Water Air Soil Pollut (2010) 207:391401 DOI 10.

1007/s11270-009-0144-3

Mercury and Methylmercury in Freshwater Fish and Sediments in South Korea Using Newly Adopted Purge and Trap GC-MS Detection Method
Jae-Sung Park & Jung-Sub Lee & Gun-Bae Kim & Jun-Seok Cha & Sun Kyoung Shin & Hak-Gu Kang & Eun-Jin Hong & Gi-Taeg Chung & Young-Hee Kim

Received: 22 September 2008 / Accepted: 7 July 2009 / Published online: 26 July 2009 # Springer Science + Business Media B.V. 2009

Abstract The use of purge and trap gas chromatography mass spectrometry (GC-MS) technique for the determination of methylmercury in biological and sediment samples was described. The GC-MS detection system was combined with the dithizone extraction method for biological samples and the distillation method for sediment samples to alleviate matrix interference problems. The method was validated by analysis of CRMs such as SRM 966 (human blood), BCR 463 (tuna fish), IAEA 407 (fish), ERM CC580 (estuarine sediment), and IAEA 405 (sediment). The performance of the purge and trap GC-MS method was also tested on field samples of freshwater fish and sediment. The results were compared with those of the GC-ECD and the GCCVAFS, which were used widely for methylmercury analysis. Additionally, total mercury and methylmercury levels in freshwater fish and sediments from various reservoirs and streams in Korea were measured to understand mercury contamination status in Korean peninsula. Methylmercury concentrations in freshwater fish were found to correlate with body weight, diet habit,

and food availability. In sediment, total mercury concentrations correlated with methylmercury concentrations and organic matter such as %C and %S. However, no significant relationships between methylmercury and sediment organic matter have been found. Keywords Methylmercury . Mercury . Dithizone extraction . Distillation technique . Purge and Trap GC-MS . Sediment . Freshwater fish

1 Introduction Among many pollutants, methylmercury has been concerned as the most harmful mercury species due to its high toxicity, high solubility in lipids, which increases the potential for biological uptake and bioconcentration, and common occurrences in the environment (Horvat 1996). Some inorganic mercury released into the aquatic system is known to be converted to methylmercury via biotic production by sulfate-reducing bacteria in sediment (Mason et al. 1999; Mason and Lawrence 1999). These methylmercury are uptaken by benthic organisms and accumulated in fish through the food chain, which is the major pathway for human exposure to methylmercury (Boening 2000; Henny et al. 2002; and Mergler et al. 2007). Generally, the concentration of total mercury is used as a biomarker for evaluation of the exposure level of methylmercury in blood because more than

J.-S. Park : J.-S. Lee : G.-B. Kim : J.-S. Cha : S. K. Shin : H.-G. Kang : E.-J. Hong : G.-T. Chung : Y.-H. Kim (*) Chemicals Behavior Research Division, National Institute of Environmental Research, Environmental Research Complex, Kyungseo-dong, Seo-gu, Incheon, Republic of Korea 404-708 e-mail: heek89@korea.kr

392

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80% of mercury in human blood exists as methylmercury forms and determination of total mercury is simple compared to that of methylmercury (Akagi and Naganuma 2000; Rice 2004). However, recent studies showed that the mercury speciation in human blood is needed to evaluate the sources of exposure, especially at low exposure levels. Lindberg et al. (2004) revealed that the proportions of methylmercury to total mercury in blood of non-fish-eating people are relatively lower than those of fish-eating people and inorganic mercury in blood has a significant association with the number of dental amalgam fillings. Thus, there is a growing need for a more simplified and popularized analytical method for the determination of methylmercury in environmental species (Vidler et al. 2007). For methylmercury analysis, a succession of analytical stages is required. The main steps to speciate mercury are digestion, clean-up, separation, and mercury-specific detection. Coupled techniques including separation by gas chromatography (GC) or liquid chromatography (LC) and detection by electron capture dissociation (ECD), atomic absorption spectroscopy (AAS), atomic fluorescence spectroscopy (AFS), and inductively coupled plasmamass spectroscopy (ICPMS) have been widely used (West 1966; Bloom 1989; Blanco et al. 2000; Ullrich et al. 2001; Ignacio et al. 2000). For methylmercury clean-up process, a solvent extraction technique and a distillation technique are commonly used (West 1966; Horvat et al. 1993). However, the extraction of methylmercury from biological samples, especially in blood has been a difficult task because of severe matrix interferences. The solvent extraction method using toluene or dichloromethane generally gave low extraction efficiencies in certain matrix (Akagi and Nishimura 1991). Additionally, the distillation technique has a drawback such as codistillation of large amounts of volatile compounds in blood and these volatile compounds transferred to the distillate can interfere with the ethylation reaction and/or deposit on the GC column leading to inaccurate determinations (Liang et al. 2000; Baxter et al 2007). Thus, in this study, it was considered appropriate to develop the accurate and simplified methylmercury analytical method using popularized analytical instrument such as purge and trap GC-MS. The GC-MS detection system was combined with the dithizone extraction method and clean-up process by Na2S for biological samples since the dithizone extraction process showed improved extraction efficiencies by

the complexation between dithizone and methylmercury (Akagi and Nishimura 1991; Logar et al. 2002). However, for the analysis of sediment, the distillation technique with H2SO4/KCl was preferred due to easy preparation and no use of harmful organic solvents. In this study, the distillation conditions were controlled with great cautions to prevent the production of methylmercury artifacts by over-distillation (Horvat et al 1993; Quevauviller et al. 1997; Hammerschmidt and Fitzgerald 2001). The performance of the purge and trap GC-MS method was validated by analyzing CRMs. Additionally, total mercury and methylmercury levels in freshwater fish and sediments from various reservoirs and streams in Korea were measured to understand mercury contamination status in Korean peninsula and the analytical results were compared with those of the GC-ECD and the GC-cold vapor atomic fluorescence spectrometry (CVAFS), respectively.

2 Experimental Section 2.1 Sample Collection and Preparation From June to September 2006, 57 freshwater fish samples were collected from reservoirs, lakes, and streams in Korea (Fig. 1). The water characteristics of the sites are various with chlorophyll a concentrations and chemical oxygen demand ranging 1.115.6 mg/m3 and 1.87.2 mg/L, respectively. The filet of fish samples were cut into small pieces with dissection scissors and homogenized to a pastry state. The samples were kept frozen until further analysis. Eighty-one sediment samples were also sampled from June to September 2007 by using a Ponar grab sampler (Fig. 1). The sediment were categorized into groups by sampling sites as lake, reservoirs, river 1 (with biochemical oxygen demand (BOD) lower than 3), river 2 (with BOD higher than 3), urban stream, and plant effluents. All the samples were kept frozen until further analysis. 2.2 Experimental Materials and Apparatus All reagents used were of ACS grade and all water was used as doubly distilled and de-ionized water obtained from Barnsted UC/A56220-8 (IA, USA). Methylmercury standard stock solution (1 mgmL1) was prepared by dissolving the appropriate amount of CH3HgCl (Aldrich, MO, USA) in toluene. Purified

Water Air Soil Pollut (2010) 207:391401 Fig. 1 Sampling sites of sediments and freshwater fish in Korean peninsula

393

S-River S-Lake & Reservoir S-Urban stream S-Plant effluent S Freshwater Fish

0.02% dithizone solution was prepared by dissolving 0.011 g of diphenylthiocarbazone in 100 mL toluene. Alkaline sodium sulfide stock solution was prepared by dissolving 0.15 g of Na2S9H2O in 10 mL of distilled water. At each use, 0.1 mL of stock solution was diluted with 50 mL of 0.1 N NaOH and 50 mL of ethanol. Walpoles buffer was prepared by mixing 200 mL of 1 M CH3COONa and about 200 mL of 1 N HCl to adjust to pH 3.0. Sodium acetate buffer (0.2 M) was prepared by dissolving 1.64 g of CH3COONa in distilled water and added with acetic acid to adjust pH at 4.9. Ethylating reagent, 2% sodium tetraethylborate, was prepared by dissolving with 0.2 g of sodium tetraethylborate [NaB(C2H5)4] powder in 10 mL of 1% w/v KOH solution and was kept in ice and darkness after preparation and throughout the analysis. For purge and trap GC-MS method, the volatile derivatized ethylmethylmercury were concentrated and injected using TekmarDohrmann purge-andtrap (Mason, OH, USA) with a Tenax A trap (Suppleco, MO, USA) as adsorbent trap. The sample was purged with helium at 40 mLmin1 for 15 min at 40C and followed by desorption at 200C for 3 min.

Chromatographic analysis was performed with Agilent 6890N GC (CA, USA) equipped with Agilent 5973N MS operating in selected ion monitoring (SIM) mode. The DB-5MS capillary column (5% phenyl95% dimethylpolysiloxane; 30 m0.5 mm I.D., 0.25 m) was used with helium as carrier gas at a flow rate of 1 mLmin1. The column temperature was programmed as follows: 40C for 4 min, increasing to 280C at 15Cmin1 then holding for 5 min. The injection port and detector were operated at 220C and 230C, respectively. 2.3 Determination of Methylmercury in Biological Samples Using Dithizone Extraction Followed by the Purge and Trap GC-MS Method Details of dithizone extraction procedures for methylmercury analysis are given elsewhere (Akagi and Ikingura 1999). Approximately 0.51 g of biological sample and 10 mL of 1 N KOHethanol solution were placed in a 40-mL screw-capped conical centrifuge tube and heated at 100C for 1 h. After cooling to room temperature, 10 mL of 1 N HCl was added followed by washing with 5 mL of n-hexane,

394 Fig. 2 GC chromatogram obtained from CRM IAEA 407 sample by the purge and trap GC-MS method

Water Air Soil Pollut (2010) 207:391401

and then, 2 mL of 20% EDTA4Na solution was added into the extracted aqueous phase to mask other metal ions contained in the samples. Five milliliters of purified 0.01% dithizone-toluene was added and the aqueous phase was discarded. The remaining excess dithizone in toluene phase was removed by washing with 5 mL of 1 N NaOH. A fixed volume of the toluene phase (7 mL) was transferred into 10-mL centrifuged tube with glass stopper and 2 mL of Na2S solution was added to back-extract the methylmercury into aqueous phase. The solution was acidified with 35 drop of 1 N HCl and aerated with N2 at 50 mLmin1 for 3 min to expel the excess sulfide ions. Lastly, 0.1 mL (0.051 mL) of the aerated solution was added into 10 mL of distilled water and 5 mL of sodium acetate buffer in a 20-mL syringe, followed by adding 0.2 mL of sodium tetraethylborate solution. Blanks and standard solutions for a calibration curve were treated in a similar manner. The combined solution in the syringe was injected into the sparser connected on the purge and trap sampler. During MS detection, the following ions were monitored using SIM mode: m/z 202, 217, and 246 for CH3HgC2H5; m/z 202, 231, and 260 for Hg (C2H5)2. Between two consecutive analyses, the distilled water was analyzed in order to clean the

system and eliminate carryover effects. The calibration curve was evaluated in the range from 0.1 to 5 ng (as Hg) and obtained with a determination coefficient, r2 >0.995 and less than 7% of RSD of calibration factors. The method detection limit was defined as the concentration equivalent to three times standard deviation of concentrations of spiked methylmercury solutions and was found to be 0.1 ngg1 for biological samples. For quality control purpose, CRMs of IAEA 407 (IAEA, Vienna, Austria) and BCR 463 (ERM, Brussels, Belgium) for fish and standard reference material (SRM) 966 (NIST, MD, USA) for blood were analyzed. The commercially available blood samples were obtained from Centre de Toxicologie du Qubec (Qubec, Canada). 2.4 Determination of Methylmercury in Sediment Using Distillation Technique Followed by the Purge and Trap GC-MS Method Details of distillation protocols for methylmercury analysis are given elsewhere (Mason et al. 1999). Briefly, 0.51 g of solid samples was distilled with a 1.5 mL of 50% sulfuric acid and 0.6 mL of 20% KCl solution in 22.9 mL of distilled water to isolate methylmercury from constituents. One milliliter of

Table 1 Determination of methylmercury in blood CRM and commercially available blood materials Materials SRM 966 (n =5) M 0605 (n =3) M 0618 (n =3)
a

Certified value (ngg1) 16.41.4 7.1a (4.69.5) 26.3 (20.032.3)

a

Determined value (ngg1) 16.61.6 5.80.8 23.21.6

RSD (%) 4.9 3.3 6.4

Recovery (%) 93105 8693 7991

Data from the total mercury analysis and the materials were spiked with methylmercury

Water Air Soil Pollut (2010) 207:391401 Table 2 Determination of methylmercury in fish CRMs CRMs IAEA 407 (n =7) BCR 463 (n =7) Certified values (ngg1) 0.200.012 2.830.16 Determined values (ngg1) GC-MS GC-ECD GC-MS GC-ECD 0.190.016 0.200.022 2.890.26 2.760.32 RSD (%) 3.9 5.7 4.3 5.9

395

Recovery (%) 8595 92101 98108 91107

distillate was added into 10 mL of distilled water and 5 mL of sodium acetate buffer in a 20-mL syringe, followed by adding 0.2 mL of sodium tetraethylborate solution. The combined solution in the syringe was injected into the sparser connected on the purge and trap sampler. The conditions of separation and detection system were identical with those for biological samples. The method detection limit was found to be 0.05 ngg1 for sediment samples. For quality control purpose, CRMs of IAEA 405 (IAEA, Vienna, Austria), and BCR CC580 (ERM, Brussels, Belgium) were analyzed. The commonly used distillation/GC-CVAFS method (USEPA 2001) was used as a control method to compare the results obtained by the GC-MS method. 2.5 Ancillary Chemical Measurements for Sediment Analysis Weighed sediment samples were heated at 105C for 4 h and reweighed in order to calculate the water content. After determining the water content of the sediment, the samples were heated at 550C overnight. The samples were then reweighed and the percent organic matter in the sediment was determined by the difference as loss on ignition. The total sulfur contents were determined using PE 2400 series II analyzer (Perkin Elmer, USA). Total mercury for sediment samples were analyzed by EPA method 7473 (DMA-80, Milestone Srl) and CRM of MESS-3 (NRC, Canada) were analyzed for quality control. For
Table 3 Determination of methylmercury in sediment CRMs CRMs ERM CC 580 (n =7) IAEA 405 (n =7) Certified values (ngg1) 75.04.5 5.490.53

other metal analysis in sediment, the samples were digested using EPA method 3051 with HNO3/HCl and analyzed by ICP-OES (Optimer 5000DV, Perkin Elmer). ERA Soil CRM (ERA, USA) was used for quality control materials. 2.6 Statistical Analyses SPSS version 10.0 for Windows (SPSS Inc., Chicago, IL, USA) was used for all statistical analysis. The t test and ANOVA were used to compare blood mercury and methylmercury concentrations between groups. The significant level was set to p 0.05. Pearsons correlation analysis to determine statistical significance between total mercury and methylmercury concentrations was also undertaken. A multiple regression statistical analysis of the sediment dataset was used to access the primary correlations between total mercury, methylmercury, %C, %S, and other metals (Fe, Mn, Cu, Cr, Ni, Zn, and Pb).

3 Results and Discussion 3.1 Optimization and Validation of Purge and Trap GC-MS Method Due to polarity of methylmercury compound, adsorption processes of methylmercury occur on the stationary phase during the chromatographic analysis, causing peak broadening and ghost peaks (Caricchia

Determined values (ngg1) GC-MS GC-CVAFS GC-MS GC-CVAFS 74.51.0 73.11.1 5.270.28 5.410.31

RSD (%) 14.2 13.8 5.5 5.76

Recovery (%) 85108 83111 89102 90109

396 Table 4 Comparison of total mercury and methylmercury concentrations (ngg1) in freshwater fish Species Mandarin fish Korean piscivorous chub Skin carp Catfish Skygager Sharpbelly Northern snake head Largemouth bass Carssius cuvieri Crusian carp Common carp Leather carp Japanese dace No of samples 2 5 4 7 6 1 6 9 1 2 11 2 1

Water Air Soil Pollut (2010) 207:391401 T-Hg 413.157.8 357.975.7 220.490.3 216.1106.2 191.8117.6 153.4 136.562.4 116.658.8 151.8 59.93.0 49.234.4 35.116.7 183.16 MeHg 219.045.7 254.268.2 206.1159.9 140.882.3 175.7118.7 77.0 102.371.7 89.853.3 125.1 42.90.7 50.341.1 24.210.2 141.4 %MeHg 53 86 88 68 90 50 69 84 82 74 69 72 77

et al. 1997). Polar methylmercury compounds are needed to convert into nonpolar methylmercury compounds before chromatographic separation (Young and Josep 1995). In this study, sodium tetraethylborate, NaBEt4, was used for the derivatization of polar methylmercury compounds to nonpolar ethylated methylmercury compounds. After the derivatization reaction, the analytes are purged with helium at 40 mLmin1 to adsorb methylmercury on the trap. The most consistent sensitivity was obtained from 15 min of purging by monitoring the changes of analytical signals. The concentrated methylmercury in Tenax trap was introduced to GC-MS and was analyzed by using selected ion monitoring mode. In the SIM mode the ions of m/z 202, 217, and 246 were monitored for CH3HgC2H5 and m/z 202, 231, and 260 for Hg(C2H5)2. The spectrum in Fig. 2 is the GC chromatogram of CRM IAEA 407 sample, showing
Fig. 3 Correlations between methylmercury concentrations and fish weight of specific species
[MeHg] (ng/g) 500 400 300 200 100 0

CH3HgC2H5 and Hg(C2H5)2 peaks which are contributed from CH3Hg+ and Hg2+ in the sample. The accuracy of the purge and trap GC-MS method was evaluated by analysis of CRMs such as SRM 966 (human blood), BCR 463 (tuna fish), IAEA 407 (fish), ERM CC580 (estuarine sediment), and IAEA 405 (sediment). As shown in Table 1, obtained results of SRM 966 were 16.61.6 ngg1 (95% confidence interval with n =5), which were within the certified range and the average RSD was 4.9%. High and low concentrations of commercially available blood samples were also analyzed. Although not certified for their methylmercury concentrations, these material was expected to be available for methylmercury analysis because they were spiked with methylmercury. The analytical results of fish CRM and sediment CRM were shown in Tables 2 and 3, which were in good agreement with the certified values.

Snake head Skygager Korean piscivorius chub Common carp

500

1000 weight (g)

1500

2000

Water Air Soil Pollut (2010) 207:391401

250 200 [MeHg] (ng/g) 150 100 50 0 100 Bass 1 Bass 2

397

300 weight (g)

500

700

Fig. 4 Correlations between methylmercury concentrations and fish weight of Largemouth bass: Bass 1 from Ju-Nam reservoir and Bass 2 from Dam-Yang reservoir

Additionally, methylmercury concentrations of freshwater fish and sediment were investigated by the purge and trap GC-MS method and the analytical results were compared with those of the GC-ECD for freshwater fish and the GC-CVAFS for sediment. For freshwater fish analysis, methylmercury concentrations between two methods were significantly related with Persons coefficient of 0.88 (p <0.05) and no statistically significant differences were found by the paired t test (p =0.37> 0.05). Subsequently, for the sediment analysis, the average ratio of methylmercury concentrations between the methods ([MeHg]CVAFS/[MeHg]GC-MS) was 0.9 and its correlation coefficient was 0.79 (p <0.05). Paired t test results showed no statistically significant differences (p =0.22>0.05). Thus, the GC-MS method was used as an alternative for a commonly used the GCECD and GC-CVAFS method. 3.2 Analysis of Total Mercury and Methylmercury in Freshwater Fish For 57 freshwater fish analysis, total mercury concentrations were in the range of 20.4454 ngg1 (mean 175.1 ngg1) and methylmercury concentrations were in the range of 12.9424 ngg1 (mean 143.2 ngg1) (Table 4). The relatively high methylmercury concen-

trations (216 to 424 ngg1) were found in predatory species such as Korean piscivorous chub, while lower concentrations (12.9 to 59.9 ngg1) were found in polyphagia species such as common carp. Generally, methylmercury concentrations in fish are expected to be proportional to trophic level and its size while methylmercury bioaccumulation is a function of several factors such as uptake (diet) and elimination pathways (excretion, growth dilution) (Bloom 1992; Wagemann et al. 1997; Sveinsdottir and Mason 2005). It is interesting to note that Korean piscivorous chub showed statistically high methylmercury concentrations, while their body weight was much less than that of other species. Korean piscivorous chub are actually the top predator and long-lived fish with small body size. Thus, it is likely that Korean piscivorous chub can accumulate methylmercury over their life span with minimal growth dilution, resulting in high methylmercury body burden. Overall, while methylmercury concentrations increased as fish weight increased, different species showed different patterns and methylmercury concentration was significantly correlated with fish body weight (R =0.580.88, p < 0.05) except Largemouth bass (Fig. 3). Largemouth bass are carnivores and their food preference is crayfish, minnows, and frogs. Despite the small number of samples, the relationship between methylmercury concentration and body weight were divided into two groups. As seen in Fig. 4, the result clearly showed a distinct pattern that methylmercury body burden was much higher in Bass 2, compared to Bass 1, even though their body weights were comparable. Additionally, two groups were collected from different locations, i.e. Bass 1 from Ju-Nam reservoir and Bass 2 from Dam-Yang artificial reservoir. The average concentrations of water quality-related parameters from 2002 to 2006 were obtained from the data of water quality monitoring networks in Korea. As shown in Table 5, Ju-Nam reservoir showed more eutrophic characters with high

Table 5 Water quality of Ju-Nam and Dam-Yang reservoirs during 2002~2006 Reservoirs Temp (C) pH DO (mg/L) Conditions COD (mg/L) SS (mg/L) TN (mg/L) TP (mg/L) Chlorophyll A (mho/cm) (mg/m3) 21025 6415 7.60.5 2.50.4 11.54.0 2.00.6 1.280.24 1.650.71 0.070.02 21.78.0 0.040.01 6.04.0

Ju-Nam

171

7.60.2 8.50.6 7.50.3 9.20.8

Dam-Yang 132

398 Fig. 5 Comparison of total mercury, methylmercury, and %MeHg in sediments

[T-Hg] (ng/g)
500

Water Air Soil Pollut (2010) 207:391401

5 [T-Hg] [MeHg] %MeHg 10

400

200

100

0
Plant effluent (n=8) Urban Stream (n=5) River 2 Lake & River 1 (BOD<3) (BOD>3) Reservoir (n=23) (n=13) (n=23)

concentrations of total phosphorous, chlorophyll a, and chemical oxygen demand than Dam-Yang reservoir and these indicators are generally related with fish growth rates. Lower mercury level in fish from Ju-Nam reservoir might be related to dilution effects with faster fish growth and longer food chain due to easy food availability than Dam-Yang reservoir, even though it is difficult to conclude without comparison of mercury levels in water and sediment (Sveinsdottir and Mason 2005; Hutchson et al. 2008). 3.3 Analysis of Total Mercury and Methylmercury in Sediment Eighty-one sediment samples were collected from various sites in Korea to understand the impact of
Fig. 6 Plots of the correlation between total mercury and methylmercury concentrations in sediments
100

contaminated sediments on regional scale. Total mercury concentrations in sediment were in the range of 2.431,564.17 ngg1 and methylmercury concentrations were in the range of N.D.5.95 ngg1. Methylmercury were found below the detection limit of 0.05 ngg1 in 13 sediment samples. The proportion of methylmercury to total mercury in sediment samples (n =68) were 2.522.39%. Methylmercury concentrations and %MeHg in sediment are useful indicators reflecting the changes of mercury loading, bioaccessibility of inorganic mercury, and changes in bacterial activities. %MeHg is generally known to be less than 5% and the range in %MeHg observed in this study (0.863.27%) is similar to values reported by Stephenson et al. (2008) and Krabbenhoft et al. (2006).

10

[MeHg] (ng/g)

0.1

Plant effulent Urban stream River 1(BOD<3) River 2(BOD>3) Lake & Reservoir

0.01 1 10 100 1000 10000

[T-Hg] (ng/g)

%MeHg

300

[MeHg] (ng/g)

Water Air Soil Pollut (2010) 207:391401 Table 6 Correlation table for the sediment dataset; values listed are correlation coefficients obtained from a multivariate analysis Parameter T-Hg MeHg Organic content, % %S Cr Ni Zn Pb Fe NC no correlation Mn T-Hg 1 0.410 0.408 0.481 0.388 0.455 0.667 0.509 NC NC MeHg 0.410 1 0.283 NC NC NC NC NC 0.285 NC Organic content, % 0.408 0.283 1 0.667 0.504 0.622 0.587 0.648 NC NC

399 %S 0.481 NC 0.667 1 0.773 0.893 0.720 0.911 NC NC

However, there was also clear differences in %MeHg observed in different sampling sites. As shown in Fig. 5, the lake and reservoir sites had comparatively high methylmercury concentration (0.94 ngg1) and low total mercury concentrations (27.73 ngg1), whereas sediments from the effluent sites were relatively high in total mercury (433.4 ngg1) but only moderate in methylmercury (1.78 ngg1). These results are expressed as the high %MeHg of lake and reservoir sites (3.27%) compared to plant effluent sites (0.86%). Recent studies showed that atmospheric total gaseous mercury levels were around 2.634.37 ng/m3 in Korean Peninsula and were enhanced by longrange transport from East Asia (Hong et al. 2007; Choi et al. 2009). Sulfate ions, one of the limiting factors for methylation, were found to be the major contributors of the wet deposition in Korea, which was mainly contributed by acidic precipitation (Han et
Fig. 7 Heavy metal concentrations in catergorized sediment (mean1SD)
1800 1600 1400 1200 1000 800 600 400 200 0 Cr

al. 2008). Thus, methylation in sediment was possibly enhanced by newly uploaded mercury and high sulfate concentrations from atmospheric deposition. The concentrations of total mercury and methylmercury showed statistically significant correlation (RT-Hg-MeHg =0.41, p <0.05), which was a similar value (RT-Hg-MeHg =0.40) of previous study (Benoit et al. 2003). However, the relationship between ecosystem (categorized as plant effluent, urban stream, river and lake, and reservoir) was not included in this study (Fig. 6). To access the interrelationships between methylmercury and other chemical parameters in sediment, organic contents, sulfur contents, and other metals (Cu, Ni, Pb, Zn, Fe, and Mn) were examined and the correlations matrix was summarized in Table 6. While there was an indication of a correlation between total mercury and sediment organic content, it appeared to
Plant effluent Urban stream River 1 River 2 Lake & Reservoir

[Metal] (mg/kg)

Ni

Cu

Zn

Pb

Fe(x100)

Mn

400

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be nonlinear. The correlation coefficients, r of this study (RT-Hg-%C =0.48; RMeHg-%C =0.28) showed similar values with those (RT-Hg-%C =0.49; RMeHg-%C =N.C.) found by Mason et al. (Mason and Lawrence 1999). Generally, recent researches suggested that the organic carbon contents control mercury concentrations and organic matter complexation/adsorption is the most important phase controlling mercury distribution in surface sediments, except at elevated mercury concentration. At high mercury concentration sites, the effect by input sources is likely more important than sediment characteristics (Benoit et al. 1998; Mason and Lawrence 1999). Other metals concentrations of sediment samples are given in Fig. 7, showing that total metal concentrations of plant effluent were also relatively high compared to those of other sites except for the concentrations of Fe and Mn. Cr, Ni, Zn, Pb, and Hg are all co-correlated and all correlate with %S and % organic contents while methylmercury does not correlate significantly with either. These results implied that multielement anthropogenic sources are controlling the metal concentrations as well as total mercury to a large degree whereas methylmercury has different source inventories than other metals.

References
Akagi, H., & Ikingura, J. R. (1999). Methylmercury production and distribution in aquatic systems. Science of the Total Environment, 234, 109118. Akagi, H., & Naganuma, A. (2000). Human exposure to mercury and the accumulation of methylmercury that is associated with gold mining in the Amazon basin, Brazil. Journal of Health Science, 46(50), 323328. Akagi, H., & Nishimura, H. (1991). Advances in Mercury Toxicology. New York: Plenum Press. Baxter, D. C., Rodushkin, I., Engstrom, E., Klockare, D., & Waara, H. (2007). Methylmercury measurement in whole blood by isotope-dilution GC-ICPMS with 2 sample preparation methods. Clinical Chemistry, 53(1), 111116. Benoit, J. M., Gilmour, C. C., Mason, P. R., Riedel, G. S., Reidel, G. F., & Sullivan, K. A. (1998). Sources and cycling of mercury in the Patuxent estuary. Biogeochemistry, 40, 249265. Benoit, J. M., Gilmour, C. C., Heyes, A., Mason, P. R., & Miller, C. (2003). Geochemical and biological controls over methylmercury production and degradation in aquatic systems. In Y. Chai & O. C. Braids (Eds.), Biochemistry of environmental important trace elements (pp. 262297). Washington, DC: American Chemical Society. Blanco, R. M., Villanueva, M. T., Uria, J. E. S., & Sanz-Medel, A. (2000). Field sampling, preconcentration and determination of mercury species in river waters. Analytica Chimica Acta, 419, 137144. Bloom, N. S. (1989). Determination of picogram levels of methylmercury by aqueous phase ethylation, followed by cryogenic gas chromatography with cold vapor atomic fluorescence detection. Canadian Journal of Fisheries and Aquatic Sciences, 46, 11311140. Bloom, N. S. (1992). On the chemical form of mercury in edible fish and marine invertebrate tissue. Canadian Journal of Fisheries and Aquatic Sciences, 49, 10101017. Boening, D. W. (2000). Ecological effects, transport, and fate of mercury: A general review. Chemosphere, 40, 13351351. Caricchia, A. M., Minervini, G., Soldati, P., Chiavarini, S., Ubaldi, C., & Morabito, R. (1997). GC-ECD determination of methylmercury in sediment samples using a SPB-608 capillary column after alkaline digestion. Microchemical Journal, 55, 4455. Choi, E. M., Kim, S. H., Holsen, T. M., & Yi, S. M. (2009). Total gaseous concentrations in mercury in Seoul, Korea: Local sources compared to long-range transport from China and Japan. Environmental Pollution, 157(3), 816822. Hammerschmidt, C. R., & Fitzgerald, W. F. (2001). Formation of Artifact Methylmercury during extraction from a sediment reference material. Analytical Chemistry, 73, 59305936. Han, J. S., Hong, Y. D., Ahn, J. Y., Chung, I. R. (2008). Acid deposition monitoring and impact assessment. National Institute of Environmental Research Report, NIER NO. 2008-78-1028. Henny, C. J., Hill, E. F., Hoffman, D. J., Spalding, M. G., & Grove, R. A. (2002). Nineteenth Century mercury: Hazard to wading birds and cormorants of the Carson River, Nevada. Ecotoxicology, 11, 213231.

4 Conclusion This study showed that the newly adapted purge and trap GC-MS method provided a reliable measurement of methylmercury in biological and sediment samples and can be successfully used as an alternative method for commonly used GC-CVAFS or ECD detection method. The present work also described the current status of total mercury and methylmercury levels in freshwater fish and sediment from Korean peninsula. Even though, the current study is preliminary and cannot represent the mercury contamination status in Korea, it is worthy to evaluate the degree of methylmercury contaminations in sediment and freshwater fish since recent study showed the total mercury level of blood in Korean was much higher than those in other countries (investigated by Korea Ministry of Environment in 2005). Thus, much more in-depth and intensive mercury monitoring studies are required to examine important factors controlling methylmercury production in sediments and accumulation in fish and human.

Water Air Soil Pollut (2010) 207:391401 Hong, Y. D., Han, J. S., Lee, Y. K., Ahn, J. Y., Chung, I. R. (2007). Study on the atmospheric distribution characteristics and test method of mercury. National Institute of Environmental Research Report, NIER NO. 2007-81-997. Horvat, M. (1996). Mercury analysis and speciation in environment samples. In W. Beayens (Ed.), Global and regional mercury cycles: Sources (fluxes and mass balances, pp. 131). The Netherlands: Kluwer Academic. Horvat, M., Liang, L., & Bloom, N. S. (1993). Comparison of distillation with other current isolation methods for the determination of methylmercury compounds in low level environmental samples. Analytica Chimica Acta, 282, 153168. Hutchson, M. S., Smith, C. M., Wallace, G. T., Rose, J., Eddy, B., Sullivan, J., et al. (2008). Freshwater fish mercury concentrations in a regionally high mercury deposition area. Water, Air, and Soil Pollution, 191, 1531. Ignacio, J., Alone, G., & Sanz-mede, A. (2000). Comparison of different derivatization approach for mercury speciation in biological tissues by gas chromatography/inductively coupled plasma mass spectrometry. Journal of Mass Spectrometry, 35, 639646. Krabbenhoft, D., Engstrom, D., Gilmour, C., Harris, H., Hurley, J., & Mason, R. (2006). Monitoring and evaluation trends in sediment and water indicators. In R. Harris, et al. (Eds.), Ecosystem responses to mercury contamination (pp. 47 86). NY: CRC Press. Liang, L., Evens, C., Lazoff, S., Woods, J. S., Cernichiari, E., Horvat, M., et al. (2000). Determination of methyl mercury in whole blood by ethylation-GC-CVAFS after alkaline digestionsolvent extraction. Journal of Analytical Toxicology, 24, 328332. Lindberg, A., Bjrnberg, A., Vahter, M., & Berglund, M. (2004). Exposure to methylmercury in non-fish-eating people in Sweden. Environmental Research, 96, 2833. Logar, M., Horvat, M., Akagi, H., & Pihlar, B. (2002). Simultaneous determination of inorganic mercury and methylmercury compounds in natural waters. Analytical and Bioanalytical Chemistry, 374, 10151021. Mason, P. R., & Lawrence, A. L. (1999). Concentration, distribution, and bioavailability of mercury and methylmercury in sediments of Baltimore Harbor and Chesapeake Bay, Maryland, USA. Environmental Toxicology and Chemistry, 18(11), 24382447. Mason, R. P., Lawson, N. N., Lawrence, A. L., Leaner, J. J., Lee, J. G., & Sheu, G.-R. (1999). Mercury in the Chesapeake Bay. Marine Chemistry, 65, 7796.

401 Mergler, D., Anderson, H. A., Chan, L. H. M., Mahaffey, K. R., Murray, M., & Sakamoto, M. (2007). Methylmercury exposure and health effects in humans: A worldwide concern. Ambio, 36, 311. Quevauviller, Ph., Fortunati, G. U., Filippelli, M., & Muntau, H. (1997). The certification of the contents (mass fractions) of total mercury and methylmercury in estuarine sediment CRM 580. EUR 17658 EN. Rice, D. C. (2004). The US EPA reference dose fir methylmercury: Sources of uncertainty. Environmental Research, 95, 406413. Stephenson, M., Landing, M., Foe, C., Gill, G. A., & Coale, K. H. (2008). Transport, Cycling, and Fate of Mercury and Monomethyl Mercury in the San Francisco Delta and Tributaries: An Integrated Mass Balance Assessment Approach, CALFED Mercury Project Final Report. Sveinsdottir, A. Y., & Mason, R. P. (2005). Factors controlling mercury and. methylmercury concentrations in largemouth bass (Micropterus. salmoides) and other fish from Maryland reservoirs. Archives of Environmental Contamination and Toxicology, 49, 528545. Ullrich, S. M., Tanton, T. W., & Abdrashitova, S. W. (2001). Mercury in the aquatic environment: A review factors affecting methylation. Critical Reviews in Environmental Science and Technology, 31(3), 241293. USEPA (2001). EPA method 1630. Methylmercury in water by distillation, aqueous ethylation, purge and trap, and CVAFS. Vidler, D., Jenkins, R., Hall, J., & Harrington, C. (2007). The determination of methylmercury in biological samples by HPLC coupled to ICP-MS detection. Applied Organometallic Chemistry, 21, 303310. Wagemann, R. E., Trebacz, R., & Hunt, R. (1997). Percent methylmercury and organic mercury in tissues of marine mammals and fish by different experimental and calculation methods. Environmental Toxicology and Chemistry, 16(9), 18591866. West, G. (1966). Determination of methylmercury compounds in food stuffs. I. Methylmercury compounds in fish, identification and determination. Acta Chemica Scandinavica, 21, 21312137. Young, C., & Josep, M. B. (1995). Determination of methylmercury in fish and river water samples using in situ sodium tetraethylborate derivatization following by solid-sample microextraction and gas chromatographymass spectrometry. Journal of Chromatography A, 113112.

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