Lipid Recovery Research Articles

Recovery of Omega-3-Rich Lipids: Toward the Sustainable Valorization of Sea-Bass Industry Side Streams

This study pertains to the selection of the appropriate process parameters of lipid extraction aimed at the valorization of sea-bass gutting and filleting side stream biomass. A comparative kinetic study between an environmentally friendly polar (ethanol) and a conventional non-polar solvent (hexane) was implemented to determine the effect of temperature, solvent to solid ratio and time, providing insights in terms of the yield recovery and quality of omega-3 fatty acids. Maximum recovery yield (43.8 and 34.0 g/100 g of dried gutting and filleting by-products, respectively) was achieved within 10 min. In gutting by-products, ethanol achieved full lipid recovery at 35 °C and a 50:1 ratio, while for hexane, the maximum yield was 83%. In the case of filleting side-streams, hexane achieved exhaustive recovery at a 50:1 ratio and 20 °C compared with ethanol, where a higher temperature (50 °C) was needed for exhaustive recovery to be achieved. Differences were attributed to the diverse fatty acid profiles among the two by-products, with 27.3% and 40.8% polyunsaturated fatty acids in viscera and fish frames, respectively. All extracts at 20 °C and 35 °C remained below the oxidation limit set by the Codex Alimentarius Commission (peroxide value < 5 meqO2/kgoil; p-anisidine value < 20). Therefore, green solvents could assist conventional extraction, combining high efficiency and an improved environmental impact.

Valorization of fat balls and primary scum from wastewater treatment: a promising renewable lipid feedstock for biodiesel production.

We investigated the potential of waste materials from wastewater treatment plants (WWTPs) to serve as an alternative lipid feedstock for biodiesel production. The average lipid recoveries from fat balls (46.4%) and primary scum (49.5-54.5%) were higher than the lipid recovery of primary sludge (15.8-16.4%). The yield of biodiesel produced from the extracted lipids ranged from 5.7 to 20.1%. There were considerable site- and season-dependent variations in the characteristics of the lipid waste materials. Radiocarbon analysis indicated the presence of fossil-derived carbon (26.0-42.0%) in the biodiesel obtained from wastewater lipids. Finally, we estimated the potential for biodiesel production from WWTP-derived lipids; about 333.0 metric tons of biodiesel per year could be produced from fat balls and primary scum in Japan. The results indicate that lipid-rich materials from WWTPs represent a valuable alternative feedstock for biodiesel production.

Sustainable co-production of plant lipids and cellulosic sugars from transgenic energycane at an industrially relevant scale: A proof of concept for alternative feedstocks

Development of sustainable and scalable technologies to convert lignocellulosic biomass to biofuels is critical to achieving carbon neutrality. The potential of transgenic bioenergy crops as a renewable source of sugars and lipids has been demonstrated at bench-scale. However, scaling up these processes is important for holistic analysis. Here proof-of-concept for chemical-free hydrothermal pretreatment of transgenic energycane-oilcane line L13 at an industrially relevant scale to recover vegetative lipids along with cellulosic sugars is presented. Pilot-scale processing of 97 kg of transgenic energycane-oilcane L13 stems and high solids pretreatment of bagasse enhanced the recovery of cellulosic glucose and xylose by 5-fold as compared to untreated bagasse and helped in the enrichment of vegetative lipids in the biomass residues which allowed its recovery at the end of the bioprocess. Palmitic and oleic acids were the predominant fatty acids (FAs) extracted from stems and leaves. The processing did not affect lipid composition. The efficiency of lipid recovery from untreated biomass was 75.9% which improved to 88.7% upon pretreatment. The vegetative tissues of transgenic energycane-oilcane L13 contained 0.42 metric tons/hectare of lipids. Processing vegetative tissues yielded 0.38 metric tons/hectare of lipids. This approaches an oil yield similar to soybean (global average 0.44 metric tons/hectare) and is almost twice as high as the oil yield from sugarcane engineered to hyperaccumulate lipids (0.20 metric tons/hectare). The study suggests that further optimization by state-of-the-art metabolic engineering and biomass processing can establish transgenic bioenergy crops for commercial drop-in fuel production.

Comparative analysis of process intensification technologies (PIT) for improved cell disruption and lipid recovery in Aurantiochytrium sp. SW1 microalgae

SummaryThis study explores enhancing lipid recovery from the lipid‐rich marine microalgae Aurantiochytrium sp. SW1, known for its robust cell wall posing challenges, which necessitates energy‐intensive cell‐disruption for efficient intracellular lipid extraction. Despite advancements in other microalgae, there is lack of reports on efficient cell disruption for Aurantiochytrium sp. SW1 which is vital to reduce the energy costs and minimising the downstream process while maximising lipid yields. Three process intensification technologies (PIT) – microwave treatment, autoclave treatment and ultrasonication were employed on wet Aurantiochytrium sp. SW1 cells to improve the lipid extraction efficiency. Assessing the disruption effectiveness via lipid content, suspension turbidity and particle size distribution, alongside scanning electron microscopy (SEM) for cellular morphology changes post‐PIT treatments, this study indicates all three methods reduced particle size with prolonged treatment time, implying successful cell wall disruption and intracellular component release. Ultrasonication, especially at 50 W power for 10 min, exhibited the highest efficacy, yielding lipid content of 67.76 ± 0.03%. Microscopic analysis unveiled ultrasonication‐induced cell shrinkage and increased membrane permeability, supporting its effectiveness in enhancing solvent penetration for improved lipid recovery. This research underscores PIT techniques' potential as sustainable, energy‐efficient strategies for lipid extraction from Aurantiochytrium sp. SW1 microalgae.

The Plasma Lipidomic Landscape in Patients with Sepsis due to Community-acquired Pneumonia.

Rationale: The plasma lipidome has the potential to reflect many facets of the host status during severe infection. Previous work is limited to specific lipid groups or was focused on lipids as prognosticators.Objectives: To map the plasma lipidome during sepsis due to community-acquired pneumonia (CAP) and determine the disease specificity and associations with clinical features.Methods: We analyzed 1,833 lipid species across 33 classes in 169 patients admitted to the ICU with sepsis due to CAP, 51 noninfected ICU patients, and 48 outpatient controls. In a paired analysis, we reanalyzed patients still in the ICU 4 days after admission (n = 82).Measurements and Main Results: A total of 58% of plasma lipids were significantly lower in patients with CAP-attributable sepsis compared with outpatient controls (6% higher, 36% not different). We found strong lipid class-specific associations with disease severity, validated across two external cohorts, and inflammatory biomarkers, in which triacylglycerols, cholesterol esters, and lysophospholipids exhibited the strongest associations. A total of 36% of lipids increased over time, and stratification by survival revealed diverging lipid recovery, which was confirmed in an external cohort; specifically, a 10% increase in cholesterol ester levels was related to a lower odds ratio (0.84; P = 0.006) for 30-day mortality (absolute mortality, 18 of 82). Comparison with noninfected ICU patients delineated a substantial common illness response (57.5%) and a distinct lipidomic signal for patients with CAP-attributable sepsis (37%).Conclusions: Patients with sepsis due to CAP exhibit a time-dependent and partially disease-specific shift in their plasma lipidome that correlates with disease severity and systemic inflammation and is associated with higher mortality.

Sequential Extraction of Carbohydrates and Lipids from Chlorella vulgaris Using Combined Physical and Chemical Pre-Treatments

A key focus of microalgae-based fuels/chemicals research and development has been on the lipids that many strains generate, but recent studies show that solely recovering these lipids may not be cost competitive with fossil-derived processes. However, if the carbohydrates can also be recovered and ultimately converted into useful chemical intermediates, this may improve the economics for microalgae-based sustainable product technologies. In the present work, physical and chemical pre-treatments were performed on the Chlorella vulgaris microalgae strain to recover the carbohydrates from the biomass primarily in the form of glucose and galactose. The effects of temperature, acid concentration, microalgae solid-to-liquid loading, and hydrolysis time on carbohydrate hydrolysis and recovery was explored to identify optimum conditions. The highest recovery of total carbohydrates, 90 ± 1.1 wt% at 95% confidence which represents 40 wt% of the initial biomass, was obtained using temperature-assisted weak-acid extraction. Sequential extraction of carbohydrates and lipids was then explored. The highest recovery of total lipids was 71 ± 1.8 wt%, which represents 22 ± 0.9 wt% of the initial biomass. The sequential extraction of carbohydrates followed by lipids resulted in an overall recovery of 60 ± 1.6 wt% of the initial biomass, which is higher than current single product recovery strategies. These results suggest that adding carbohydrate recovery may be a viable strategy for overcoming a major economic hurdle to microalgae-derived chemical and fuel production by significantly increasing the yield of usable materials from microalgae biomass.

Neochloris oleoabundans cell wall rupture through melittin peptide: a new approach to increase lipid recovery.

Microalgae cell wall affects the recovery of lipids, representing one of the main difficulties in the development of biofuel production. This work aimed to test a new method based on melittin peptide to induce a cellular disruption in N. oleoabundans. Neochloris oleoabundans cells were grown at 32°C in the presence of a high concentration of nitrate-phosphate, causing a cell disruption extent of 83.6%. Further, a two-fold increase in lipid recovery following melittin treatment and solvent extraction was observed. Additionally, it was possible to verify the effects of melittin, both before and after treatment on the morphology of the cells. Scanning electron microscopy (SEM) and confocal images of the melittin-treated microalgae revealed extensive cell damage with degradation of the cell wall and release of intracellular material. Melittin produced a selective cell wall rupture effect in N. oleoabundans under some culture conditions. These results represent the first report on the effect of melittin on lipid recovery from microalgae.

Integrated municipal wastewater treatment and lipid accumulation by a self-flocculating/floating microalga Limnothrix sp.

The present study evaluated the growth, self-flocculation, lipid content, and pollutants removal by Limnothrix sp. BASMWW-9 isolated from municipal wastewater treatment system and cultivated in municipal wastewater. The biomass yield and lipid content after 6 days of cultivation were 1.07 g dw/L and 27.34 %dw, respectively. In addition, its self-flocculating ability reached up to 90 % after harvesting time of 180 min. Moreover, COD,NH3-N, TN, and TP removalefficiencies were 71.65 %, 81.89 %, 74.64 %, and 80.16 %, respectively. The self-flocculation performance of Limnothrix sp. was greatly associated to its morphology and production of extracellular polymeric substances (EPS), with significant positive impact of the high calcium and magnesium content in municipal wastewater. Interestingly, blue light irradiation during harvest enhanced the aggregation and floc formation as a floating biomat, which was attributed to enhanced polysaccharides production. This study provides innovative harvest method for Limnothrix sp. BASMWW-9 cultivated in wastewater using blue light for enhanced lipid recovery.

Application of adaptive laboratory evolution to improve the tolerance of Rhodotorula strain to methanol in crude glycerol and development of an effective method for cell lysis.

Rhodotorula toruloides can utilize crude glycerol as the low-cost carbon source for lipid production, but its growth is subjected to inhibition by methanol in crude glycerol. Here, transcriptome profiling demonstrated that 1004 genes were significantly regulated in the strain R. toruloides TO2 under methanol stress. Methanol impaired the function of membrane transport and subsequently weakened the utilization of glycerol, activities of the primary metabolism and functions of nucleus and ribosome. Afterwards the tolerance of TO2 to methanol was improved by using two-round adaptive laboratory evolution (ALE). The final strain M2-ale had tolerance up to 3.5% of methanol. 1 H NMR-based metabolome analysis indicated that ALE not only improved the tolerance of M2-ale to methanol but also tuned the carbon flux towards the biosynthesis of glycerolipid-related metabolites. The biomass and lipid titer of M2-ale reached 14.63±0.45 g L-1 and 7.06±0.44 g L-1 at 96 h in the crude glycerol medium, which increased up to 17.69% and 31.39%, respectively, comparing with TO2. Afterwards, an effective method for cell lysis was developed by combining sonication and enzymatic hydrolysis (So-EnH). The lytic effect of So-EnH was validated by using confocal imaging and flow cytometry. At last, lipid recovery rate reached 95.4±2.7% at the optimized condition.

Enhancing biodiesel production from urban sewage sludge: A novel industrial configuration and optimization model

In this work, a novel industrial configuration for the lipid extraction from sewage sludge was proposed. In detail, respect to conventional scheme based on direct extraction of lipids from wet sedimented sewage sludge using hexane, a preliminary centrifugation of sewage sludge prior to the extraction process was introduced. Economic and energetic balances for both configurations were evaluated using Aspen Plus simulator, including the post-treatment of exhausted sewage sludge obtained after the process. By centrifuging sewage sludge prior to the extraction process, overall costs can be reduced (27 % less), while preserving the same extraction efficiency for lipid recovery (80 %). Initial capital investments, amounts of reactants and total energy demand were sensibly reduced to achieve BEP of extracted lipids of 784 € ton-1 (vs 1.291 € ton-1 for conventional route). Finally, the economic feasibility of biodiesel production from lipids obtained from centrifuged sludge was evaluated by applying direct esterification with methanol and AlCl3·6H2O BEP of biodiesel produced of 1.186 € ton-1 was obtained with NPV of 27.5 M€ and DPP of 2.5 years. These values, show that biodiesel production from sewage sludge could be profitable considering only the proposed approach, fully unlocking the potential of waste in coherence with circular economy principles.

Curcumin inhibits the activity and induces apoptosis of activated hepatic stellate cell by suppressing autophagy.

Curcumin, a kind of natural compound, has been previously proven to inhibit the autophagy in hepatic stellate cells (HSCs) and induce their apoptosis. However, it is not clear whether the enhanced apoptosis of activated HSCs (aHSCs) caused by curcumin depends on autophagy inhibition. We aim to verify this hypothesis and explore the potential mechanisms in this study. Immortalized human HSC line LX-2 was used as an experimental specimen and pretreated with transforming growth factor β1(TGF-β1) for 24 h to activate it before drug application. The levels of autophagy, apoptosis, cell activity, lipid metabolism, and the activity of the PI3K/Akt/mTOR signal pathway were evaluated by multiple methods, such as Western blotting, mcherry-EGFP-LC3B adenoviruses transfection, immunofluorescence, Nile Red staining, flow cytometry among others. Our results showed that rapamycin, an autophagy activator, could partly offset the effects of curcumin on autophagy and apoptosis of LX-2 cells, while 3-Methyladenine (3-MA), an autophagy inhibitor, could enhance these effects. Furthermore, curcumin could promote the activity of the PI3K/Akt/mTOR signal pathway in LX-2 cells, while PI3K inhibitor could partly offset this effect and increase the autophagy level. Overall, we demonstrated that curcumin could inhibit the activity and promote LX-2 cells apoptosis by suppressing autophagy by activating the PI3K/Akt/mTOR signal pathway. In addition, lipid recovery and energy deprivation due to autophagy inhibition may be the exact mechanism by which curcumin attenuates the pro-fibrotic activity of LX-2.

Lipid recovery from deep eutectic solvents by polar antisolvents

Deep eutectic solvents (DES) can be a green alternative extraction technology for microalgae lipids, acting simultaneously as a pre-treatment agent and solvent. However, due to the low vapor pressure of both DES and lipid solute, the recovery of lipids and solvent regeneration remains difficult. In this study, we developed a novel strategy to separate the dissolved sunflower and model algae oil from imidazole/hexanoic acid DES by using polar antisolvents (water, methanol, and ethanol). The polarity and the amount of antisolvent influenced the lipid solubility in DES. While the water was the strongest antisolvent, the alcohols were easier to evaporate, ensuring easy DES regeneration. By adding small amounts of water and methanol, more than 90% of the lipids were recovered in the form of high-purity oils (>90%). In the case of ethanol, a large amount of ethanol was required, which diluted the solvent-rich phase and solubilized more lipids in it. Based on three repeated cycles with the selected antisolvent methanol, > 90% of the eutectic solvent could be regenerated.

Evaluation of Lipid Extraction Protocols for Untargeted Analysis of Mouse Tissue Lipidome.

Lipidomics refers to the full characterization of lipids present within a cell, tissue, organism, or biological system. One of the bottlenecks affecting reliable lipidomic analysis is the extraction of lipids from biological samples. An ideal extraction method should have a maximum lipid recovery and the ability to extract a broad range of lipid classes with acceptable reproducibility. The most common lipid extraction relies on either protein precipitation (monophasic methods) or liquid-liquid partitioning (bi- or triphasic methods). In this study, three monophasic extraction systems, isopropanol (IPA), MeOH/MTBE/CHCl3 (MMC), and EtOAc/EtOH (EE), alongside three biphasic extraction methods, Folch, butanol/MeOH/heptane/EtOAc (BUME), and MeOH/MTBE (MTBE), were evaluated for their performance in characterization of the mouse lipidome of six different tissue types, including pancreas, spleen, liver, brain, small intestine, and plasma. Sixteen lipid classes were investigated in this study using reversed-phase liquid chromatography/mass spectrometry. Results showed that all extraction methods had comparable recoveries for all tested lipid classes except lysophosphatidylcholines, lysophosphatidylethanolamines, acyl carnitines, sphingomyelines, and sphingosines. The recoveries of these classes were significantly lower with the MTBE method, which could be compensated by the addition of stable isotope-labeled internal standards prior to lipid extraction. Moreover, IPA and EE methods showed poor reproducibility in extracting lipids from most tested tissues. In general, Folch is the optimum method in terms of efficacy and reproducibility for extracting mouse pancreas, spleen, brain, and plasma. However, MMC and BUME methods are more favored when extracting mouse liver or intestine.

Valorization of palm oil mill effluent via enhanced oil recovery as an alternative feedstock for biodiesel production.

Residual oil from palm oil mill effluent (POME) can be valorized into value-added products like biofuel. However, the complex structure in POME limits the full recovery of intracellular lipids. To address this challenge, low-frequency ultrasonication was used as a pre-treatment prior to oil recovery to improve the yield by liberating the entrapped oil via the cell disruption technique. This study focused on optimizing the ultrasound conditions (i.e., ultrasonication amplitude, ultrasonication duration, and probe immersion depth) to maximize the improvement of oil recovery yield using response surface methodology. The optimized conditions were 30.074% ultrasonication amplitude, 0.167 min ultrasonication duration, and 2 cm probe immersion depth. This resulted in an additional 42.50% improvement in oil recovery yield over non-ultrasonicated POME, which is in close agreement with the model prediction. Additionally, a cost-benefit analysis was incorporated to determine the feasibility of ultrasonication for enhancing oil recovery. The study also explored the synthesis of biodiesel from POME-recovered oil and characterized the fuel attributes according to American Society for Testing and Materials- and European Standards-prescribed procedures. The attributes of biodiesel produced from POME-recovered oil are comparable to those of palm-based biodiesel in Malaysia, demonstrating its potential as an alternative source for biodiesel production.

Strategy for lipid extraction and transesterification to enhance Chlorella sorokiniana strain BV 12 feedstock utilization

Efficient production and recovery of fatty acid methyl esters (FAMEs) from lipid containing feedstock is the key step of biodiesel production. This enhances the utilization of feedstock and improves the final economics of the process. Currently, direct and indirect transesterification strategies are being employed for biodiesel production from microalgae with few reports of tailoring the process to a specific feedstock. In this study, Chlorella sorokiniana strain BV 12 was grown in modified BG-11 medium and biomass was harvested. This was subjected to different physical, chemical and physico-chemical strategies for increasing the transesterification yield. The transesterified lipid was evaluated for various biodiesel properties like iodine value, saponification value, acid value, peroxide value and cetane number. After harvesting, gravimetric lipid yield of 161.7 mg/g and 4117.5 µg/g of FAME were obtained. Extraction of lipid was dependent on the cell disruption technique as it is accumulated intracellularly as droplets. Sonication resulted in maximum FAME production using either dry (4724.2 µg/g) and wet (4835.8 µg/g) biomass which shows that wet biomass is more suitable for processing. Gravimetric results favour the extraction by sonication showing a 63.22% increase in lipid recovery. Testing of different solvents for extraction showed methanol as most favourable for FAME analysis with acidic catalyst like HCl and H2SO4 giving FAME with better ignition value (∼51, Cetane number).

Demonstration of n-dodecane suitability for milking lipids from Chlorella vulgaris for the production of biodiesel

In this research a biocompatible solvent was employed for milking of lipid from Chlorella vulgaris sp. in order to recover the microalgal lipid in-situ. Milking is an integrated approach for the simultaneous generation and removal of intracellular lipids within an aqueous biphasic system using the organic solvent dodecane. The purpose of this study was to investigate the viability of non-destructive lipid extraction from C. vulgaris for up to four cycles. The maximum amount of lipid was produced (1.069 ± 0.003 g L−1) in the first cycle, which utilized a solvent containing 20 % dodecane and had a maximum lipid recovery ratio of 38.33 ± 1.154 %. The maximum growth rate (0.832 ± 0.036 d−1) and dehydrogenase activity of cell (4.346 ± 0.280 OD492 g−1 cells) were also reported at 20 % dodecane. Dodecane, a biocompatible solvent, promoted lipid buildup in along with microalgal growth. The current findings lead to the possible proposal of a milking method using biocompatible organic solvents.

Optimizing the subcritical water valorization of insect (Hermetia illucens l.) farming waste for biodiesel production

To counter the proliferation of secondary waste streams generated on insect (black soldier fly) farms, the present study integrated green water technology and acid-catalyzed transesterification as a basis to assess further value extraction potentials. By valorizing the waste streams, a process was developed to promote the circular economy paradigm. In this regard, wastes generated from a local insect farm were initially subjected to subcritical water extraction (SWE) for lipid recovery. The SWE process was optimized such that an enhanced lipid yield of 13.31 wt% was obtained at temperature, time, and solid loading conditions of 236.8 °C, 10 min of extraction, and 1 g/ 100 mL respectively. The lipids recovered were then subjected to an acid-catalyzed transesterification reaction for the production of a fatty acid methyl ester (FAME) mixture. A preliminary economic assessment of the process was also undertaken. The results showed that the wastes had the potential to be economically employed in the production of biodiesel that satisfied fuel property standards. In addition, the potential of employing the side streams as an animal feed was also highlighted due to the determined dominance of oligopeptides, which are known for their favorable bioactive properties. The results indicated that insect farming waste is a promising renewable source for high-quality biodiesel and animal feed production through the environmentally friendly biorefinery.

Optimisation of ultrasound-assisted lipid extraction in the pretreatment of purple-spotted bigeye fish skin

The present work investigated the defatting of purple-spotted bigeye fish skin to obtain lipid prior to hydrolysis for gelatine production. Lipid was extracted using ultrasound-assisted extraction (UAE). The drying temperature of the skin on lipid recovery and preliminary kinetic of UAE were studied before a two-stage extraction was optimised by employing response surface methodology. Qualities of lipids and of the remaining solids after extraction were then analysed. The optimal drying temperature was found to be 80°C. The kinetic study that followed indicated that the highest extraction rate was achieved at 30°C, and from UAE using ethanol (UAEE) and sequential UAE using hexane (UAEH), the maximum percentage of total lipid recovery was 91.326 at 30°C, 37 kHz, and 60 min. The qualities determined by peroxide, conjugated diene, thiobarbituric acid reactive substances, and acid values showed that the extracted lipid could be suitable for industry, while the remaining solid could be used as material for gelatine production.

Lipid extraction and CO2 capture with switchable polarity solvent

In this study, lipid extraction and CO2 capture are combined using N, N-dimethylcyclohexylamine (DMCHA) as switchable polarity solvent. The effects of operation parameters are discussed according to the CO2 absorption/desorption and lipid/DMCHA recovery results. A triphasic models considering lipid, water, and gas phases are established to analyze the kinetic behaviors. The results show that DMCHA is reversible through CO2 absorption/desorption, and the enhanced dispersion of droplets and bubbles in water phase improves the lipid/DMCHA recovery and CO2 absorption/desorption. The triphasic kinetic models fit well with experimental data, and gas–liquid mass transfer is regarded as the rate-determining step. The lower interfacial areas result in the poorer gas–liquid mass transfer for the DMCHA recovery than lipid recovery process.

A rapid single-phase extraction for polar staphylococcal lipids.

The lipid membrane is gaining appreciation as a critical factor in the emergence of antibiotic resistance, both for antibiotics that target lipid synthesis or the membrane directly and for cell-wall-targeting antibiotics. The methods used to study the emergence of antibiotic resistance in vitro can generate a large number of samples that may be low in volume and in cell density. As in eukaryotic/mammalian lipidomics, two-phase liquid-liquid extractions are the most commonly used approach to recover lipids from bacteria. The need to separate the lipid layer is cumbersome for high-throughput applications and can be a source of poor reproducibility or contaminant introduction. While several single-phase extractions have been proposed for serum, tissue, and eukaryotic cells, there have been far fewer efforts to adapt or develop such methods for bacteria lipidomics. Here, we describe a simple, single-phase lipid extraction method based on methanol, acetonitrile, and water-the MAW method. The merits of the MAW method are evaluated against the Bligh & Dyer (B&D) method for the recovery of the major membrane lipids (phosphatidylglycerols, diglycosyldiacylglycerols, and lysyl-phosphatidylglycerols) in the Gram-positive pathogen Staphylococcus aureus. We demonstrate that the MAW method achieves recoveries that are comparable to that of the B&D extraction (≥ 85% for PG 15:0/d7-18:1). The benefits of the MAW method enable the detection of lipids from lower amounts of bacteria than the B&D method (0.57 vs 0.74 McFarlands for PG 32:0, respectively) and is easily scaled down to microplate volumes to facilitate high-throughput studies of bacterial lipids.