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Characterization of algal organic matter and formation of DBPs from chlor(am)ination

Jingyun Fang a,b, Xin Yang c,d, Jun Ma a,b,*, Chii Shang d,**, Quan Zhao d
a

State Key Laboratory of Urban Water Resources and Environment, Harbin Institute of Technology, Harbin 150090, China National Engineering Research Center of Urban Water Resources, Harbin 150090, China c School of Environmental Science and Engineering, Sun Yat-Sen University, Guangzhou 510275, China d Department of Civil and Environmental Engineering, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong
b

article info
Article history: Received 27 December 2009 Received in revised form 5 July 2010 Accepted 6 July 2010 Available online 13 July 2010 Keywords: Algae Disinfection Organic nitrogen By-products Chlorine Chloramine

abstract
The frequent occurrence of algal blooms in drinking water reservoirs causes problems to water supply, one of which is the release of algal organic matter in high concentrations to affect drinking water quality. Algal organic matter, including extracellular organic matter (EOM) and intracellular organic matter (IOM), was characterized. The formation of a variety of disinfection by-products (DBPs) in chlorination and chloramination of EOM, IOM and algal cells was evaluated. Natural organic matter (NOM) isolated from Suwannee River was also studied for comparison. EOM and IOM were rich in organic nitrogen, which consisted of high (over 10 kDa) and low (70e1000 Da) molecular weight (MW) organic matter, whilst the MW of organic carbon in EOM and IOM was relatively lower. IOM had a higher fraction of total organic nitrogen, with larger proportions of higher MW and more hydrophobic contents than did EOM. IOM also contained higher fractions of free amino acids but lower fractions of aliphatic amines than did EOM. During chlorination of EOM and IOM, organic chloramines were rst formed and then became undetectable after 1 d. Chlorination of EOM and IOM produced more nitrogenous DBPs (N-DBPs) and haloaldehydes and less carbonaceous DBPs (C-DBPs) than did chlorination of NOM. Organic chloramines were found after 3-d chloramination of EOM and IOM. The amounts of N-DBPs and C-DBPs formed from chloramination of EOM or IOM were much less than that from NOM. EOM produced less DBPs (except for trichloronitromethane) than did IOM and algal cells in chlorination and chloramination. 2010 Published by Elsevier Ltd.

1.

Introduction

The frequent occurrence of algal blooms in drinking water reservoirs causes problems to water supply. One major problem is the release of algal organic matter, including extracellular organic matter (EOM) and intracellular organic

matter (IOM), in high concentrations to water sources. EOM are the metabolites excreted from algal cells into the surrounding environment (Paralkar and Edzwald, 1996). IOM can be released to the environment when algal cells die and subsequently lyse (Thurman, 1985), or after an algae-laden water goes through water treatment processes, such as

* Corresponding author at. State Key Laboratory of Urban Water Resources and Environment, Harbin Institute of Technology, Harbin 150090, China. Tel.: 86 45186 282 292; fax: 86 45182 368 074. ** Corresponding author at. Tel.: 852 2358 7885; fax: 852 2358 1534. E-mail addresses: majun@hit.edu.cn (J. Ma), cechii@ust.hk (C. Shang). 0043-1354/$ e see front matter 2010 Published by Elsevier Ltd. doi:10.1016/j.watres.2010.07.009

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ozonation or chlorination (Ma et al., 2006; Plummer and Edzwald, 2002). Neither EOM nor IOM is readily removed by coagulation and pre-oxidation enhanced coagulation processes (Widrig et al., 1996; Ma et al., 2006). Their presence affects drinking water quality. The characterization of algal organic matter is important for understanding their fates and treatability in water supplies. Several methods for the characterization of algal organic matter have been reported in the literature, which include UVevisible absorbance, uorescence excitation emission matrix (EEM), high pressure size exclusive chromatography (HPSEC) coupled with UV/uorescence/DOC detection, afnity chromatography, and those using the Fourier transform infrared (FTIR) spectrophotometer, solidstate 13C NMR spectroscopy and Pyrolysis-GCeMS (Her et al., 2004; Nguyen et al., 2005; Pivokonsky et al., 2006; Widrig et al., 1996). Algal organic matter appears to contain more organic nitrogen (org-N), more hydrophilic content, and less aromatic carbon content. It has much lower specic UV absorbance (SUVA) values (<2 L/mg/m) and higher heterogeneity as compared with natural organic matter (NOM) (Widrig et al., 1996; Nguyen et al., 2005; Her et al., 2004). Both EOM and IOM contain biopolymers, such as proteins, peptides and amino acids, while it has been reported that the portion of protein in IOM is larger than that in EOM (Pivokonsky et al., 2006). However, less is known about the characteristics, such as molecular weight (MW) distribution, polarity distribution, and specic composition of org-N in IOM and EOM. The high org-N content of algal organic matter affects the disinfection efciency and the formation of disinfection byproducts (DBPs) in chlorination and chloramination. The former is due to the formation of less germicidal organic chloramines and the latter is due to the formation of more toxic nitrogenous DBPs (N-DBPs). The roles of algal cells, EOM and IOM in the formation of N-DBPs and carbonaceous DBPs (C-DBPs) in chlor(am)ination are unclear. Nevertheless, some information about the reactions between aqueous chlorine and various org-N model compounds is available. The most reactive compounds are aliphatic amines, free amino acids and some nitrogen-heterocyclic aromatics (Scully et al., 1994). Amide linkages in combined amino acids do not participate signicantly in the chlorine demand (Hureiki et al., 1994). Free amino acids and aliphatic amines have been reported to be the precursors of some N-DBPs (e.g. halonitriles, halonitroalkanes and nitrosamines) (Joo and Mitch, 2007; Mitch and Schreiber, 2008). However, extrapolation of the knowledge from chlor(am)ination of the model org-N compounds to that of algal cells and algal organic matter is not possible, partially due to the unclear characteristics of the latter two. The aims of this work were 1) to characterize the org-N contents in both EOM and IOM produced from Microcystis aeruginosa (the most widely occurring blue-green algae) using EEM uorescence, HPLC/uorescence, and through analyses of amino acids and aliphatic amines, and 2) to assess the roles of algal cells, IOM and EOM in the formation of organic chloramines and a number of commonly found DBPs (particularly N-DBPs) in chlorination and chloramination. The results were also compared with those of NOM.

2.
2.1.

Materials and methods

Solution preparation

Axenic cultures of M. aeruginosa (blue-green algae, Collection No. HB909) were obtained from the Culture Collection of Algae at the Institute of Hydrobiology, Chinese Academy of Sciences, China. M. aeruginosa was cultivated for 42 d to allow algal cells to grow into the stationary growth phase. The cultivation procedures can be found in Fang et al. (2010). Algal cells were then separated from the algal suspensions by centrifugation. The supernatant was collected and ltered through a GF/F membrane (Whatman), which is hereafter referred to as the EOM solution. The deposited algal cells in the centrifuge tube were collected and washed with 200-mL Milli-Q water (Milli-Q biocel), followed by two cycles of centrifugation and supernatant removal. IOM was extracted from the deposited algal cells by physically grinding the cells with a mortar and pestle in Milli-Q water, followed by ltration through a GF/F membrane. The ltrate is hereafter referred to as the IOM solution. Suwannee River NOM (Cat. No. 1R101N) obtained from International Humic Substances Society was dissolved into Milli-Q water and ltered through a 0.45-mm membrane to make a stock NOM solution. All chemical solutions were prepared from reagent-grade chemicals and Milli-Q water. A free chlorine stock solution (2600 mg/L as Cl2) was prepared from 4% sodium hypochlorite (NaOCl) (SigmaeAldrich) and periodically standardized by DPD/FAS titration (APHA, AWWA, WEF 1998). A preformed monochloramine (NH2Cl) solution was freshly prepared prior to experiments by adding an aliquot of the free chlorine solution to an ammonium chloride solution at a chlorine to NH 4 -N molar ratio of 0.8:1 for 30 min with rapid stirring (Yang et al., 2007). Buffer solutions at pH 7 and 8.5 were prepared with phosphate and borate salts, respectively. A mixed standard kit of USEPA Method 551A, 551B and chloral hydrate (Product No. 40846) and a mixture of HAA9 standards (Product No. 47787) were obtained from Supelco. A dichloroacetaldehyde (DCA) standard was purchased from TCI, USA. 9-Fluorenylmethyl chloroformate (FMOC) was obtained from Fluka.

2.2.

Experimental procedures

Chlorination/chloramination experiments were carried out using 100-mL solutions of organic matter diluted to 5 mg/L TOC and buffered at pH 7.0 (phosphate, 10 mM). The chlorine or preformed monochloramine dosage added to IOM, algal cell and NOM solutions was 15 mg/L. The dosage added to EOM was 25 mg/L to compensate for the rapid chlorine consumption by the 2.09 mg/L nitrite in the EOM solution (Johnson and Margerum, 1991; Margerum et al., 1994). The reacting samples were incubated in amber glass bottles capped with Teon-faced septa at room temperature (22 1  C) in the dark for 2 h, 1 d and 3 d. After a scheduled period of time, samples were divided into four portions. The rst portion was analyzed by membrane introduction mass spectrometry (MIMS) for the determination of inorganic chloramine residuals and

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cyanogen chloride (CNCl). The second portion was analyzed by DPD/FAS titration for the determination of free chlorine residuals and total chlorine residuals. The third and fourth portions were quenched with ascorbic acid for analyses of HAAs and volatile DBPs. N-nitrosodimethylamine (NDMA) measurement was conducted separately using the protocol described below.

Table 1 e The properties of various algal materials and NOM. NH SUVA TOC TON TOC/TON NO 2 4 (mg/L) (mg/L) (L/mg/m) (mg/L) (mg/L)
EOM Algae IOM NOM 5.0 5.0 5.0 5.0 0.899 1.111 1.149 0.056 5.4e5.7 4.3e4.8 4.35 89.83 2.09 n.d. n.d. n.d. n.d. n.d. n.d. n.d. 1.38 n.a. 0.58 4.79

2.3.

Analytical methods

Note: EOM was pretreated by dialysis to remove the ions from the cultivation media before the determination of TON; n.d. e not detectable; n.a. e not available.

Total organic carbon (TOC) and total nitrogen (TN) concentrations were measured using a TOC and TN analyzer (TOCVCPH, Shimadzu). Concentrations of nitrite and nitrate were measured using an ion chromatograph (Dionex DX 500) equipped with an anionic column (IonPac AS9-HC). Concentrations of ammonia were measured using a ow injection analyzer (QuickChem FIA, 8000 Series). Concentrations of total organic nitrogen (TON) were then estimated by subtracting the nitrite, nitrate, and ammonia concentrations from the total nitrogen concentrations. The EOM solution was pretreated with dialysis (Lee and Westerhoff, 2004) to reduce

interferences from inorganic ions in the cultivation media (Table S1) in the TON measurement. Fluorescence EEM spectra were recorded on a Hitachi F4500 uorescence spectrometer following the procedures developed by Chen et al. (2003). Details are described in the Supporting Information (Text S1). MW distribution of samples was obtained using a high pressure size exclusive chromatography (HPSEC) (LC-10A, Shimadzu) coupled with a uorescence detector (Hitachi F-4500). The separation was achieved

Fig. 1 e Fluorescence EEMs of EOM, IOM, algal cells and NOM, (a) EOM-dialysis; (b) algal cell; (c) IOM; (d) NOM. The maximum uorescence intensity of each EEM (Max. F.I., unit: AU/5.0 mg C/L) was divided into 20 contour intervals. (EOM was pretreated by dialysis to eliminate the interference of inorganic ions from the cultivation media to the EEM spectra.)

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Fig. 2 e HPSEC chromatograms of EOM, IOM and NOM with uorescence detector at different Ex/Em wavelength pairs, (a) HPSEC-FL 280/340, (b) HPSEC-FL 275/450. All the chromatograms were normalized to the DOC of 1 mg/L.

) in by a Macrosphere GPC column (Alltech) with 6 nm (60 A pore size and 250 mm 4.6 mm in dimensions and packed with 7 mm spherical particles. The uorescence detector was set at the excitation/emission (Ex/Em) wavelength pairs of 230/340, 280/340, 275/450 and 350/450 nm/nm. Other operating details can be found in the Supporting Information (Text S2). Polarity distribution was measured using a reverse-phase HPLC (RP-HPLC) (LC-10A, Shimadzu) (Namjesnik-Dejanovic and Cabaniss, 2004) coupled with a uorescence detector (RF-10AXL). The separation was achieved with an Atlantis C18 column (5 mm 4.6 mm 250 mm), with 60% methanol and 40% pH 4.0 aqueous acetic acid/acetate buffer (0.01 M) as the mobile phase. The Ex/Em wavelength pairs of the uorescence detector were set to the same as those in the HPSEC analysis. Analysis of amino acids and aliphatic amines with precolumn derivatization by uorenylmethyl chloroformate (FMOC) was conducted using HPLC (LC-10A, Shimadzu) with a C18 column (5 mm 4.6 mm 250 mm, Atlantis) and a uorescence detector (RF-10A XL) (Molna-Perl, 2003). Compounds analyzed included 16 amino acids: histidine (His), lysine (Lys), arginine (Arg), asparagine (Asn), glutamine (Gln), serine (Ser), glycine (Gly), alanine (Ala), tyrosine (Tyr), proline (Pro), methionine (Met), valine (Val),

phenylalanine (Phe), isoleusine (Ile), leucine (Leu), and cysteine (Cys); and 5 aliphatic amines: methylamine (MA), dimethylamine (DMA), ethylamine (EA), methylethylamine (MEA), and diethylamine (DEA). Free chlorine and total combined chlorine were measured by DPD/FAS titration (APHA , AWWA, WEF 1998). Concentrations of inorganic chloramines, including mono-, di- and tri-chloramines, were analyzed by the MIMS method as described elsewhere (Shang and Blatchley, 1999). Concentrations of organic chloramines were calculated by subtracting total inorganic chloramines from total combined chloramines. Analyses of trichloromethane (TCM), chloral hydrate (CH), dichloroacetaldehyde (DCA), 1,1-dichloro-2-propanone (1,1-DCP), 1,1,1-trichloro-2-propanone (1,1,1-TCP), dichloroacetonitrile (DCAN), trichloroacetonitrile (TCAN), trichloronitromethane (TCNM) and nine haloacetic acids (HAA9) were carried out with a gas chromatograph (Agilent 6890) with an electron capture detector (ECD), based on USEPA Method 551.1 (USEPA, 1995) and USEPA Method 552.3 (USEPA, 2003). The column used was an HP-5 fused silica capillary column (30 m 0.25 mm I.D. with 0.25 mm lm thickness). NDMA was analyzed by isotope dilution analysis

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Fig. 3 e RP-HPLC chromatograms of EOM, IOM and NOM with uorescence detector at different Ex/Em wavelength pairs, (a) RP-HPLC-FL 280/340, (b) RP-HPLC-FL 275/450. All the chromatograms were normalized to the DOC of 1 mg/L.

as described in Yang et al. (2008). CNCl was analyzed by the MIMS method (Yang and Shang, 2005).

3.
3.1.

Results
Characterization of EOM, IOM and algal cells

Table 1 shows the TOC, TON, TOC/TON ratios and the SUVA values of EOM, IOM, algal cells and NOM. The SUVA value of IOM was lower than that of EOM and both were much lower than that of NOM, suggesting relatively fewer aromatic moieties in IOM than in EOM and than in NOM. The TOC/TON ratios followed the order: NOM >> EOM > IOM z algal cells. The much lower TOC/TON ratios of EOM, IOM and algal cells indicated that they were rich in org-N. Fig. 1 shows the EEM spectra of EOM, IOM, algal cells and NOM solutions. EOM, IOM and algal cells had uorophores that centered in regions II and IV, which represent the protein-like organic matter or org-N rich compounds (Chen et al., 2003). NOM had uorophores that primarily centered in regions III and V, which represent the humic-like and fulvic-like organic matter rich in org-C (Chen et al., 2003). The results of EOM are consistent with the literature (Henderson et al., 2008). Based on the results as well as the literature (Chen et al., 2003),

uorescence peaks at the Ex/Em wavelength pairs of 280/340, 230/340, 350/450 and 275/450 nm/nm were selected for characterization of the MW and polarity distributions of organic matter. The former two represent org-N rich substances and the latter two represent org-C rich substances (Chen et al., 2003). Fig. 2(a) shows the MW distribution of org-N rich substances in EOM, IOM and NOM at the Ex/Em wavelength pair of 280/340 nm/nm. EOM and IOM showed multiple org-N peaks (heterogeneity) representing both high-MW (over 10 kDa) and low-MW (from 70 to 1000 Da) org-N matter. The high-MW org-N matter could be proteins or proteinaceous materials, while the low-MW org-N matter could be free amino acids, aliphatic amines, or peptides, with MW of around several hundred Da. The results also show that IOM consisted of more high-MW org-N substances than low-MW substances, while the results of EOM display the opposite trend, which is in agreement with the ndings in Pivokonsky et al. (2006). Pivokonsky et al. (2006) also reported that EOM and IOM of M. aeruginosa contained proteins with MW higher than 60 kDa, which cannot be detected in the current study due to the size exclusion limits of the column used. Fig. 2(b) shows the MW distribution of org-C substances in EOM, IOM and NOM at the Ex/Em wavelength pair of 275/450 nm/nm. EOM and IOM contained relatively lower MW org-C substance with multiple peaks at around several hundred and several

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8 Concentration (ug N/mg DOC)

a
6

Free amino acid

EOM IOM

14 12

a
2h

Cl NH Cl NHCl NCl Org-chloramine

Residuals (mg/L as Cl )

10 8 6 4 2 0 14 12

1d

2 EOM IOM 0 1.6 His Lys Arg Asn Gln Ser Gly Ala Tyr Pro Met Val Phe Ile Leu Cys

3d

Concentration (ug N/mg DOC)

1.4 1.2 1.0 .8

b
EOM IOM

Aliphatic amine

b
NH Cl Org-chloramine

.4 .2 0.0 MA DMA EA MEA DEA

Residuals (mg/L as Cl )

.6

10 8 6 4 2 0

3d

Fig. 4 e The composition of organic nitrogen in EOM and IOM, (a) free amino acids, (b) aliphatic acids. The specic orgN concentrations were normalized to the DOC of 1 mg/L.

EOM

Cell

IOM

NOM

dozen Da. The MW distributions obtained at Ex/Em wavelength pairs of 230/340 and 275/450 nm/nm were similar to those at Ex/Em 280/340 and 350/450 nm/nm, respectively. Comparing the distributions of EOM and IOM to those of NOM, org-N fractions in EOM and IOM consisted of organic matter with much higher MW than that in NOM, but the org-C contents showed the opposite trends. Fig. 3 shows the polarity distributions of EOM, IOM and NOM, measured with a uorescence detector at wavelength pairs of Ex/Em 280/340 and 275/450 nm/nm. The RP-HPLC combined with uorescence provides quantitative information on the hydrophobicity of org-N and org-C. The retention time correlates with the octanolewater partition coefcient (log Kow) in the RP-HPLC chromatography (Braumann et al., 1983). As retention time increases, polarity decreases and hydrophobicity increases. As shown in Fig. 3(a), the org-N components in EOM and IOM were relatively more hydrophobic than those in NOM. There were some especially hydrophobic org-N components in EOM and IOM, the fractions of which were larger in IOM than in EOM. On the other hand, the org-C components in EOM showed a narrower range of polarity than those in NOM. The peaks for org-C in IOM were unclear. The org-N contents of EOM and IOM were further characterized. IOM contained a higher concentration of TON (180 mg N/mg DOC) than did EOM (230 mg N/mg DOC). The concentrations of the measured free amino acids and aliphatic amines only constituted z2.5% and 11.3% of the TON in EOM and IOM, respectively. Fig. 4 presents the breakdown of free amino acids and aliphatic amines in EOM and IOM. IOM contained higher concentrations of the 16 measured amino acids per mg DOC than did EOM. The amino acids in high abundance in IOM included arginine (Arg), lysine (Lys), and glycine (Gly). EOM had higher concentrations of the measured

Fig. 5 e The concentrations of free chlorine, inorganic chloramines and organic chloramines in EOM, IOM, algal cells and NOM (5 mg/L as TOC) after chlorination/ chloramination at pH 7.0 with different reaction time, (a) chlorination with the reaction time of 2 h, 1 d and 3 d, and (b) chloramination with the reaction time of 3 d.

aliphatic amines per mg DOC than did IOM, among which diethylamine (DEA) and ethylamine (EA) were the most abundant. Nevertheless, dimethylamine (DMA) and methylamine (MA), which are the major precursors of NDMA and CNCl, respectively (Mitch et al., 2003; Yang et al., 2007), were not detectable in IOM and EOM. The low concentrations of free amino acids and aliphatic amines detected in EOM cannot explain the two large org-N peaks in the HPSEC chromatogram at the Ex/Em pair of 280/340 nm/nm shown in Fig. 2(a). These two large peaks with MW of a hundred and a few hundred Da belonged to other unidentied org-N components. In summary, EOM and IOM were rich in org-N components, with higher heterogeneities in MW and polarity distributions. The high MW org-N components corresponded to the hydrophobic org-N contents in EOM and IOM. In comparison with EOM, IOM had higher concentrations of TON and free amino acids, lower concentrations of aliphatic amines, and larger proportions of higher MW and hydrophobic contents.

3.2. Chlorine residuals and DBP formation during chlorination and chloramination
Fig. 5(a) and (b) displays the concentrations of residual free chlorine, inorganic chloramines and organic chloramines

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400

Concentration (g/L)

Concentration (g/L)

120

TCM
Chlorination Chloramination

100 80 60 40 20 0

DCAA

300

200

100

Concentration (g/L)

Concentration (g/L)

1,1,1-TCP

8 6 4 2 0

1,1-DCP

Concentration (g/L)

80 60 40 20 0

Concentration (g/L)

CH

DCA

EOM

Cell

IOM

NOM

EOM

Cell

IOM

NOM

Fig. 6 e C-DBP formation in EOM, IOM, algal cells and NOM (5 mg/L as TOC) after chlorination and chloramination at pH 7.0 with the reaction time of 3 d.

after chlorination and chloramination, respectively. In general, in addition to the large chlorine consumption, due to high org-N contents in the algal materials, organic chloramine concentrations in chlorination and chloramination of EOM, IOM and algal cells were much higher than those of NOM. The organic chloramines formed in 2-h chlorination were unstable in the oxidative environment and thus they became undetectable after 1 d. The release of ammonia (Hawkins et al., 2003) from organic chloramine decomposition reacted with residual free chlorine to form inorganic chloramines, which showed maximum concentrations after 1 d. Transformation of preformed monochloramine to organic chloramines at concentrations of about 1.0 mg/L (as Cl2) was obvious, after 3-d chloramination of EOM, IOM and algal cells. Fig. 6 shows C-DBP formation after 3-d chlorination/ chloramination of EOM, IOM, algal cells and NOM. Most C-DBP yields, such as those of TCM, HAAs and HKs, were lower in chlorination/chloramination of EOM, IOM and algal cells than those of NOM. However, the production of haloacetaldehydes (HAs), such as chloral hydrate (CH) and dichloroacetaldehyde (DCA), was higher when the algal materials were the precursors. EOM formed less C-DBPs than did IOM and algal cells. Most trichloro C-DBP yields, such as those of TCM, CH and

1,1,1-TCP, in chloramination were much lower than those from chlorination. 1,1-DCP and DCA yields were higher in chloramination than in chlorination. Fig. 7 presents N-DBP formation after 3-d chlorination/ chloramination of EOM, IOM, algal cells and NOM. Concentrations of DCAN, CNCl and TCNM were much higher in chlorination of the algal materials than in chlorination of NOM. However, it was the opposite in chloramination. EOM formed smaller quantities of DCAN and CNCl but a larger quantity of TCNM than did IOM and algal cells. The 3-d NDMA formation in chlorination and chloramination of algal cells was 35.1 and 66.4 ng/L, respectively. The DBP formation from chlor(am) ination of IOM and algal cells was in similar quantity, which showed that the IOM retained most of the DBP precursors in algal cells. However, it should be noted that the characteristics of IOM depend on the extraction solvent (Her et al., 2004).

4.

Discussion

Org-N reacts with chlorine or monochloramine to form organic chloramines and it is the precursor of N-DBPs and C-DBPs (Lee and Westerhoff, 2009; Joo and Mitch, 2007). Amine

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30

Concentration (g/L)

25 20 15 10 5 0

Chlorination Chloramination

DCAN

Concentration (g/L)

CNCl

0 1.6 1.4 1.2 1.0 .8 .6 .4 .2 0.0

Concentration (g/L)

TCNM

EOM

Cell

IOM

NOM

Fig. 7 e N-DBP formation in EOM, IOM, algal cells and NOM (5 mg/L as TOC) after chlorination and chloramination at pH 7.0 with the reaction time of 3 d.

functional groups in aliphatic amines and free amino acids rapidly react with chlorine to form organic chloramines (Scully et al., 1994). Fig. 8 was drawn to illustrate the reaction pathways of these reactions, based on the general agreement in the literature (Hawkins et al., 2003; Hureiki et al., 1994; Scully et al., 1994; Joo and Mitch, 2007; Yang et al., 2010) and the results from this study, with amino acids taken as the model org-N in algal materials. Primary amines share similar pathways in chlor(am) ination, except for the omission of the decarboxylation step (Hawkins et al., 2003; Joo and Mitch, 2007). The formation of organic chloramines and inorganic chloramines during chlorination and chloramination of algal materials can be interpreted with Fig. 8. Different organic chloramines, including mono-(RNHCl) and di-chloramines (RNCl2), formed in chlorination, depend on the chlorine to orgN molar ratios. As shown in Table 1, DON concentrations in EOM, IOM and algal cells were 0.90, 1.11 and 1.15 mg/L (as N), and the initial molar ratios of chlorine to org-N were 3.29, 2.57 and 2.66, respectively. At these ratios, RNCl2 was the major form of organic chloramines in chlorination, which is much less stable than RNHCl (Vit and Barer, 1976) and can selfdecompose or be oxidized by chlorine. Therefore, organic chloramines were only found after 2-h chlorination. The RNCl2, after decarboxylation and the chlorine attack, forms TCNM (Joo and Mitch, 2007). It also undergoes decarboxylation to form nitriles or aldehydes, the latter with the loss of ammonia. The released ammonia reacts with excess free chlorine to form inorganic chloramines (Shang and Blatchley, 1999), such as mono-, di-, and tri-chloramines, depending on

the molar ratio of ammonia to free chlorine. Hence, inorganic chloramines were found after chlorination in the current study. Transferring chlorine from NH2Cl to org-N also formed organic chloramines in chloramination. Chlorination of org-N is known to produce haloaldehydes (e.g. DCA, CH), DCAN and CNCl (Joo and Mitch, 2007; Yang et al., 2010). The higher concentrations of these by-products in chlorination of algal materials than those of NOM chlorination can be explained by the higher concentrations of org-N, such as free amino acids and aliphatic amines in algal materials. As shown in Fig. 8, nitriles and aldehydes are the major intermediate products, the former react with chlorine to form halonitriles (e.g. DCAN and CNCl) and the latter react with chlorine to form haloaldehydes (e.g. DCA and CH). Therefore, the higher org-N concentrations in algal materials contributed to the formation of DCA, CH, DCAN and CNCl in larger quantities in chlorination. The org-N concentration in EOM was lower than that in IOM, which correlated with the lower yields of haloacetaldehydes (e.g. CH, DCA) and halonitriles (e.g. DCAN, CNCl) from EOM chlorination than from IOM chlorination. Formation of N-DBPs in chloramination showed an opposite trend, which was higher in quantities in chloramination of NOM than in chloramination of algal materials. As shown in Fig. 8, aldehydes react with NH2Cl to form N-DBPs, which is one major pathway for NH2Cl to contribute N to the formation of N-DBPs. Therefore, org-C, instead of org-N, is suggested to play an important role in the formation of these N-DBPs during chloramination. One isotopic study using model org-N compounds and NOM has shown that N in N-DBPs originated from both org-N and NH2Cl, and NH2Cl was sometimes the major contributor (Yang et al., 2010). The lower concentrations of some C-DBPs, such as TCM, HAAs and HKs, produced during chlorination and chloramination of algal materials, compared with those produced from NOM may be due to the fact that the org-N in algal materials consumes chlorine and hinders the reactions between chlorine and org-C to form these C-DBPs. The different C-DBP yields can also be explained by the different nature of the organic materials. As shown in Table 1, the SUVA values of the algal materials were much lower than that of NOM. SUVA is a good indicator of the aromatic content of organic matter and it has been found to correlate with THM and HAA formation in chlorination of organic matter (Reckhow et al., 1990). The pathways of NDMA and TCNM formation are less clear. NDMA was detected in chlorination and chloramination of algal cells, though one important precursor, dimethylamine (DMA), was not found in algal cells. It is likely that other org-N precursors, such as tertiary alkylamines or others, formed the DMA intermediate to contribute to the NDMA formation. It has been reported that TCNM formed in chlorination or chloramination of aliphatic amines and amino acids (Joo and Mitch, 2007; Hu et al., 2010). TCNM formation may involve chlorination of nitrite to form an intermediate, ClNO2, followed by its reactions with phenols (Thibaud et al., 1987) and humic acids (Choi and Richardson, 2004). The higher production of TCNM from chlor(am)ination of EOM than that of IOM and algal cells may be partially due to the high concentration of nitrite in the EOM solution. Results from this study reveal the importance of algal removal in drinking water treatment plants. Many solideliquid

w a t e r r e s e a r c h 4 4 ( 2 0 1 0 ) 5 8 9 7 e5 9 0 6

5905

Algal organic matter (N-organics)

COOH R-CH-NH2
+Cl2/NH2Cl

COOH R-CH-NCl2
Oxidation -CO2 -2HCl -CO2 +H2O -HCl -CO2 +H2O -NH3

COOH R-CH-NHCl
-HCl -CO2

R-CH2-NO2
+Cl2/NH2Cl

R- C N
+Cl2/NH2Cl (R=CH3) (R=H)

R-CH=NH
+H2O -RCHO

Cl Cl C-NO2 R
R=H

O DCAN C N-Cl R-C-H

+Cl2/NH2Cl Cl Cl +Cl2 O Cl C-C-H (CH) Cl Cl (R=CH3) O CH-C-H (DCA) +NH2Cl

NH3

Cl Cl C-NO2 Cl

R- C N

NH2Cl NHCl2 NCl3

+Cl2/NH2Cl (R=H)

DCAN

C N-Cl

Fig. 8 e Proposed pathways for the formation of organic chloramines and N-DBPs from chlorination or chloramination of algal organic matter.

separation techniques are available to remove algae, such as coagulation, clarication, media ltration, and microltration/ ultraltration. However, strong oxidants such as ozone and chlorine cause the release of IOM from algal cells into water. The algal materials then serve as the precursor of higher yields of CH, DCA, DCAN and CNCl in later chlorination or chloramination. Therefore, effective removal of intact algal cells by coagulation, clarication, media ltration and membrane ltration translates into effective control of DBPs in drinking water treatment.

haloaldehydes (e.g. DCA and CH), but smaller quantities of CDBPs (e.g. TCM, HAAs, and HKs) than did NOM. During chloramination, formation of most DBPs was smaller in quantities in EOM and IOM than in NOM. In general, EOM formed smaller quantities of DBPs (except for TCNM) than did IOM and algal cells, in chlorination and chloramination. It should be noted that the ndings were obtained with M. aeruginosa. Verication is needed to generalize the ndings to other algal species and other algal organic matter, with techniques demonstrated in this study.

Acknowledgements 5. Conclusions
This study was supported by the Natural Science Foundation of China under the scheme of National Creative Research Groups, 50821002. It was also supported by the Science and Technology Ministry of China (2008ZX07421-002) and the Education Ministry of China (705013). We gratefully acknowledge Drs. Hui Tao and Lei Yang for their help on some experiments.

EOM, IOM and NOM displayed different characteristics in carbon/nitrogen content, EEM uorescence spectra, MW and polarity distributions. EOM and IOM were nitrogen-rich materials and contained uorophores in the regions representing protein-like organics, primarily with MW over 10 kDa and of 70e1000 Da. The org-C in EOM and IOM consisted of relatively lower MW organic matter, with MW from several dozen to several hundred Da. Analysis of specic aliphatic amines and free amino acids showed that they constituted 2.5% and 11.3% of the TON in EOM and IOM, respectively. EOM, IOM and NOM had different DBP yields in chlorination and chloramination. During chlorination, EOM and IOM formed larger quantities of N-DBPs (e.g. DCAN, CNCl, and TCNM) and

Appendix. Supplementary material

Supplementary data associated with this article can be found in the online version, at doi:10.1016/j.watres.2010.07.009.

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