Trace determination of macrolide and sulfonamide antimicrobials, a human sulfonamide metabolite, and trimethoprim in wastewater using liquid chromatography coupled to electrospray tandem mass spectrometry

An analytical method has been developed and validated for the simultaneous trace determination of four macrolide antibiotics, six sulfonamides, the human metabolite N4-acetylsulfamethoxazole, and trimethoprim in wastewater. The method was validated for tertiary, secondary, and-unlike in previously published methods-also for primary effluents of municipal wastewater treatment plants. This wide range of application is necessary to thoroughly investigate the occurrence and fate of chemicals in wastewater treatment. Wastewater samples were enriched by solid-phase extraction, followed by reversed-phase liquid chromatography coupled to tandem mass spectrometry using positive electrospray ionization. Recoveries from all sample matrixes were generally above 80%, and the combined measurement uncertainty varied between 2 and 18%. Concentrations measured in tertiary effluents ranged between 10 ng/L for roxithromycin and 423 ng/L for sulfamethoxazole. Corresponding levels in primary effluents varied from 22 to 1450 ng/L, respectively. Trace amounts of these emerging contaminants reach ambient waters, since all analytes were not fully eliminated during conventional activated sludge treatment followed by sand filtration. In the case of sulfamethoxazole, the amount present as human metabolite N4-acetylsulfamethoxazole had to be taken into account in order to correctly assess the fate of sulfamethoxazole in wastewater treatment.     

Determination of steroid estrogens in wastewater by immunoaffinity extraction coupled with HPLC-electrospray-MS

A new method, based on immunoaffinity extraction coupled with liquid chromatography/electrospray mass spectrometry (LC/ESI-MS) is described for the determination of the steroid estrogens beta-estradiol (E2), estrone (E1), and alpha-ethynylestradiol (E2) in wastewater. The use of highly selective immunosorbents in sample preparation prior to analysis allows the removal of interfering sample matrix compounds present in the wastewater extracts that would otherwise cause severe ionization suppression of the estrogens during the electrospray process. In addition, immunoextraction removes much of the isobaric noise from the selected ion monitoring chromatograms, increasing the signal-to-noise ratios for analytes, and contributing to the low detection limits (0.18 and 0.07 ng/L for E2 and E1, respectively) achieved by the current method. The method was applied to analysis of estrogens in two wastewater effluents. Recoveries of E2 and E1 were excellent (>90%), while the nonimmunogen (but structurally related) analyte EE2 was not retained (recovery <2%) from effluent extracts by the immunosorbent. This illustrates the extreme selectivity of the immunoextraction purification step. Precision of the method was high, with relative standard deviations below 5%. Concentrations of E2 in wastewater varied from 0.77 to 6.4 ng/L, while concentrations of E1 were greater (1.6-18 ng/L).     

Identification and measurement of illicit drugs and their metabolites in urban wastewater by liquid chromatography-tandem mass spectrometry

Residues of illicit drugs and their metabolites that are excreted by humans may flow into and through wastewater treatment plants. The aim of this study was to develop a method for the determination of cocaine, amphetamines, morphine, cannabinoids, methadone, and some of their metabolites in wastewater. Composite 24-h samples from urban treatment plants were enriched with deuterated internal standards before solid-phase extraction. High-pressure liquid chromatography tandem mass spectrometry with multiple reaction monitoring was used for quantitation. Recoveries were generally higher than 80%, and limits of quantifications were in the low nanograms-per-liter range for untreated and treated wastewater. The overall variability of the method was lower than 10% for untreated and 5% for treated wastewater. The method was applied to wastewater samples coming from two treatment plants in Italy and Switzerland. Quantification ranges were found to be 0.2-1 microg/L for cocaine and its metabolite benzoylecgonine, 80-200 ng/L for morphine, 10 ng/L for 6-acetylmorphine, 60-90 ng/L for 11-nor-9-carboxy-Delta9-tetrahydrocannabinol, 10-90 ng/L for methadone and its main metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine, and lower than 20 ng/L for amphetamines. As previously reported for cocaine, this method could be useful to estimate and monitor drug consumption in the population in real time, helping social scientists and authorities to combat drug abuse.     

Trace analysis of antidepressant pharmaceuticals and their select degradates in aquatic matrixes by LC/ESI/MS/MS

Treated wastewater effluent is a potential environmental point source for antidepressant pharmaceuticals. A quantitative method was developed for the determination of trace levels of antidepressants in environmental aquatic matrixes using solid-phase extraction coupled with liquid chromatography-electrospray ionization tandem mass spectrometry. Recoveries of parent antidepressants from matrix spiking experiments for the individual antidepressants ranged from 72 to 118% at low concentrations (0.5 ng/L) and 70 to 118% at high concentrations (100 ng/L) for the solid-phase extraction method. Method detection limits for the individual antidepressant compounds ranged from 0.19 to 0.45 ng/L. The method was applied to wastewater effluent and samples collected from a wastewater-dominated stream. Venlafaxine was the predominant antidepressant observed in wastewater and river water samples. Individual antidepressant concentrations found in the wastewater effluent ranged from 3 (duloxetine) to 2190 ng/L (venlafaxine), whereas individual concentrations in the waste-dominated stream ranged from 0.72 (norfluoxetine) to 1310 ng/L (venlafaxine).     

Determination of polar drug residues in sewage and surface water applying liquid chromatography-tandem mass spectrometry

A simple and rapid method is presented for the trace-level analysis of 10 polar pharmaceutical residues in various types of water samples from the aquatic environment. Using this method, the pharmaceuticals and several drug metabolites can be analyzed in drinking and surface waters and in wastewater (treated and untreated sewage) at concentrations down to 0.01 microg/L. Samples are prepared by a simple in situ derivatization enabling the preconcentration of very polar metabolites by automated solid-phase extraction. The analytes were separated by liquid chromatography with tandem mass spectrometric detection and quantified by comparison with an internal standard. Limits of quantification were between 0.01 and 0.02 microg/L for three phenazone-type pharmaceuticals, six of their metabolites, and the antiepileptic drug carbamazepine. Except for dimethylaminophenazone, recoveries for all analytes were between 87 and 117% for raw and purified sewage, groundwater, and surface and drinking water. Investigations of some environmental samples revealed that sewage and surface water treatment causes a slight reduction of the concentrations of some analytes whereas other compounds were persistent during water treatment. Thus, some compounds were detected at the low-microgram per liter level in sewage effluents of wastewater treatment plants in Berlin (Germany) and were also found at high-nanogram per liter concentrations in Berlin surface water samples.     

Multiresidue determination of ultratrace levels of fluoroquinolone antimicrobials in drinking and aquaculture water samples by automated online molecularly imprinted solid phase extraction and liquid chromatography

The present work describes the development of a sensitive and highly selective innovative method for the simultaneous detection of six fluoroquinolone (FQ) antimicrobials (enrofloxacin, ciprofloxacin, norfloxacin, levofloxacin, danofloxacin, and sarafloxacin) in water samples. This detection is based on online solid phase extraction, coupled to liquid chromatography (LC), using for the first time tailor-made molecularly imprinted microspherical polymer particles prepared via precipitation polymerization. Various parameters affecting the extraction efficiency of the polymer have been optimized to reduce nonspecific interactions and to achieve selective uptake of the antibiotics from real samples. The method shows good recoveries ranging between 62% and 102% (V = 25 mL) for the different FQs tested and excellent interday and intraday precision with relative standard deviation (RSD) values between 2-5% and 2-6%, respectively. The detection limits were between 1-11 ng L(-1) (drinking water) and 1-12 ng L(-1) (fish farm water) when 25 mL samples were processed. The polymer showed selectivity for FQs containing a piperazine moiety whereas no retention was found for other antibiotics or nonrelated compounds. The method has been applied to the analysis of trace amounts of the FQs tested in drinking and fish farm water samples with excellent recoveries (>91%) and good precision (RSDs <5%).     

Determination of dissolved and adsorbed EDTA species in water and sediments by HPLC

This paper describes a method for determining EDTA species in various environmental samples at low molar concentrations by high-performance liquid chromatography (HPLC). Distinction between Fe(III)EDTA and all the other species can be made. NiEDTA can be detected semiquantitatively. The fraction of EDTA adsorbed to suspended particles or to sediments can be determined after desorption with phosphate. After complexation with Fe(III), the EDTA is detected by reversed-phase ion-pair liquid chromatography as the Fe(III)EDTA complex at a wavelength of 258 nm. The behavior of a variety of metal-EDTA complexes during analysis was checked. Determination of different EDTA species (Fe(III)EDTA, NiEDTA, and adsorbed EDTA) is possible in river water, groundwater, and effluents from wastewater treatment plants. Fe(III)EDTA was found to be the main species, at 30-70%; NiEDTA was <10% in most of the samples. Adsorbed EDTA was detected in suspended particles from rivers and wastewater treatment plants and in sediment cores from a lake. The method is suitable for a variety of different samples with different concentration ranges.     

Automatic searching and evaluation of priority and emerging contaminants in wastewater and river water by stir bar sorptive extraction followed by comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry

A new analytical method based on stir bar sorptive extraction (SBSE), followed by comprehensive two-dimensional gas chromatography (GCxGC-TOF-MS), has been developed for the automatic searching and evaluation of nonpolar or semipolar contaminants in wastewater and river water. The target compounds selected were 13 personal care products (PCPs), 15 polycyclic aromatic hydrocarbons (PAHs) and 27 pesticides. Excellent results have been obtained in terms of separation efficiency and also in terms of compound identification. Exceptional method detection limits were achieved applying the optimized method, at or below 1 ng/L for most of the compounds in real samples. The reliable confirmation of analyte identity was possible at this low concentration level, even for typically troublesome compounds such as the PAHs. The other validation parameters were good. In addition to obtaining analytical information such as identification and quantification of target analytes, it is also possible to screen for nontarget compounds or unknowns. New contaminants have been identified in the wastewater effluents and river water samples, such as cholesterol and its degradation products, pharmaceuticals, industrial products, other pesticides, and PCPs. The multidimensional information generated by the instrument can also be used by the researchers for contrasting samples and identifying, much more easily, the major differences between samples. We have used this feature to propose studies of comparison between the fingerprinting of different water samples, such as the contamination variation along a river affected by the discharge of urban wastewaters and also the contamination variation over a period of time in the effluent. Results show that the most frequently detected contaminants (and the contaminants detected at higher concentrations) were the PCPs. The musk fragrances galaxolide and tonalid were the most concentrated compounds in the samples. The pesticides and PAHs were present at much lower concentration than PCPs.     

Application of liquid chromatography/quadrupole-linear Ion trap mass spectrometry and time-of-flight mass spectrometry to the determination of pharmaceuticals and related contaminants in wastewater

This paper describes an enhanced liquid chromatography-mass spectrometry (LC-MS) strategy for the analysis of a selected group of 56 organic pollutants in wastewater. This group comprises 38 pharmaceuticals and 10 of their most frequent metabolites, 6 pesticides, and 2 disinfectants. The LC-MS methodology applied is based in the use of a hybrid triple-quadrupole linear ion trap mass spectrometer (QTRAP) in combination with time-of-flight mass spectrometry (TOF-MS). The join application of both techniques provided very good results in terms of accurate quantification and unequivocal confirmation. Quantification was performed by LC-QTRAP-MS operating under selected reaction monitoring (SRM) mode in both positive and negative electrospray ionization. Unequivocal identification was provided by the acquisition of three SRM transitions per compound in most of the cases and by LC-TOF-MS analysis, which allows obtaining accurate mass measurements of the identified compounds with errors lower than 2 ppm. Additionally, the use of TOF-MS permits retrospective analysis, since the full spectrum is recorded at all times with a high sensitivity. Thus, review of recorded chromatograms looking for new compounds or transformation products suspected to be present in the samples is feasible allowing one to increase the scope of the method along the monitoring program. The analytical performance of the quantitative LC-QTRAP-MS method was evaluated in effluent wastewater samples. Linearity of response over 3 orders of magnitude was demonstrated for most compounds (R(2) > 0.99). Method limits of detection were between 0.04 and 50 ng L(-1). Finally, the methodology was successfully applied to a monitoring study intended to characterize wastewater effluents of six sewage treatment plants in Spain. The presence of most of compounds was detected at concentrations ranging from 9 ng L(-1) (atrazine) to 15 microg L(-1) (paraxanthine).     

Analysis of androgenic steroids in environmental waters by large-volume injection liquid chromatography tandem mass spectrometry

A new method was developed for the analysis of natural and synthetic androgenic steroids and their selected metabolites in aquatic environmental matrixes using direct large-volume injection (LVI) high-performance liquid chromatography (HPLC) tandem mass spectrometry (MS/MS). Method accuracy ranged from 87.6 to 108% for analytes with well-matched internal standards. Precision, quantified by relative standard deviation (RSD), was less than 12%. Detection limits for the method ranged from 1.2 to 360 ng/L. The method was demonstrated on a series of 1 h composite wastewater influent samples collected over a day with the purpose of assessing temporal profiles of androgen loads in wastewater. Testosterone, androstenedione, boldenone, and nandrolone were detected in the sample series at concentrations up to 290 ng/L and loads up to 535 mg/h. Boldenone, a synthetic androgen, had a temporal profile that was strongly correlated to testosterone, a natural human androgen, suggesting its source may be endogenous. An analysis of the sample particulate fraction revealed detectable amounts of sorbed testosterone and androstenedione. Androstenedione sorbed to the particulate fraction accounted for an estimated 5 to 7% of the total androstenedione mass.     

Analysis of nitrosamines in wastewater: exploring the trace level quantification capabilities of a hybrid linear ion trap/orbitrap mass spectrometer

A method was developed to determine nine N-nitrosamines in wastewater on the basis of solid-phase extraction and liquid chromatography mass spectrometry using a linear ion trap-orbitrap hybrid instrument at high mass resolution. Analytes and five deuterated internal standards were preconcentrated by solid-phase extraction. Positive electrospray ionization resulted in protonated molecular ions of all nitrosamines. One to three product ions were formed by collision-induced dissociation or higher energy C-trap dissociation. The signal intensity of the product ions differed up to a factor of 3 between the two techniques. The molecular ions were usually used for quantification, because of the better sensitivity, and the product ions for confirmation. An actual mass resolving power of 25 000-40 000 ensured a sufficient selectivity to distinguish all molecular and product ions from interfering background ions. Only for N-nitrosomorpholine was a coeluting isobaric molecular ion detected in wastewater samples, which, however, formed different product ions. The mass accuracy was between -12 ppm at m/z 55 and 0 ppm at m/z 205 and did not change for more than 5 ppm over a sample sequence of 20 h analysis time. The optimized method allowed quantifying nine N-nitrosamines in drinking water and wastewater samples down to method detection limits of 0.3-3.9 ng/L at instrumental detection limits of 2-14 pg on column. Recoveries over the whole method were between 75 and 125% for six compounds, but considerably lower for three compounds, probably due to strong matrix effects causing a signal suppression of up to 95% in wastewater samples. N-Nitrosodimethylamine and N-nitrosomorpholine were the most abundant compounds (3-22 ng/L) in samples from two wastewater treatment plants, another four nitrosamines (N-nitrosopyrrolidone, -piperidine, -diethylamine, and -dibutylamine) were also detected. Our study demonstrates that the LTQ Orbitrap is a powerful instrument to quantify low molecular weight compounds at the picogram level in complex matrixes with both a high sensitivity and selectivity.     

Analysis of sulfonated naphthalene-formaldehyde condensates by ion-pair chromatography and their quantitative determination from aqueous environmental samples

An ion-pair solid-phase extraction (IPE), ion-pair chromatography (IPC) procedure with fluorescence detection for the quantitative analysis of sulfonated naphthalene-formaldehyde condensates (SNFC) was developed, which provides full resolution of SNFC up to a degree of condensation n = 5 and partial resolution up to n = 15. Liquid chromatography-electrospray ionization-mass spectrometry confirmed that SNFC elute in the order of condensation. Response factors in fluorescence detection proved to be mass-constant, thereby allowing us to determine total SNFC amounts. With this IPC method, the weight- and the number-average molecular weights of these high-volume production chemicals (kiloton per annum), used as synthetic tanning agents, concrete plasticizers, and dispersants, can be determined. Recoveries in IPE range from 73 to 85% in river Rhine water and from 79 to 93% in tap water for n = 2 to n = 7 with limits of detection of 3-8 ng/L for individual homologues from 500 mL of water. The IPE-IPC procedure was applied to samples of secondary industrial effluents, river Rhine water, a river bank filtrate, and a groundwater sample. SNFC up to n = 6 were detected in the treated effluents. Total concentrations ranged from 208 micrograms/L in a secondary treated SNFC production effluent to < 1.4 micrograms/L in groundwater. These first analyses suggest a widespread occurrence of the lower oligomers of SNFC in the aquatic environment.     

Determination of benzotriazole corrosion inhibitors from aqueous environmental samples by liquid chromatography-electrospray ionization-tandem mass spectrometry

The first method for the determination of commonly used corrosion inhibitors in environmental water samples by liquid chromatography-electrospray ionization-tandem mass spectrometry is presented. Benzotriazole (BTri) and the two isomers of tolyltriazole (5- and 4-TTri) are separated in an isocratic run. By gradient elution, BTri, 4-TTri, 5-TTri, and xylyltriazole can be determined simultaneously with three benzothiazoles, but here TTri isomers coelute. The instrumental detection limit of 2 pg allows the determination of the three most important benzotriazoles from municipal wastewater and most surface waters by direct injection into the HPLC system without previous enrichment. When solid-phase extraction is employed with mean recovery rates of 95-113%, the limit of quantification for benzotriazoles range from 10 ng/L in groundwater to 25 ng/L in untreated wastewater. BTri and TTri were determined in municipal wastewater in microgram per liter concentrations. Elimination in wastewater treatment appears to be poor, and BTri and TTri can be followed through a water cycle from treated municipal wastewater through surface water to bank filtrate used for drinking water production. The TTri isomers show markedly different biodegradation behavior with 4-TTri being more stable.     

Liquid chromatography-tandem mass spectrometry determination of nonionic organophosphorus flame retardants and plasticizers in wastewater samples

A comprehensive method for the determination of nonionic organophosphorus flame retardants/plasticizers in wastewater samples by solid-phase extraction (SPE) with liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) is presented. It allows the determination of 11 organophosphorus compounds, trialkyl and trichloralkyl phosphates, triaryl phosphate, and biphosphates together with triphenylphosphine oxide (TPPO). Limits of quantification after SPE of 100 mL of water are between 3 and 80 ng/L, which are adequate for most aqueous samples. The sensitivity of the LC-ESI-MS approach allows direct injection of aqueous sample without preceding extraction for concentrations in the low microg/L range. The method was finally applied to municipal wastewater samples, showing the occurrence of six phosphoric acid triesters and TPPO in both raw and treated municipal wastewaters.     

Determination of fluoroquinolone antibacterial agents in sewage sludge and sludge-treated soil using accelerated solvent extraction followed by solid-phase extraction

A method for the quantitative determination of humanuse fluoroquinolone antibacterial agents (FQs) ciprofloxacin and norfloxacin in sewage sludge and sludge-treated soil samples was developed. The accelerated solvent extraction was optimized with regard to solvents and operational parameters, such as temperature, pressure, and extraction time. A 50 mM aqueous phosphoric acid/ acetonitrile mixture (1:1) was found to be optimum in combination with an extraction temperature of 100 degrees C at 100 bar, during 60 and 90 min for sewage sludge and sludge-treated soil samples, respectively. A cleanup step using solid-phase extraction substantially improved the selectivity of the method. Overall recovery rates for FQs ranged from 82 to 94% for sewage sludge and from 75 to 92% for sludge-treated soil, with relative standard deviations between 8 and 11%. Limits of quantification were 0.45 and 0.18 mg/kg of dry matter for sewage sludge and sludge-treated soils, respectively. The presented method was successfully applied to untreated and anaerobically digested sewage sludges and sludge-treated soils. Ciprofloxacin and norfloxacin were determined in sewage sludges from several wastewater treatment plants with concentrations ranging from 1.40 to 2.42 mg/kg of dry matter. Therefore, contrary to what may be expected for human-use pharmaceuticals, FQs may reach the terrestrial environment as indicated by the occurrence of FQs in topsoil samples from experimental fields, to which sewage sludge had been applied.     

Determination of phosphoric acid mono- and diesters in municipal wastewater by solid-phase extraction and ion-pair liquid chromatography-tandem mass spectrometry

The first analytical method for the determination of 13 phosphoric acid mono- and diesters from aqueous samples is presented. The method consists of solid-phase extraction (SPE) and ion-pair liquid chromatographic separation with tri-n-butylamine coupled to electrospray ionization tandem mass spectrometry in the negative ion mode. Due to a lack of pure standards, only 3 of the 13 esters could be quantified. SPE recoveries ranged from 71 to 112% for di-n-butyl phosphate, diphenyl phosphate, and di-(2-ethylhexyl) phosphate (DEHP) with limits of quantification from 7 to 14 ng/L for 100-mL samples. At analyte concentrations >or=1 microg/L, aqueous samples can be analyzed by direct injection without extraction. In municipal wastewater, six diesters and one monoester were unambiguously identified by comparison with synthesized reference material. DEHP showed highest concentrations of 60 and 5 microg/L in raw and treated wastewater, respectively. The detection of monoethylhexyl phosphate was confirmed by LC-Q-TOF-MS analysis, and it was found at a concentration level comparable to DEHP. Laboratory degradation tests show that phosphoric acid diesters can be formed as intermediates in the microbial degradation of trialkyl phosphates that are being used as flame retardants and plasticizers.     

Quantitative analysis of polycyclic aromatic hydrocarbons in water in the low-nanogram per liter range with two-step laser mass spectrometry

Polycyclic aromatic hydrocarbons (PAHs) are of major concern in all environmental compartments due to the mutagenic and carcinogenic properties of many PAHs. Two-step laser mass spectrometry (L2MS) is a sensitive and selective method to measure PAHs in complex solid matrixes. However, in most studies, L2MS was used for qualitative or semiquantitative analyses. Here we present for the first time a quantitative method analyzing PAHs in water at the nanogram per liter level. PAHs are extracted from a 30-mL water sample with a solid PVC membrane, which is then directly measured by L2MS without further treatment. Detection limits are in the low-nanogram per liter range (2-125 ng/L) for skeletal three- to six-ring PAHs. Extraction efficiencies of this method are between 75 and 90%. In a first application, samples from a wastewater treatment plant were measured, showing that microbial activities efficiently decrease PAH concentrations by 75-90%.     

Determination of basic antidepressants and their N-desmethyl metabolites in raw sewage and wastewater using solid-phase extraction and liquid chromatography-tandem mass spectrometry

A novel analytical method has been developed for the determination of six basic antidepressants (venlafaxine, sertraline, paroxetine, citalopram, amitriptyline, and fluoxetine) and four of their metabolites (O-desmethylvenlafaxine, desmethylsertraline, nortriptyline, and norfluoxetine) in raw sewage and roughly primary-treated wastewater. For analytical development purposes, two ion exchange solid-phase extraction cartridges were compared. Extracts were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with positive-mode electrospray (+ESI) and selected reaction monitoring transitions. The choice of a basic mobile phase significantly improved the instrumental sensitivity (by up to 14-fold for norfluoxetine) relative to common +ESI acidic mobile phases. In addition to the remarkable gain in sensitivity, negligible matrix effects were also observed in the raw sewage samples. Analyte recoveries ranged from 80 to 103% and effluent detection limits from 0.048 to 0.10 ng/L. Samples collected at the Montreal Wastewater Treatment Plant showed the unequivocal presence of all the target compounds at concentrations of 2-346 ng/L. The target antidepressants were also detected in samples taken from the effluent receiving waters (i.e., the St. Lawrence River) but at lower concentrations (0.41-69 ng/L). The highly sensitive proposed method constitutes one of the best means for monitoring the environmental occurrence of tricyclic antidepressants, selective serotonin reuptake inhibitors (SSRIs), and some of their metabolites.     

Understanding the operational parameters affecting NDMA formation at Advanced Water Treatment Plants

N-nitrosodimethylamine (NDMA) can be formed when secondary effluents are disinfected by chloramines. By means of bench scale experiments this paper investigates operational parameters than can help Advanced Water Treatment Plants (AWTPs) to reduce the formation of NDMA during the production of high quality recycled water. The formation of NDMA was monitored during a contact time of 24h using dimethylamine as NDMA model precursor and secondary effluent from wastewater treatment plants. The three chloramine disinfection strategies tested were pre-formed and in-line formed monochloramine, and pre-formed dichloramine. Although the latter is not employed on purpose in full-scale applications, it has been suggested as the main contributing chemical generating NDMA during chloramination. After 24h, the NDMA formation decreased in both matrices tested in the order: pre-formed dichloramine>in-line formed monochloramine?pre-formed monochloramine. The most important parameter to consider for the inhibition of NDMA formation was the length of contact time between disinfectant and wastewater. Formation of NDMA was initially inhibited for up to 6h with concentrations consistently <10 ng/L during these early stages of disinfection, regardless of the disinfection strategy. The reduction of the contact time was implemented in Bundamba AWTP (Queensland, Australia), where NDMA concentrations were reduced by a factor of 20 by optimizing the disinfection strategy.