Folch Extraction Method Research Articles

Unwrapping the (glyco-)lipidome in the microalgae Microchloropsis gaditana: Effects of eco-friendly extraction methods

Microalgae have recently captivated worldwide attention because of their extraordinary lipid composition, which includes essential omega-3 and omega-6 long-chain polyunsaturated fatty acids (LC-PUFA), neutral lipids, and more complex lipids (glycolipids and phospholipids), as well as eicosanoids. To explore this further, we present a novel (glyco-)lipidomics strategy, including a green chemistry extraction approach, liquid chromatography high-resolution mass spectrometry (LC-HRMS) lipid profiling, and a comprehensive data evaluation workflow. Using untargeted lipidomics based on LC-HRMS approaches, we studied the lipid composition up to the molecular species level of the microalgae Microchloropsis gaditana (formerly Nannochloropsis gaditana), focusing on the distribution of saturated, monounsaturated, and polyunsaturated fatty acids in different lipid classes. Our analysis successfully identified 520 lipid molecular species from 18 lipid classes. Regarding fatty acid composition, our primary focus was on elucidating the distribution of LC-PUFA across the different microalgal lipid classes. This particular strategy holds potential as it may unravel the use of microalgal lipids as innovative sources of food ingredients. Our findings show that LC-PUFA are relatively elevated in membrane lipids, in which cholesterol esters (ChE) have the highest PUFA share (94 %), followed by polar membrane lipids (glycolipids and phospholipids), with phosphatidylethanolamines (PE) up to 67 %, phosphatidylglycerols (PG) up to 47 %, and monogalactosyldiacylglycerols (MGDG) up to 47 % PUFA content. This highlights the high percentage of PUFA in the structural parts of microalgae and the importance of these lipid classes as a reservoir for PUFA. Finally, we also compared a classical Folch extraction method with environmentally friendly ultrasound-assisted extraction approaches and enzymatic pre-treatments to investigate the lipid composition and enrichment of valuable lipid classes. Thus, this extensive analysis of the (glyco-)lipidome of Microchloropsis gaditana provides valuable insights into the existing knowledge about this microalgae species, enhancing our understanding of the complex lipid composition and opening up new opportunities for potential applications in the biotechnology and food industry.

Untargeted hair lipidomics: comprehensive evaluation of the hair-specific lipid signature and considerations for retrospective analysis.

Lipidomics investigates the composition and function of lipids, typically employing blood or tissue samples as the primary study matrices. Hair has recently emerged as a potential complementary sample type to identify biomarkers in early disease stages and retrospectively document an individual's metabolic status due to its long detection window of up to several months prior to the time of sampling. However, the limited coverage of lipid profiling presented in previous studies has hindered its exploitation. This study aimed to evaluate the lipid coverage of hair using an untargeted liquid chromatography-high-resolution mass spectrometry lipidomics platform. Two distinct three-step exhaustive extraction experiments were performed using a hair metabolomics one-phase extraction technique that has been recently optimized, and the two-phase Folch extraction method which is recognized as the gold standard for lipid extraction in biological matrices. The applied lipidomics workflow improved hair lipid coverage, as only 99 species could be annotated using the one-phase extraction method, while 297 lipid species across six categories were annotated with the Folch method. Several lipids in hair were reported for the first time, including N-acyl amino acids, diradylglycerols, and coenzyme Q10. The study suggests that hair lipids are not solely derived from de novo synthesis in hair, but are also incorporated from sebum and blood, making hair a valuable matrix for clinical, forensic, and dermatological research. The improved understanding of the lipid composition and analytical considerations for retrospective analysis offers valuable insights to contextualize untargeted hair lipidomic analysis and facilitate the use of hair in translational studies.

Возможности липидомного анализа семенной плазмы в прогнозировании успеха microTESE у мужчин с азооспермией

The aim of this work is to assess the possibility of metabolomic (specifically lipidomic) analysis of seminal plasma to identify patients with preserved focal spermatogenesis in the testes who may have a reasonable chance of sperm retrieval during the microTESE procedure.lipid composition of semen plasma samples from 64 men with azoospermia and 24 fertile control men was analyzed. Lipids were isolated from semen by a modified Folch extraction method. Lipid extracts were analyzed by reverse phase liquid chromatography coupled to mass spectrometry. Lipidomic data were compared with the results of the microTESE procedure.Comparison of two groups revealed a statistically significant difference in concentration for 23 lipids detected in positive ion mode and 37 lipids detected in negative ion mode. Those lipids mainly belong to hexosylceramides, sphingomyelins and phosphatidylcholines, phosphatidylethanolamines and their ethers. In multivariate analysis content of SM d16: 1/18:0 lipid (beta coefficient: -7.23; 95% confidence interval [95% CI]: -11.93 to - 2.53; odds ratio: 7.23e-04; CI for odds ratio: 6.59e-06 to 7.93e-02; Walds test: -3.02; p=0.003), content of TG 14: 1_16 : 0_18: 3 lipid (beta 2.95; 95% CI 0.98 to 4.93; odds ratio: 1.92e + 01; CI for odds ratio: 2.66e + 00 to 1.39e + 02 ; Walds test: 2.93; p=0.003) and testicular volume (beta: 0.14; 95% CI: 0.04 to 2.45; odds ratio: 1.15e + 00; CI odds ratio: from 1.04e + 00 to 1.27e + 00; Walds test: 2.65; p=0.008) were significantly associated with positive MicroTESE outcome. The sensitivity of this regression model was 61%, the specificity was 83%, and the AUC was 0.75.seminal plasma serves as a rich source of biological markers for identifying patients with preserved focal spermatogenesis in the testes. Seminal plasma lipidomic profile of the of patients in the control group with normal spermatogenesis clearly differs from the profile of patients with azoospermia, also there was a significant difference in content of a difference in lipids between patients with positive and negative microTESE outcomes. These are preliminary results and further research is needed to confirm the validity of the resulting lipid panel.

NMR spectroscopy as a “green analytical method” in metabolomics and proteomics studies

Nuclear magnetic resonance (NMR) is one of two methods (in addition to MS) that predominate in metabolomics and structural analysis. Due to the great demand for this type of research, there is a very high consumption of solvents. Fortunately, most biological studies can be performed using simple protocols that include 10% D2O and 90% H2O. More complex metabolomics studies with polar compound extraction require the use of MeOH or MeCN as reagents for protein precipitation. The most extensive Folch extraction method requires a mixture of solvents (MeOH/H2O/CHCl3) for tissue metabolites and sometimes biofluid extraction. In this review, we show that NMR can be a green method for most biological studies that yields useful information where the overall approach is in line with the GSK (GlaxoSmithKline) solvent sustainability guide.

Comprehensive and High-Coverage Lipidomic Analysis of Oilseeds Based on Ultrahigh-Performance Liquid Chromatography Coupled with Electrospray Ionization Quadrupole Time-of-Flight Mass Spectrometry.

Oilseeds are an important source of dietary lipids, and a comprehensive analysis of oilseed lipids is of great significance to human health, while information about the global lipidomes in oilseeds was limited. Herein, an ultrahigh-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry method for comprehensive lipidomic profiling of oilseeds was established and applied. First, the lipid extraction efficiency and lipid coverage of four different lipid extraction methods were compared. The optimized methyl tert-butyl ether extraction method was superior to isopropanol, Bligh-Dyer, and Folch extraction methods, in terms of the operation simplicity, lipid coverage, and number of identified lipids. Then, global lipidomic analysis of soybean, sesame, peanut, and rapeseed was conducted. A total of 764 lipid molecules, including 260 triacylglycerols, 54 diacylglycerols, 313 glycerophospholipids, 36 saccharolipids, 35 ceramides, 30 free fatty acids, 21 fatty esters, and 15 sphingomyelins were identified and quantified. The compositions and contents of lipids significantly varied among different oilseeds. Our results provided a theoretical basis for the selection and breeding of varieties of oilseed as well as deep processing of oilseed for the edible oil industry.

Quantitative Analysis of Polyphosphoinositide, Bis(monoacylglycero)phosphate, and Phosphatidylglycerol Species by Shotgun Lipidomics After Methylation.

Phospholipids play important roles in biological process even at a very low level. For example, bis(monoacylglycerol)phosphate (BMP) is involved in the pathogenesis of lysosomal storage diseases, and polyphosphoinositides (PPI) play critical roles in cellular signaling and functions. Phosphatidylglycerol (PG), a structural isomer of BMP, mediates lipid-protein and lipid-lipid interactions, and inhibits platelet activating factor and phosphatidylcholine transferring. However, due to their low abundance, the analysis of these phospholipids from biological samples is technically challenging. Therefore, the cellular function and metabolism of these phospholipids are still elusive. This chapter overviews a novel method of shotgun lipidomics after methylation with trimethylsilyl-diazomethane (TMS-D) for accurate and comprehensive analysis of these phospholipid species in biological samples. Firstly, a modified Bligh and Dyer procedure is performed to extract tissue lipids for PPI analysis, whereas modified methyl-tert-butylether (MTBE) extraction and modified Folch extraction methods are described to extract tissue lipids for PPI analysis. Secondly, TMS-D methylation is performed to derivatize PG/BMP and PPI, respectively. Then, we described the shotgun lipidomics strategies that can be used as cost-effective and relatively high-throughput methods to determine BMP, PG, and PPI species and isomers with different phosphate position(s) and fatty acyl chains. The described method of shotgun lipidomics after methylation achieves feasible and reliable quantitative analysis of low-abundance lipid classes. The application of this novel method should enable us to reveal the metabolism and functions of these phospholipids in healthy and disease states.

Optimization of a Method for the Simultaneous Extraction of Polar and Non-Polar Oxylipin Metabolites, DNA, RNA, Small RNA, and Protein from a Single Small Tissue Sample.

A more comprehensive picture of tissue biology can be obtained through the application and integration of multiple omic technologies. However, the common challenge in working with a precious sample is having a sample too small to separately extract analytes of interest for each experiment. Considering the high heterogeneity that can be present in a single tissue sample, extracting all biomolecules from a single and undivided tissue is preferable because it allows direct comparison of results. Here, we combined a modified Folch extraction method with DNA, RNA, small RNA, and protein extraction using two commercial kits, which allowed us to extract polar metabolites and non-polar oxylipin metabolites, DNA, RNA, small RNA, and protein simultaneously from a small tissue sample. The method was validated in terms of quantity and quality of analytes for downstream analyses.

Black soldier fly (Hermetia illucens L.): effect on the fat integrity using different approaches to the killing of the prepupae

The increase of waste, due to the rise of the world population, renders the use of the black soldier fly (BSF;Hermetia illucens) as agri-food leftover biotransformer very attractive. Indeed, feeding on these substrates, BSF is capable of transforming them into valuable fat, proteins, and chitin. The present study is aimed at evaluating different approaches to the killing of the prepupae in order to assess which is associated with the lowest production of artefacts, notably free fatty acids. Folch extraction method, via an Ultra-Turrax homogeniser, was selected to isolate and analyse prepupal fat. The same method was also applied for a direct grinding method to kill black soldier fly prepupae. The integrity of fat originated from this direct grinding approach to the killing was compared in terms of fatty acid profile with the samples obtained with two other killing procedures, i.e. freezing and blanching. Direct grinding proved to be a better preservative of fat integrity in terms of lowest amount of free fatty acids (about 2.5%) in comparison with freezing (about 15%), but also with blanching (about 10%). Furthermore, the direct grinding is a killing strategy that reduces energetic demand and process time and helps lower analytical costs.

Lipid Coverage in Nanospray Desorption Electrospray Ionization Mass Spectrometry Imaging of Mouse Lung Tissues

Lipids are a naturally occurring group of molecules that not only contribute to the structural integrity of the lung preventing alveolar collapse but also play important roles in the anti-inflammatory responses and antiviral protection. Alteration in the type and spatial localization of lipids in the lung plays a crucial role in various diseases, such as respiratory distress syndrome (RDS) in preterm infants and oxidative stress-influenced diseases, such as pneumonia, emphysema, and lung cancer following exposure to environmental stressors. The ability to accurately measure spatial distributions of lipids and metabolites in lung tissues provides important molecular insights related to lung function, development, and disease states. Nanospray desorption electrospray ionization (nano-DESI) and other ambient ionization mass spectrometry techniques enable label-free imaging of complex samples in their native state with minimal to absolutely no sample preparation. However, lipid coverage obtained in nano-DESI mass spectrometry imaging (MSI) experiments has not been previously characterized. In this work, the depth of lipid coverage in nano-DESI MSI of mouse lung tissues was compared to liquid chromatography tandem mass spectrometry (LC-MS/MS) lipidomics analysis of tissue extracts prepared using two different procedures: standard Folch extraction method of the whole lung samples and extraction into a 90% methanol/10% water mixture used in nano-DESI MSI experiments. A combination of positive and negative ionization mode nano-DESI MSI identified 265 unique lipids across 20 lipids subclasses and 19 metabolites (284 in total) in mouse lung tissues. Except for triacylglycerols (TG) species, nano-DESI MSI provided comparable coverage to LC-MS/MS experiments performed using methanol/water tissue extracts and up to 50% coverage in comparison with the Folch extraction-based whole lung lipidomics analysis. These results demonstrate the utility of nano-DESI MSI for comprehensive spatially resolved analysis of lipids in tissue sections. A combination of nano-DESI MSI and LC-MS/MS lipidomics is particularly useful for exploring changes in lipid distributions during lung development, as well as resulting from disease or exposure to environmental toxicants.

Effect of modified Soxhlet (Soxtec) and Folch extraction method selection on the total lipid determination of aged beef.

This study compared the modified Soxhlet (Soxtec) and Folch extraction methods for determining the total lipid or intramuscular fat (IMF) content of aged beef samples. Ageing periods tested were 0, 5, 8, 10 and 12weeks during which samples were held in vacuo and at ~ 1.0°C. Prior to solvent extraction, all samples were freeze-dried and ground. The Folch method was found to result in higher IMF values and therefore higher extraction efficiency when compared to the Soxtec. A linear relationship was evident between the two methods that described 80.9% of the variation between IMF values. An interaction between ageing period and extraction method was identified, although ageing period did not independently impact on averaged IMF findings. Based on these observations and reagent toxicity not being a limiting factor, it is recommended that the Folch extraction method be used to determine beef IMF, although it is acknowledged that the Soxtec method IMF results were strongly aligned with those found using the Folch extraction method and tended towards comparability for beef aged < 5weeks.

Inflammatory and Nutritional Biomarkers in the Plasma of Women in Zambia Reflect Low DHA and HIV Status

Docosahexanenoic Acid (DHA) is a fatty acid that is abundant in the brain, retina, and cerebral cortex. It is critical in fetal brain development and is implicated in resolution of inflammation. In recent years, there has been a growing concern that low dietary levels of DHA in pregnancy may lead to poor birth outcomes and delayed or reduced cognitive function in children after birth. Uncontrolled HIV infection may lead to cognitive impairments as well as reduced immune function. Therefore, for pregnant women living with food insecurity and HIV infection, these problems may be compounded and health risks to their infants high. To evaluate these concerns, a pre‐clinical trial was conducted in pregnant women in Zambia, a country where malnutrition and HIV are endemic. The study participants included 91 women in the second trimester of pregnancy, 46 of which were HIV seropositive and 45 HIV seronegative. Among the HIV positive participants 11 had high viral load. The overarching study goal was to determine if plasma DHA levels were low relative to global averages and if they correlated with inflammation and HIV status. Hematological assessment included measurement of biomarkers for nutritional status, HIV status and viral load, as well as inflammation. Plasma lipids were assessed using the Folch extraction method, fatty acid methyl esters were prepared and analyzed by gas chromatography/mass spectrometry. Protein biomarkers data were measured using commercial ELISA kits. Overall, DHA levels were low at 2.16±0.75 mass percent of total fatty acids but did not vary significantly with HIV status. Averages of the nutritional biomarkers insulin and leptin were low and high, respectively, compared to global averages when comparing all participants. Insulin was not significantly different based on HIV status. Leptin was reduced in HIV seropositive participants compared to those who were seronegative and this correlated, as expected, with pre‐pregnancy BMI. For the inflammatory markers CRP and LCN2, only LCN2 was significantly different dependent on HIV status, and levels were on average lower in seropositive individuals. Within the HIV positive participants, the 11 with high vial load did not have levels of insulin, leptin, CRP or LCN2 that were significantly different from those with low viral load. These data demonstrate that select protein biomarkers for inflammation and nutrition are variably affected by HIV and nutritional status. Currently, these studies are being expanded to understand the role of dietary fatty acids in contributing to health outcomes in pregnancy and dependent upon HIV status.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Fatty acids composition in South African freshwater fish as indicators of food quality

Lipid classes and fatty acid composition of three commercially important freshwater fish species Oreochromis mossambicus (Mozambique tilapia), Clarias gariepinus (African catfish) and Cyprinus carpio (carp) obtained from an aquaculture, different river systems and fish markets from different provinces in South Africa were investigated. Fatty acids were extracted from the fish fillets through the Folch extraction method (using chloroform: methanol at the ratio of 2:1). Generally, tilapia fish species was found to be the richest in fatty acid composition. In all fish species analysed, palmitic acid (16:0) was found to be the most abundant fatty acid ranging from 18.24 to 21.84%. Appreciable quantities of essential polyunsaturated fatty acid such as docosahexaenoic (DHA) (22:6 n-3, 3.92 to 6.16%), eicosapentaenoic acid (EPA) (20:5 n-3, 1.91 to 2.92%) and arachidonic acid (20:4 n-6, 7.19 to 8.50%) were also found. Observations show that fish species obtained from Gauteng Province are richer in fatty acids compared to those in Limpopo Province. The study points out that all fish species investigated contain appreciable levels of Omega-3 (n-3) polyunsaturated fatty acids (PUFA) and are therefore suitable for an unsaturated low-fat diet. This is important especially for poor communities who cannot afford to get a balanced diet, rich in some essential fatty acids.Therefore, it is important to determine the nutritional value of local fish, since it significantly contribute to a healthy diet in rural communities.

Sponges from North Borneo and their bioactivity against human colorectal cancer cells

Invertebrates are a major reservoir for numerous cytotoxic compounds that are used to defence themselves against prey and adaptation towards the environment. Throughout the years, numerous studies discovered from sponge extracts were effective against a wide range of cancer cells. In this study, 23 sponges comprising of some 19 species were collected from Northeast Borneo. Sponges were treated and extracted using modified Folch extraction method, followed by cytotoxicity assay to determine their effectiveness against different colorectal cancer cells. From the results, Monanchora clathrate, Dysidea sp., and Jaspis sp. were found to possess different degrees of cytotoxicity against wide range of human colorectal cancer cell. Monanchora clathrate (KDT07), Dysidea sp. (KDT09), and Jaspis sp. (KDT18) are among the demosponges which possess significant cytotoxicity against colorectal cancer cell lines, including HCT116, LoVo, SW480, and SW620. Besides, we found that environmental factors tend to alter chemical yield from the different species under the same genera. Among them, KDT08 and KDT21 which fall under same genus Dysidea, yet possess insignificant cytotoxicity against colorectal cancer cells. This research article provides a preliminary test for cytotoxicity activity of a wide range of marine sponges. Throughout this study, acquired results could provide useful information to determine the worthiness to further isolate nor purify the nature product from these sponges.

The potential use of infrared spectroscopy and multivariate analysis for differentiation of beef meatball from dog meat for Halal authentication analysis

Objective: The objective of this research was to assess the suitability of FTIR spectroscopy coupled with multivariate analysis of partial least square regression (PLSR) along with pattern recognition technique of principal component analysis (PCA) for rapid quantitative and qualitative (identification) analysis of dog meat in beef meatball formulation. Materials and Methods: The lipid fraction of meatball was obtained by employing two different extraction techniques, namely Bligh-Dyer and Folch method. FTIR spectral bands correlated with beef fat, pork fat, chicken fat and rat fat were measured, interpreted, and qualitatively analyzed. The small variations among spectra were exploited as a basis tools to differentiate between dog fat and other animal fats. Results : PCA at combined wavenumber regions of 1700-700 cm -1 was capable of identifying dog meat in meatball. These wavenumbers were also used for quantitative analysis of dog meat in meatball using PLSR model. Based on statistical parameters used, namely R 2 , RMSEC and RMSEP, Folch extraction method offered higher R 2 and lower RMSEC and RMSEP than Bligh-Dyer. PCA is succesfully applied for classification between meatball containing dog meat and other meats. Conclusion: FTIR spectroscopy coupled with multivariate analyses of PLSR and PCA was effective means for rapid screening of dog meat in meatball products. http://doi.org/10.5455/javar.2018.e281

Diagnostic performance of cytology for assessment of hepatic lipid content in dairy cattle

The objective of our study was to characterize the diagnostic performance of cytology for assessing hepatic lipid content (HLC) in dairy cows by comparing microscopic evaluation of lipid vacuolation in touch imprint slide preparations of liver biopsies with quantitative measurement of triglyceride concentration ([TG]; mg/mg of wet weight) in paired biopsy samples. Our study also sought to compare the diagnostic performance of liver cytology, plasma nonesterified fatty acid concentration ([NEFA]), and plasma β-hydroxybutyrate concentration ([BHB]) derived from a measurement performed on whole blood, for assessing HLC. Chemical extraction of TG from liver tissue remains the gold standard for quantifying HLC, largely because available blood tests, although useful for detecting some types of pathology, such as increased lipid mobilization, ketosis, or hepatocellular injury, are nonspecific as to etiology. Veterinary practitioners can sample bovine liver for cytological evaluation in a fast, minimally invasive, and inexpensive manner. Thus, if highly predictive of HLC, cytology would be a practical diagnostic tool for dairy veterinarians. In our study, liver biopsy samples from Holstein cows (219 samples from 105 cows: 52 from cows 2 to 20 d prepartum, 105 from cows 0 to 10 d in milk, 62 from cows 18 to 25 d in milk) were used to prepare cytology slides and to quantify [TG] using the Folch extraction method followed by the Hantzch condensation reaction and spectrophotometric measurement. An ordinal scale (0-4) based on amount of hepatocellular cytoplasm occupied by discrete clear vacuoles was used by 3 blinded, independent observers to rank HLC in Wright-Giemsa-stained slides. Interobserver agreement in cytology scoring was good. Corresponding plasma [NEFA] and [BHB] measurements were available for 187 and 195 of the 219 samples, respectively. Liver [TG] correlated more strongly with cytology score than with NEFA or BHB, and receiver operating characteristic curve analysis showed that cytology had better diagnostic performance than either NEFA or BHB for correctly categorizing [TG] at thresholds of 5, 10, and 15%. Hepatic lipidosis in high-producing dairy cows is of major clinical and economic importance, and this study demonstrates that cytology is an accurate means of assessing HLC. Additional work is indicated to evaluate the diagnostic utility of liver cytology.

Bitter gourd (Momordica charantia L.) seed oil as a naturally rich source of bioactive compounds for nutraceutical purposes

Abstract Background Characterization of food lipids has triggered the development of applications for the food and health industries. Thus, the lipid profiles of an increasing number of fruits and their seeds have been characterized and numerous bioactive components have been isolated. The bitter gourd seed oil has aroused great interest because it contains relevant amounts of conjugated fatty acid α-eleostearic acid (C18:3 9c11t13t), a positional and geometric isomer of α-linolenic acid. The aim of this paper was to evaluate the phytochemical composition and quality parameters of the seed oil of bitter gourd grown in Brazil. Methods Bitter gourd was purchased from CEAGESP (Brazilian company of fresh food storages and warehouses). Seeds were lyophilized, and their oil was extracted using the Soxhlet and Folch extraction methods. The profiles of fatty acids and phytosterols were analyzed by gas chromatography, and the acidity and peroxide values were evaluated by methods of the American Oil Chemists’ Society (AOCS). Results The Soxhlet extraction (40 % w/w) resulted in a higher yield of bitter gourd seed oil than the Folch extraction (16 % w/w). For both methods, α-eleostearic acid (56 and 58 %) was the major lipid in bitter gourd seed oil, followed by stearic acid (C18:0; 32 and 27 %). The oil displayed high content of phytosterols (886 mg/100 g), mainly β-sitosterol, and low acidity and peroxide values. Conclusions Bitter gourd seed oil from Brazil is an oil of good quality and its high contents of α-eleostearic acid and phytosterols with potential health-beneficial properties make it an attractive plant byproduct.

Abstract 202: Dysregulation of the Small Organic Carnitine Transporter and Carnitine in Heart Failure

Introduction: Advanced heart failure (HF) is characterized by metabolic abnormalities. We have recently shown myocardial lipid species to be significantly decreased in non-diabetic HF. The finding of deficient carnitine species has directed us to examine a key gene SLC22A5 the high-affinity carnitine transporter. Known mutations of this gene lead to primary carnitine deficiency and cardiomyopathy. Methods: Left ventricular samples procured at the time of orthotopic heart transplantation (OHT) from non-diabetic idiopathic dilated cardiomyopathy (DCM) n=16, and brain-dead organ donors without a history of diabetes or HF (NF) n=18 formed the study sample. All hearts received in situ cold cardioplegia. Lipids including carnitines were quantitated with liquid chromatography and high-resolution mass spectrometry (LC-HRMS) following the Folch extraction method. mRNA expression data of target genes including SLC22A5 from a separate study of n=117 DCM patients and n=67 NF patients p≤0.05 were analyzed. Results: Representative trends for all carnitine species assayed were shown to be significantly reduced in DCM. The figure below represents the mRNA expression between DCM and NF patients. There was a statistically significant reduction of SLC22A5 expression in the DCM population (Figure: ***p≤ 0.0001, dots 95 th percentiles). Conclusions: A statistically significant deficiency of carnitine species and a reduction in the expression of the carnitine transporter SLC22A5 was demonstrated in non-diabetic HF. Future studies will attempt to resolve the mechanisms behind these observations.

Fresh water green microalga Scenedesmus abundans: A potential feedstock for high quality biodiesel production

Present investigation studied the potential of fresh water green microalga Scenedesmus abundans as a feedstock for biodiesel production. To study the biomass and lipid yield, the culture was grown in BBM, Modified CHU-13 and BG-11 medium. Among the tested nitrogen concentration using Modified CHU-13 medium, the highest biomass and lipid yield of 1.113±0.05g/L and 489±23mg/L respectively was found in the culture medium with 0.32g/L of nitrogen (KNO3). Different lipid extraction as well as transesterification methods were also tested. Fatty acid profile of alga grown in large scale indigenous made photobioreactor has shown abundance of fatty acids with carbon chain length of C16 and C18. Various biodiesel properties such as cetane number, iodine value and saponification value were found to be in accordance with Brazilian National Petroleum Agency (ANP255) and European biodiesel standard EN14214 which makes S. abundans as a potential feedstock for biodiesel production.

Abstract 206: Intramyocardial Lipid Dysregulation in End Stage Heart Failure

Background: Advanced heart failure (HF) is characterized by metabolic abnormalities but studies conflict about the nature of these defects. Methods: We investigated the presence or absence of toxic intramyocardial lipid species using carefully procured human myocardium from lean, non-diabetic end-stage heart failure patients. Left ventricular myocardial samples were obtained at the time of heart transplant. Cases included idiopathic dilated (n=5) and ischemic (n=2) cardiomyopathy (n=7), and brain-dead organ donors without any history of heart failure (n=12). The failing hearts were examined before and after support with a left ventricular assist device (LVAD, n=7). Frozen samples were evaluated by liquid chromatography and high-resolution mass spectrometry (LC-HRMS) to identify lipid species and further quantitate their abundance. Samples for LC-HRMS were prepared using the Folch extraction method, and the samples were normalized to protein concentration via BCA protein assay. We selected for masses with p-values less than or equal to (p= .005) and a fold change greater than or equal to 2.5 Results: Diacylglycerols (DAGs), ceramides, sphingosines, and cardiolipins (CLs) were lower in failing samples than in non failing. The figure below for L-Palmitoylcarnitine is representative of the quantitative changes in lipids observed between failing and non-failing groups. Conclusion: Excess intramycellular lipid accumulation is not present in advanced heart failure. A significant deficiency in cardiac lipid species has been identified in end stage non-diabetic heart failure. Future studies in diabetic and obese patients with advanced heart failure are warranted.

Optimization of supercritical carbon dioxide removal of lipid and cholesterol from goat placenta using response surface methodology

Supercritical CO 2 (SC-CO 2) extraction was applied to remove lipid and cholesterol from freeze-dried goat placenta. A response surface methodology (RSM) was employed to optimize the extraction parameters. The effects of pressure, temperature, flow rate of CO 2 and extraction time on the yields of lipid and cholesterol were investigated. Response surface analysis showed that the data were adequately fitted to second-order polynomial model. The independent variables, quadratics of pressure and extraction time, and the interaction between pressure and temperature had significant effects on the yields of lipid and cholesterol, respectively. The optimum parameters within the experimental range of the variables were 34.6 MPa, 35.3 °C, 29.1 min with a CO 2 flow rate of 18.2 L/h. Under such condition, the yields of lipid and cholesterol were predicted to be 21.02% and 8.46 mg/g, respectively. Furthermore, the removal efficiency of cholesterol by SC-CO 2 was higher than those achieved by Soxhlet and Folch extraction methods.