(17) Dong, M.; Locke, D. C.; Ferrand, E. Anal. Chem. 1976, 48, 368. (18) Daisey, J. M.; Leyko, M. A. Anal. Chem. 1979,51,24. (19) Brockhaus, A.; Tomingas, R., Staub-Reinhalt. Luft 1976,36, 96. (20) Grimmer, G.; Bohnke, H.; Glaser, A. Zbl. Baht. Hyg., I. Abt. Orig. B 1977,164,218. (21) Grimmer, G.; Naujack, K.-W.; Schneider, D. "Polynuclear Aromatic Hydrocarbons: Chemistry and Biological Effects"; Bjorseth,
A., Dennis, A. J., Eds.; Battelle Press: Columbus, OH, 1980; pp 107-25. (22) Greenberg, A.; Giorgio, P., unpublished observations.
Received for review July 30,1980. Accepted December 17,1980. This work was supported by the New Jersey Department of Enuironmental Protection and the New Jersey Institute of Technology Research Foundation.
Analysis of Organic Substances in Highly Polluted River Water by Mass Spectrometry Akio Yasuhara,* Hiroaki Shiraishi, Masahiko Tsuji,? and Toshihide Okunot Division of Chemistry and Physics, National Institute for Environmental Studies, Yatabe, Tsukuba-gun, lbaraki 305, Japan
atile compounds or thermally unstable compounds. This paper also describes the environmental application of FD-MS to the analysis of trace amounts of nonvolatile organics in river water. Volatile and nonvolatile compounds were separated by vacuum distillation and were identified by GC/MS and FD-MS, respectively.
m Volatile and nonvolatile organics in Hayashida River water which contained effluents from the leather industry were separated and analyzed by mass spectrometry. Volatile components were concentrated by steam distillation using a Nickerson-Likens apparatus or by vacuum distillation. The extracts were analyzed with a computerized gas chromatography/mass spectrometry system using a glass-packed column with high resolution. The results of the analyses indicated particularly the presence of large amounts of ethanediol monoalkyl ethers. The nonvolatile organics in the residue from vacuum distillation were extracted and analyzed by field desorption-mass spectrometry. The results showed the presence of polyethylene glycols, many kinds of poly( oxyethylene) alkylphenyl ethers, and some free fatty acids. Introduction I t is very important to determine various kinds of organic substances in river water from the viewpoint of water pollution. Many techniques have been developed for extraction and identification of organic compounds present in river water. For separation of very volatile compounds the head-space gas has been used (1-9). Adsorption methods with resin such as XAD-2 have also given good results ( 1 0 , l l ) .Recently several hundreds of compounds have been detected in the retained entities of reverse osmosis (12).Although direct extraction with organic solvents is very popular for the analysis of organics, direct extraction of organic substances from highly polluted water is very difficult because emulsions form by mixing with organic solvents and because hydrophilic compounds are only slightly transferred to the organic layer. From the point of view of gas-chromatographic analysis, the inclusion of nonvolatile organics is very undesirable because of a decrease in column resolution. Steam distillation or vacuum distillation is suitable for the separation of volatile and nonvolatile compounds. The usual technique for steam distillation is unfavorable because the volume of distillate is large and because extraction efficiency is bad. In this study the Nickerson-Likens apparatus (13),which provides a combination of cyclic steam distillation and continuous extraction, was used for improved extraction efficiency and to avoid the contamination potential from steam and solvent. Identification was carried out by gas chromatography/mass spectrometry (GC/MS). Recently field desorption-mass spectrometry (FD-MS) has shown great success for the analysis of nonvol-
t The Environmental Science Institute of Hyogo Prefecture, Suma-ku,Kobe, Hyogo 654, Japan. 570
EnvironmentalScience & Technology
Experimental Section Sampling and Separation Procedure. Water was sampled from the Hayashida River a t the Matsubara area in Tatsuno City, Hyogo Prefecture, on April 2,1980. The water was divided into two parts. One part (1.7 L) was steam-distilled with a Nickerson-Likens apparatus with diethyl ether (100 mL) for 3 days. The ether solution was dried on anhydrous sodium sulfate and then concentrated with a Kuderna-Danish concentrator under atmospheric pressure. The concentrated solution was analyzed by GC/MS. The other part (2 L) of the water was vacuum-distilled a t Torr after freezing, and the receiver was cooled a t -80 "C. From the trapped water in the receiver, organic substances were extracted with ether (300 mL) for 24 h with a continuous liquid-liquid extractor. The extracted solution was concentrated with a Kuderna-Danish concentrator under atmospheric pressure. The concentrated solution was analyzed by GC/MS. Nonvolatile organics in the residue after vacuum distillation were extracted with ethyl acetate (400 mL) a few times. The solution was concentrated by a rotary evaporator under reduced pressure. The concentrated solution was analyzed by FD-MS. Analytical Procedure. Gas chromatograms and mass spectra were measured with a JEOL Model JMS-D 100 mass spectrometer connected with a JEOL JGC-BOK gas chromatograph and a JEOL JMA-2000 mass data analysis system. The glass column (3 m X 2 mm i.d.) was packed with Thermon-1500, which is a packing material coated with 5% polyester plus 5% starch plus 0.3% phenolic resin on 80-100-mesh, acid-washed, DMCS-treated Chromosorb-W and was obtained from Shinwa Kako Co., Ltd. (Japan). The column temperature was set at 50 "C for 2 min, followed by an increase to 210 "C at a rate of 4 "C/min, and then held at 210 "C until completion of analysis. The injector temperature was 240 "C, and the helium carrier gas flow rate was 40 mL/min. The separator temperature for GC/MS was 240 "C. The gaschromatographic conditions for the analysis of fatty acids were as follows: column, glass column (2 m X 2 mm i.d.) packed with 5% FFAP on Uniport S (60-80 mesh); column temperature, 240 "C; injector temperature, 300 "C; helium carrier gas rate, 40 mL/min. The mass-spectrometric conditions were as follows: ionizing current, 3 X lo-* A; ionizing energy, 25 eV; accelerating voltage, 3 kV; scan range, m/z 10-400; scan speed, 2.6 s/scan; scan interval, 5 s. FD-MS was performed on a
0013-936X/81/0915-0570$01.25/0 @ 1981 American Chemical Society
JEOL Model JMS-OlSG2 double-focusing mass spectrometer with a combined field desorptionlfield ionizationlelectron impact source, JMA-2000 mass data analysis system, and an emitter current programmer ( 1 4 ) which was constructed with minor modifications according to Maine et al. (15). The emitter was tungsten wire (10 pm in diameter) with carbon needles (20-40 pm) made in a manner similar to that of Schulten and Beckey (16),with a JEOL MS-FDA 01 activator. The anode potential was 10 kV, and the cathode potential -3 kV. The emitter current was linearly increased by the emitter current programmer a t a rate of 3 mA/min. The sample was loaded by the microsyringe technique (17).FD mass spectra were obtained by integration of repetitive scans over the range from mlz 200 to 1000 a t an interval of 12 s (the individual FD mass spectrum obtained by repeated scans during an emitter current programming was integrated more than 15 times by computer). NMR spectra were measured by a Bruker Model SXP 4-100 Fourier transform nuclear magnetic resonance spectrometer; 89.99-MHz frequency was used for l H NMR, and CDC13 containing a small amount of Me&i was the solvent. IR spectra were measured with a Hitachi Model 285 grating infrared spactrophotometer, and the sample was coated on a KBr plate.
N
c a C 0 0
a
9,
a L
0
+
N
U
L
9,
e 9,
n
0
Time ( m i n ) Figure 1. Gas chromatogram of the volatile extract using the Nickerson-Likens apparatus. N
Results and Discussion The Hayashida River studied here is one of the branches of the Ibo River, which is a large river in Japan and runs through Tatsuno City, a city famous for the manufacture of leather goods. Although the river has been polluted only by effluents from the leather factories, it is one of the most polluted rivers in Japan. The annual averages of COD and BOD in the last few years have been ca. 200 and 380 ppm. The results for basic measurements of the river-water quality are shown in Table I. Figures 1and 2 show the gas chromatograms of the volatile extracts by the Nickerson-Likens apparatus and vacuum distillation, respectively. The results for identification and quantification are shown in Table 11.It may be concluded that vacuum distillation was better than steam distillation, since concentrations by vacuum distillation were higher in most components. Some compounds were detected only in one method, but they might be unimportant in the interpretation of water pollution in this river. Aliphatic hydrocarbons are usually detected as the main components in water of a typical river, since they are very easily extracted by direct solvent extraction. In this study the compounds, which distribute more in water than in organic solvents such as ether or dichloromethane, were extracted effectively by the use of the Nickerson-Likens apparatus or the continuous extraction apparatus. The presence of 2-alkoxyethanols is very interesting, since these compounds had not yet been detected in rivers in Japan. Alcohols were readily detected by these techniques and also by selection of a good packing material for GC. Resolution of the packed column used in this study
Table 1. Basic Water-Quality Data for Hayashlda River Water atmospheric temperature water temperature transparency
2.5 cm
COD BOD dry residue PH
365 ppm 489 ppm 4.65 g/L 9.20
14 OC 17 OC
9,
a C
0
P
a
e
a L
0
+ .
V
c c
c
0
0
I
I
4
I
I
8
I
I
12
I
I
16
I
I
20
I
I
24
I
28
I
I
32
I
I
36
I
I
I
40
Time ( m i n ) Figure 2. Gas chromatogram of the volatile extract using vacuum distillation. is comparable to that of a capillary column. 2,6-Di-tertbutyl-4-methylphenol, which is well known as an antioxidant, was detected in fairly large amounts. The concentrations of phenol and p-cresol were higher than in other river waters. 1,5-Di-tert-butyl-3,3-dimethylbicyclo[3.l.0]hexan-2-one has already been detected in the water of the Kanzaki River (18). 1,3-Hexanediol was assigned tentatively. Ethyl acetate, 4methyl-2-pentanone, toluene, butanol, 2-ethoxyethanol, and 2-butoxyethanol have already been determined in paint thinners used for dying leather (19).Therefore, the detection of these compounds was not unexpected. The residue after vacuum distillation contained fewer organics than inorganics. The organic extract contained various kinds of components, so the mixture was directly analyzed by FD-MS without separation of the components. The mass spectra, which are shown in Figures 3 and 4, had a characteristic pattern. The NMR and IR spectra of the extract were measured and are shown in Figures 5 and 6. The presence of carboxylic acids was considered from the IR spectrum. Judging from the results of measurements by FD-MS, NMR, and IR, we assigned compounds corresponding to the peaks a t mlz 256,282, and 284 in Figure 3 as tetradecanoic, 9-octadecenoic, and stearic acids, respectively. These results were also confirmed by GCIMS analysis, which showed the presence of the corresponding fatty acids. Usually each peak in the FD mass spectra represents each molecular ion or each Volume 15, Number 5, May 1981 571
~
639
Table II. Identified Compounds and Concentrations (ppb) in Hayashida River Water peak no.
retention time, min
compd
1 ethyl acetate 2 ethanol
3.00 3.49
4-methyl-2-pentanone toluene 2-methylpropanol butanol 7 2-ethoxyethanol 8 tridecane 9 2-butoxyethanol
5.20 6.00
3 4 5 6
Ib
60 1970 232 99 142 87
250 9 1310 NDd
16.04 18.56
ether e tetradecane 1,3-hexanediole 2-ethyl-1-hexanol pentadecane 15 hexadecane 16 heptadecane 17 a,a-dimethylbenzylalcohol 11 12 13 14
18.64
298
19.60 21.48
251 66 104
22.44 25.48 28.48 30.92 31.52 32.28 34.00
18 2-(2'-butoxyethoxy)ethanol 19 hexyglycol monoisobutyrate e 20 2,6-di-tert-butyl-4-
551
h
concn a method method
8.00 9.80 12.44 15.32
10 bis(2-dimethylaminoethyl)
683
595
56 27 NDd
35 12 201
IIC
585
4020 535
.-
1002 727
0,
NDd
771
685 318 1200 34
800
1000
900
Mass Number ( m k ) Figure 4. FD mass spectrum of the nonvolatile extract at high emitter current (8-13 mA).
5680 143 NDd NDd 111 208 114 32 119 240 N D ~ 2095
methylphenol 21 phenol 22 p-cresol 23 1,5-di-tert-butyl-3,3dimethylbicyclo[3.1.O]-
35.72 37.76 40.40
306
895
8
1
52
204 N D ~ 35
6 5 4 3 2 1 Chemical S h i f t ( p p t )
0
Figure 5. NMR spectrum of the nonvolatile extract.
hexan-2-one a These values were not corrected with recovery coefficients. Method i represents steam distillation using the Nickerson-Likens apparatus. Method II represents vacuum distillation. ND means "not detected". e These compounds were tentatively assigned.
371
'"1
I
>
' I c
C
282
327
~
,
41,5
1
459
503
I
547
I
I . . " 1 ~ ' . . 1 ' " ' I " " I
4000
3500
3000
2500
'
2000
I
1600
Wavenumber
'
I
(
cm-'
'
I
1200 )
'
I
'
I
800
'
I
I
Figure 6. IR spectrum of the nonvolatile extract.
Figure 3. FD mass spectrum of the nonvolatile extract at low emitter
current (0-6 mA). quasi-molecular ion. I t was very interesting that the difference between the corresponding peaks was always 44 mass units. Otsuki and Shiraishi (20) have already reported similar FD mass spectra, where 44 mass units corresponds to the molecular weight of ethylene oxide. The peaks a t mlz 327,371,415, 459,503, and 541 in Figure 3 may be due to polyethylene glycols, where the number of oxyethylene units is 7-12. From the investigation of FD mass spectra of various kinds of reference materials, the FD mass spectrum of poly(oxyethy1ene) alkyl ether shows a quasi-molecular ion a t M (molecular weight) 1 mass number, and that of poly(oxyethy1ene) alkylphenyl ether shows the molecular ion at M mass number. But further investigation of FD mass spectra of various poly(oxyethy1ene)
+
572
Environmental Science & Technology
derivatives revealed that the spectra are significantly affected by the coexistence of salts and that quasi-molecular ions of the type (M alkali metal)+ are major ion species for both poly(oxyethy1ene) alkyl ether and poly(oxyethy1ene) alkylphenyl ether when the spectra are measured in the presence of salts of alkali-metals. The extracts from river water were found to contain sufficient amounts of salts of alkali metals to form quasi-molecular ions (M alkali-metal)+.The peaks a t m/z 507,551,595,639,683,727,771,815,859, and 903 in Figure 4, which have greater intensities than peaks in other series, may be (M Na)+ ions of poly(oxyethy1ene) nonylphenyl ethers with a degree of polymerization of 6-15 oxyethylene units. The peaks a t mlz 567,611,655,699,743,787, 831, and 875 may be (M K)+ ions of poly(oxyethy1ene) nonylphenyl ethers. The other peaks a t mlz 493,537,581,625, 669,713,757,801,845, and 889 may be (M Na)+ ions and indicate the presence of poly(oxyethy1ene) octylphenyl ethers with a degree of polymerization of 6-15 oxyethylene units. The
+
+
+
+
+
+
Hiroyasu Ito for the measurement of fatty acids by GCI MS.
corresponding (M K)+ ions of poly(oxyethy1ene) octylphenyl ethers may overlap with isotope ions of quasi-molecules (M Na) of poly(oxyethy1ene) nonylphenyl ethers a t mlz 509,553,597,641,685,729,773,817,861,and 905. The relative intensities of these poly(oxyethy1ene) alkylphenyl ethers are considered to reflect their distributions in the extract fairly exactly, assuming that each poly(oxyethy1ene) alkylphenyl ether could have the same ionization efficiency in the FD-MS. These results of identification agree with an interpretation of the NMR spectrum which shows the presence of a -CHZO-group at -3.7 ppm. These poly(oxyethy1ene) alkylphenyl ethers including polyethylene glycols may probably come from detergents used for tanning leather. The same compounds were determined from the water sampled on March 31, 1979, a t the same station. I t is interesting that the NMR signal arising from the oxyethylene unit (3.7 ppm) is remarkably small when compared to that of fatty acids, while poly(oxyethy1ene)alkylphenyl ethers gave predominant ion peaks in the FD-MS. This may be due to large differences in the ionization efficiency between the poly(oxyethy1ene) alkylphenyl ethers and the fatty acids. However, no attempt was made to quantify these poly(oxyethy1ene) alkylphenyl ethers by FD-MS at the present time, but studies are now in progress. Some compounds identified in this study are very soluble in water and pose difficulties for direct extraction by organic solvents. The successful determination of both volatile and nonvolatile compounds in this study is due mainly to the use of vacuum distillation under a frozen state and the appropriate combination of GC/MS and FD-MS.
+
Literature Cited (1) McAuliffe, C. CHEMTECH. 1971,1,46. (2) Rook, J. J. Water Treat. Exam. 1972,21, 259. (3) Mieure, J. P.; Dietrich, M. W. J . Chromatogr. Sci. 1973, 11, 559. (4) Zlatkis, A.; Lichtenstein, H. A.; Tishbee, A. Chromatographia 1973,6, 67. (5) Grob, K. J. Chromatogr. 1973,84,255. (6) Zlatkis, A.; Lichtenstein, H. A,; Tishbee, A.; Bertsch, W.; Shunbo, F.; Liebich, H. M. J. Chromatogr. Sci. 1973,11, 299. (7) Bellar, T. A,; Lichtenberg, J. J. J. Am. Water Works Assoc. 1974, 66,739. ( 8 ) Dowty, B.; Carlisle, D.; Laseter, J. Science 1975,187, 75. (9) Dowty, B.; Carlisle, D.; Laseter, J. Enuiron. Sci. Technol. 1975, 9, 762. (10) Burnham, A. K.; Calder, G. V.; Fritz, J. S.; Junk, G. A.; Svec, H. J.; Willis, R. Anal. Chem. 1972,44, 139. (11) Junk, G. A.; Richard, J. J.; Grieser, M. D.; Witiak, D.; Witiak, J. L.; Arguello, M. D.; Vick, R.; Svec, H. J.; Fritz, J. S.; Calder, G. V. J. Chromatogr. 1974,99,745. (12) Coleman, W. E.; Melton, R. G.; Kopfler, F. C.; Barone, K. A.; Aurand, T. A.; Jellison, M. G. Enuiron. Sci. Technol. 1980, 14, 576. (13) Nickerson, G. B.; Likens, S. T. J. Chromatogr. 1966,21,1. (14) Shiraishi, H.; Otsuki, A.; Fuwa, K. Bull. Chem. SOC. Jpn. 1979, 52,2903. (15) Maine, J. W.; Soltmann, B.; Holland, J. F.; Toung, N. D.; Gerber, J. N.; Sweeley, C. C. Anal. Chem. 1976,48,427. (16) Schulten, H.-R.; Beckey, H. D. Org. Mass Spectrom. 1972,6, 885. (17) Beckey, H. D.; Heindrichs, A.; Winkler, H. U. Int. J . Mass Spectrom. Ion Phys. 1970,3,11. (18) Yasuhara, A.; Fuwa, K. Chemosphere 1977,6,179. (19) Okuno, T.; Tsuji, M.; Yamazaki, T. “Report on Odor Pollution in Hyogo Prefecture”; The Environmental Science Institute of
Acknowledgment
Hyogo Prefecture: Kobe, Japan, 1977; pp 22-33. (20) Otsuki, A.; Shiraishi, H. Anal. Chem. 1979,51,2329.
We thank Mr. Tomio Yamazaki and Mr. Hideo Yasuhara for the sampling of the Hayashida River water, Mr. Masayuki Kunugi for the measurement of the NMR spectrum, and Mr.
Received for review July 31,1980. Accepted January 5,1981
Effects of Dechlorination on Early Life Stages of Striped Bass (Morone saxatilis) Lenwood W. Hall, Jr.,*t Dennis T. Burton,? William C. Graves,t and Stuart L. Margreyt Academy of Natural Sciences of Philadelphia, Benedict Estuarine Research Laboratory, Benedict, Maryland 20612
Effects of sulfur dioxide (S02) dechlorination on estuarine striped bass (Morone sazatilis) eggs and larvae were evaluated by exposing the organisms to the following conditions: total residual chlorine (TRC); S02; TRC-SO2 dechlorination, and control. Continuous exposure to TRC concentrations ranging from 0.06 to 2.0 mg/L were lethal to both life stages over a 96-h exposure period. The same range of SO2 (sulfite) concentrations caused an effect on the eggs after 36 h; however, percent mortality did not increase with concentration of S02. Few mortalities occurred a t exposures less than 36 h. Mortality of larvae was higher than the the controls at all SO2 conditions after 96 h although the mean mortality a t all concentrations was only 20% greater than the controls. Minimal mortality occurred at shorter exposure intervals. Mean mortality of test organisms exposed to TRC-SO2 dechlorination conditions was only 11%higher for eggs (36 h) and 22% higher for larvae (96 h) than controls. Dechlorination caused significant reductions in TRC toxicity a t all exposure periods less than 36 and 96 h for eggs and larvae, respectively.
+ Present address: The Johns Hopkins University, Applied Physics Laboratory, Aquatic Ecology Section, Shady Side, MD 20867. 0013-936X/81/0915-0573$01.25/0
Introduction
Dechlorination has been used as a means to reduce the residual toxicity of chlorinated industrial effluent to aquatic life. In recent years, the following methods of chemical dechlorination have been used: (1)sodium bisulfite ( 1 , 2 ) ; (2) sulfur dioxide (3-6); (3) sodium sulfite (7,8); and (4) sodium thiosulfate (9-12). Although all of the preceding chemical agents have been used to dechlorinate water used in industrial operations, sulfur dioxide (SO2) is considered one of the most feasible methods to use for large volumes of water ( 1 3 , 1 4 ) . This chemical agent provides an excellent means of dechlorination because its handling and metering are similar to those of chlorine. Various investigators (3-6) have evaluated the response of aquatic biota exposed to SO2 dechlorination in freshwater systems; however, the use and the possible ecological effect of this chemical in estuarine waters are limited. This study was designed to evaluate the effects of SO2 dechlorination on striped bass (Morone s a r a t i l i s ) eggs and larvae in estuarine water. These developmental stages of striped bass were selected for this investigation because of their occurrence in estuaries where chlorinated industrial effluent is discharged.
@ 1981 American Chemical Society
Volume 15, Number 5, May 1981 573
