Sub-ppm determination of perfluorinated carboxylic acids in solution by UV-vis high-performance liquid chromatography through solid phase extraction.

This study investigated a novel and sensitive analytical method based on a simple heat-based derivatization using 3-bromoacetyl coumarin as the reagent and analysis with a HPLC-UV system or just a UV-vis spectrometer to allow the sub-ppm determination of PFCAs in water solution with the potential for utilization in simple laboratories and field laboratory scenarios. A Strata-X-AW cartridge was used for the solid phase extraction (SPE) procedure and higher than 98% recoveries were obtained. The derivatization condition showed that a high efficiency of peak separation was obtained with obviously different retention time among various PFCAs derivatives using HPLC-UV analysis. The derivatization stability and repeatability showed favorable results with stable derivatized analytes for ≤12 h and a relative standard deviation (RSD) of <2% for all repetitions. The limit of detection for the HPLC-UV analysis was between 0.1 ppm and 0.5 ppm. A satisfactory linearity response was found with R2 >0.998 for all individual PFCA compounds. The limit of detection for simple UV-Vis analysis was <0.0003 ppm to measure the presence of PFCAs. Contamination of standards with humic substances and measurement of industrial samples in a complex wastewater matrix showed no adverse effects on the accuracy of PFCA determination by using the developed methodology.

Ozone combined with ceramic membranes for water treatment: Impact on HO radical formation and mitigation of bromate.

The beneficial effect of combining ozone with ceramic membrane filtration (CMF) to enhance membrane flux performances during water treatment (e.g., wastewater and drinking water) could be related to the formation of hydroxyl (HO) radicals from the interaction of ozone with ceramic membrane. To explore this effect, para-chlorobenzoic acid was used to probe HO radical activity during a combined ozone/CMF process using a 0.1 µm pore size membrane supplied by Metawater, Japan. Tests were then extended to explore the impact on bromate formation downstream CMF, a well-known undesirable by-product from ozone use in water treatment. Ozone reduction by the membrane and its module appeared to be more associated with physical degassing, but a noticeable formation of HO radicals was observed during the interaction of ozone with the ceramic membrane. CMF treatment of ozonated potable water containing bromide showed a reduced bromate formation of 50% when the water was recirculated to the filtration module containing the ceramic membrane, compared to the experiment performed with an empty module. Single pass experiments showed bromate mitigation of around 10%. The mitigation of bromate formation was attributed to reduced overall ozone exposure by deagassing effect, but also potentially from suppression of the oxidation of Br- and HOBr/BrO- to BrO3- due to the catalytic degradation of ozone via a HO radical pathway.

Computational models of populations of bacteria and lytic phage.

The use of phages to control and reduce numbers of unwanted bacteria can be traced back to the early 1900s, when phages were explored as a tool to treat infections before the wide scale use of antibiotics. Recently, phage therapy has received renewed interest as a method to treat multiresistant bacteria. Phages are also widely used in the food industry to prevent the growth of certain bacteria in foods, and are currently being explored as a tool for use in bioremediation and wastewater treatment. Despite the large body of biological research on phages, relatively little attention has been given to computational modeling of the population dynamics of phage and bacterial interactions. The earliest model was described by Campbell in the 1960s. Subsequent modifications to this model include partial or complete resistance, multiple phage binding sites, and spatial heterogeneity. This review provides a general introduction to modeling of the population dynamics of bacteria and phage. The review introduces the basic model and relevant concepts and evaluates more complex variations of the basic model published to date, including a model of disease epidemics caused by infectious bacteria. Finally, the shortcomings and potential ways to improve the models are discussed.

Nitrogen deprivation of microalgae: effect on cell size, cell wall thickness, cell strength, and resistance to mechanical disruption.

Nitrogen deprivation (N-deprivation) is a proven strategy for inducing triacylglyceride accumulation in microalgae. However, its effect on the physical properties of cells and subsequently on product recovery processes is relatively unknown. In this study, the effect of N-deprivation on the cell size, cell wall thickness, and mechanical strength of three microalgae was investigated. As determined by analysis of micrographs from transmission electron microscopy, the average cell size and cell wall thickness for N-deprived Nannochloropsis sp. and Chlorococcum sp. were ca. 25% greater than the N-replete cells, and 20 and 70% greater, respectively, for N-deprived Chlorella sp. The average Young's modulus of N-deprived Chlorococcum sp. cells was estimated using atomic force microscopy to be 775 kPa; 30% greater than the N-replete population. Although statistically significant, these microstructural changes did not appear to affect the overall susceptibility of cells to mechanical rupture by high pressure homogenisation. This is important as it suggests that subjecting these microalgae to nitrogen starvation to accumulate lipids does not adversely affect the recovery of intracellular lipids.

A quantitative analysis of microalgal lipids for optimization of biodiesel and omega-3 production.

The lipid characteristics of microalgae are known to differ between species and change with growth conditions. This work provides a methodology for lipid characterization that enables selection of the optimal strain, cultivation conditions, and processing pathway for commercial biodiesel production from microalgae. Two different microalgal species, Nannochloropsis sp. and Chlorella sp., were cultivated under both nitrogen replete and nitrogen depleted conditions. Lipids were extracted and fractionated into three major classes and quantified gravimetrically. The fatty acid profile of each fraction was analyzed using GC-MS. The resulting quantitative lipid data for each of the cultures is discussed in the context of biodiesel and omega-3 production. This approach illustrates how the growth conditions greatly affect the distribution of fatty acid present in the major lipid classes and therefore the suitability of the lipid extracts for biodiesel and other secondary products.

A chemical credit framework to predict the removal performance of organic chemicals of concern from water through an ozonation process.

Ozonation is an effective barrier for removing a wide spectrum of organic Chemicals of Concern (CoC) in a water treatment process. In this study, bench- and full-scale tests were conducted on a secondary treated effluent at the Eastern Treatment Plant (ETP) of Melbourne Water to probe the performance of ozonation in removing CoC in a wastewater discharge. From the secondary treated effluent as the feed to the ozone process, 58 organic chemicals were measured out of a possible 949 compounds by using the AIQS-DB analytical method. A chemical credit framework for the ozonation process has been established according to the bench-scale results. Chemical classifications based on the chemical structures (aromatics, aliphatic and halogenated aliphatic compounds) and reaction rate constants with O3 (KO3) and the ∙OH radical (K∙OH) and a combined O3/TOC ratio and O3 CT value as operating parameters were confirmed to be useful and important in determining whether a chemical would be removed by ozone. It is shown that an O3/TOC ratio of >0.404 was shown to be necessary to overcome the instantaneous ozone demand (IOD) to guarantee enough ozone to oxidise CoC. For CoC with KO3 >105M - 1s - 1 and K∙OH >109M - 1s - 1, an O3/TOC ratio of ≥0.461 or a measurable O3 CT of ≥0.063 mg min/L can achieve log reduction values (LRVs) of ≥1, these are chemicals with aromatic structures; For CoC with low KO3 and high KOH, a combined O3/TOC ratio and O3 CT value inclusive of a chemical structure classification is indicated as necessary criteria to evaluate the removal. UV254 and TOC were demonstrated to be good online surrogates of ozone barrier performance in the absence of continuous O3/TOC ratio and O3 CT value measurements. Full-scale operational results confirm the effective predictions of the chemical credit framework, which highlights the necessity and importance of monitoring both the O3/TOC ratio and O3 CT values to predict the removal efficiency of a given chemical with a known response to O3 or a known chemical structure.

Demonstrating removal credits for contaminants of emerging concern in recycled water through a reverse osmosis barrier-A predictive framework.

The performance of individual reverse osmosis (RO) systems varies significantly with different contaminants of emerging concern (CECs). As such, log reduction values (LRVs) of the concentration of these chemicals cannot be arbitrarily credited in water treatment and water recycling. This study looks to present an approach to the management of chemical risks by providing a systematic validation of RO barrier performance with respect to LRV credits for various classes of CECs. In this work, a one-off sampling campaign across five treatment barriers (strainer filtration, ultrafiltration, RO, ion exchange, chlorination) of a full-scale water recycling plant was conducted, followed by a systematic sampling campaign for a period of six weeks across just the RO barrier. The CECs screening methodology used GC-MS for quantification of 948 trace organic chemicals along with specific 44 per- and polyfluoroalkyl substances (PFAS) screening using LC-MS/MS to demonstrate the removal credits of the RO barrier to a wide spectrum of CECs. The work was used to validate an LRV barrier credit framework so as to predict the performance of a polyamide RO membrane for removal of a range of chemical classes, under typical operational conditions. Conductivity was validated as an efficient surrogate for membrane integrity and RO performance, along with specified operational conditions associated with permeate flux and recovery rate. A bioassay method (photobacterium test) showed good potential to be used as a quick measure to indicate the general toxicity of a sample caused by chemical contamination, because of its high detection sensitivity and time and cost efficiency.

Effects of solvency and interfacial nanobubbles on surface forces and bubble attachment at solid surfaces.

The forces between a carbon sphere and highly ordered pyrolytic graphite (HOPG) surface were measured in ethanol aqueous solutions. A long-range attractive interaction was observed in ethanol aqueous solutions with less than 60% ethanol on a volume basis. The magnitude and the range of the attractive interaction decreased with an increase in ethanol concentration in solution. An air bubble was found to attach to the HOPG surface in solutions with an ethanol concentration less than 20 vol %. The presence of nanobubbles at the HOPG surface, made possible through a surface washing process and confirmed through in situ imaging, extended the range of both the long-range attraction and the bubble attachment regime to ethanol concentrations 60-80 vol %. The shear yield stress of suspensions of carbon black in ethanol-aqueous solutions was used as an indicator of the presence of a net attractive interaction between particles. The yield stress was observed to increase rapidly for ethanol concentrations less than 60 vol %. Contact angle analysis for ethanol-aqueous solutions in air showed that an angle of greater than 40° was required to induce a long-range attraction. Modeling indicated that the range and form of the interaction was consistent with a capillary cavitation force. The data clearly shows that surface nanobubbles are not critical to the development of a long-range attractive force, bubble pickup on surfaces, or enhanced suspension rheology, but that their presence greatly aids these processes.

Interplay between interfacial behaviour, cell structure and shear enables biphasic lipid extraction from whole diatom cells (Navicula sp.).

HYPOTHESIS: Bacillariophyceae (i.e., diatoms) are an important class of algae with potential use in the production of proteins and lipids including long-chain ω-3 polyunsaturated fatty acids. Biphasic extraction of microalgae lipids using water-immiscible solvents such as hexane, can avoid the excessive energy required to distil solvents from water, but generally requires energy-intensive rupture of the cells. The unique cell structure and surface chemistry of diatoms compared to other microalgae species might allow biphasic lipid extraction without prior cell rupture. EXPERIMENTS: The kinetics of biphasic lipid extraction from intact Navicula sp. cells was investigated during low-shear and high-shear mixing, and with prior or simultaneous application of ultrasound (20 kHz at 0.57 W/mL). Dynamic interfacial tension measurements and electron microscopic analysis were used to investigate lipid extraction in relation to interfacial behaviour and cell structure. RESULTS: High yields (>80%) of intracellular lipids were extracted from intact cells over the course of hours upon low-shear contacting with hexane. The cells associated with and stabilised the hexane-water interface, allowing hexane to infiltrate pores in the frustule component of the cell walls and access the intracellular lipids. It was shown that mucilaginous extracellular polymeric substances (EPS) bound to the cell walls acted as a barrier to solvent penetration into the cells. This EPS could be removed by prior ultrasonication. Biphasic extraction was greatly accelerated by shear applied by rotor-stator mixing or ultrasound. High-shear could remove mucilaginous EPS from the cell surfaces to facilitate direct contact of the cell surface with hexane and produced smaller emulsion droplets with increased surface area. The combination of high-shear in the presence of hexane resulted in the in-situ rupture of the cells, which greatly accelerated lipid extraction and allowed high yields of neutral lipid (>95%) to be recovered from freshly harvested cells within less than 5 min. The study demonstrated the ability of shear to enable simultaneous cell rupture and lipid extraction from a diatom alga based on its cell structure and interfacial behaviour.

Wastewater treatment using filamentous algae - A review.

Wastewater treatment using algae is a promising approach for efficient removal of contaminating nutrients and their conversion into useful products. Monocultures of filamentous algae provide easier harvesting compared to microalgae, and better control of biomass quality than polyculture systems such as algal turf scrubbers. In this review, recent research into wastewater treatment using freshwater filamentous algae is compiled and critically analysed. Focus is given to filamentous algae monocultures, with key relevant findings from microalgae and polyculture systems discussed and compared. The application of monocultures of filamentous algae is an emerging area of research. Gaps are identified in our understanding of key aspects important to large-scale system design, including criteria for species selection, influence of nutrient type and loading, inorganic carbon supply, algae-bacteria interactions, and parameters such as pond depth, mixing and harvesting regimes. This technology has much promise, however future research is needed to maximise productivity and wastewater treatment efficiency.

A critical control point approach to the removal of chemicals of concern from water for reuse.

The reuse of water in a range of potable and non-potable applications is an important factor in the augmentation of water supply and in improving water security and productivity worldwide. A key hindrance to the reuse of water is the cost of compliance testing and process validation associated with ensuring that pathogen and chemicals in the feedwater are removed to a level that ensures no acute or chronic health and/or environmental effects. The critical control point (CCP) approach is well established and widely adopted by water utilities to provide an operational and risk management framework for the removal of pathogens in the treatment system. The application of a CCP approach to barriers in a treatment system for the removal of chemicals is presented. The application exemplar is to a small community wastewater treatment system that aims to produce potable quality water from a secondary treated wastewater effluent, however, the concepts presented are generic. The example used seven treatment barriers, five of which were designed and operated as CCP barriers for pathogens. The work demonstrates a method and risk management framework by which three of the seven barriers could also include a CCP approach for the removal of chemicals. Analogous to a CCP approach for pathogens, the potential is to reduce the use of chemical analysis as a routine determinant of performance criteria. The operational deployment of a CCP approach for chemicals was augmented with the development of a decision tree encompassing the classification of chemicals and the total removal credits across the treatment train in terms of the mechanistic removal of chemicals for each barrier. Validation of the approach is shown for an activated sludge, ozone and reverse osmosis barrier.

Conversion and recovery of saponifiable lipids from microalgae using a nonpolar solvent via lipase-assisted extraction.

A single-step method for transesterifying and recovering lipids in concentrated slurries (ca 20% w/w solids) of ruptured microalgae is presented. A soluble Rhizomucor miehei lipase (RML) was used to directly transesterify the lipids in the marine microalgae Nannochloropsis salina. This allowed both triglycerides (TAG) and polar saponifiable lipids to be recovered as fatty acid methyl esters (FAME) using a nonpolar solvent (hexane). Up to 90 wt% of the total saponifiable lipids (SL) were converted to FAME within 24 h, approximately 75% of which was recovered in the hexane by centrifugation. Two pathways for the conversion and recovery of polar lipids were identified. The water in the slurry buffered against potential lipase inhibition by methanol, but necessitated a high methanol dose for maximal FAME conversion. Nonetheless the method enables the recovery of polar lipids as FAME while avoiding the need for both drying of the biomass and a downstream transesterification step.

Emulsifying properties of ruptured microalgae cells: Barriers to lipid extraction or promising biosurfactants?

A systematic investigation of the emulsifying properties of ruptured algae cells was performed for the first time. The slurry of ruptured algae cells was separated into different biomass fractions, namely the cell debris, the delipidated debris, the serum, and the lipid. The interfacial interactions of these biomass fractions with a nonpolar solvent (e.g. hexane or hexadecane) were characterized using pendant drop tensiometry and interfacial shear rheology. The stability of the different emulsions (formed by the different biomass fractions) was tested using analytical centrifugation. The extracted lipid was an excellent surfactant that reduced the interfacial tension, however, it was not effective at stabilizing the emulsions. The protein-rich serum produced a strong interfacial film that stabilized the emulsions against coalescence during centrifugation. The cell debris stabilized the emulsions to a lesser extent by adsorbing to the droplet surface, presumably via interactions with hydrophobic extracellular polymeric substances (EPS). However, neither the serum nor the cell debris were very effective surfactants, and required the presence of the lipid fraction to produce small emulsion droplets. When present together, the components exhibited competitive interfacial adsorption, which influenced emulsion stability. In particular, the interruption of the protein film by the presence of lipid or cell debris reduced the stability of the emulsions. This study provides a new mechanistic understanding of emulsification during wet lipid extraction from microalgae that will be useful for determining strategies to improve solvent recovery. The results also suggest potential for developing effective bioemulsifiers or biosurfactants from fractionated microalgae biomass for commercial application.

Compartmentalization of extracellular polymeric substances (EPS) solubilization and cake microstructure in relation to wastewater sludge dewatering behavior assisted by horizontal electric field: Effect of operating conditions.

Sludge treatment and disposal have become important environmental issues in China. Mechanical dewatering is widely used to reduce the amount of sludge to be disposed and relieve the rapid growth pressure of waste sludge. In comparison to traditional sludge dewatering processes, pressure electro-osmotic dewatering has many advantages on sludge dewatering efficiency, low conditioner dosage and concentrated cake are both beneficial to further recycling of waste sludge. In general, complex electrochemical effects (eg. electrochemical oxidation, ohmic heating and pH gradient effect) are accompanied by the pressure electro-osmotic dewatering process. These electrochemical effects will inevitably cause solubilization and/or degradation of key constituents of wastewater sludge - extracellular polymeric substances (EPS). In this study, the effects of voltage, pH and ionic strength on sludge electro-osmotic dewatering performance and electrochemical effects were investigated. The solubilization and degradation of EPS were analyzed by examining the variation of dissolved organic matter (DOM) in the filtrate, and the relationships between microstructural properties of sludge cake and DOM and electro-osmosis dewatering performance in electro-dewatering process was examined. It was found that electro-dewatering properties were improved by raising the operating voltage or decreasing the pH value, while dewatering rate initially increased at low ionic strength it decreases with increased ionic strength. In addition, the porous structure of cathodic cake was more plentiful than that at the anode. At the cathode, the EPS dissolution was mainly related to alkalization, while the oxidation and acidification were responsible for release of EPS at the anode. Meanwhile, electrophoresis effect was able to promote migration of EPS toward the anode. The average electro-osmotic dewatering rate at the anode (R2.>0.79, p < 0.02) and at the cathode (R2.>0.87, p < 0.03) strongly correlated with the volume of pore of sludge cake. There was no correlation between the total content of anodic DOM (R2<0.31, p>0.08) and electro-osmotic dewatering rate at the anode, however, the content of cathodic DOM (R2 > 0.62, p < 0.09) negatively correlated with average electro-osmosis dewatering rate of cathode. Since cathode is the main water-permeable side in sludge electro-dewatering, and the sticky biopolymers (proteins and humic subtances) could not be converted into small molecules, higher EPS release was associated with worse sludge filterability. As for the anode, the biopolymers were degraded into small molecules due to electrochemical oxidation, which greatly reduced the impact of DOM on dewatering effect. Therefore, the operating conditions (voltage, pH and ionic strength) caused changes in electrochemical effects, which played a crucial role in compartmentalization of sludge EPS dissolution and consequently sludge electro-dewatering behavior.

The influence of protruding filamentous bacteria on floc stability and solid-liquid separation in the activated sludge process.

Filamentous bacteria can impact on the physical properties of flocs in the activated sludge process assisting solid-liquid separation or inducing problems when bacteria are overabundant. While filamentous bacteria within the flocs are understood to increase floc tensile strength, the relationship between protruding external filaments, dewatering characteristics and floc stability is unclear. Here, a quantitative methodology was applied to determine the abundance of filamentous bacteria in activated sludge samples from four wastewater treatment plants. An automated image analysis procedure was applied to identify filaments and flocs and calculate the length of the protruding filamentous bacteria (PFB) relative to the floc size. The correlation between PFB and floc behavior was then assessed. Increased filament abundance was found to increase interphase drag on the settling flocs, as quantified by the hindered settling function. Additionally, increased filament abundance was correlated with a lower gel point concentration leading to poorer sludge compactability. The floc strength factor, defined as the relative change in floc size upon shearing, correlated positively with filament abundance. This influence of external protruding filamentous bacteria on floc stability is consistent with the filamentous backbone theory, where filamentous bacteria within flocs increase floc resistance to shear-induced breakup. A qualitative correlation was also observed between protruding and internal filamentous structure. This study confirms that filamentous bacteria are necessary to enhance floc stability but if excessively abundant will adversely affect solid-liquid separation. The tools developed here will allow quantitative analysis of filament abundance, which is an improvement on current qualitative methods and the improved method could be used to assist and optimize the operation of waste water treatment plants.

Energy evaluation of algal cell disruption by high pressure homogenisation.

The energy consumption of high pressure homogenisation (HPH) was analysed to determine the feasibility of rupturing algal cells for biodiesel production. Experimentally, the processing capacity (i.e. flow rate), power draw and cell disruption efficiency of HPH were independent of feed concentration (for Nannochloropsis sp. up to 25%w/w solids). Depending on the homogenisation pressure (60-150 MPa), the solids concentration (0.25-25%w/w), and triacylglyceride (TAG) content of the harvested algal biomass (10-30%), the energy consumed by HPH represented between 6% and 110-times the energy density of the resulting biodiesel. Provided the right species (weak cell wall and high TAG content) is selected and the biomass is processed at a sufficiently high solids concentration, HPH can consume a small fraction of the energy content of the biodiesel produced. This study demonstrates the feasibility of process-scale algal cell disruption by HPH based on its energy requirement.

Quantification of wastewater sludge dewatering.

Quantification and comparison of the dewatering characteristics of fifteen sewage sludges from a range of digestion scenarios are described. The method proposed uses laboratory dewatering measurements and integrity analysis of the extracted material properties. These properties were used as inputs into a model of filtration, the output of which provides the dewatering comparison. This method is shown to be necessary for quantification and comparison of dewaterability as the permeability and compressibility of the sludges varies by up to ten orders of magnitude in the range of solids concentration of interest to industry. This causes a high sensitivity of the dewaterability comparison to the starting concentration of laboratory tests, thus simple dewaterability comparison based on parameters such as the specific resistance to filtration is difficult. The new approach is demonstrated to be robust relative to traditional methods such as specific resistance to filtration analysis and has an in-built integrity check. Comparison of the quantified dewaterability of the fifteen sludges to the relative volatile solids content showed a very strong correlation in the volatile solids range from 40 to 80%. The data indicate that the volatile solids parameter is a strong indicator of the dewatering behaviour of sewage sludges.

Particulate hematite diffusion in sodium polyacrylate solutions. The effect of ionic strength.

The diffusion coefficients of hematite particles in polyelectrolyte solution have been investigated using dynamic light scattering. Two apparent diffusion coefficients, a fast and a slow diffusional mode, are observed for the hematite particles in high-molecular-weight sodium polyacrylate solution at pH 10.5. The slow diffusion coefficient (Dslow) shows a decrease with increase in polyelectrolyte concentration. The fast diffusion coefficient (Dfast) shows an increase to a maximum with increasing polyelectrolyte concentration and then a rapid decrease as the polyelectrolyte concentration increases further. With an increase in ionic strength from 10(-4) to 0.1 M NaNO3, the maximum value of Dfast increased in magnitude, while the polyacrylate concentration at which the maximum occurs is seen to increase. The dependence of Dfast on the measurement angle indicates that it is coupled to the fluctuations of the chains. The observed behavior is attributed to the hematite probe particle sensing both macroscopic (viscous) and elastic fluctuations associated with the polyelectrolyte motion.

PAA/PEO comb polymer effects on rheological properties and inter-particle forces in aqueous silica suspensions.

The effects of a poly(acrylic acid) (PAA)-poly(ethylene) (PEO) comb polymer dispersant on the rheological properties and inter-particle forces in aqueous silica suspensions have been studied under varying pH conditions. The comb polymer was found to adsorb more strongly under acidic than basic conditions, indicating that the PAA backbone of the copolymer preferentially adsorbs onto silica surfaces with the PEO "teeth" extending out from the surface into the solution. In the presence of low concentrations of copolymer, the silica suspensions were stable due to electrostatic repulsions between the silica surfaces. At higher copolymer concentrations and under neutral and basic conditions, where the copolymer interacted only weakly with silica, the suspensions showed a transition from a dispersed to weakly flocculated state and attractive forces were measured between silica surfaces. Under acidic conditions, the silica dispersion also destabilized at intermediate copolymer adsorbed density and then was re-stabilized at higher adsorbed coverage. The silica suspensions were stable at high copolymer coverage due to steric repulsions between the particles. The destabilization at intermediate coverage is thought to be due to polymer bridging between particles or possibly depletion forces.

Lipid profile remodeling in response to nitrogen deprivation in the microalgae Chlorella sp. (Trebouxiophyceae) and Nannochloropsis sp. (Eustigmatophyceae).

Many species of microalgae produce greatly enhanced amounts of triacylglycerides (TAGs), the key product for biodiesel production, in response to specific environmental stresses. Improvement of TAG production by microalgae through optimization of growth regimes is of great interest. This relies on understanding microalgal lipid metabolism in relation to stress response in particular the deprivation of nutrients that can induce enhanced TAG synthesis. In this study, a detailed investigation of changes in lipid composition in Chlorella sp. and Nannochloropsis sp. in response to nitrogen deprivation (N-deprivation) was performed to provide novel mechanistic insights into the lipidome during stress. As expected, an increase in TAGs and an overall decrease in polar lipids were observed. However, while most membrane lipid classes (phosphoglycerolipids and glycolipids) were found to decrease, the non-nitrogen containing phosphatidylglycerol levels increased considerably in both algae from initially low levels. Of particular significance, it was observed that the acyl composition of TAGs in Nannochloropsis sp. remain relatively constant, whereas Chlorella sp. showed greater variability following N-deprivation. In both algae the overall fatty acid profiles of the polar lipid classes were largely unaffected by N-deprivation, suggesting a specific FA profile for each compartment is maintained to enable continued function despite considerable reductions in the amount of these lipids. The changes observed in the overall fatty acid profile were due primarily to the decrease in proportion of polar lipids to TAGs. This study provides the most detailed lipidomic information on two different microalgae with utility in biodiesel production and nutraceutical industries and proposes the mechanisms for this rearrangement. This research also highlights the usefulness of the latest MS-based approaches for microalgae lipid research.