Bligh-Dyer Method Research Articles

Rapid and simple dual extraction for the analysis of lipids and autoantigenic peptides within phosphatidylserine-liposomes

Autoimmune diseases are a major health concern in developed countries. Currently, only palliative treatments based on anti-inflammatories and immunosuppressors are available. A novel antigen-specific therapy that uses a physiological process of tolerance generation is being developed. This is plausible by using phosphatidylserine rich liposomes (PS-liposomes), which bio-mimic apoptotic cells, encapsulated with the autoantigen responsible of generating the autoimmunity. In this way, tolerance against the own cells or tissues that were considered hostile can be achieved. In addition, only by changing the encapsulated peptide, different autoimmune diseases can be treated. Efficacy of this approach was demonstrated in type I diabetes, rheumatoid arthritis, multiple sclerosis, and myasthenia gravis.In the regulatory pre-clinical phase, analytical methodologies to evaluate the quality of the product need to be developed. In this regard, identification and quantification of the encapsulated peptide and lipids are considered critical quality attributes. In this study, a rapid and simple liquid–liquid extraction procedure, based on Bligh-Dyer method, is described for dual extraction of peptides and lipids within PS-liposomes formulation. This single step allows the separation of lipids and the encapsulated peptide in two different phases. For the subsequent analysis, two different HPLC methods were developed. The organic phase, which contains the lipids was analysed by HPLC-ELSD, while the aqueous phase, containing the encapsulated peptide, was analysed by HPLC-UV. Both methods were also validated in terms of accuracy, precision, linearity, LOD and LOQ. The extraction procedure has demonstrated highly efficient separation of lipids and peptides, avoiding interferences between them in the quantification.

Composition of fatty acids of higher aquatic plants in Lake Kotokel as valuable biological raw materials

Fatty acids, as part of cell membranes, determine their fluidity and dielectric constant. In addition, they play an important role in human and animal nutrition. This work examines the composition and fatty acid content of dried higher aquatic plants from Lake Kotokel (Republic of Buryatia, Russia) and their potential as a raw material for the production of feed and biologically active food supplements. A modified Bligh-Dyer method was used to extract lipid fraction. Fatty acids in the form of their methyl esters were analysed by gas chromatography–mass spectrometry. Palmitic C16:0 acid was the main saturated fatty acid found in Myriophyllum sibiricum, Elodea canadensis, Persicaria amphibia and Potamogeton perfoliatus in Lake Kotokel. The main unsaturated acid found in plants, regardless of their type, was the α-linolenic C18:3-ω3 acid. In addition, a relatively high content of stearic C18:0, myristic C14:0 and pentadecanoic C15:0 acids was noted. Branched acids (i-C14:0, i-C15:0, i-C16:0, i-C17:0, i-C20:0, a/i-C15:0, a/i-C17:0 and 10Me-C16:0) and odd carbon numbered acids (C13:0, C15:0, C17:0, C19:0, C21:0 and C23:0) were identified. An analysis of the authors’ and literature data revealed that the studied species of higher aquatic plants in reservoirs in the Asian part of Russia are characterised by a high content of α-linolenic C18:3-ω3 (up to 56%) and a low content of oleic C18:1-ω9 (up to 1.81%) acids. The content of α-linolenic C18:3-ω3 acid in plants from reservoirs in the European part of Russia was lower (up to 42%), while that of oleic acid C18:1-ω9 was higher (up to 14%).

RAPID DETECTION OF RAT MEAT ADULTERATION IN BEEF SAUSAGES USING FTIR‑ATR SPECTROSCOPY AND CHEMOMETRICS FOR HALAL AUTHENTICATION

Objective: The objective of this study was to employ Fourier Transform Infrared-Attenuated Total Reflectance (FTIR‑ATR) spectroscopy in combination with chemometrics for the analysis of rat meat adulteration in beef sausages.
 Methods: Lipid components in sausages were extracted using three extraction methods, namely Bligh and Dyer, Folch, and Soxhlet methods. The lipid components extracted were then analysed using FTIR‑ATR spectroscopy, and their spectra obtained were used as variables during chemometrics modeling. Samples were prepared by mixing beef with adulterant of rat meat in the concentration range of 0-100% of rat meat. Each sample was scanned using FTIR-Attenuated Total Reflectance (ATR) spectroscopy in three replicates at 4000-650 cm-1 wavenumber region.
 Results: The absorbance values at wavenumbers regions of 3100-700 cm-1 were used to discriminate lipid components extracted by the Bligh Dyer, Folch, and Soxhlet Method with an accuracy level of 100%. The prediction of rat sausages was successfully determined using multivariate calibrations of Partial Least Square (PLS) and Principle Component Regression (PCR) using optimised conditions.
 Conclusion: FTIR-ATR spectroscopy coupled with chemometrics is a rapid and accurate method for detecting and quantifying rat meat in beef sausages for halal authentication.

Comparing the Extraction Performance in Mouse Plasma of Different Biphasic Methods for Polar and Nonpolar Compounds.

Many metabolomic studies are interested in both polar and nonpolar analyses. However, the available sample volume often precludes multiple separate extractions. Therefore, there are major advantages in performing a biphasic extraction and retaining both phases for subsequent separate analyses. To be successful, such approaches require the method to be robust and repeatable for both phases. Hence, we determined the performance of three extraction protocols, plus two variant versions, using 25 μL of commercially available mouse plasma. The preferred option for nonpolar lipids was a modified diluted version of a method employing methyl tert-butyl ether (MTBE) suggested by Matyash and colleagues due to its high repeatability for nonpolar compounds. For polar compounds, the Bligh-Dyer method performs best for sensitivity but with consequentially poorer lipid performance. Overall, the scaled-down version of the MTBE method gave the best overall performance, with high sensitivity for both polar and nonpolar compounds and good repeatability for polar compounds in particular.

Development of FTIR Spectroscopy and multivariate calibration for authentication beef meatballs from pork meat

In several countries, especially Indonesia, meatballs are one of the favorite foods made from meat. The high price of beef is a reason for adulterated beef meat with pork to gain a significant profit. All food products must not contain pig derivatives such as pork in Muslim societies. This study aimed to develop Fourier Transform Infrared (FTIR) spectroscopy in combination with linear discriminant analysis (LDA) for classifying pork in beef meatballs and multivariate calibration for qualitative and quantitative analysis methods using partial least square regression (PLSR) and principal component regression (PCR). The lipid component from meatballs containing pork and beef in different concentrations was obtained by employing three lipid extraction methods, namely BlighDyer, Folch, and Soxhlet methods, and subjected to attenuated total reflectance (ATR) spectral measured at wavenumbers of 4000-650 cm-1 . LDA at 3800-800 cm-1 was able to differentiate between beef meatballs and meatballs containing pork meat (PM). Additionally, PLSR using second derivative FTIR spectra at wavenumbers of 3100-900 cm-1 delivered great region for predicting pork levels in beef meatballs extracted using Folch methods with coefficient determination (R2 ) values was > 0.99 and 0.02% for root mean square error of calibration (RSMEC) and root mean square error of prediction (RSMEP) values, respectively. PCR using absorbance values of 1st derivative FTIR spectra at wavenumbers 1400-800 cm-1 for Bligh-Dyer method and 3800-800 cm-1 for Soxhlet was offered the R2 value was> 0.99 with low RSMEC and RSMEP values for prediction of pork fat in beef meatballs, respectively. It can be concluded that FTIR spectroscopy on the Savitzy-Golay derivatization method in combination with chemometrics was an accurate and quick approach for authenticating pork adulteration in beef meatballs.

Sugar Cane Vinasse as a Renewable and Sustainable Feedstock for Lipid Production from Aspergillus Niger and Aspergillus flavus

Background and Aims: In the search for sustainability and the use of renewable energies, biodiesel has stood out as a notable alternative to the growing energy demand of modern society. Microbial fat synthesis for lipids has been a possibility to produce this biofuel. In this context, the research work aimed to cultivate Aspergillus niger and Aspergillus flavus fungi in commercial and homemade broth, plus variable concentrations of sugarcane vinasse as a substrate to extract lipids produced using different extraction techniques and solvents.
 Methodology: The fungi were grown in a commercial medium and a homemade one made with potato extract. After cultivation, they were submitted to two pre-treatments: the Bead Mill technique and the Ultrasound technique. The extraction of lipids produced in a commercial medium was performed via Soxhlet with a mixture of solvents. Therefore, the lipids produced in a homemade medium were extracted with the adapted Bligh-Dyer method.
 Results: The species grew to the different culture media submitted, but the home culture medium increased the reproductive. The industrial residue proves to be a good alternative as a supplement to the culture medium, helping to reduce costs and preserve the environment. The extraction by Soxhlet showed satisfactory results when the fungi were grown in a commercial medium, with Aspergillus niger being extracted from the ethyl acetate solvent, obtaining a yield of 18.78%, and for A. flavus extracted with dichloromethane/methanol (9:1) and chloroform, bringing yield of 32.98% and 32%, respectively. Although extraction by Soxhlet showed some excellent results, extraction with the Bligh-Dyer method demonstrated better lipid fractions ranging from 25% to 67% and reduced energy expenditure, indicating a promising use, cost, and benefit.
 Conclusion: Filamentous fungi of the genus Aspergillus can be used to produce biodiesel as they proliferate and can produce the same amount of lipids as a vegetable, which requires a much larger area of soil, for example.

Accurate LC-MS/MS Analysis of Diacylglycerols in Human Plasma with Eliminating Matrix Effect by Phospholipids Using Fluorous Biphasic Extraction.

We developed an accurate method for determining diacylglycerols (DAGs) in human plasma using a fluorous biphasic liquid-liquid extraction method, followed by liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis. The lipid mixture in the plasma was first extracted with chloroform by using the Bligh-Dyer method. The resulting solution was subjected to fluorous biphasic liquid-liquid extraction to remove phospholipids, which are known to cause matrix effects during the LC-MS/MS analysis. In this method, phospholipids in a lipid mixture solution (nonfluorous solvent) were selectively extracted to tetradecafluorohexane (fluorous solvent) via the specificity of fluorous affinity by forming a complex with a perfluoropolyethercarboxylic acid-lanthanum(III) salt. The remaining DAGs in the nonfluorous solvent could be directly injected into the LC system through the positive electrospray ionization-MS/MS mode. The removal rate of the phospholipids through the fluorous biphasic extraction was more than 99.9%; thus, the matrix-effect-eliminating analysis of DAGs in human plasma with LC-MS/MS was enabled. Furthermore, the applicability of this method and the possibility of using DAGs as biomarkers were evaluated by applying this method to human plasma samples obtained from major depressive disorder as a related disease.

Nutritional evaluation and bioactive compounds of flours of unconventional edible roots

The bakery sector has increasingly sought flour alternatives, such as unconventional edible root flours, with technologically viable characteristics for industrial production, with greater nutritional value and pleasing consumers in their sensory aspects. So, this work aims to evaluate the nutritional value and bioactive compounds of unconventional edible root meals: purple yam (Dioscorea alata), Jerusalem artichoke (Helianthus tuberosus) and ariá (Calathea allouia). Moisture, protein and ash were determined by standardized bromatological methods, lipids by the Bligh-Dyer method, total crude fiber (TGF) by means of the Fiber Analyzer-Ankom 220 device and carbohydrates by difference. The determination of the total phenolic compounds and tannins in the samples were performed using colorimetric analysis methods. A significant variation (p<0.05) was observed between all samples for the humidity (8.31 to 9.37%); proteins (7.47 to 8.93%); ash (3.15 to 4.42%); lipids (0.13 at 1.76%); fiber (0.92 to 4.85%) and carbohydrates (71.52 to 78.18%). For the bioactive compounds, the variation of total phenolics was from 7.35 to 100.77 mg EAG/100g, and for the tannins 0.28 to 1.41 mg EAT /100g. The study showed that the analyzed samples have significant nutritional value, as well as bioactive compounds that can be used in food product formulations.

Development of a screening method for the determination of carotenoids in ham sausage, juice, and cookies by an improved Bligh-Dyer method and LC-MS/MS.

A screening method of 23 carotenoids in foods, such as ham sausage, juice, and cookies, was proposed using an improved Bligh-Dyer method that can satisfy the extraction requirements of common carotenoids with different physicochemical properties, including free and esterified carotenoids, by collecting the aqueous and organic phases simultaneously. Purification was then performed by loading the aqueous phase onto a hydrophile-lipophile balance (HLB) column; a methanol-water solution was used for washing, and the organic phases and additional chloroform were used for elution. By optimizing this step, interference from hydrophilic compounds and neutral triglycerides was effectively eliminated, and the matrix suppression effect after purification was greater than -16.3%. Finally, the extract was analyzed by ultrahigh performance liquid chromatography-quadrupole-orbitrap high-resolution mass spectrometry. The results showed that the 23 carotenoids showed a good linear relationship in the range of 0.01-0.2µg/mL, and the limits of quantification (signal-to-noise ratio≥10) were from 0.02 to 0.05mg/kg. The average recoveries were 80.1%-98.7%, with relative standard deviation ≤10%. The proposed method can simultaneously identify and quantify 23 carotenoids in foods with high throughput, sensitivity, and reliability.

AUTHENTICATION OF RATTUS NORVEGICUS FAT AND OTHER ANIMAL FATS USING GAS CHROMATOGRAPHY-MASS SPECTROMETRY (GC-MS) AND PRINCIPAL COMPONENT ANALYSIS (PCA)

Objective: The objective of this study was to analyze fatty acids using Gas Chromatography-Mass Spectrometry (GC-MS) in combination with chemometric Principal Component Analysis (PCA) for the authentication of Rattus norvegicus fat from other animal fats.
 Methods: Extraction of fat from raw meat of Rattus norvegicus, beef, chicken, pork, and dogs using the Bligh Dyer method, then derivatized with 0.2 N NaOCH3, precipitation of sodium glycerol was carried out by adding saturated NaCl to obtain methyl esters which were then injected into the GC-MS instrument. The GC-MS data were then processed using chemometric Principal Component Analysis (PCA) to group Rattus norvegicus fat with other animal fats (beef, chicken, pork, and dog).
 Results: The results of the study revealed that fatty acids in Rattus norvegicus using GC-MS produced eleven types of fatty acids, namely: Lauric acid (1,1%), Myristic acid (1,15%), Palmitic acid (21,12%), Palmitoleic acid (2,06%), Stearic acid (8,23%), Vaccenic acid (2,43%), Oleic acid (26,51%), Linoleic acid (19,19%), Arachidic acid (0,09%), and Eucosatrienoic acid (0,39%). Chemometrics Principal Component Analysis (PCA) of Rattus norvegicus fat allows it to be classified with other animal fats.
 Conclusion: The Gas Chromatography-Mass Spectrometry (GC-MS) method, in combination with chemometric Principal Component Analysis (PCA), offered effective tools for the authentication of fatty acid of Rattus norvegicus.

The anti-Alzheimer compounds from tempeh oil in LPS-induced neuronal Schwann cells

AbstractTempeh is a traditional fermented Indonesian food from white soybean. Tempe has better nutritional value than non-fermented white soybean. The aim of this study was to extract tempeh oil and analyze the inhibitory potency of Alzheimer-related gene expression in LPS-induced neuronal Schwann cells. Tempeh oil was extracted with Bligh Dyer method and was analyzed with PUFA identification, anticholinesterase activity, antioxidant activity, and quantitative PCR. Tempeh oil had a total yield of 7.14%, and PUFA identification found 8.37% omega-3. The anti-acetylcholinesterase activity showed that tempeh oil 25 µg/mL had the highest activity and 500 µg/mL in anti-butyrylcholinesterase activity. The quantitative PCR showed that tempeh oil had downregulated the gene expression of PSEN1, Gsk3b, cdk5, and TNF. From this study, tempeh oil may have the potential to lower the risk of Alzheimer’s disease by regulating certain gene traits.

245 Level of Colostrum Intake Affects Vaginal Lipidome of Gilts at Three Weeks Postnatal

Abstract Colostrum is the first milk and available to neonatal pigs the first 24-36 h after birth. Colostrum provides immunity, nutrients, essential fatty acids, and bioactive factors. Colostrum consumption is related to long-term fertility. Previously, vaginal lipidome of female piglets (gilts) on d 21 postnatal (PN) was found to be different between animals that became highly fertile production sows versus infertile animals. Arachidonic acid, a derivative of the essential fatty acids was less in infertile animals. We hypothesized that differences in vaginal lipidome profiles of gilts were due to differing levels of colostrum intake, thus we hypothesized gilts with greater immunocrit (IM) ratios, and greater 24-hour weight gain (measures of colostrum intake) will have greater levels of essential fatty acids and derivatives. Here we determined the relationship of IM and 24 h gain to vaginal lipids on d 21 PN. Gilts (n = 60) were weighed at birth and 24 h PN, and blood sampled to measure IM. On d 21 PN vaginal swabs were taken from surviving gilts (n = 53). Lipids were extracted from swabs using the Bligh-Dyer method and measured using multiple reaction monitoring profiling). MetaboAnalyst 5.0 was used for statistical analysis. Phosphatidyl choline (PC) (36:3), PC(36:2),and arachidonic acid positively (P < 0.05) correlated with immunocrit. PCo(38:6) and cholesterol ethers negatively (P < 0.05) correlated with IM and 24 h gain. Overall, our hypothesis was supported as greater colostrum intake is related to greater levels of essential fatty acids and derivatives in vaginal lipids at 21d PN.

High level of phosphatidylcholines/lysophosphatidylcholine ratio in urine is associated with prostate cancer.

The altered levels of phospholipids (PLs) and lysophospholipids (LPLs) in prostate cancer (CaP) and benign tissues in our previous findings prompted us to explore PLs and LPLs as potential biomarkers for CaP. Urinary lipidomics has attracted increasing attention in clinical diagnostics and prognostics for CaP. In this study, 31 prostate tissues obtained from radical prostatectomy were assessed using high‐resolution matrix‐assisted laser desorption/ionization imaging mass spectrometry (HR‐MALDI‐IMS). Urine samples were collected after digital rectal examination (DRE), and urinary lipids were extracted using the acidified Bligh‐Dyer method. The discovery set comprised 75 patients with CaP and 44 with benign prostatic hyperplasia (BPH) at Kyoto University Hospital; the validation set comprised 74 patients with CaP and 59 with BPH at Osaka University Hospital. Urinary lipidomic screening was performed using MALDI time‐of‐flight MS (MALDI‐TOF/MS). The levels of urinary lysophosphatidylcholine (LPC) and phosphatidylcholines (PCs) were compared between the CaP and BPH groups. The (PC [34:2] + PC [34:1])/LPC (16:0) ratio was significantly higher (P < .001) in CaP tissues than in benign epithelial tissues. The urinary PCs/LPC ratio was significantly higher (P < .001) in the CaP group than in the BPH group in the discovery and validation sets.

Changes in sow milk lipidome across lactation occur in fatty acyl residues of triacylglycerol and phosphatidylglycerol lipids, but not in plasma membrane phospholipids

Milk fats are vital to neonate survival and development, but vary highly by diet, maternal metabolic state and stage of lactation. To gain a better understanding of changes in lipid composition of sow milk across lactation, milk was collected from nine multiparous sows on days 0, 3, 7, and 14, relative to birth and lipids were profiled using multiple reaction monitoring (MRM) profiling. Percent fat was determined by creamatocrit, and found to be different (P < 0.05) between day 0 (12.36 ± 5.90%) and day 3 (16.22 ± 3.65%) but not between day 7 (13.13 ± 2.19%) and day 14 (12.13 ± 2.45%). Fat was extracted from milk (n = 6/day) using the Bligh-Dyer method and profiled using tandem mass spectrometry MRM to determine the abundance of lipids defined by class and fatty acyl residue composition. Lipid species relative concentration was calculated from internal standards, and data analysis was performed using Metaboanalyst 4.0. Concentration of phosphatidyl-choline, -serine, -ethanolamine, -inositol, cholesteryl ester and sphingomyelin did not vary across lactation days, nor did the distribution of associated fatty acyl residues. The total abundance of triacylglycerides (TGs) and phosphatidylglycerols (PGs) increased (P < 0.05) from colostrum (day 0) to transitional (days 3 and 7) and mature milk (day 14). As lactation days increased from day 0 to day 14, the number of carbons and unsaturation within fatty acyl residues decreased (P < 0.05) in both TGs and PGs. The proportion of TGs and PGs increased (P < 0.05) relative to other lipid classes. Changes in composition of milk triglycerides and phosphatidylglycerols likely reflect the metabolic activity of the mammary gland and developmental needs of neonates.

Chemical Composition, Antioxidant, Antimicrobial and Cytotoxic/Cytoprotective Activity of Non-Polar Extracts of Grape (Vitis labrusca cv. Bordeaux) and Blackberry (Rubus fruticosus) Seeds.

The aim of this study was to compare the influence of the extraction method, chemical composition, antimicrobial effects, antioxidant activity, and cytotoxicity on human cells of the non-polar extracts of grape (Vitis labrusca) and blackberry (Rubus fruticosus) seeds. The Soxhlet (Sox), Bligh–Dyer (BD), and ultrasound (US) methods were used for extractions. For blackberry non-polar seed extract, extraction via the BD method showed the highest mean values of total phenolic content (TPC), expressed in milligrams of gallic acid equivalent per 100 mL of non-polar seed extracts (102.37 mg GAE/100 mL), and higher antioxidant activity in relation to the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, expressed in milligrams of gallic acid equivalent per 100 mL of non-polar seed extracts (11.50 mg AAE/100 mL), if compared with the Sox and US extractions. Similar results were obtained for the non-polar grape seed extracts, where BD extraction obtained the highest values for TPC (28.61 mg GAE/100 mL) and DPPH (35.36 mg AAE/100 mL). The type of extraction method had an impact on the composition of fatty acids. Only the non-polar blackberry and grape seed extracts obtained via the Sox method showed some in vitro inhibitory effect against Escherichia coli (IAL 2064) and Staphylococcus aureus (ATCC 13565). Regardless of the extraction method used, the non-polar blackberry and grape seed extracts did not decrease the cell viability (IC50 >1000 µg/mL) of cancer and normal cell lines, thus indicating the relative safety of the extracts. All the seed extracts decreased the generation of reactive oxygen species in the cell lines. Blackberry and grape seed lipid fractions can be utilized as antioxidants, and the extraction methods used cause significant changes in relation to their bioactivity and chemical composition.

55 Sow Milk Lipidome Study Reveals Changes in Fatty Acyl Residues in Triglycerides and Phosphatidylglycerol, but Not in Plasma Membrane Phospholipids Across Lactation

Abstract Sow milk fat content is crucial to neonatal survival, as it is utilized for thermogenesis and nutrition. However, fat is the most variable component of milk in concentration and lipid species. Characterizing lipid changes across the course of a sow’s lactation may help identify molecules or systems to target to help enhance milk fat quality and quantity for neonatal survival and growth. Percent fat variation is greatest in colostrum, the first milk. Little is known regarding colostrum synthesis, other than it accumulates in the gland beginning in mid-late pregnancy, which is prior to the initiation of fatty acid synthesis in lactocytes. The objective of this study was to characterize changes in lipid composition of milk across the course of lactation and determine if there was a relationship between fat percent and lipid species in colostrum and mature milk. Milk was collected from 9 multiparous sows on days 0, 3, 7, and 14 relative to birth. Percent fat was determined by creamatocrit, and found to be different (p&amp;lt; 0.05) between day 0 (12.36 ± 5.90%) and day 3 (16.22 ± 3.65%) but not between day 7 (13.13 ± 2.19%) and 14 (12.13 ± 2.45%). Fat was extracted from milk using the Bligh-Dyer method and profiled using multiple reaction monitoring. Amounts of lipid species were calculated relative to standards and data analysis was performed using Metaboanalyst 4.0. Principle component analysis revealed lactation day had a significant effect on distribution of fats. Triacylglycerides (TAG), phosphatidylglycerol (PG), and plasma membrane lipids were modified from colostrum to mature milk, with a significant increase in PGs and TAGs across the course of lactation. Correlation analysis of percent fat with lipid concentration indicated strong relationships (P &amp;lt; 0.05; |r| &amp;gt;0.80) with eight lipids. No differences are found in the abundance of plasma membrane phospholipids, sphingomyelin, or cholesterol esters across lactation days.

Untargeted lipidomic approach in studying pinot noir wine lipids and predicting wine origin

An untargeted lipidomic profiling approach based on ultra - performance liquid chromatography – time-of-flight tandem mass spectrometry (UPLC-TOF-MS/MS) was successfully used to study the origin of commercial Pinot noir wines. The total wine lipids were extracted using a modified Bligh-Dyer method. In all wine samples, the total lipids were less than 0.1% (w/w) of wine. The wines analyzed consisted of 222 lipids from 11 different classes. 48 commercial Pinot noir wine samples were collected from producers in Burgundy, California, Oregon, and New Zealand. Lipidomic data was studied using advanced multivariate analysis methods, random forest, k-nearest neighbor (k-NN), and linear discriminant analysis. The overall classification accuracy was 97.5% for random forest and 90% for k-NN. Wine lipids showed a strong potential for classifying wines by origin, with the top 58 lipids contributing to the discrimination. This information could potentially be used for further study of the impacts of lipids on wine characteristics and authenticity.

60 High lipid exposure during invitro maturation alters the lipid profile of bovine oocyte and benefits blastocyst development

Lipid metabolism provides a potent source of energy and has an important role in the acquisition of oocyte competence. However, there are conflicting reports about how lipid exposure during invitro maturation (IVM) impacts the gamete and further embryo development. In this study, we performed IVM of oocytes in the presence of lipid-rich culture media and used a broad lipid screening to accurately map the impact on the lipid profile and developmental potential. For that, nonpolar lipids were extracted from fetal bovine serum (FBS) with organic solvents (Bligh-Dyer method) and then used to supplement IVM medium (TCM-199 bicarbonate+10% FBS, hormones, pyruvate and antibiotics). COCs obtained from abattoir ovaries were submitted to IVM (4 biological replicates) in 2 groups: OC (control; IVM medium) and OHL (high lipid; IVM medium supplemented with extra 10% FBS nonpolar lipids). After 24h, we collected mature oocytes and those remaining followed to IVF and then to IVC (synthetic oviductal fluid with amino acids, SOFaa, with 5% FBS) for 7 days at 38.5°C, 20% O2, and 5% CO2 in air in high humidity. Expanded blastocysts were collected (BC and BHL) and blastocyst rates were assessed. Lipid extracts of individual oocytes and embryos (n=10/group) were analysed by multiple reaction monitoring (MRM)-profiling mass spectrometry. A total of 379 lipids from 10 classes were investigated [triacylglycerol (TAG), cholesteryl esters (CE), free fatty acids (FFA), acyl-carnitine, sphingomyelin (SM) and phospholipids derived from choline (PC), ethanolamine (PE), glycerol (PG), serine (PS), and inositol (PI)]. Exploratory data analysis was performed by principal component analysis (PCA; Metaboanalyst 4.0), and fold-change (FC) values were calculated based on the relative intensity of lipid ions (FC &amp;gt; 2 and P&amp;lt;0.05). IVC rates were compared by t-test (α=5%). PCA revealed a clear distinction in the lipid content for both oocytes and blastocysts (control vs. treated). More specifically, there was 2-fold enrichment for total TAG and CE in control groups and a 1.5-fold enrichment for total FFA in the treated groups at the oocyte and the blastocyst stages. Surprisingly, the average blastocyst rate was higher in the group derived from oocytes exposed to a high-lipid environment (41.56±7.73 vs. 22.62±1.67; P=0.003), which led us to investigate specific lipid ions. Groups OHL and BHL had increased contents of structural and signalling phospholipids (PC, SM, PE, and PS) and up to 3 times more oleic and linoleic acids, which have been associated with improved oocyte maturation and blastocyst development. Here, we demonstrate how distinct lipid exposures during IVM can robustly alter the lipid profile of oocytes. But more interestingly, it is clear that these are long-term effects, still observed at the blastocyst stage. More studies are required to verify the metabolic impact of this alternative lipid supplementation; however, these results indicate that high lipid exposure is not necessarily detrimental and, at a certain point, may even counteract lipid accumulation commonly observed during invitro embryo production.

RELATIONSHIP BETWEEN THE DIET QUALITY INDEX IN NURSING MOTHERS AND THE FATTY ACID PROFILE OF MATURE BREAST MILK.

ABSTRACTObjective: To evaluate the relationship between the maternal diet quality and the fatty acid composition of breast milk in the first trimester of lactation.Methods: This is an observational cross-sectional epidemiological study of nursing mothers. The data collection instruments were a semi-structured questionnaire for sample characterization and a recall of usual intake. Diet quality was assessed based on the healthy eating index (HEI). Samples of mature breast milk were obtained by hand milking. Milk fat was extracted using the Bligh-Dyer method and methylated with 0.25 mol/L sodium methoxide in methanol diethyl ether. A gas chromatograph equipped with a flame ionization detector determined the milk fatty acid profile. Pearson’s and Spearman’s correlation tests evaluated association between the variables. Subsequently, a multiple linear regression model was built and multivariate regression analysis was applied.Results: Our findings revealed an inverse relationship between the consumption of total fruits and the polyunsaturated fatty acid profile and a direct association of the intake of total fruits and total grains with monounsaturated and saturated fatty acids.Conclusions: The results of this study suggest that maternal diet quality affects the fatty acid composition of breast milk.

PLFA analysis of the microbial community structure at the points of the temporary localization of radioactive waste

Aim. PTLRW are the trenches and bursts for the localization of radioactive waste that were created during the first priority measures for elimination of the Chornobyl accident. The aim of the presented work was to characterize the microbial community structure on PTLRW. Methods. To describe the influence of environmental factors on the soil microflora, the agrochemical parameters of soil (pH, carbon, nitrogen, mobile potassium and phosphorus) were evaluated. Dose loading was calculated using the ERICA tool software package. The total lipid fraction was extracted with a modified Bligh-Dyer method. Results. The pH of the soil ranged from 3.0 to 3.9. The carbon content ranged from 0.95% to 2.11%. The exception was Red Forest from the trench/outside the trench where the carbon content was 2.52 and 1.98% and with a pH 4.5. Nitrogen content ranged from 33.6 mg / kg to 74.2 mg / kg. The PLFA content ranged from 15 μg / g to 18.9 μg / g, except Novoshepelychi and Zalissia (33.3 μg / g and 23 μg / g). Conclusions. In terms of the structural composition of the microorganisms, the PTLRW points were more homogeneous compared to the contaminated radionuclide ecosystems. In natural ecosystems, gram-positive bacteria were the main dominant group, unlike PTLRW where there were several groups.&#x0D; Keywords: PTLRW, microbial community structure, PLFA, biomarkers, ERICA tool.