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Fate of glutaraldehyde in hospital wastewater and combined effects of

glutaraldehyde and surfactants on aquatic organisms

Article in Environment International · May 2005

DOI: 10.1016/j.envint.2004.08.011 · Source: PubMed

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 Environment International 31 (2005) 399 – 406
www.elsevier.com/locate/envint

Fate of glutaraldehyde in hospital wastewater and combined effects of

glutaraldehyde and surfactants on aquatic organisms
Evens Emmanuela,b,*, Khalil Hannab, Christine Bazinc, Gérard Keckd,
Bernard Clémenta, Yves Perrodina
a
Laboratoire des Sciences de l’Environnement, École Nationale des Travaux Publics de l’État,
Rue Maurice Audin, 69518 Vaulx-en-Velin, France
b
Laboratoire d’Analyse Environnementale des Procédés et Systèmes Industriels, Institut National des Sciences Appliquées de Lyon,
20 avenue Albert Einstein, 69621 Villeurbanne Cedex, France
c
Insavalor-Division Polden, Institut National des Sciences Appliquées de Lyon, 20 avenue Albert Einstein, 69621 Villeurbanne Cedex, France
d
Unité d’Ecotoxicologie, Ecole Nationale Vétérinaire de Lyon, BP 83, 69280 Marcy l’Etoile, France

Received 18 March 2004; accepted 31 August 2004

Available online 13 October 2004

Abstract

Glutaraldehyde (GA), an aliphatic dialdehyde disinfectant, and surfactants, one of the major components of detergents, are widely used
in hospitals in order to eliminate pathogenic organisms causing nosocomial infectious diseases. After their use, disinfectants and
surfactants reach the wastewater network together. The discharge of chemical compounds from hospital activities into wastewater is also a
well-known problem, causing pollution of water resources and constituting an ecological risk for aquatic organisms. In this study, the
chemistry and toxicology of GA and surfactant mixtures were reviewed in order to estimate their fate in aquatic ecosystems. Furthermore,
their joint effects on aquatic organisms were experimentally assessed in the laboratory. A simple model of the additive joint action of
toxicants was used to determine combined acute toxicity effects on the bacteria luminescence and Daphnia mobility of three mixtures
containing GA at 1.5
EC50 24 h [in mg/L] on Daphnia and anionic, cationic and nonionic surfactants at twice their critical micellar
concentration (CMC). The mixture of GA and a cationic surfactant gave an EC50 30 min on Vibrio fischeri of 0.158%, with a
concentration of 0.04 mg GA/L and 1.04 mg CTAB/L, which provided an additive action. The interaction between GA and an anionic
surfactant on V. fischeri produced an antagonistic joint action with an EC50 30 min of 3.95%, containing 1.06 mg GA/L and 33.2 mg
SDS/L. A synergistic action with an EC50 30 min of 8.4% on V. fischeri was observed for the mixture containing GA and a nonionic
surfactant. Antagonistic interactions were observed for the joint action between GA and the surfactants studied on Daphnia. The mixture
of GA and CTAB was more toxic (EC50 24 h=0.02%) than the two other mixtures (EC50 24 h GA+SDS=6%; EC50 24 h GA+TX
100=10%). This study provides new data on the toxicity of certain hospital pollutants entering the aquatic environment and detected in
surface and groundwaters. It is necessary to study the joint effects of GA and surfactant mixtures following chronic and sublethal standard
bioassays in order to estimate the contribution of the additive joint action models in assessing the environmental risk of hospital
wastewater (HW).
D 2004 Elsevier Ltd. All rights reserved.

Keywords: Glutaraldehyde; Surfactants; Bioassays; Additive joint action; Hospital wastewater

1. Introduction

Disinfectants, highly complex products or mixtures of

* Corresponding author. Laboratoire des Sciences de l’Environnement,
École Nationale des Travaux Publics de l’État, Rue Maurice Audin, 69518
active substances (Kümmerer, 2001), and surfactants, sur-
Vaulx-en-Velin, France. Tel.: +33 4 72 04 72 89; fax: +33 4 72 04 77 43. face-active agents with a polar head group and a nonpolar
E-mail address: evemm1@yahoo.fr (E. Emmanuel). chain, are recognized as toxic to aquatic organisms
0160-4120/$ - see front matter D 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.envint.2004.08.011
400 E. Emmanuel et al. / Environment International 31 (2005) 399–406
(Schwarz and Vaeth, 1987; Talmage, 1994). As a result of
widespread nosocomial infectious diseases, i.e., diseases
contracted during hospitalisation, hospitals use large
amounts of disinfectants and detergents, in order to
eliminate pathogenic organisms (Leprat, 1998). Since
patients and medical staff spend most of their time indoors,
a healthy indoor environment in hospitals is critical to
maintaining human health (Berry, 1994). After their use,
disinfectants and surfactants reach the wastewater network
together (Kümmerer, 2001). The discharge of chemical
compounds from hospital activities into wastewater is also a
well-known problem, causing pollution of water resources
and ecological risks for aquatic organisms (Richardson and
Bowron, 1985; Gartisser et al., 1996; Kümmerer et al.,
1997; Halling-Sbrensen et al., 1998; Sprehe et al., 1999;
Emmanuel et al., 2001; Jolibois et al., 2002).
Polluted hospital wastewater generally contains hun-
dreds of chemicals. U.S. EPA (1989) has detected 400
toxic and hazardous pollutants in hospital wastewater. The
fate of several chemical substances released by hospital
activities has been reported in the literature, such as
analysis of radionuclides used in nuclear medicine
(Rodier, 1971; Erlandsson and Matsson, 1978), monitor-
ing of antibiotics in the aquatic environment (Hirsch et
al., 1999), release of disinfecting solutions containing
glutaraldehyde, a volatile, irritant and toxic chemical
compound (Jolibois et al., 2002), and surfactants (Singer
and Tjeerdema, 1993). However, very few studies have
dealt with the combined effects of pollutants contained in
hospital solutions. Indeed, the release of complex mixtures
by hospital departments may represent a real environ-
mental problem, thus it is very important to predict the
fate of hospital pollutants after their discharge into the
environment.
Chemical and physicochemical processes have been
identified as one of the important steps resulting from the
emission of a toxic substance into the aquatic environment,
that is to say the interaction of the substance discharged
with other constituents in water and the process by which
it becomes more or less available to organisms (Calamari
and Alabaster, 1980). By-products of chemical and
physicochemical processes are sometimes more toxic than
initial substances. Furthermore, the effect of a mixture of
chemicals can be simply additive, more than additive
(synergistic) or less than additive (antagonistic) (Calamari
and Alabaster, 1980; Hermens et al., 1984). Since
glutaraldehyde (GA) is a widely used disinfectant, we felt
it necessary to study the type of relation that this chemical
and surfactants may produce when present together in
hospital wastewater (HW). Three types of surfactants were
chosen for this study: sodium dodecyl sulfate (SDS—
anionic), cetyltrimethylammonium bromide (CTAB—cati-
onic) and Triton X-100 (TX-100—nonionic). The aim of
this study was: (i) to review the chemistry and toxicology
of GA and surfactant mixtures in order (ii) to estimate their
fate in an aquatic ecosystem, and (iii) to evaluate their
joint effects on aquatic organisms experimentally in the
laboratory.
2. Theory
2.1. Chemistry and toxicology of GA and surfactant
mixtures
2.1.1. Chemistry and toxicology of GA
Glutaraldehyde (GA) solutions, a cool sterilant, are used
in hospitals to disinfect and clean heat-sensitive equipment
such as dialysis instruments, surgical instruments, suction
bottles, bronchoscopes, endoscopes, and ear, nose and
throat instruments (HSDB, 1996; NIOSH, 2001). This
chemical is also used as a tissue fixative in histology and
pathology laboratories and as a hardening agent in the
development of X-rays (NIOSH, 2001). Table 1 shows
certain physical and chemical properties of GA.
GA is classified according to Appendix 1 of Council
Directive 67/548/EEC as T (toxic) and assigned as RT25
based on oral LD50 in rats of 100 mg/kg (EC, 1967). GA is
an active biocidal substance effective against aerobic and
anaerobic bacteria, moulds and yeast, and algae (FEI,
2001). The effects of GA on human beings have been
reported in the literature. Cases of proctitis (Burtin et al.,
1993), colitis (Asselah et al., 1996), and rectitis (Ledinghen
et al., 1996) have been observed in patients who underwent
medical examinations with materials disinfected with GA
that had been insufficiently rinsed. Due to its volatility and
irritating nature, occupational asthma has also been reported
among workers repeatedly exposed to glutaraldehyde
(Cullinan et al., 1992; Chan-Yeung et al., 1993; Stenton
et al., 1994; Gannon et al., 1995). Other evidence of the
toxicity of GA to humans is limited to reports of occupa-
tional exposure due to its use as a disinfectant and
sterilizing agent. Frequently observed effects of exposure
include skin sensitivity resulting in dermatitis, cutaneous
disorders like allergic eczema (Foussereau, 1985), and
irritation of the eyes and nose accompanied by rhinitis
(Jordan et al., 1972; Corrado et al., 1986; Hansen, 1983;
Wiggins et al., 1989).
Table 1
Physical and chemical properties of glutaraldehyde (HSDB, 1996)
Molecular formula C5H802
Structural formula (CHO-(CH2)3-CHO)
Molecular weight 100.12 g/mol
Melting point 14 8C
Boiling point 188 8C
Vapor pressure 0.6 Torr at 25 8C
Specific gravity 0.7 25 8C/25 8C
Water solubility 100 g/100 g H2O at 25 8C
Solubility soluble in water, alcohol, benzene
Henry’s law constant 1.1e7 atm/m3 mol
kOH 2.4e11 cm3/molecule s
log K ow 0.18
Conversion factor 4.1 Ag/m3 per ppb at 25 8C
 E. Emmanuel et al. / Environment International 31 (2005) 399–406
2.1.2. Chemical structures and toxicological effects of
surfactants
A surfactant molecule is amphiphilic, having two distinct
structural moieties, one polar and the other nonpolar. The
polar moiety of the molecule has an affinity for water and
other polar substances, while the nonpolar moiety is
hydrophilic (Tanford, 1980; Clint, 1992). Anionic, nonionic,
and cationic surfactants are widely used in the production of
cleaning products. These three main classes of surfactants
correspond to the charge of the polar portion of the
surfactant (Shinoda et al., 1963).
The presence of surfactants in the environment is
reported in the literature (Swisher, 1991). They were
identified in domestic and municipal sewage (Swisher,
1991), and in sediments (CCPCT, 2000). Some of these
compounds could be found in surface water and ground-
water (Tabor and Barber, 1996), and in drinking water
(CCPCT, 2000) due to their high polarity.
Some surfactants may irritate the human skin, mucous
membranes and eyes (Bartnik and Kunstler, 1987; Berry,
1994). Sodium dodecyl sulfate is listed as a skin sensitizer
(HSDD, 1996). The presence of surfactants in aquatic
ecosystems highlights a danger that can reach levels harmful
to aquatic life. What is more, the toxicity of surfactants on
the three first organization levels of food chains (algae,
crustacean, fish) has been established (Schwarz and Vaeth,
1987; Talmage, 1994).
2.2. GA partitioning behaviour
Information on the physical and chemical properties of a
compound, such as its octanol/water, air/water and surfac-
tant micelle/water partition coefficients, may help to
determine whether it is likely to concentrate in the aquatic,
terrestrial, or atmospheric environment (Valsaraj, 1995).
Certain equations can be used to calculate the partition
coefficients of GA based on their octanol/water distribution
coefficient (K ow). These parameters are determined in order
to estimate the proportion of GA that can partition with
phases (solid, micellar or air).
2.2.1. Solid/water
The adsorption of a compound on the natural organic
matter of soils can be estimated by calculating the organic
carbon/water partition coefficient (K oc). To do this, we used
the semiempirical correlation proposed by Valsaraj and
Thibodeaux (1989):
logKoc ¼ 0:92logKow  0:23 ð3Þ
Kd ¼ foc Koc ð4Þ
where K d is the solid/water distribution coefficient and f oc is
the fraction of organic carbon content of the sediment or
soil. According to Eq. (4), K d depends on K oc and the nature
of the soil, i.e., solid organic matter. Based on the K oc value
401
Table 2
Estimated partition coefficients of GA at 298 K
Compound log K ow log K oc log K mw K aw
GA 0.18 0.39 0.65 4.44
106
(Table 2), weak sorption of GA in the solid phase is
expected. Thus, glutaraldehyde is not considered to adsorb
on sediment in significant amounts except possibly as the
result of chemisorption in the biomass occurring as part of
its metabolism during rapid biodegradation (FEI, 2001).
2.2.2. Micelle/water
The theoretical approach described by Valsaraj (1995)
was used to estimate the partition coefficient of GA between
micelles (cationic, anionic or nonionic) and water:
logKmw ¼ 0:858logKow þ 0:807 ð5Þ
where, K mw=X m/X w is the micelle/water partition coeffi-
cient. X m: molar fraction in micelle and X w: molar fraction
in aqueous phase.
Depending on its micelle/water partition coefficient
(Table 2), the partition of GA with micelles is low, since a
large proportion of the molecules are not incorporated inside
the micelles.
2.2.3. Air/water
On the basis of Henry’s law constant (H), the air/water
partition coefficient can be determined by
P
vg
Hx ¼ H ð6Þ
RT
where v g is the partial molar volume of the gas phase, R is
the gas constant, T=298 K.
There are at least two ways for organic molecules to pass
into the atmosphere. The first is via an equilibrium
phenomenon: as the chemical potential of organic molecules
must be the same in water and in vapor at thermodynamic
equilibrium, molecules pass into the atmosphere until vapor
pressure is reached. The second cause is dynamic. Wind
stirring the water surface generates small drops, forming an
aerosol whose molecules can pass into the air phase.
It is interesting to state that the solution chemistry (pH,
ionic strength) and temperature of a given medium and the
presence of organic additives in it can influence the GA
partition coefficients (Table 2).
3. Experimental section
In this study, the acute toxicity of GA on aquatic
organisms was experimentally determined. A 50% commer-
cial GA solution (CEETAL Laboratories of France) was
used. The determination of the 24-h CE50 of GA 50% on
Daphnia magna, with three replicates of definitive assay,
402 E. Emmanuel et al. / Environment International 31 (2005) 399–406
was carried out to obtain an average value in mg/L for the
24-h EC50, represented in this study by the symbol bc mg/
LQ. The study of toxic effects of the GA and surfactant
mixtures on aquatic organisms (Vibrio fischeri, D. magna)
was carried out by using a certain number of experimental
solutions formulated for this study by following two
different steps. Experimental solutions were fabricated on
the day of the preliminary assay and used within 72 h.
Distilled water was used for the main solutions and
synthetic ISO medium was applied for the dilutions.
3.1. Step 1
This step was based on the determination of EC50 24 h
on Daphnia of the different substances at certain concen-
trations, i.e., GA at [1.5
c mg/L] and surfactants at twice
their CMC. SDS, CTAB and TX-100 (anionic, cationic and
nonionic surfactants) obtained from Sigma Aldrich were
used in this study. The physicochemical properties of these
surfactants are shown in Table 3.
3.2. Step 2
In this step, three experimental solutions containing GA
and surfactants were produced (Table 4) with the objective
of determining the EC50 of the different mixtures on the
aquatic organisms selected (V. fischeri and D. magna).
3.3. Toxicity test procedures
The bioassay on bacteria luminescence was carried out
with a LUMIStox system (Dr. Lange, Duesseldorf,
Germany) in conformity with the procedure of European
standard NF EN ISO 11348-3. The tests were performed
using gram negative marine bioluminescent bacteria of the
species V. fischeri NRRL-B-11177 of the Vibrionaceae
family. In order to prevent TSS interference on bacteria
luminescence, the samples were filtered using a 0.45 Am
pore size membrane. The samples were treated with 20 g/L
of NaCl solution and brought to 50 mS/cm conductivity
before analysis. Eight consecutive dilutions of the con-
centrated sample were tested (dilution factor 1:2); the
inhibition of bioluminescence was measured at a wave-
length of 490 nm, with readings after 5 and 15 min of
incubation at 15 8C.
Determining the inhibition of D. magna Straus mobility
was done by carrying out an acute toxicity assay with the
aim of identifying the initial concentration of a pollutant in
Table 3
A few physicochemical proprieties of surfactants used in this study (Shinoda et al., 1963)
Surfactant Symbol
Cationic (cetyltrimethylammonium bromide) CTAB
Anionic (sodium dodecyl sulfonate) SDS
Nonionic (Triton X-100) TX-100
Table 4
Characteristics of the experimental solutions
Mixture Composition of the mixtures
(1) GA+SDS GA50% at [1.5 c mg/L]+SDS at [0.84 g/L]
(anionic surfactant)
(2) GA+TX-100 GA50% at [1.5 c mg/L]+TX-100 at [0.37 g/L]
(nonionic surfactant)
(3) GA+CTAB GA50% at [1.5 c mg/L]+CTAB at [0.66 g/L]
(cationic surfactant)
solution or an aqueous mixture capable of immobilizing
50% of daphnia exposed to toxicants within 24 or 48 h. In
conformity with European standard NF EN ISO 6341, the
different assays were carried out on D. magna maintained
in parthenogenetic culture at the POLDEN laboratory
(National Institute of Applied Sciences of Lyon—INSA
de Lyon). The sensitivity of the laboratory species was
controlled by regular tests with potassium dichromate.
Only young female Daphnia aged less than 24 h were
used. There were 20 daphnia per concentration distributed
in four tubes. The normal medium, without EDTA, was
also used. The assays were carried out at 20F2 8C in the
dark.
The three conditions required for assay validity were
observed: (i) the concentration of dissolved oxygen (DO) in
the control group was z2 mg/L at the end of each assay; (ii)
the observed percentage of immobilization in the control
group vessels was V10%; (iii) 24 h EC50 of potassium
dichromate was from 0.6 to 1.7 mg/L.
3.4. EC50 calculations and analysis of combined toxicity of
mixtures
The EC50 values for the bioassay on bacteria lumines-
cence were calculated as reported by Bulich (1979). The
EC50 values for the Daphnia mobility inhibition assays
were determined by using the Litchfield–Wilcoxon statis-
tical method and probit analysis (Finney, 1971).
In this study, the combined toxicity of mixtures was
evaluated using the simplest model bTIQ Toxicity Index of
the additive joint action of toxicants in a mixture (Plackett
and Hewlett, 1948; Birky, 1976; Belkadir, 1979): the
interactions were determined by dividing the concentration
of the compound in the mixture at EC50 (mg/L) by the
EC50 of the compound and then totalled to obtain TI. If
TI=1, the interaction is additive; if TIb1, the interaction is
synergistic; and if TIN1, the interaction is antagonistic
(Belkadir, 1979).
Formula MW (g/mol) CMC (mmol/L)
[CH3(CH2)15N(CH3)3]Br 364 0.9
C12H25SO4Na 288.38 1.45
C8H17C6H4O(CH2CH2O)9.5H 625 0.27
 E. Emmanuel et al. / Environment International 31 (2005) 399–406 403

4. Results Table 6
Toxicity of mixtures on V. fischeri
4.1. Toxicity of GA and surfactants Mixtures EC50 30 Concentration of TI
min Vibrio compound in
fischeri mixture at EC50
The results shown in Table 5 summarize the acute (mg/L)
toxicity of single GA and single surfactants on the aquatic
GA Surfactant
organisms selected in this study.
GA+CTAB 0.158% 0.04 1.04 1
[26.85 mg GA/L+660
4.2. V. fischeri mg CTAB/L]
GA+SDS 3.95% 1.06 33.2 120.12
Generally, cationic and anionic surfactants are more toxic [26.85 mg GA/L+840
than nonionic surfactants (Rouse et al., 1994; Park and mg SDS/L]
GA+TX 100 8.4% 2.26 31.08 0.74
Bielefeldt, 2003). This information has been proved in this
[26.85 mg GA/L+370
study (Table 5). The EC50 on V. fischeri obtained for SDS is mg TX 100/L]
lower than the EC 50 determined for the two other
surfactants. Its represents 3% of the acute toxicity of GA
on the same organism. This result indicates that the toxicity The toxicity of surfactants towards invertebrates and
of a mixture containing SDS and GA is influenced only by crustaceans has been studied (Cserháti et al., 2002). The
the action of this surfactant. information in the literature reported that the sensitivity of
The acute and chronic toxicities of SDS towards micro- crustaceans towards SDS shows high variations: blue crab
organisms were compared with the toxicities of nonionic (Callinectes sapidus) 9.8 mg/L; grass shrimp 34 mg/L and
surfactants (Cserháti et al., 2002). The effects of SDS and mysids 48 mg/L (Whiting et al., 1996). The sensitivity of
Tween 80 on the growth of the cyanobacterium, Gloeo- aquatic invertebrates showed high variations according to
capsa, and its capacity to fix nitrogen is reported in the both the species and the type of surfactants. D. magna was
literature (Tozum-Calgan and Atay-Guneyman, 1994). Both found to be the most sensitive (Cserháti et al., 2002). Acute
growth and nitrogen fixing were inhibited at 50 ppm SDS toxicity values of surfactants on aquatic invertebrates varied
and 500 ppm Tween 80. The results clearly show that the from 1.7 to 270 mg/L for linear alkylbenzene sulfonate, 1.0
toxicity of SDS is considerably higher than that of the and 6.8 mg/L for nonionic alkylethoxylate, and 0.1 and 58
nonionic surfactant (Tozum-Calgan and Atay-Guneyman, mg/L for the cationic cetyltrimethylammonium chloride
1994). We also established that anionic surfactants influence (Lewis and Surprenant, 1983; Verge and Moreno, 2000).
the activity of microorganisms. Thus, they stimulate Among the acute toxicity values on Daphnia of the selected
respiration rate at low concentrations and inhibit it at higher surfactants obtained in the study, only the EC50 24 h (0.024
concentrations (Cserháti et al., 2002). Anionic surfactants mg/L) value for CTAB was outside the range of values
influence not only the enzyme’s respiration but also its reported in the literature. Indeed, CTAB appears to be more
activity and the growth of bacteria (Proksová et al., 1998). toxic for Daphnia than CTAC.
In this study, the results showed that SDS toxicity (0.276
mg/L) on V. fischeri, a marine species, was more toxic than 4.4. Toxicity studies of mixtures
the toxicity of both CTAB (1.02 mg/L), a cationic
surfactant, and TX-100 (63.6 mg/L) on the same organism. 4.4.1. V. fischeri
The results obtained for the acute toxicity of mixtures on
4.3. D. magna V. fischeri and the joint effects observed are summarized in
Table 6.
Of the four pollutants selected, CTAB had the highest An additive interaction was obtained for the interaction
acute toxicity on Daphnia. The EC50 24 h on Daphnia of GA and CTAB. This result is probably due to the fact that
obtained (0.024 mg/L) was only 0.1% of the acute toxicity both compounds are antimicrobial agents. However, it has
determined for GA. This result indicates that the toxicity of been demonstrated that solutions of GA can be inactivated
a mixture containing CTAB and GA is influenced only by by ammonium compounds (Gorman and Scott, 1980) or by
the action of this surfactant. reaction with sodium bisulfite (Jordan et al., 1996). The
joint action observed on GA and SDS mixtures indicated
Table 5 antagonistic action between these compounds. This infor-
Acute toxicities of GA and surfactants on V. fischeri and Daphnia mation confirms the prediction made on the nominal or
Parameters Units GA CTAB TX 100 SDS independent EC50 concentration of SDS on V. fischeri. The
EC50 30 min mg/L 9.06 1.02 63.6 0.276 only synergistic TI observed was for the GA and TX 100
Vibrio fischeri mixtures. This result could be attributed to the nominal EC50
EC50 24 h mg/L 19.03 0.024 38.05 29.21 concentration of this surfactant on V. fischeri. Indeed, of the
Daphnia
three surfactants studied, TX 100 revealed itself to be the
404 E. Emmanuel et al. / Environment International 31 (2005) 399–406

chemical with the lowest acute toxicity on bacteria
luminescence.
4.4.2. D. magna
The results obtained for the acute toxicity of mixtures on
Daphnia and the joint effects observed are presented in
Table 7. Antagonistic interactions were observed for the
joint action between GA and the surfactants studied.
5. Discussion
5.1. Fate and effects of glutaraldehyde
The results obtained from this study indicate that GA is
acutely toxic to aquatic organisms. The chemical’s inhib-
ition of bioluminescence was studied using a solution
containing 26.85 mg GA/L and in conformity with the
MICROTOX bioassay. An EC50 of 9.06 mg/L was obtained.
Besides bioluminescence inhibition, GA can also inhibit
the growth and metabolism of bacteria. Leung (2001a)
studied the aquatic metabolism of GA using a solution of
9.45 mg GA/L. The inhibition of bacteria was observed in
the solution after 4 h of incubation (2.5
103 CFU), showing
a decrease in comparison to the presiding measurement
(1
105 CFU). The metabolism of glutaraldehyde was quite
rapid under aerobic conditions, with a half-time of 10.6 h
based on the disappearance of the parent compound from
the water phase. Glutaraldehyde was ultimately metabolized
to CO2, achieving a yield of 68% after 30 days. Its
metabolism under anaerobic conditions was also rapid (t 1/2
of 7.7 h). However, anaerobic metabolism did not ultimately
result in methane but terminated with the production of 1,5-
pentanediol (Leung, 2001a).
Since these results on the metabolism of GA and its effects
on bacteria were achieved by using a solution of 9.45 mg
GA/L, and since the difference between this GA concen-
tration in this solution and the EC50 (9.06 mg GA/L) obtained
in the present study is insignificant, it seems evident that the
results of the acute bioassay on V. fischeri could permit
predicting organo toxicant effects not only on marine
ecosystems, but also on artificial wastewater ecosystems
(wastewater treatment) and plant and freshwater ecosystems.
The results obtained from the environmental portioning
studies (Table 2) indicate that glutaraldehyde tends to
remain in the aquatic compartment. Leung (2001b) noted
that GA has little tendency to bioaccumulate. Indeed,
bioaccumulation is a selective process by which a chemical
is concentrated in an organism in quantities greater than the
surrounding medium. The bioconcentration factor (BCF)
has been defined as the ratio of the concentration of a
chemical in the whole organism under steady-state con-
ditions, so it can be an indicator of the accumulation of a
chemical in the lipid compartment of an organism.
Bioaccumulation studies can be performed in situ by
measuring the chemical compound in an organism exposed
to effluents. Consequently, BCF can be easily determined by
a semiempirical first-order relationship between BCF and
log K ow reported by Neely et al. (1974):
BCF ¼ 0:542logKow þ 0:124 ð7Þ
Existing models of bioaccumulation and bioconcentra-
tion processes correlate well with the physicochemical value
of the partition coefficient of the compound in an octanol/
water system. Based on Eq. (7), the BCF calculated for GA
is 0.026. Thus, GA is not a bioaccumulate chemical in the
same way as the hydrophobic organic compounds (HOC).
As a biocide, this chemical is selectively toxic to a
variety of aquatic organisms. The NOEC for bacterial
inhibition has been determined at about 5 mg/L (UCC,
1994). Discharge of GA in excess of this level into a
wastewater treatment plant may inhibit the sewage micro-
organisms and adversely impact treatment performance
(Leung, 2001b). Concentrations of GA ranging from 0.50
to 3.72 mg/L have been detected in hospitals (Jolibois et al.,
2002); these concentrations were lower than its EC50 (9.06
mg/L) on V. fischeri and its EC50 24-h on D. magna. In the
absence of other toxicants, it appears that GA concentrations
in hospital wastewater may not have major effects on the
biological process of wastewater treatment.

5.2. Combined effects of GA and surfactants on aquatic

Table 7 organisms
Toxicity of mixtures on Daphnia
Mixtures EC50 24 h Concentration of TI The information on the toxicity of GA in hospital
Daphnia compound in mixture wastewater to aquatic life stems exclusively from laboratory
at EC50 24 h (mg/L) and field studies on this chemical (NICNAS, 1994; Jolibois
GA Surfactant et al., 2002). Information reported in the literature revealed
GA+CTAB 0.02% 0.005 0.132 5.55 the presence of other substances in hospital effluents that
[26.85 mg GA/L+660 may contribute to a combined toxic action with GA. In
mg CTAB/L]
France, various solutions containing GA and surfactants are
GA+SDS 6% 1.61 50.4 1.81
[26.85 mg GA/L+840 used in hospital activities. In this study, the acute toxicity on
mg SDS/L] Daphnia of Pyosynthene EA 20R, a disinfectant solution
GA+TX 100 10% 2.69 37 1.11 containing GA and a cationic surfactant widely used in
[26.85 mg GA/L+370 France to disinfect surfaces and medical equipment, was
mg TX100/L]
carried out and an EC50 24 h of 1.4 mg/L was obtained.
 E. Emmanuel et al. / Environment International 31 (2005) 399–406
The concentrations of compounds in the three mixtures at
EC50 (mg/L) showed that all the surfactants’ toxicities were
lower than their CMC. This information proved the absence
of micelles and permitted the assumption that the majority
of GA molecules were in aqueous phase.
Contrary to GA, the toxicity of mixtures containing
surfactants has received considerable attention since they
are present in several combinations in commercial deter-
gents and household cleaning products (Lewis, 1992).
Surfactants have been more frequently combined in toxicity
tests with pesticides, metals, oil and other surfactants
(Turner et al., 1985; Versteeg and Woltering, 1990). The
information observed in this study shows that the surfactants
associated with GA used in hospitals have several consid-
erable acute toxicity effects on aquatic organisms (Tables 6
and 7). The results indicate that the acute toxicity of the
mixtures is greater than the sum of the simple acute toxicity
of the different chemicals constituting the mixture.
When formulating water criteria, No Observed Effect
Concentration (NOEC) is of more interest than EC50
values (Hermens et al., 1984). In this study, the different
joint toxicity effects determined were calculated using the
results of acute toxicity tests. It would be necessary to
study the joint effects of GA and surfactant mixtures
according to chronic and sublethal standard bioassays, in
order to estimate the adequacy of additive joint action
models in assessing the environmental risks of hospital
wastewater.
6. Conclusion
The different partition coefficients estimated for GA in
environmental compartments prove the high solubility of
this chemical in water. GA, SDS, CTAB and TX-100
(anionic, cationic and nonionic surfactants) showed acute
toxicity to V. fischeri and Daphnia. The results calculated
and determined for GA were close to the available data
reported in the literature. Due to the similarity between the
results of this study and the data in the literature, the toxic
index obtained for interactions between GA and surfactants
could be helpful for assessing the real environmental risk
and life cycle of these hospital pollutants. Since No
Observed Effect Concentration (NOEC) is of more interest
than EC50 acute toxicity in the formulation of water criteria,
it will now be necessary to study the joint effects of GA and
surfactant mixtures according to chronic and sublethal
bioassay standards.
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