawa Prefecture
NA
860
NA
450
<75
21
34
Black-eared kite (n ¼ 1) M. lineatus
Sep-99
Haneda Airport, Tokyo
M
966
605
180
<75
<19
<7.5
Gray heron (n ¼ 2) Ardea cinerea
Oct-97 and Jul-98
Gyotoku bird observatory
M
395 (368–422)
738 (495–980)
50 (49–52)
<75
<19
<7.5
Common cormorant (n ¼ 2) Phalacrocorax carbo
Feb-99
Sagami River, Kanagawa Prefecture
M
2144 (2039–2249)
820 (810–830)
390 (330–450)
115 (100–130)
<19
<7.5
Common cormorant (n ¼ 8) P. carbo
Feb-99
Sagami River, Kanagawa Prefecture
F
1873 (1619–2130)
762 (740–816)
385 (170–650)
162 (110–215)
<19
<7.5–10a
K. Kannan et al. / Chemosphere 49 (2002) 225–231
Species
All cormorants are adults except one female, which was a juvenile. a Only one sample above detection limit.
227
228
Table 2 Concentrations (ng/g, wet weight) of PFOS in livers of birds collected from Koreaa Scientific name
N
Sampling date
Sex
Length (cm)
Weight (g)
Feeding habits
PFOS
Bar-tailed godwit
Limosa lapponica
3
Mar-93
F
46 (42–51)
255 (210–300)
Crustaceans, insects, small fish
148 (22–310)
Black-headed gull
L. ridibundus
1 4
Feb-94 Dec-92–Jan-94
F M
38 43 (42–45)
250 304 (240–340)
Omnivorous Omnivorous
292 296 (148–500)
Black-tailed gull
Larus crassirostris
7
May–Dec-93
NA
NA
533 (480–600)
Omnivorous
112 (36–215)
Black-tailed gull Black-tailed gullb
L. crassirostris L. crassirostris
1 1
1997 1997
NA NA
NA NA
NA NA
Omnivorous Omnivorous
74 71
Common gull
Larus canus
1 2
Nov-93 Dec-92–Feb-94
F M
55 40–49
550 250–550
Omnivorous Omnivorous
28 29–63
Black-necked grebe
Podiceps nigricollis
1
Mar-93
F
35
400
Insects, small fish, shrimp
10
Common tern
Sterna hirundo
1 1
May-93 May-93
F M
35 30.6
123 104
Fishes Fishes
11.2 <10
Great knot
Calidris tenuirostris
1
Aug-93
M
27.5
138
Invertebrates, shellfish, crustaceans
13.5
Greenshank
Tringa nebularia
1 2
Nov-93 Aug–Nov-93
F M
19.9 24–33
230 143–190
Invertebrates, shellfish, crustaceans Invertebrates, shellfish, crustaceans
13.8 61–112
Herring gull
Larus argentatus
10
Feb–Dec-93
6M, 1F, 3NA
1000–1400
62–71
Omnivorous
49.6 (<10–116)
Sanderling
Crocethia alba
2
May-94
F
20–22
60–62
Invertebrates, crustaceans, shellfish
21–112
Little egret
Egretta garzetta
4
NA
NA
NA
NA
Fish, frogs
24.8 (19–30)
a
FOSA, PFOA and PFHS were not detected in any of the birds from Korea at a quantitation limit of 38, 36 and 36 ng/g, wet weight. a From Hong Do. b From Nan Do.
K. Kannan et al. / Chemosphere 49 (2002) 225–231
Species
K. Kannan et al. / Chemosphere 49 (2002) 225–231
column with a 2 mM ammonium acetate/methanol mobile phase starting at 10% methanol at a flow rate of 300 ll/min, to 100% methanol at 11.5 min before reverting to original conditions at 13 min. Column temperature was maintained at 25 °C. For quantitative determination, the HPLC system was interfaced to a Micromassâ (Beverly, MA) Quattro II atmospheric pressure ionization tandem mass spectrometer operated in the electrospray negative mode. Instrumental parameters were optimized to transmit the [M-K] ion for all analytes before fragmentation to one or more product ions. When possible, multiple daughter ions were monitored, but quantitation was based on a single product ion. In all cases, the capillary was held between 1.6 and 3.2 kV. Primary and product ions monitored for PFOS, PFOA, FOSA and PFHS determinations were 499 > 99, 413 > 169, 498 > 78 and 399 > 80, respectively. Product ions 99, 169, 78 and 80 correspond to FSO 3 , C3 F7 , SO2 N , SO3 , respectively. Recoveries of 250 ng of PFOS, FOSA, PFOA and PFHS spiked into livers of sea gulls were 98%, 44%, 100% and 56% respectively, while recoveries from samples of cormorants were 230%, 107%, 95% and 59%, respectively, and those from black-tailed gulls were 121%, 63%, 136% and 25%, respectively. Causes of great recoveries of PFOS (230%) spiked to cormorant livers are unknown. Recoveries of PFHS from bird livers were low (25–59%). Relative standard deviations of replicate analyses of spiked tis-
229
sues were less than 15%. Concentrations of fluorochemicals were not corrected for recoveries. For the estimation of the limit of quantitation (LOQ), the tissue samples were compared to an unextracted standard calibration curve. For instance, if 5 ng/ml standard is the lowest acceptable standard, and sample had been diluted by a factor of 7, the LOQ is reported as 35 ng/ml. LOQs for fluorochemicals varied from 10 to 75 ng/g, wet weight. Representative HPLC-ESMSMS chromatograms of liver extracts of a black-eared kite that contained 450, 21 and 34 ng/g, wet weight, PFOS, PFOA and PFHS, respectively, is presented (Fig. 2).
3. Results and discussion 3.1. Birds from Japan PFOS was found at concentrations greater than the LOQ of 19 ng/g, wet weight, in 38 of the 40 bird livers analyzed from Japan (Table 1). The greatest concentration of PFOS of 650 ng/g, wet weight, was found in the liver of a common cormorant from the Sagami River. Concentrations of PFOS in birds from Japan were within the range of values found in the livers of birds collected across the United States (Kannan et al., 2001a). However, the greatest concentration of 650 ng PFOS/g, wet weight, found in cormorant liver from the
Fig. 2. Representative HPLC-ESMSMS chromatograms of PFOS (ions 499 > 80 and 499 > 99) and PFOA (ions 413 > 169) and PFHS (ions 399 > 99) in sample extracts of black-eared kite liver. Concentrations of PFOS, PFOA and PFHS in this sample were 451, 21 and 34 ng/g, wet weight, respectively.
230
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Sagami River was 3-fold less than the highest concentrations of 1780 ng/g, wet weight, found in Brandt’s cormorants from San Diego, United States (Kannan et al., 2001a). Mean concentrations of PFOS in livers of cormorants from Japan were 6-fold greater than those found in livers of cormorant from Sardinia Island in Italy (Kannan et al., 2002b). Similarly, concentrations of PFOS in cormorants were 5–10-fold greater than those found in white-tailed sea eagles from eastern Germany (Kannan et al., 2002b). Mean concentrations of PFOS in cormorants were greater than those found in other species of birds with the exception of a black-eared kite (450 ng/g, wet weight) collected from Atsugi city in Kanagawa Prefecture. Great concentrations of PFOS in cormorants and blackeared kite collected from Kanagawa can be explained by heavy industrialization in this region. A sea gull collected from Haneda airport in Tokyo contained 5–6-fold greater concentrations of PFOS than those collected from remote areas such as Rishiri Island in Hokkaido. These results suggest that urbanized and industrialized areas are major sources of exposure of birds to PFOS. FOSA and PFHS were detected in 5–10% of the bird livers analyzed. While PFOS was not found in blackheaded gull at the LOQ of 19 ng/g, PFOA was detected in this individual at a concentration of 21 ng/g, wet weight. This indicates that fluorochemicals other than PFOS can be prevalent in some locations, although at lesser frequencies. PFHS was found in individual sea gulls, black-eared kite and common cormorants with a maximum concentration of 34 ng/g, wet weight. FOSA was found in all the livers of cormorants from the Sagami River. Concentrations of FOSA in cormorant livers were approximately 3-fold less than those of PFOS (Table 1). Concentrations of FOSA in cormorant livers were not significantly (p > 0:05; r2 ¼ 0:12) correlated with concentrations of PFOS (Fig. 3). Similarly, concentrations of FOSA in livers of cormorants were not correlated with concentrations of PFHS or PFOA. Concentrations of PFHS and PFOA in cormorants were generally less
than the LOQ. FOSA was not found in the livers of other species of birds analyzed at the LOQ of 75 ng/g, wet weight. These results suggest that the exposure of birds to FOSA is relatively less than to PFOS and that it is localized. Furthermore, these results indicate that the sources of FOSA are not directly associated with those of PFOS. Similarly, exposures of PFHS and PFOA are relatively less and localized compared to those of PFOS. No sex or length/weight associated variations in concentrations of fluorochemicals were observed in sea gulls or cormorants. This is similar to those observed for marine mammals and birds from several locations (Kannan et al., 2001a,b). These results suggest that accumulation of fluorochemicals in biota is different from those observed for lipophilic pollutants such as PCBs. The accumulation features of fluorochemicals are similar to those of contaminants like tributyltin, which bind to proteins (Kannan et al., 1997). PFOS-based fluorochemicals have been used in a wide variety of products such as textiles including carpets, paper products to impart oil, soil and water resistance and in fire-fighting foams. PFOS is sparingly volatile (3:3 104 Pa; USEPA, 1999) and moderately water soluble (1080 mg/l; USEPA, 2000). Ubiquitous environmental distribution of PFOS in biota suggests that neutral, precursor compounds such as n-ethyl perfluorooctanesulfonamidoethanol (n-EtFOSEA; C8 F17 SO2 N(CH2 CH3 )CH2 CH2 OH) and n-methyl perfluorooctanesulfonamidoethanol (n-MeFOSEA; C8 F17 SO2 N(CH3 )CH2 CH2 OH) have the potential to volatilize and yield PFOS upon metabolism (Giesy and Kannan, 2002). The polar functional group of these neutral molecules can be metabolized in animals to yield PFOS (USEPA, 2000). Occurrence of volatile precursors of PFOS has been shown in air collected over the Great Lakes (Martin et al., 2002). PFHS is an impurity in PFOS-based technical formulations. PFOA is not only an impurity in PFOS mixtures, but also is used as a plasticizer, corrosion inhibitor and anti-wetting agent. PFOA has been identified in areas where aqueous film fire-fighting foams have been used to control fuel fires (Moody and Field, 2000). FOSA is a metabolite of nethyl FOSA (Sulfluramid), which is used as an insecticide to control roaches, ants and termites (Manning et al., 1991). FOSA is also found as an impurity in technical mixtures of fluoroorganic compounds. 3.2. Birds from Korea
Fig. 3. Relationship between PFOS and FOSA concentrations in livers of cormorants from the Sagami River, Kanagawa Prefecture, Japan.
PFOS was found in 95% of the birds analyzed from Korea at concentrations greater than 10 ng/g, wet weight (Table 2). The greatest concentration of 500 ng PFOS/g, wet weight, was found in the liver of a black-headed gull. Mean concentrations of PFOS were greater than 100 ng/g, wet weight, in bar-tailed godwit, black-headed gull and black-tailed gull. PFHS, FOSA and PFOA
K. Kannan et al. / Chemosphere 49 (2002) 225–231
were not detected in the livers of birds from Korea at LOQs of 36, 38 and 36 ng/g, wet weight, respectively. Although most of the species analyzed from Korea were migratory birds, comparison of residue concentrations between migratory and resident birds can be confounded by several factors including age, sex, time spent in wintering grounds, feeding habits, breeding location, etc. Sampling was not designed to compare the residue concentrations between migratory and resident birds and therefore such interpretation has not been made. In general, results of this study suggest widespread occurrence of PFOS in birds collected from Japan and Korea. The measured concentrations are within the range of values reported for birds from the United States and Europe. PFOA and PFHS are found sporadically in livers of birds. FOSA is found in Japanese birds from certain regions. There was no significant correlation between the concentrations of PFOS and FOSA in livers of cormorants from the Sagami River in Japan. Concentrations of FOSA were 3-fold less than those of PFOS in cormorant livers from Japan.
Acknowledgements This study (analysis) was supported by 3M Company, St. Paul, Minnesota.
References Giesy, J.P., Kannan, K., 2001. Global distribution of perfluorooctane sulfornate and related perfluorinated compounds in wildlife. Environ. Sci. Technol. 35, 1339–1342. Giesy, J.P., Kannan, K., 2002. Perfluorochemical surfactants in the environment. Environ. Sci. Technol. 36, 147A–152A. Hansen, K.J., Clemen, L.A., Ellefson, M.E., Johnson, H.O., 2001. Compound-specific, quantitative characterization of organic fluorochemicals in biological matrices. Environ. Sci. Technol. 35, 766–770. Kannan, K., Senthilkumar, K., Loganathan, B.G., Takahashi, S., Odell, D.K., Tanabe, S., 1997. Elevated accumulation
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of tributyltin and its breakdown products in bottlenose dolphins (Tursiops truncatus) found stranded along the U.S. Atlantic and Gulf coasts. Environ. Sci. Technol. 31, 296– 301. Kannan, K., Hansen, S.P., Franson, C.J., Bowerman, W.W., Hansen, K.J., Jones, P.D., Giesy, J.P., 2001a. Perfluorooctane sulfonate in fish-eating water birds including bald eagles and albatrosses. Environ. Sci. Technol. 35, 3065– 3070. Kannan, K., Koistinen, J., Beckmen, K., Evans, T., Gorzelany, J., Hansen, K.J., Jones, P.D., Giesy, J.P., 2001b. Accumulation of perfluorooctane sulfonate in marine mammals. Environ. Sci. Technol. 35, 1593–1598. Kannan, K., Hansen, K.J., Wade, T.L., Giesy, J.P., 2002a. Perfluorooctane sulfonate in oysters, Crassostrea virginica, from the Gulf of Mexico and Chesapeake Bay, USA. Arch. Environ. Contam. Toxicol. 42, 313–318. Kannan, K., Corsolini, S., Falandysz, J., Oehme, G., Focardi, S., Giesy, J.P., 2002c. Perfluorooctane sulfonate and related fluorinated hydrocarbons in marine mammals, fish and birds from coasts of the Baltic and the Mediterranean Seas. Environ. Sci. Technol. 36. Kannan, K., Newsted, J., Halbrook, R.S., Giesy, J.P., 2002b. Perfluorooctane sulfonate and related fluorinated hydrocarbons in mink and river otters from the United States. Environ. Sci. Technol. 36, 2566–2571. Manning, R.O., Bruckner, J.V., Mispagel, M.E., Bowen, J.M., 1991. Metabolism and disposition of sulfluramid, a unique polyfluorinated insecticide, in the rat. Drug Metab. Dis. 19, 205–211. Martin, J.W., Muir, D.C.G., Kwan, W.C., Moody, C.A., Ellis, D.A., Solomon, K.A., Mabury, S.A., 2002. Collection of airborne fluorinated organics and analysis by gas chromatography-chemical ionization mass spectrometry. Anal. Chem. 74, 584–590. Moody, C.A., Field, J.A., 2000. Perfluorinated surfactants and the environmental implications of their use in fire-fighting foams. Environ. Sci. Technol. 34, 3864–3870. US Environmental Protection Agency, 1999. Determination of the vapor pressure of PFOS using the spinning rotor gauge method. AR226-0048; Wildlife International Limited, Easton, MD. US Environmental Protection Agency, 2000. Sulfonated fluorochemicals in the environment: sources, dispersion, fate and effects. AR226-0620; 3M Company, St. Paul, MN.
Concentrations of perfluorinated acids in livers of birds from Japan and Korea Kurunthachalam Kannan a,*, Jae-Won Choi b, Naomasa Iseki c, Kurunthachalam Senthilkumar c, Dong Hoon Kim a, Shigeki Masunaga c, John P. Giesy a a
c
National Food Safety and Toxicology Center, Department of Zoology, Institute for Environmental Toxicology, Michigan State University, East Lansing, MI 48824, USA b National Institute for Environmental Studies, Onogawa 16-1, Tsukuba, Ibaraki 305-8506, Japan Graduate School of Environment and Information Sciences, Yokohama National University, 79-7 Tokiwadai, Hodogaya-ku, Yokohama 240-8501, Japan Received 1 March 2002; received in revised form 23 May 2002; accepted 30 May 2002
Abstract Livers of birds collected from Japan and Korea (n ¼ 83) were analyzed to determine the concentrations of perfluorooctanesulfonate (PFOS), perfluorooctanesulfonamide (FOSA), perfluorooctanoic acid (PFOA) and perfluorohexanesulfonate (PFHS). PFOS was found in the livers of 95% of the birds analyzed at concentrations greater than the limit of quantitation (LOQ) of 10 ng/g, wet weight. The greatest concentration of PFOS of 650 ng/g, wet weight, was found in the liver of a common cormorant from the Sagami River in Kanagawa Prefecture. Concentrations of PFOS in bird livers from Japan and Korea were within the ranges of values reported for those from the United States and certain European countries. PFOA and PFHS were found in 5–10% of the samples analyzed. The greatest concentrations of PFOA and PFHS in bird livers were 21 and 34 ng/g, wet weight, respectively. FOSA was found in all the samples (n ¼ 10) of cormorants collected from the Sagami River in Japan. The greatest concentration of FOSA in cormorant liver was 215 ng/g, wet weight. There was no significant correlation between the concentrations of PFOS and FOSA in cormorants collected from the Sagami River. These results suggested that the distribution of FOSA is localized. No ageor gender-specific differences in fluorochemical concentrations could be discerned in birds. Ó 2002 Elsevier Science Ltd. All rights reserved. Keywords: PFOS; Perfluorinated compounds; Birds; Asia
1. Introduction Concern about fluorinated organic compounds, particularly perfluorinated (fully fluorinated) compounds, is increasing. Perfluorooctanesulfonate (PFOS) and related perfluorinated acids are shown to be globally
*
Corresponding author. Tel.: +1-517-432-6321; fax: +1-517432-2310. E-mail address: [email protected] (K. Kannan).
distributed, environmentally persistent and bioaccumulative (Giesy and Kannan, 2001, 2002). PFOS, perfluorooctanesulfonamide (FOSA), perfluorooctanoic acid (PFOA) and perfluorohexanesulfonate (PFHS) have been reported to occur in blood sera of humans (Hansen et al., 2001). Similarly, PFOS, FOSA, PFOA and PFHS have also been identified in tissues of wildlife from various parts of the world (Giesy and Kannan, 2001; Kannan et al., 2001a,b, 2002a,b,c). Earlier studies have reported the occurrence of these compounds in wildlife collected from North America and certain European
0045-6535/02/$ - see front matter Ó 2002 Elsevier Science Ltd. All rights reserved. PII: S 0 0 4 5 - 6 5 3 5 ( 0 2 ) 0 0 3 0 4 - 1
226
K. Kannan et al. / Chemosphere 49 (2002) 225–231
countries. In this study, concentrations PFOS, FOSA, PFOA and PFHS in livers of birds collected from Japan and Korea are reported to provide information on the extent of contamination in these industrialized Asian countries.
2. Materials and methods Liver samples from forty individuals belonging to six species of birds were collected from Japan during June 1998 to February 1999 except gray herons, which were collected during 1997–1998. Samples of Japanese birds were from Rishiri Island (Hokkaido), Haneda airport (Tokyo), Atsugi city and the Sagami River in Kanagawa Prefecture and Gyotoku wild bird observatory in Chiba, Tokyo. Kanagawa Prefecture is located just south of highly urbanized areas such as Tokyo and Yokohama is one of the major industrial cities located in Kanagawa Prefecture (Fig. 1). Sea gulls were collected from Rishiri Island and common cormorants were collected from the Sagami River. Gyotoku wild bird observatory is a rehabilitation center for birds. Forty three individuals belonging to 10 species of birds were collected from the Nakdong River estuary in Korea from 1993 to 1994, except two black-tailed gulls, which were collected in 1997. The Nakdong River estuary is located along the east coast of Korea near Pusan (Fig. 1). Several industrial complexes including chemical manufacturers, oil refineries and heavy industries are located near this estuary. The estuary has been designated as a natural conservation area to protect wildlife. Among several species of birds analyzed from Korea,
the black-tailed gull is a resident, piscivorous bird. Black-tailed gulls were also collected in Nan Do, which is located on the west coast of Korea and Hong Do is located south of the Nakdong estuary. Most other species studied are migratory birds that winter in the Nakdong River estuary. Bird species analyzed in this study are listed in Tables 1 and 2. Prior to dissection, body weight, length and sex of birds were assessed. Livers were stored at )20 °C until analysis. Concentrations of PFOS, FOSA, PFOA and PFHS in liver were measured using high performance liquid chromatography (HPLC) with electrospray tandem mass spectrometry (Hansen et al., 2001). A liver homogenate of 1 g of liver in 5 ml of high purity Milli-Q water was prepared. One ml of the homogenate, 1 ml of 0.5 M tetrabutyl ammonium hydrogen sulfate solution (adjusted to pH 10), and 2 ml of 0.25 M sodium carbonate buffer were added to a 15-ml polypropylene tube for extraction. After thorough mixing, 5 ml of methyl-tertbutyl ether (MTBE) was added, and the mixture was shaken for 20 min. The organic and aqueous layers were separated by centrifugation, and an exact volume of MTBE (4 ml) was removed from the solution. The aqueous mixture was rinsed with MTBE and separated twice. The solvent was allowed to evaporate under nitrogen before being reconstituted in 0.5 or 1 ml of methanol. The sample was vortexed for 30 s and passed through a 0.2 lm nylon mesh filter into an autosampler vial. Analyte separation was performed using a HewlettPackard HP1100 liquid chromatograph modified with low dead-volume internal tubing. Ten ll of extract was injected onto a 50 2 mm (5 lm) Keystone Betasilâ C18
Fig. 1. Map of Japan and Korea showing sampling locations.
Table 1 Concentrations (ng/g, wet weight) of perfluorinated acids in livers of birds from Japan Date of collection
Location
Sex
Body weight (g)
Length (cm)
PFOS
FOSA
PFOA
PFHS
Sea gull (n ¼ 14) Larus crassirostris
Jun-98
Rishiri Island, Hokkaido
M
658 (586–706)
487 (468–521)
53 (23–89)
<75
<19
<7.5–34a
Sea gull (n ¼ 7) L. crassirostris
Jun-98
Rishiri Island, Hokkaido
F
504 (470–570)
468 (435–510)
40 (<19–53)
<75
<19
<7.5
Sea gull (n ¼ 1) L. crassirostris
Sep-98
Haneda Airport, Tokyo
NA
789
535
230
<75
<19
<7.5
Spot-billed duck (n ¼ 1) Anas poecilorhyncha
Jun-98
Gyotoku bird observatory
F
722
500
160
<75
<19
<7.5
Black-headed gull (n ¼ 1) Larus ridibundus
Jun-98
Gyotoku bird observatory
M
196
401
<19
<75
21
<7.5
Black-eared kite (n ¼ 1) Milvus lineatus
Jul-99
Atsugi city, Kanagawa Prefecture
NA
860
NA
450
<75
21
34
Black-eared kite (n ¼ 1) M. lineatus
Sep-99
Haneda Airport, Tokyo
M
966
605
180
<75
<19
<7.5
Gray heron (n ¼ 2) Ardea cinerea
Oct-97 and Jul-98
Gyotoku bird observatory
M
395 (368–422)
738 (495–980)
50 (49–52)
<75
<19
<7.5
Common cormorant (n ¼ 2) Phalacrocorax carbo
Feb-99
Sagami River, Kanagawa Prefecture
M
2144 (2039–2249)
820 (810–830)
390 (330–450)
115 (100–130)
<19
<7.5
Common cormorant (n ¼ 8) P. carbo
Feb-99
Sagami River, Kanagawa Prefecture
F
1873 (1619–2130)
762 (740–816)
385 (170–650)
162 (110–215)
<19
<7.5–10a
K. Kannan et al. / Chemosphere 49 (2002) 225–231
Species
All cormorants are adults except one female, which was a juvenile. a Only one sample above detection limit.
227
228
Table 2 Concentrations (ng/g, wet weight) of PFOS in livers of birds collected from Koreaa Scientific name
N
Sampling date
Sex
Length (cm)
Weight (g)
Feeding habits
PFOS
Bar-tailed godwit
Limosa lapponica
3
Mar-93
F
46 (42–51)
255 (210–300)
Crustaceans, insects, small fish
148 (22–310)
Black-headed gull
L. ridibundus
1 4
Feb-94 Dec-92–Jan-94
F M
38 43 (42–45)
250 304 (240–340)
Omnivorous Omnivorous
292 296 (148–500)
Black-tailed gull
Larus crassirostris
7
May–Dec-93
NA
NA
533 (480–600)
Omnivorous
112 (36–215)
Black-tailed gull Black-tailed gullb
L. crassirostris L. crassirostris
1 1
1997 1997
NA NA
NA NA
NA NA
Omnivorous Omnivorous
74 71
Common gull
Larus canus
1 2
Nov-93 Dec-92–Feb-94
F M
55 40–49
550 250–550
Omnivorous Omnivorous
28 29–63
Black-necked grebe
Podiceps nigricollis
1
Mar-93
F
35
400
Insects, small fish, shrimp
10
Common tern
Sterna hirundo
1 1
May-93 May-93
F M
35 30.6
123 104
Fishes Fishes
11.2 <10
Great knot
Calidris tenuirostris
1
Aug-93
M
27.5
138
Invertebrates, shellfish, crustaceans
13.5
Greenshank
Tringa nebularia
1 2
Nov-93 Aug–Nov-93
F M
19.9 24–33
230 143–190
Invertebrates, shellfish, crustaceans Invertebrates, shellfish, crustaceans
13.8 61–112
Herring gull
Larus argentatus
10
Feb–Dec-93
6M, 1F, 3NA
1000–1400
62–71
Omnivorous
49.6 (<10–116)
Sanderling
Crocethia alba
2
May-94
F
20–22
60–62
Invertebrates, crustaceans, shellfish
21–112
Little egret
Egretta garzetta
4
NA
NA
NA
NA
Fish, frogs
24.8 (19–30)
a
FOSA, PFOA and PFHS were not detected in any of the birds from Korea at a quantitation limit of 38, 36 and 36 ng/g, wet weight. a From Hong Do. b From Nan Do.
K. Kannan et al. / Chemosphere 49 (2002) 225–231
Species
K. Kannan et al. / Chemosphere 49 (2002) 225–231
column with a 2 mM ammonium acetate/methanol mobile phase starting at 10% methanol at a flow rate of 300 ll/min, to 100% methanol at 11.5 min before reverting to original conditions at 13 min. Column temperature was maintained at 25 °C. For quantitative determination, the HPLC system was interfaced to a Micromassâ (Beverly, MA) Quattro II atmospheric pressure ionization tandem mass spectrometer operated in the electrospray negative mode. Instrumental parameters were optimized to transmit the [M-K] ion for all analytes before fragmentation to one or more product ions. When possible, multiple daughter ions were monitored, but quantitation was based on a single product ion. In all cases, the capillary was held between 1.6 and 3.2 kV. Primary and product ions monitored for PFOS, PFOA, FOSA and PFHS determinations were 499 > 99, 413 > 169, 498 > 78 and 399 > 80, respectively. Product ions 99, 169, 78 and 80 correspond to FSO 3 , C3 F7 , SO2 N , SO3 , respectively. Recoveries of 250 ng of PFOS, FOSA, PFOA and PFHS spiked into livers of sea gulls were 98%, 44%, 100% and 56% respectively, while recoveries from samples of cormorants were 230%, 107%, 95% and 59%, respectively, and those from black-tailed gulls were 121%, 63%, 136% and 25%, respectively. Causes of great recoveries of PFOS (230%) spiked to cormorant livers are unknown. Recoveries of PFHS from bird livers were low (25–59%). Relative standard deviations of replicate analyses of spiked tis-
229
sues were less than 15%. Concentrations of fluorochemicals were not corrected for recoveries. For the estimation of the limit of quantitation (LOQ), the tissue samples were compared to an unextracted standard calibration curve. For instance, if 5 ng/ml standard is the lowest acceptable standard, and sample had been diluted by a factor of 7, the LOQ is reported as 35 ng/ml. LOQs for fluorochemicals varied from 10 to 75 ng/g, wet weight. Representative HPLC-ESMSMS chromatograms of liver extracts of a black-eared kite that contained 450, 21 and 34 ng/g, wet weight, PFOS, PFOA and PFHS, respectively, is presented (Fig. 2).
3. Results and discussion 3.1. Birds from Japan PFOS was found at concentrations greater than the LOQ of 19 ng/g, wet weight, in 38 of the 40 bird livers analyzed from Japan (Table 1). The greatest concentration of PFOS of 650 ng/g, wet weight, was found in the liver of a common cormorant from the Sagami River. Concentrations of PFOS in birds from Japan were within the range of values found in the livers of birds collected across the United States (Kannan et al., 2001a). However, the greatest concentration of 650 ng PFOS/g, wet weight, found in cormorant liver from the
Fig. 2. Representative HPLC-ESMSMS chromatograms of PFOS (ions 499 > 80 and 499 > 99) and PFOA (ions 413 > 169) and PFHS (ions 399 > 99) in sample extracts of black-eared kite liver. Concentrations of PFOS, PFOA and PFHS in this sample were 451, 21 and 34 ng/g, wet weight, respectively.
230
K. Kannan et al. / Chemosphere 49 (2002) 225–231
Sagami River was 3-fold less than the highest concentrations of 1780 ng/g, wet weight, found in Brandt’s cormorants from San Diego, United States (Kannan et al., 2001a). Mean concentrations of PFOS in livers of cormorants from Japan were 6-fold greater than those found in livers of cormorant from Sardinia Island in Italy (Kannan et al., 2002b). Similarly, concentrations of PFOS in cormorants were 5–10-fold greater than those found in white-tailed sea eagles from eastern Germany (Kannan et al., 2002b). Mean concentrations of PFOS in cormorants were greater than those found in other species of birds with the exception of a black-eared kite (450 ng/g, wet weight) collected from Atsugi city in Kanagawa Prefecture. Great concentrations of PFOS in cormorants and blackeared kite collected from Kanagawa can be explained by heavy industrialization in this region. A sea gull collected from Haneda airport in Tokyo contained 5–6-fold greater concentrations of PFOS than those collected from remote areas such as Rishiri Island in Hokkaido. These results suggest that urbanized and industrialized areas are major sources of exposure of birds to PFOS. FOSA and PFHS were detected in 5–10% of the bird livers analyzed. While PFOS was not found in blackheaded gull at the LOQ of 19 ng/g, PFOA was detected in this individual at a concentration of 21 ng/g, wet weight. This indicates that fluorochemicals other than PFOS can be prevalent in some locations, although at lesser frequencies. PFHS was found in individual sea gulls, black-eared kite and common cormorants with a maximum concentration of 34 ng/g, wet weight. FOSA was found in all the livers of cormorants from the Sagami River. Concentrations of FOSA in cormorant livers were approximately 3-fold less than those of PFOS (Table 1). Concentrations of FOSA in cormorant livers were not significantly (p > 0:05; r2 ¼ 0:12) correlated with concentrations of PFOS (Fig. 3). Similarly, concentrations of FOSA in livers of cormorants were not correlated with concentrations of PFHS or PFOA. Concentrations of PFHS and PFOA in cormorants were generally less
than the LOQ. FOSA was not found in the livers of other species of birds analyzed at the LOQ of 75 ng/g, wet weight. These results suggest that the exposure of birds to FOSA is relatively less than to PFOS and that it is localized. Furthermore, these results indicate that the sources of FOSA are not directly associated with those of PFOS. Similarly, exposures of PFHS and PFOA are relatively less and localized compared to those of PFOS. No sex or length/weight associated variations in concentrations of fluorochemicals were observed in sea gulls or cormorants. This is similar to those observed for marine mammals and birds from several locations (Kannan et al., 2001a,b). These results suggest that accumulation of fluorochemicals in biota is different from those observed for lipophilic pollutants such as PCBs. The accumulation features of fluorochemicals are similar to those of contaminants like tributyltin, which bind to proteins (Kannan et al., 1997). PFOS-based fluorochemicals have been used in a wide variety of products such as textiles including carpets, paper products to impart oil, soil and water resistance and in fire-fighting foams. PFOS is sparingly volatile (3:3 104 Pa; USEPA, 1999) and moderately water soluble (1080 mg/l; USEPA, 2000). Ubiquitous environmental distribution of PFOS in biota suggests that neutral, precursor compounds such as n-ethyl perfluorooctanesulfonamidoethanol (n-EtFOSEA; C8 F17 SO2 N(CH2 CH3 )CH2 CH2 OH) and n-methyl perfluorooctanesulfonamidoethanol (n-MeFOSEA; C8 F17 SO2 N(CH3 )CH2 CH2 OH) have the potential to volatilize and yield PFOS upon metabolism (Giesy and Kannan, 2002). The polar functional group of these neutral molecules can be metabolized in animals to yield PFOS (USEPA, 2000). Occurrence of volatile precursors of PFOS has been shown in air collected over the Great Lakes (Martin et al., 2002). PFHS is an impurity in PFOS-based technical formulations. PFOA is not only an impurity in PFOS mixtures, but also is used as a plasticizer, corrosion inhibitor and anti-wetting agent. PFOA has been identified in areas where aqueous film fire-fighting foams have been used to control fuel fires (Moody and Field, 2000). FOSA is a metabolite of nethyl FOSA (Sulfluramid), which is used as an insecticide to control roaches, ants and termites (Manning et al., 1991). FOSA is also found as an impurity in technical mixtures of fluoroorganic compounds. 3.2. Birds from Korea
Fig. 3. Relationship between PFOS and FOSA concentrations in livers of cormorants from the Sagami River, Kanagawa Prefecture, Japan.
PFOS was found in 95% of the birds analyzed from Korea at concentrations greater than 10 ng/g, wet weight (Table 2). The greatest concentration of 500 ng PFOS/g, wet weight, was found in the liver of a black-headed gull. Mean concentrations of PFOS were greater than 100 ng/g, wet weight, in bar-tailed godwit, black-headed gull and black-tailed gull. PFHS, FOSA and PFOA
K. Kannan et al. / Chemosphere 49 (2002) 225–231
were not detected in the livers of birds from Korea at LOQs of 36, 38 and 36 ng/g, wet weight, respectively. Although most of the species analyzed from Korea were migratory birds, comparison of residue concentrations between migratory and resident birds can be confounded by several factors including age, sex, time spent in wintering grounds, feeding habits, breeding location, etc. Sampling was not designed to compare the residue concentrations between migratory and resident birds and therefore such interpretation has not been made. In general, results of this study suggest widespread occurrence of PFOS in birds collected from Japan and Korea. The measured concentrations are within the range of values reported for birds from the United States and Europe. PFOA and PFHS are found sporadically in livers of birds. FOSA is found in Japanese birds from certain regions. There was no significant correlation between the concentrations of PFOS and FOSA in livers of cormorants from the Sagami River in Japan. Concentrations of FOSA were 3-fold less than those of PFOS in cormorant livers from Japan.
Acknowledgements This study (analysis) was supported by 3M Company, St. Paul, Minnesota.
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