<Supplemental Materials>

Perfluorinated Compounds in Soils from Liaodong Bay with

Concentrated Fluorine Industry Parks in China

Pei Wang a, b, Tieyu Wang a, *, John P. Giesy c, Yonglong Lu a, *

* Corresponding author: Tel: +86 10 62849466; Fax: +86 10 62918177

E-mail address: yllu@rcees.ac.cn (Y. LU)；wangty@rcees.ac.cn (T. Wang)

Analytical methods

Standards and reagents

Extraction and clean-up

Analysis of Organic Carbon (OC) and pH

Instumental analysis

References

Supplemental table

Table S1 Target analytes of 22 PFCs measured in the present study with QA/QC information including monitoring transitions (MT), limit of quantification (LOQ), matrix spike recovery (MSR) for soil samples (Mean±SD), method detection limit (MDL) and sample detected ratio (DR) within 14 sampling sites.

Table S2 Sampling information including location, Organic Carbon (OC) and pH.Table S3 Spearman rank correlation coefficients among PFCs, EOF, TF, OC and

pH.

Table S4 The result of PCA analysis.

Supplemental figures

Figure S1 The component plot of individual PFC, EOF, TF, OC and pH.

Analytical methods

Standards and reagents

The 22 PFCs, including Perfluorobutanoic acid (PFBA) [1,2,3,4 13C],

Perfluorohexanoic acid (PFHxA) [1,2 13C], PFOA [1,2,3,4 13C], Perfluorononanoic

acid (PFNA) [1,2,3,4,5 13C], Perfluorodecanoic acid (PFDA) [1,2 13C],

Perfluoroundecanoic acid (PFUdA) [1,2 13C], Perfluorododecanoic acid (PFDoA) [1,2

13C], Sodium Perfluorohexanesulfonate [18O] and Sodium Perfluorooctanesulfonate

[1,2,3,4 13C] that were used as either external or surrogate standards (Table S1), had

purities of >98% (Wellington Laboratories, Guelph, Ontario, Canada). HPLC grade

methanol, methyl tert-butyl ether (MTBE) and ammonium acetate were purchased

from J.T. Baker (Phillipsburg, NJ, USA). Analytical grade sodium thiosulfate was

purchased from EMD Chemicals (Gibbstown, NJ, USA). Nano-pure water was

obtained from a Milli-Q gradient A-10 (Millipore, Bedford, MA, USA).

Extraction and clean-up

2.5 g of soil was weighed into a 50 mL PP centrifuge tube, and then moistened with 2

mL cleaned Milli-Q water while vortex mixing. 1 mL of 0.5 M TBAHS and 2 mL of

25 mM sodium acetate buffer were added for extraction. Samples were spiked with

internal standards that were labeled with 13C or 18O with shaking for 5 min at 700 rpm.

Subsequently, 5 mL MTBE was added and shaken for 20 min at 400 rpm. After

centrifuging for 20 min at 3500 rpm, the supernatant of MTBE was collected. The

remnant aqueous mixture was rinsed with 5 mL MTBE, followed by shaking and

centrifuging, the two supernatants were combined in a 15 mL PP centrifuge tube. The

MTBE solvent was evaporated to dryness under a gentle flow of high purity nitrogen,

and reconstituted in 1 mL methanol. After 0.2 mL solution was taken for

quantification of EOF, the remaining 0.8 mL was added with 40 mg Envi-carb and 1

mL methanol, followed by sonication and centrifuging for 10 min at 3500 rpm. The

supernatant was transferred to a new 15 mL PP tube. The remnant was rinsed twice

with 1 mL methanol, followed by sonication and centrifuging, and the supernatants

combined. The solution was reduced to 0.5 mL under nitrogen. Then filtered through

a 13 mm/0.2 um nylon filter, and transferred into a 1.5 mL PP snap top vial with

polyethylene (PE) cap.

Analysis of Organic Carbon (OC) and pH

OC in soil was determined using external heating potassium dichromate method

according to the Agricultural Standard of China (NY/T 1121.6-2006) with some

modifications. Briefly, 0.3g soil ground through 0.15mm sieve was weighed into a

150mL triangular flask, with 5mL 0.8mol/L potassium dichromate solution and 5mL

concentrated sulfuric acid added. After shaking the mixture well and put a crookneck

funnel over the flask, heat the flask to 170-180℃ and kept boiling 5 min and then

cooled off. Washing the funnel with Milli-Q water to keep the volume of solution 60-

70mL, here the color of the solution should be orange yellow or jasmine. Then

Phenanthroline indicator 3-4 drops were added and titrated with 0.198mol/L green

copperas solution to turn the color of the solution to green, pea green and finally

redbrown. Two blanks were necessary for each set of samples and 0.5g mealiness

silicon-dioxide was used for surrogate. The OC content was calculated using the

following formula:

OC ( g/kg )=C × (V 0−V ) ×3 ×1.1 ×10−3

m

Where C is 0.198mol/L green copperas solution, V0 is the volume (mean value) of

blanks used to titrate green copperas solution (mL), V is the volume of samples used

to titrate green copperas solution (mL), 3 stands for a quarter mole mass of a carbon

atom (g/mol), 1.1 is oxidation correction factor and m is the weight of a sample (kg).

pH in soil was determined using potentiometry according to the Agricultural Standard

of China (NY/T 1377-2007) with some modifications. Briefly, 10g soil ground

through 2mm sieve was weighed into a 50mL beaker and added 25mL CO2 free Milli-

Q water, followed by vigorous agitation using a glass rod for 1-2 min, then left for

30min. Corrected the pH meter using buffer solution and cleaned it. Put the bulb of

glass electrode underneath the suspension of the sample with slightly shaking and the

saturated calomel electrode supernatant fluid, recorded when reading is stable.

Recorrected the pH meter after 5-6 samples.

Instumental analysis

The CIC was a combination of an automated combustion unit (AQF-100 type AIST;

Dia Instruments Co., Ltd.) and an ion chromatography system (ICS-3000 type AIST;

Dionex Corp., Sunny-vale, CA). The details for analytical conditions are given

elsewhere (Miyake et al. 2007). 0.2g of soil or 0.2mL of extract was set on a slica

boat and then combusted in a furnace at 900-1000℃, where organic and inorganic

fluoride were converted into hydrogen fluoride (HF). The HF was then absorbed into

sodium hydroxide solution (0.2 mmol/L). The concentration of F- was determined

using IC. Sodium fluoride (99% purity; Sigma-Aldrich Co. LLC, St. Louis, United

States) was used as a standard for the quantification.

The HPLC was fitted with a Thermo Scientific Betasil C18 (100×2.1 mm, 5 μm

particle size) analytical column, and a guard column was used for separation of

background from analytes in samples. An aliquant of 2 mM ammonium acetate was

used as an ionization aid. Water and methanol were used as mobile phases. Gradient

conditions were: 300 ml/min flow rate and 10 μl of the sample was injected, starting

with 60% A (2mM ammonium acetate) and 40% B (100% methanol). Initial

conditions were held for 2 min and then ramped to 20% A at 18 min, held until 20

min, decreased to 0% A at 21 min, increased to 100% A at 22 min, held until 22.5

min, returned to initial conditions at 23 min, and finally held constant until 26min.

The temperature of the column oven was kept constant at 35 ℃.

Mass spectra were collected using an Applied Bioscience SCIEX 3000 (Foster City,

CA) tandem mass spectrometer, fitted with an electrospray ionization source, operated

in negative ionization mode. Chromatograms were recorded in multiple reaction

monitoring mode (MRM) with a dwell time of 40 ms. The following instrument

parameters were used: desolvation temperature (450 ℃), desolvation (curtain) gas 6.0

arbitrary units (AU), nebulizer gas flow5 AU, ion spray voltage−3500 V, and

collision gas 12 AU. Optimal settings for collision energies and declustering potential

were determined for each analyte's MRM transitions. Quantification using these

transitions was performed using Analyst 1.4.1 software (Applied Bioscience, Foster

City, CA).

References

Naile, J.E., Khim, J.S., Wang, T.Y., Chen, C.L., Luo, W., Kwon, B.O., Park, J.,Koh,

C.H., Jones, P.D., Lu, Y.L., Giesy, J.P., 2010. Perfluorinated compounds in water,

sediment, soil and biota from estuarine and coastal areas of Korea. Environmental

Pollution 158 (5), 1237-1244.

Higgins, C.P., Field, J.A., Criddle, C.S., Luthy, R.G., 2005. Quantitative

Determination of Perfluorochemicals in Sediments and Domestic Sludge.

Environmental Science and Technology 39 (11), 3946-3956.

Miyake, Y., Yamashita, N., Rostkowski, P., So, M. K., Taniyasu, S., Lam,

P.K.S.,Kannan, K., 2007. Determination of trace levels of total fluorine in water

using combustion ion chromatography for fluorine: A mass balance approach to

determine individual perfluorinated chemicals in water. Journal of

Chromatography A 1143 (1-2), 98-104.

Supplemental table

Table S1 Target analytes of 22 PFCs measured in the present study with QA/QC information including monitoring transitions (MT), limit of quantification(LOQ), matrix spike recovery (MSR) for soil samples (Mean±SD), method detection limit (MDL) and sample detected ratio (DR) within 14 sampling sites.

Analyte Acronym MT MSR%

LOQ (ng/g

)

MDLa(ng/

g)

DRb

%

Perfluioroalkyl compounds

Perfluoro-n-butanoic acid (C4) PFBA

213→16

9 76±7 0.6 0.6 0(0)

Perfluoro-n-pentanoic acid (C5) PFPeA

263→21

9 94±11 1.0 1.0 0(0)

Perfluoro-n-hexanoic acid (C6) PFHxA

313→26

9 83±5 0.1 0.1 0(0)

Perfluoro-n-heptanoic acid (C7) PFHpA

363→31

9 96±10 0.1 0.1 0(0)

Perfluoro-n-octanoic acid (C8) PFOA

413→36

9 89±3 0.5 0.5 7(50)

Perfluoro-n-nonanoic acid (C9) PFNA

463→41

9 92±5 1.0 1.0 0(0)

Perfluoro-n-decanoic acid (C10) PFDA

513→46

9 86±4 0.1 0.1 1(7)

Perfluoro-n-undecanoic acid (C11) PFUdA

563→51

9 88±8 0.5 0.5 9(71)

Perfluoro-n-dodecanoic acid (C12) PFDoA

613→56

9 92±7 0.1 0.1 0(0)

Perfluoro-n-tridecanoic acid (C13) PFTrDA

663→61

9 93±11 1.0 1.0 3(21)

Perfluoro-n-tetradecanoic acid (C14) PFTeDA

713→66

9 72±4 1.0 1.0 0(0)

Perfluoro-n-hexadecanoic acid (C16) PFHxDA

813→76

9

107±1

4 0.5 0.3 0(0)

Perfluoro-n-octadecanoic acid (C18) PFODA

913→86

9 ISc IS IS IS

Potassium Perfluoro-1-butanesulfonate (C4) PFBS 299→99 76±9 1.0 1.0 0(0)

Sodium Perfluoro-1-hexanesulfonate (C6) PFHxS 399→99 94±8 0.5 0.5 0(0)

Sodium Perfluoro-1-octanesulfonate (C8) PFOS 499→99 97±9 0.5 0.5 2(14)

Sodium Perfluoro-1-decanesulfonate (C10) PFDS 599→99 95±11 0.5 0.5 0(0)

Perfluorinated precursors

2-N-ethylperfluoro-1-octanesulfonamido-ethanlo N-EtFOSE 616→59 85±6 1.0 1.0 0(0)

2-N-methylperfluoro-1-octanesulfonamido-

ethanlo N-MeFOSE 630→59 75±8 0.8 0.8 0(0)

Perfluoro-1-octanesulfonamidoacetic FOSAA

556→49

8

107±1

2 0.6 0.6 1(7)

N-methylperfluoro-1-octanesulfonamidoacetic

N-

MeFOSAA

570→41

9 91±4 1.0 1.0 0(0)

N-ethylperfluoro-1-octanesulfonamidoacetic N-EtFOSAA

584→41

9 93±8 1.5 1.5 0(0)

Table S2. Sampling information including location, Organic Carbon (OC) and pH.

Sample ID Location OC(g/kg) pH

LS1

LS2

LS3

LS4

LS5

LS6

LS7

LS8

LS9

LS10

LS11

LS12

LS13

LS14

Agriculture fields(corn), small River, 5km from sea, lots of trash

Agriculture fields(corn), 1km to River Wangbao

Agriculture fields(corn), near river Wuli, many factories around, lots of air pollution

On cliff, Beach(open sea, close to harbor), near downtown

Agriculture fields(corn), close to salt/shrimp ponds

Green area, downtown river bank

Agriculture fields(corn), Daling River (upstream)

Agriculture fields(corn), Daling River(middle stream), papermill around

Agriculture fields(corn), Daling River(downstream), lots of oil wells

Agriculture fields(rice), close to downtown, factories in the distance upstream

Reclaimed land, close to open sea (harbor)

Tree farm, Daliao River(midstream), near a heavy traffic bridge, large construction

Tree farm, Daliao River(downstream), downtown, lots of industry, busy harbor

Agriculture fields, beach(open sea), downtown, huge plants around

10.46

11.17

7.77

27.99

9.53

7.93

5.13

3.67

4.75

5.94

3.23

5.25

3.32

7.67

5.36

5.03

6.91

6.09

5.07

7.34

7.29

7.73

7.95

7.55

7.84

8.12

6.55

5.67

Table S3 Spearman rank correlation coefficients among PFCs, EOF, TF,

OC and pH.

EOF TF OC pH

PFCs 0.013 -0.339 0.013 -0.022

EOF 0.301 -0.064 0.305

TF 0.530 -0.552*

OC -0.697**

*. Correlation is significant at the 0.05 level (2-tailed).

**. Correlation is significant at the 0.01 level (2-tailed).

Table S4 The result of PCA analysis.

Component

1 2 3

PFOA .526 .535 .550

PFUdA .743 .333 -.039

PFCs -.050 .765 -.048

EOF -.280 -.391 .761

TF .674 -.661 .155

OC .894 .043 .264

pH -.724 .267 .539

Eigenvalue

Percentage of variance

Cumulative percentage

2.688

38.406

38.406

1.644

23.492

61.898

1.270

18.144

80.042

Extraction Method: Principal Component Analysis.a

a. 3 components extracted.

Supplemental figures

Fig.S1. The component plot of individual PFC, EOF, TF, OC and pH.