Cholesterol Oxidation Products Research Articles

Cholesterol and Its Oxidation Derivatives Content in Market Dairy Products.

Cholesterol oxidation products (COPs) are contaminants of food of animal origin. Increased levels of these compounds in the human body are associated with an increased risk of many non-communicable diseases. Dairy products are mentioned among the main sources of these compounds in the diet. The objective of this study was to evaluate the contents of cholesterol and its oxidized derivatives in eleven groups of dairy products, willingly consumed in European countries. The levels of COPs were determined by applying the GC-TOF/MS method. In the tested products, cholesterol and its oxidation derivatives, such as 7-ketocholesterol, 7α-hydroxycholesterol, 7β-hydroxycholesterol, 5,6β-epoxycholesterol and 5,6α-epoxycholesterol, were determined. The studied dairy products differed in their contents and profiles of oxysterols. The highest contents of COPs were found in cheese with internal mold (13.8 ± 2.5 mg kg-1) and Cheddar (11.7 ± 3.5 mg kg-1), while the lowest levels were detected in yoghurt (0.94 ± 0.30 mg kg-1) and kefir (0.57 ± 0.11 mg kg-1). 7-ketocholesterol and 5,6β-epoxycholesterol were the dominant oxysterols. The ratio of oxidized derivatives to total cholesterol was on average 1.7%. Our results confirmed that dairy products are an important dietary source of COPs. Their levels should be monitored in dairy products to provide the best health quality.

Egg vs. Oil in the Cookbook of Plasters: Differentiation of Lipid Binders in Wall Paintings Using Gas Chromatography-Mass Spectrometry and Principal Component Analysis.

Wall paintings are integral to cultural heritage and offer rich insights into historical and religious beliefs. There exist various wall painting techniques that pose challenges in binder and pigment identification, especially in the case of egg/oil-based binders. GC-MS identification of lipidic binders relies routinely on parameters like the ratios of fatty acids within the plaster. However, the reliability of these ratios for binder identification is severely limited, as demonstrated in this manuscript. Therefore, a more reliable tool for effective differentiation between egg and oil binders based on a combination of diagnostic values, specific markers (cholesterol oxidation products), and PCA is presented in this study. Reference samples of wall paintings with egg and linseed oil binders with six different pigments were subjected to modern artificial ageing methods and subsequently analysed using two GC-MS instruments. A statistically significant difference (at a 95% confidence level) between the egg and oil binders and between the results from two GC-MS instruments was observed. These discrepancies between the results from the two GC-MS instruments are likely attributed to the heterogeneity of the samples with egg and oil binders. This study highlights the complexities in identifying wall painting binders and the need for innovative and revised analytical methods in conservation efforts.

The influence of the food matrix on the content of cholesterol and oxysterols during simulated process of digestion

AbstractThe purpose of the study was to evaluate the influence of the simulated digestion process on the content of the standard cholesterol and this sterol found in fats. Additionally, the content of cholesterol oxides formed after the thermal processing of fats and standard of cholesterol was investigated in samples after performing in vitro digestion procedure. The next step was to evaluate changes in the concentration of cholesterol and cholesterol oxidation products in all analyzed samples. On that basis, the bioaccessibility of cholesterol and cholesterol oxidation products in analyzed samples was established. Cholesterol bioaccessibility ranged from 1.05% to 28.38% in thermally processed samples, while cholesterol standard samples accounted for 7.16% and 5.00%. The bioaccessibility of cholesterol oxides found in analyzed fats and fats with 0.2% addition of cholesterol standard ranged from 5.44% to 19.20%, and only in butter heated at 180°C per 1 h, it reached 47.61%. Studies show that cholesterol present in the food matrix is better protected from degradation, and oxidation during thermal treatment than the standard of cholesterol (processed without food matrix). The results of this study confirm that the bioavailability of cholesterol and its oxides are influenced by the food matrix.Practical applications: The results of our experiment can be used for further scientific analyzes related to the process of estimating the bioavailability of cholesterol and cholesterol oxidation products, which can be found in food products.

Effects of Complete and Partial Loss of the 24S-Hydroxycholesterol-Generating Enzyme Cyp46a1 on Behavior and Hippocampal Transcription in Mouse.

Brain cholesterol metabolic products include neurosteroids and oxysterols, which play important roles in cellular physiology. In neurons, the cholesterol oxidation product, 24S-hydroxycholesterol (24S-HC), is a regulator of signaling and transcription. Here, we examined the behavioral effects of 24S-HC loss, using global and cell-selective genetic deletion of the synthetic enzyme CYP46A1. Mice that are globally deficient in CYP46A1 exhibited hypoactivity at young ages and unexpected increases in conditioned fear memory. Despite strong reductions in hippocampal 24S-HC in mice with selective loss of CYP46A1 in VGLUT1-positive cells, behavioral effects were not recapitulated in these conditional knockout mice. Global knockout produced strong, developmentally dependent transcriptional effects on select cholesterol metabolism genes. These included paradoxical changes in Liver X Receptor targets. Again, conditional knockout was insufficient to recapitulate most changes. Overall, our results highlight the complex effects of 24S-HC in an in vivo setting that are not fully predicted by known mechanisms. The results also demonstrate that the complete inhibition of enzymatic activity may be needed for a detectable, therapeutically relevant impact on gene expression and behavior.

Dietary supplementation with natural antioxidants: assessment of growth performance and meat quality in broiler chickens

Exposure to air and light lowers the meat quality of chickens, which is mainly determined by the color alteration and accumulation of detrimental products from lipid oxidation. This study tests the effects of two supplements rich in polyphenols, Oxilem (OX) and OxiGem (OG), on broiler Ross 308 performance and meat quality in comparison to a control group (C). A total of 105 one-day-old Ross 308 male chicks were allocated to 21 pens and randomly assigned to one of the three treatments. The trial lasted 42 days. Individual live weight and feed intake per pen were recorded. Proximate analysis, color, cholesterol content and cholesterol oxidation products (COPs), fatty acids (FAs), volatile aldehydes profile, and secondary oxidation products of meat FAs were determined, with analyses for detecting oxidative alterations conducted on breast burgers preserved for seven days at 4°C. Birds fed OG grew 7 g/day more than those receiving OX, reaching a higher final weight. After slaughtering, meat from the OX group had a higher yellow index compared to C. After seven days of air and light exposure, the influence on the color parameters of the OG and OX burgers was significantly less pronounced than that of C. Secondary oxidation products of the FAs of the burgers were not significantly affected by the diet regimens. In addition, OX and OG burgers exhibited lower amounts of volatile aldehydes, triol and COPs. These results confirm the effectiveness of OX and OG supplementation against lipid oxidation at the inclusion level used in this trial.

Cholesterol is more readily oxidized than phospholipid linoleates in cell membranes to produce cholesterol hydroperoxides

Cholesterol is an essential component of cell membranes and serves as an important precursor of steroidal hormones and bile acids, but elevated levels of cholesterol and its oxidation products have been accepted as a risk factor for maintenance of health. The free and ester forms of cholesterol and fatty acids are the two major biological lipids. The aim of this hypothesis paper is to address the long-standing dogma that cholesterol is less susceptible to free radical peroxidation than polyunsaturated fatty acids (PUFAs). It has been observed that cholesterol is peroxidized much slower than PUFAs in plasma but that, contrary to expectations from chemical reactivity toward peroxyl radicals, cholesterol appears to be more readily autoxidized than linoleates in cell membranes. The levels of oxidation products of cholesterol and linoleates observed in humans support this notion. It is speculated that this discrepancy is ascribed to the fact that cholesterol and phospholipids bearing PUFAs are localized apart in raft and non-raft domains of cell membranes respectively and that the antioxidant vitamin E distributed predominantly in the non-raft domains cannot suppress the oxidation of cholesterol lying in raft domains which are relatively deficient in antioxidant.

Effect of plasma-activated water (PAW) soaking on the lipid oxidation of sardine (Sardina pilchardus) fillets

The efficacy of plasma-activated water (PAW) as a chemical-free and environmentally friendly preservative has been documented for a variety of foods, but the onset of lipid oxidation induced by plasma-reactive species has been less extensively studied. In this work, global indices (peroxide value, UV specific absorbance) and direct analytical determinations of volatile and non-volatile oxidation products were performed on sardine lipids extracted from fish fillets immersed in PAW (treatments) and distilled water (controls) for 10–30 min. Evidence of PAW-induced lipid oxidation was provided by higher UV specific absorbances and higher levels of C5-C9 secondary volatile oxidation products in the treated samples. However, the degree of fatty acid oxidation was not sufficient to cause a significant reduction in nutritionally valuable eicosapentaenoic acid and docosahexaenoic acid. Twelve cholesterol oxidation products (COPs) were identified in the sardine lipids, but no significant differences in total COPs content were found between PAW processed and control samples.

Impact of Oxysterols in Age-Related Disorders and Strategies to Alleviate Adverse Effects.

Oxysterols or cholesterol oxidation products are a class of molecules with the sterol moiety, derived from oxidative reaction of cholesterol through enzymatic and non-enzymatic processes. They are widely reported in animal-origin foods and prove significant involvement in the regulation of cholesterol homeostasis, lipid transport, cellular signaling, and other physiological processes. Reports of oxysterol-mediated cytotoxicity are in abundance and thus consequently implicated in several age-related and lifestyle disorders such as cardiovascular diseases, bone disorders, pancreatic disorders, age-related macular degeneration, cataract, neurodegenerative disorders such as Alzheimer's and Parkinson's disease, and some types of cancers. In this chapter, we attempt to review a selection of physiologically relevant oxysterols, with a focus on their formation, properties, and roles in health and disease, while also delving into thepotential of natural and synthetic molecules along with bacterial enzymes for mitigating oxysterol-mediated cell damage.

Cholesterol Oxidation Products: Potential Adverse Effect and Prevention of Their Production in Foods.

Cholesterol oxidation products (COPs) are a group of substances formed during food processing. COPs in diet is a health concern because they may affect human health in association with the risk of various diseases including atherosclerosis, Alzheimer's disease, age-related macular degeneration, diabetes, and chronic gastrointestinal inflammatory colitis. Production of COPs in foods can be affected by many factors such as temperature, pH, light, oxygen, water, carbohydrates, fatty acids, proteins, and metal cations. The key issue is preventing its generation in foods. Some COPs can also be produced in vivo by both nonenzymatic and enzymatic-catalyzed oxidation reactions. Currently, a number of natural antioxidants such as catechins, flavonoids, and other polyphenols have been proven to inhibit the generation of COPs. In addition, measures taken during food processing can also minimize the production of COPs, such as the Maillard reaction and marinating food with plant polyphenol-rich seasonings. In conclusion, a comprehensive approach encompassing the suppression on COPs generation and implementation of processing measures is imperative to safeguard human health against the production of COPs in the food chain.

Determination of cholesterol oxidation products in cheese under photo-oxidative stress using QuEChERS and LC–MS

Determination of cholesterol oxidation products in cheese under photo-oxidative stress using QuEChERS and LC–MS

Water thermodynamics and lipid oxidation in stored whey butter

Whey butter is the result of the rational use of the whey component, which is cream whey. It is an alternative to milk cream butter. The aim of the presented study was to analyze the effect of storage conditions on water thermodynamics and cholesterol oxidation products as reliable markers of quality and safety. After 4 mo of storage, the water loss (at 3°C and 13°C) and water activity in whey butter (only at 13°C) were reduced. Three-factorial ANOVA showed that the value of water activity was independent of the type of butter in interaction with the storage temperature. The duration of the translational movement of water molecules from the inside of whey butter was definitely longer than in butter and shortened with storage time. This was in contrast to butter. For whey butter stored at 13°C, the kinetics of the movement of water molecules was at the highest speed. In the case of whey butter and butter, the higher storage temperature almost doubled the gloss. Increasing the temperature to 13°C resulted in different yellowness index, chroma, and browning index between whey butter and butter. There were no statistically significant differences in the percentage of fatty acids and triacylglycerols in whey butter and milk cream butter during storage. In whey butter, compared with butter, the cholesterol content was higher, but the amount of cholesterol oxidation products was smaller. However, in whey butter, these amounts increased significantly. The presence of epoxides and their transformation products (i.e., triol cholesterol) was found in storage whey butter.

OR09-06 The Cholesterol Oxidation Product 7-Ketosterol Reduces Progesterone Production By Human Cytotrophoblast Cells In A Sex Dimorphic Manner

Abstract Disclosure: E. Ticiani: None. N. Pascuali: None. P. Danos: None. C. Murga-Zamalloa: None. I. Buhimschi: None. A. Veiga-Lopez: None. Many pregnancy complications including recurrent miscarriage and preterm birth are strongly associated with placental dysfunction and heightened inflammation. Diet-derived cholesterol, especially via consumption of fast foods that have undergone thermal processing or long-term storage, is highly susceptible to oxidation which results in cholesterol oxidation products, known as oxysterols. Oxysterols are known drivers of inflammation and have been implicated in chronic diseases, including atherosclerosis and neurodegeneration. We hypothesized that the prototype oxysterol 7-ketosterol (7-KT) induces reduced steroid synthesis and/or secretion in human placental cells associated with a pro-inflammatory response. To test this hypothesis, we isolated human primary cytotrophoblast cells (hCTB) from healthy term pregnancies with singleton female fetuses (n=4) and exposed them to 7-KT (2.5 µg/ml) or vehicle control (C; 0.1% DMSO) for 96h. Cells were subjected to RNA sequencing using Illumina TruSeq (p<0.05, FDR <0.05 and log2 fold change >1). Pathway analysis discovered that 10 of the most down-regulated genes (CYP51A1, MSMO1, IDI1, HMGCS1, HMGCR, ACAT2, DDX31, DHCR7, SREBF2, NLRP7) were associated with lipid metabolism. Among them, sterol regulatory element binding transcription factor 2 (SREBF2), is a known activator of cholesterol synthesis. Using female (n=4) and male hCTB (n=4), we further evaluated sex specific effects of 7-KT after 96h of exposure using RT-qPCR. We confirmed that 7-KT reduced the expression of SREBF2 in female and in male hCTB, as well as of genes responsible for cholesterol influx (LDLR) and synthesis (HMGCR and MVK), but not of the cholesterol efflux gene ABCA1. Similar gene expression patterns were observed in JEG-3 cells exposed to 7-KT, a choriocarcinoma cell line that retains the ability to synthetize progesterone. This was coupled with a ∼50% reduction in intracellular cholesterol in JEG-3 cells exposed to 7-KT. Considering that cholesterol is the precursor for progesterone, we assessed progesterone secretion in JEG-3 and in male and female-derived hCTBs cells by ELISA. 7-KT reduced progesterone secretion by JEG-3 and female hCTB by ∼40% while male hCTB were not affected. Additionally, a short exposure (24h) to 7-KT increased the expression of pro-inflammatory genes (IL1α, IL6 and TNFα) in female, but not male hCTB. Altogether, our results demonstrate that the oxysterol 7-KT induces early pro-inflammatory gene expression, followed by a reduction in cholesterol metabolism, lower intracellular cholesterol, and progesterone secretion. Future studies should further investigate the mechanism by which 7-KT drives sex-specific changes in placental cells, and the impact of excess oxysterols on pregnancy outcomes. Presentation: Friday, June 16, 2023

Assessment of lipid oxidation and microbial decontamination of sardine (Sardina pilchardus) fillets processed by plasma-activated water (PAW)

Plasma activated water (PAW) is emerging as a mild and environmentally friendly technology for microbial and chemical decontamination of food. The aim of this work was to evaluate the degree of oxidation of lipids of sardine tissue treated by PAW generated with a pulsed corona discharge. The effect of PAW on the natural microflora of the fish was also evaluated. Under the adopted experimental conditions, PAW was able to reduce the number of mesophilic aerobes and Pseudomonas spp. by 0.22 and 0.20 log units, respectively, but no increase in shelf life was observed at chilling temperatures compared to water washing. A loss of polyunsaturated fatty acids (PUFAs) and an increase in volatile oxidation products derived from the cleavage of PUFA hydroperoxides were observed at the longest treatment duration. Twelve cholesterol oxidation products (COPs) were identified in sardine lipids, but no significant differences in total COPs content were observed between PAW processed and control samples. Free radical mediated oxidation pathways led to the most abundant COPs, but a significant contribution of non-radical pathways was also observed. Further studies are needed to better understand the low efficiency of PAW in microbial decontamination of proteinaceous materials.

Concentrated Grape Powder Consumption Modulates Cholesterol Metabolism and Homeostasis in Healthy Subjects.

Four weeks' of concentrated grape powder (GP) consumption reduces circulating cholesterol in healthy free-living subjects consuming a low-fiber/low-polyphenol diet. Here, the study aims to investigate the underlying mechanisms for cholesterol reduction by evaluating biomarkers of cholesterol de novo biosynthesis, intestinal absorption, miRNA involved in transcriptional regulation of cholesterol metabolism, as well as cholesterol oxidation. Fasting plasma samples collected from 19 healthy free-living subjects at baseline and week 4 of GP consumption are used in this study. Gas chromatography-mass (GC-MS) analysis of plasma samples shows that lathosterol, a precursor of cholesterol synthesis, is significantly decreased after GP consumption indicating reduced cholesterol de novo biosynthesis. Markers of intestinal absorption, campesterol, and β-sitosterol are not changed. Realtime PCR shows that plasma exosomal miRNA-1 is increased after GP consumption. GC-MS also shows that GP consumption reduces the plasma cholesterol oxidation product 27-hydroxycholesterol (27-HC). This study enhances the understanding of the mechanisms of the cholesterol lowering effects of GP, and provides new insights into the potential health benefits of grape consumption.

Regulation of cholesterol homeostasis in osteoporosis mechanisms and therapeutics.

Osteoporosis is a metabolic bone disease that affects hundreds of millions of people worldwide and is characterized by excessive loss of bone protein and mineral content. The incidence and mortality of osteoporosis increase with age, creating a significant medical and economic burden globally. The importance of cholesterol levels has been reported in the development of diseases including osteoporosis. It is important to note that key enzymes and molecules involved in cholesterol homeostasis are closely related to bone formation. Excessive cholesterol may cause osteoporosis, cholesterol and its metabolites affect bone homeostasis by regulating the proliferation and stimulation of osteoblasts and osteoclasts. Therefore, antagonism of elevated cholesterol levels may be a potential strategy to prevent osteoporosis. There is sufficient evidence to support the use of bisphosphonates and statin drugs for osteoporosis in the clinic. Therefore, in view of the aggravation of the aging problem, we summarize the intracellular mechanism of cholesterol homeostasis and its relationship with osteoporosis (including cholesterol and cholesterol oxidation products (COPs) in osteoporosis). Furthermore, the current clinical cholesterol-lowering drugs for osteoporosis were also summarized, as are new and promising therapies (cell-based therapies (e.g., stem cells) and biomaterial-delivered target drug therapies for osteoporosis as well).

7-Ketocholesterol accelerates pancreatic β-cell senescence by inhibiting the SIRT1/CDK4–Rb–E2F1 signaling pathway

ABSTRACT Pancreatic β-cell dysfunction is a key factor in the development of type 2 diabetes. Pancreatic β-cell senescence accelerates abnormal glucose metabolism, which decreases insulin secretion and cell regeneration ability, eventually leading to diabetes. A cholesterol oxidation product, 7-ketocholesterol (7-KC) can affect pancreatic β-cell function. However, its role in pancreatic β-cell senescence has not been reported. We investigated the role of 7-KC in pancreatic β-cell senescence and its underlying molecular mechanism in MIN6 cells. MIN6 cells were treated with 25 μmol/L 7-KC for 24 h and the proportion of senescent cells was detected based on senescence-associated β-galactosidase (SA-β-gal) activity. The cell cycle, DNA damage, and the senescence-associate secretory phenotype (SASP) and protein expression were detected by flow cytometry, immunofluorescence, and western blotting, respectively. 7-KC can significantly increase SA-β-gal activity, promoted G0/G1 arrest, DNA damage, and interleukin-1β expression in MIN6 cells and significantly inhibited insulin synthesis. Further studies indicated that 7-KC induced β-cell senescence by inhibiting the SIRT1/CDK4–Rb – E2F1 signaling pathway.

The CH24H metabolite, 24HC, blocks viral entry by disrupting intracellular cholesterol homeostasis

Cholesterol-24-hydroxylase (CH24H or Cyp46a1) is a reticulum-associated membrane protein that plays an irreplaceable role in cholesterol metabolism in the brain and has been well-studied in several neuro-associated diseases in recent years. In the present study, we found that CH24H expression can be induced by several neuroinvasive viruses, including vesicular stomatitis virus (VSV), rabies virus (RABV), Semliki Forest virus (SFV) and murine hepatitis virus (MHV). The CH24H metabolite, 24-hydroxycholesterol (24HC), also shows competence in inhibiting the replication of multiple viruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). 24HC can increase the cholesterol concentration in multivesicular body (MVB)/late endosome (LE) by disrupting the interaction between OSBP and VAPA, resulting in viral particles being trapped in MVB/LE, ultimately compromising VSV and RABV entry into host cells. These findings provide the first evidence that brain cholesterol oxidation products may play a critical role in viral infection.

Effect of packaging in preventing cholesterol autoxidation in milk chocolates for a higher quality and safer shelf-life.

Non-enzymatic cholesterol oxidation products (COPs) are nowadays receiving increasing attention in food technology for their potential use as biomarkers of freshness and safety in raw materials and complex food matrices, as well as markers of cholesterol oxidation during the production and shelf-life of end products. Here reported is the investigation of how long three prototype milk chocolates containing whole milk powders (WMPs) of increasing shelf-lives (i.e. 20, 120, and 180 days), could be safely stored in the market by adopting the non-enzymatic COPs as a quality markers. In addition, the protective effect of two different primary packaging, sealed and unsealed ones, in mitigating the generation of non-enzymatic COPs in three prototype milk chocolates after 3, 6, 9, 12 months of shelf-life was assessed to simulate two real storage conditions. Quantifying oxysterols' levels by mass spectrometry, the oxygen impermeable packaging (PLUS) resulted to significantly quench the non-enzymatic COPs production up to 34% as to that found in the same product but with unsealed standard packaging (STD). This study represents one practical application of non-enzymatic COPs as a reliable tool for corrective strategies to prevent food oxidation.

Single-Cell Transcriptomics Reveals the Difference of Aortic Atherosclerosis Response to Phytosterols and Oxidation Products of Sterols.

Phytosterols (PS) and sterol oxidation products are key dietary factors influencing atherosclerosis besides cholesterol, although the mechanisms remain elusive. Recently, single-cell RNA sequencing (scRNA-seq) has revealed the heterogeneity of multiple cell types associated with complex pathogenesis in atherosclerosis development. Here, scRNA-seq is performed to investigate the alterations in the aortic cells from ApoE-/- mice induced by diet-derived PS or two sterol oxidation products, phytosterols oxidation products (POPs), and cholesterol oxidation products (COPs). The study identifies four fibroblast subpopulations with different functions, and immunofluorescence demonstrates their spatial heterogeneity, providing evidence that suggests the transformation of smooth muscle cells (SMCs) and fibroblasts in atherosclerosis. The composition and gene expression profiles of aortic cells change broadly in response to PS/COPs/POPs exposure. Notably, PS exhibits an atheroprotective effect where different gene expressions are mainly found in B cells. Exposure to COPs accelerates atherosclerosis and results in marked alternations in myofibroblast subpopulations and T cells, while POPs only alter fibroblast subpopulations and B cells. The data elucidate the effects of dietary PS/COPs/POPs on aortic cells during atherosclerosis development, especially on the newly identified fibroblast subpopulations.

Replacement of soybean oil by Hermetia illucens larvae fat in broiler diets alters the breast muscle lipidome and reduces lipid oxidation of the breast muscle during heat-processing

ABSTRACT Replacement of soybean oil by insect fat from Hermetia illucens (HI) has been reported to increase the proportions of saturated fatty acids (SFA) and decrease those of polyunsaturated fatty acids (PUFA) in total lipids of breast and thigh meat in broilers. Since the susceptibility of meat to oxidation is strongly dependent on its PUFA content, the present study hypothesised that replacement of soybean oil by HI larvae fat in broiler diets reduces the formation of lipid oxidation products, including oxidation products of cholesterol and phytosterols, in heat-processed breast muscle of broilers. To test this hypothesis, 100 male, 1-day-old Cobb 500 broilers were assigned to three groups and fed three different nutrient adequate diets, which varied only in the fat source (group HI-0: 0% HI larvae fat and 5% soybean oil; group HI-2.5: 2.5% HI larvae fat and 2.5% soybean oil; group HI-5.0: 5.0% HI larvae fat and 0% soybean oil), in a three-phase feeding system for 35 days. While the growth performance of the broilers was not different, the absolute and relative breast muscle weights were higher in group HI-5.0 than in group HI-0 (p < 0.05). The proportions of C12:0, C14:0, C14:1, C16:0, C16:1 and total SFA were higher and those of C18:1, C18:2 n-6, C18:3 n-3 and total PUFA were lower in breast muscle total lipids of group HI-5.0 than in groups HI-2.5 and HI-0 (p < 0.05). Lipidomic analysis of breast muscle revealed that the concentration of triacylglycerols was 46% and 53% lower in groups HI-2.5 and HI-5.0, respectively, than in group HI-0 (p < 0.05), whereas all other lipid classes detected did not differ among groups. Concentrations of thiobarbituric acid-reactive substances, 7α-hydroxycholesterol, 7β-hydroxycholesterol and total cholesterol oxidation products in heat-processed breast muscle were lower in group HI-5.0 than in group HI-0 (p < 0.05). Concentrations of oxidation products of phytosterols in heat-processed breast muscle were generally much lower than those of cholesterol oxidation products and did not differ between the three groups of broilers. In conclusion, complete replacement of soybean oil with HI larvae fat in broiler diets strongly alters the fatty acid composition of breast muscle total lipids and reduce lipid oxidation of the breast muscle during heat-processing.