Source Of Squalene Research Articles

Accumulation of squalene in filamentous fungi Trichoderma virens PS1-7 in the presence of butenafine hydrochloride, squalene epoxidase inhibitor: biosynthesis of 13C-enriched squalene.

Squalene is a triterpenoid compound and widely used in various industries such as medicine and cosmetics due to its strong antioxidant and anticancer properties. The purpose of this study is to increase the accumulation of squalene in filamentous fungi using exogeneous butenafine hydrochloride, which is an inhibitor for squalene epoxidase. The detailed settings achieved that the filamentous fungi, Trichoderma virens PS1-7, produced squalene up to 429.93±51.60mg/L after culturing for 7 days in the medium consisting of potato infusion with glucose at pH 4.0, in the presence of 200µm butenafine. On the other hand, no squalene accumulation was observed without butenafine. This result indicated that squalene was biosynthesized in the filamentous fungi PS1-7, which can be used as a novel source of squalene. In addition, we successfully obtained highly 13C-enriched squalene by using [U-13C6]-glucose as a carbon source replacing normal glucose.

Liposomes Loaded with Amaranth Unsaponifiable Matter and Soybean Lunasin Prevented Melanoma Tumor Development Overexpressing Caspase-3 in an In Vivo Model.

The objective of this study was to assess the effectiveness of liposomes loaded with soybean lunasin and amaranth unsaponifiable matter (UM + LunLip) as a source of squalene in the prevention of melanoma skin cancer in an allograft mice model. Tumors were induced by transplanting melanoma B16-F10 cells into the mice. The most effective treatments were those including UM + LunLip, with no difference between the lunasin concentrations (15 or 30 mg/kg body weight); however, these treatments were statistically different from the tumor-bearing untreated control (G3) (p < 0.05). The groups treated with topical application showed significant inhibition (68%, p < 0.05) compared to G3. The groups treated with subcutaneous injections showed significant inhibition (up to 99%, p < 0.05) in G3. During tumor development, UM + LunLip treatments under-expressed Ki-67 (0.2-fold compared to G3), glycogen synthase kinase-3β (0.1-fold compared to G3), and overexpressed caspase-3 (30-fold compared to G3). In addition, larger tumors showed larger necrotic areas (38% with respect to the total tumor) (p < 0.0001). In conclusion, the UM + LunLip treatment was effective when applied either subcutaneously or topically in the melanoma tumor-developing groups, as it slowed down cell proliferation and activated apoptosis.

Enhancement of Squalene Production by Constitutive Expression of the 3-Hydroxy-3-Methylglutaryl-CoA Reductase in Aurantiochytrium sp. 18W-13a.

Squalene has a wide range of applications in the industry sectors of dietary supplements, cosmetics, immunization, and pharmaceuticals. Yet, suitable organisms as the source of squalene are limited. It is reported that the thraustochytrid Aurantiochytrium sp. strain 18W-13a can accumulate high content of squalene. However, squalene production in this organism is fluctuated under various conditions and is not yet optimized for commercialization. In this organism, the mevalonate pathway supplies isopentenyl pyrophosphate, the initial substrate for squalene production. In this pathway, 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) is the rate-limiting enzyme. We found that the HMGR activity had a strong positive correlation with the squalene contents in the strain. We constitutively expressed the HMGR in this organism and found that the transformant showed increased and stable production of squalene as well as carotenoids and biomass. These results clearly indicated that the HMGR expression is the bottleneck of squalene synthesis in Aurantiochytrium sp.

The extraction and purification of squalene from Nyamplung (Calophyllum Inophyllum L) leaves

Squalene is a natural organic compound that is primarily found in shark liver oil. It can act as a skin moisturizing, anticancer, antivirus, and drug delivery agent. The advantages of people consuming healthy oil which contains squalene can lower the risk of heart disease. Due to the high market demand for squalene, it becomes a severe threat to shark existence. Therefore, investigation to find a new source of squalene is deemed necessary. Squalene is present in the leaves of nyamplung (Calophyllum inophyllum), a species of mangrove plant commonly found along Indonesia’s coast. This study aims to extract and purify squalene from nyamplung leaves. First, nyamplung leaves were crushed and extracted with n-hexane. The extract was designated as a crude extract of nyamplung leaves. After that, squalene was purified from the crude extract by the multistage adsorption-desorption method at 3°C. The crude extract was dissolved in hexane, and silica gel was then added at 30 min. The mass ratios of silica gel to crude extract were varied at 1:1, 2:1 and 4:1 (g/g). N-hexane fraction was separated from silica gel by filtration paper. N-hexane fraction was designated as a non-polar fraction (NPF), while silica gel fraction was designated as a polar fraction (PF). The dark-colored of PF was employed for the desorption process using methanol until its color became colorless. Meanwhile, NPF was extracted with fresh silica gel until it reached the 10th stage of extraction. NPF and PF were then quantitatively and qualitatively analyzed using TLC and GC, respectively. The best result was obtained from silica gel to crude extract mass ratio of 1:1 (g/g) at the 5th stage with squalene purity of 35.96% (45.82% recovery).

Liposomes Loaded with Unsaponifiable Matter from Amaranthus hypochondriacus as a Source of Squalene and Carrying Soybean Lunasin Inhibited Melanoma Cells.

Amaranthus hypochondriacus is a source of molecules with reported health benefits such as antioxidant activity and cancer prevention. The objective of this research was to optimize the conditions for preparing a liposome formulation using amaranth unsaponifiable matter as a source of squalene in order to minimize the particle size and to maximize the encapsulation efficiency of liposomes for carrying and delivering soybean lunasin into melanoma cell lines. Amaranth oil was extracted using supercritical dioxide carbon extraction (55.2 MPa pressure, 80 °C temperature, solvent (CO2)-to-feed (oil) ratio of 20). The extracted oil from amaranth was used to obtain the unsaponifiable enriched content of squalene, which was incorporated into liposomes. A Box–Behnken response surface methodology design was used to optimize the liposome formulation containing the unsaponifiable matter, once liposomes were optimized. Soybean lunasin was loaded into the liposomes and tested on A-375 and B16-F10 melanoma cells. The squalene concentration in the extracted oil was 36.64 ± 0.64 g/ 100 g of oil. The particle size in liposomes was between 115.8 and 163.1 nm; the squalene encapsulation efficiency ranged from 33.14% to 76.08%. The optimized liposome formulation contained 15.27 mg of phospholipids and 1.1 mg of unsaponifiable matter. Cell viability was affected by the liposome formulation with a half-maximum inhibitory concentration (IC50) equivalent to 225 μM in B16-F10 and 215 μM in A-375. The liposomes formulated with lunasin achieved 82.14 ± 3.34% lunasin encapsulation efficiency and improved efficacy by decreasing lunasin IC50 by 31.81% in B16-F10 and by 41.89% in A-375 compared with unencapsulated lunasin.

Low-Density Insulation Blocks and Hardboards from Amaranth (Amaranthus cruentus) Stems, a New Perspective for Building Applications

Nowadays, amaranth appears as a promising source of squalene of vegetable origin. Amaranth oil is indeed one of the most concentrated vegetable oils in squalene, i.e., up to 6% (w/w). This triterpene is highly appreciated in cosmetology, especially for the formulation of moisturizing creams. It is almost exclusively extracted from the liver of sharks, causing their overfishing. Thus, providing a squalene of renewable origin is a major challenge for the cosmetic industry. The amaranth plant has thus experienced renewed interest in recent years. In addition to the seeds, a stem is also produced during cultivation. Representing up to 80% (w/w) of the plant aerial part, it is composed of a ligneous fraction, the bark, on its periphery, and a pith in its middle. In this study, a fractionation process was developed to separate bark and pith. These two fractions were then used to produce renewable materials for building applications. On the one hand, the bark was used to produce hardboards, with the deoiled seeds acting as natural binder. Such boards are a viable alternative to commercial wood-based panels. On the other hand, the pith was transformed into cohesive and machinable low-density insulation blocks revealing a low thermal conductivity value.

Adapting of a Method for Qualitative and Quantitative Determination of Squalene in Distillation Cuts of Sunflower Oil

Squalene is a naturally-occurring dihydro-triterpene hydrocarbon (C30H50) with six double bonds, which is an intermediate in the biosynthesis of phytosterol or cholesterol in plants or animals. The sources of squalene and the main methods forsqualene production and determination are consideredin brief. Sunflower oil distillation cuts have been selected as the subject of the study, since they area promising secondary raw material for the industrial squalene production. The methods of sample preparation and quantification of squalene in sunflower oil distillation cuts applying gas chromatography in combination with mass spectrometry have been adapted. The aim of the studyis to create an integrated approach to determining the qualitative and quantitative content of squalene in distillation cuts of vegetable oils. To achieve the goal of the study, the following tasks have been solved: – Amethod of sample preparation of distillation cuts for determination of squalene has been adapted; – A method of qualitative and quantitative determination of squalene in distillation cuts has been modified. As a result of this study, a technique for sample preparation of distillation cuts was proposed as well as a method for the qualitative and quantitative (absolute calibrationmethod) determination of squalene in distillation cuts of sunflower oil. To implement the technique, a Kristall 5000 gas chromatograph equipped with a mass spectrometric detector was used. Squalene and background components were recorded using the NIST 11 mass spectral database.

Characteristics of fatty acid composition and minor constituents of red palm olein and palm kernel oil combination.

Red palm olein (RPOl) is one of the derivatives of palm oil. It contains a high composition of unsaturated fatty acids such as oleic and linoleic, whereas palm kernel oil (PKO) contains more saturated fatty acids of lauric acid. RPOl provides high nutrient contents such as squalene, Vitamin E, and carotene, whereas PKO that is rich in lauric acid can fight Gram-positive microorganisms. This research aims to study the chemical characteristics of RPOl, PKO, and the combination. A combination of RPOl with four different concentrations of PKO (20%, 50%, 80%, and 100%) was analyzed to obtain the composition of fatty acids, squalene content, Vitamin E levels, total carotene, and saponification numbers. RPOL contains high levels of squalene, Vitamin E, and total carotene, followed by RPOl and PKO combination of oil, with a higher percentage of RPOl in its composition. The increase of the PKO level added to the combination will decrease the saponification number and increasing the acid number. Therefore, it can be concluded that RPOl could be the source of squalene, Vitamin E, carotenoids, and oleic acid, whereas PKO is the largest source of lauric acid.

Squalene: More than a Step toward Sterols.

Squalene (SQ) is a natural triterpene widely distributed in nature. It is a metabolic intermediate of the sterol biosynthetic pathway and represents a possible target in different metabolic and oxidative stress-related disorders. Growing interest has been focused on SQ’s antioxidant properties, derived from its chemical structure. Strong evidence provided by ex vivo models underline its scavenging activity towards free radicals, whereas only a few studies have highlighted its effect in cellular models of oxidative stress. Given the role of unbalanced free radicals in both the onset and progression of several cardiovascular diseases, an in depth evaluation of SQ’s contribution to antioxidant defense mechanisms could represent a strategic approach in dealing with these pathological conditions. At present experimental results overall show a double-edged sword role of squalene in cardiovascular diseases and its function has to be better elucidated in order to establish intervention lines focused on its features. This review aims to summarize current knowledge about endogenous and exogenous sources of SQ and to point out the controversial role of SQ in cardiovascular physiology.

Determination of squalene in edible oils by transmethylation and GC analysis

Squalene is a triterpenoid with medicinal, therapeutic and cosmetic applications. There is interest in identifying novel sources of squalene, creating a need for a rapid and accurate method of squalene determination. Here, we describe such an approach that involves first transmethylating the oil base containing squalene using a basic catalyst, followed by addition of internal standard and quantification by gas chromatography flame ionization detection (GCIFD).•The method uses a single reference standard for quantification.•Validation showed it to be both repeatable and accurate.•It was successfully applied to nutraceutical and edible oils containing squalene.

Plant Sources, Extraction Methods, and Uses of Squalene

Squalene (SQ) is a natural compound, a precursor of various hormones in animals and sterols in plants. It is considered a molecule with pharmacological, cosmetic, and nutritional potential. Scientific research has shown that SQ reduces skin damage by UV radiation, LDL levels, and cholesterol in the blood, prevents the suffering of cardiovascular diseases, and has antitumor and anticancer effects against ovarian, breast, lung, and colon cancer. The inclusion of SQ in the human diet is recommended without causing health risks; however, its intake is low due to the lack of natural sources of SQ and efficient extraction methods which limit its commercialization. Biotechnological advances have developed synthetic techniques to produce SQ; nevertheless, yields achieved are not sufficient for global demand for industrial or food supplement purposes. The effect on the human body is one of the scientific issues still to be addressed; few research studies have been developed with SQ from seed or vegetable sources to use it in the food sector even though squalene was discovered more than half a century ago. The aim of this review is to provide an overview of SQ to establish research focus with special reference to plant sources, extraction methods, and uses.

Optimization of fermentation conditions for squalene production by heterotrophic marine microalgae Schizochytrium mangrovei PQ6

Squalene is currently known as chemopreventive agents in reducing the incident of coronary heart disease and cancer. It is also a strong antioxidant used extensively in the food and cosmetic industries. Microbial sources of squalene are being explored in recent years. The objective of this study was to investigate fermentation conditions of Schizochytrium mangrovei PQ6 for high squalene production in 30L bioreactor. Squalene production was influenced by various nutritional factors such as glucose, nitrogen, vitamins etc. and physical factors such as batch and fed-batch fermentation. Our study established that fed-batch fermentation with nitrogen source of 1.2% yeast extract and 6.4% monosodium glutamate and 0.4% mixture of vitamin B1, 6 and 12 as optimal culture condition for maximal production of squalene by S. mangrovei PQ6. In fed-batch culture, the glucose concentration feeding was added up to 22% into the culture at 36 h of fermentation. The maximum squalene production was achieved 6.9 ± 0.1 g/L at 48 h of fed-batch fermentation, respectively. The values were 2-3 times more than that in fed-batch cultivation without adding mixture of vitamins and 4-6 times more than that in batch cultivation. Our obtained results are a breakthrough for the development of alternative squalene source from heterotrophic marine microalgae of Schizochytrium mangrovei PQ6 species for health-promoting functional foods. Citation: Nguyen Cam Ha, Hoang Thi Minh Hien, Le Thi Thom, Hoang Thi Huong Quynh, Dang Diem Hong, 2017. Optimization of fermentation conditions for squalene production by heterotrophic marine microalgae Schizochytrium mangrovei PQ6. Tap chi Sinh hoc, 39(3): 349-358. DOI: 10.15625/0866-7160/v39n3.9130. *Corresponding author: hoangminhhien@yahoo.com; ddhong60vn@yahoo.com Received 6 January 2017, accepted 20 August 2017

Supercritical Extraction of Cobia (Rachycentron canadum) Liver Oil as a New Source of Squalene

Cobia is a fish cultivated worldwide and there are few studies on the potential of their waste, such as the liver, it's rich in oil and active compounds. The objective of this study was to extract cobia liver oil (CLO) using supercritical CO2 under moderate temperature conditions (50, 60 and 70 °C, 250 bar) and characterize the oil (physico-chemical analysis, fatty acids profile and squalene content). The results showed that CLO is rich in MUFA and presented squalene, suggesting that this matrix can be applied in the production of high nutritional value oil.

Biochemical Characterization of Liver Oil of Echinorhinus brucus (Bramble Shark) and Its Cytotoxic Evaluation on Neuroblastoma Cell Lines (SHSY-5Y).

The objective of the present study was to characterize the liver oil extracted from the deep sea shark, Echinorhinus brucus, caught from Central Indian Ocean and to evaluate its cytotoxic effect on neuroblastoma cell line (SHSY-5Y). Characterization of liver oil of Echinorhinus brucus revealed the presence of palmitic acid (15%), oleic acid (12%), stearic acid (8%), docosahexaenoic acid (DHA) (18%), and eicosapentaenoic acid (EPA) (16%). It was also found to be a good source of squalene (38.5%) and fat soluble vitamins such as A, D, and K (vitamin A: 17.08 mg/100 g of oil, vitamin D: 15.04 mg/100 g oil, and vitamin K: 11.45 mg/100 g oil). Since it was found to be rich in essential fatty acids, fat soluble vitamins, and squalene, it can be considered as better dietary supplement. The oil of Echinorhinus brucus also showed high in vitro cytotoxic effect against the human neuroblastoma cell line (SHSY-5Y) and the IC50 value laid between 35 and 45 ng.

Squalene emulsions for parenteral vaccine and drug delivery.

Squalene is a linear triterpene that is extensively utilized as a principal component of parenteral emulsions for drug and vaccine delivery. In this review, the chemical structure and sources of squalene are presented. Moreover, the physicochemical and biological properties of squalene-containing emulsions are evaluated in the context of parenteral formulations. Historical and current parenteral emulsion products containing squalene or squalane are discussed. The safety of squalene-based products is also addressed. Finally, analytical techniques for characterization of squalene emulsions are examined.

ELUSIDASI STRUKTUR ISOLAT SKUALENA DARI IKAN HIU CUCUT BOTOL (Centrophorus sp)

The main source of squalene was liver oil of shark. Producing of squalene this oil could be separated from other components inside. The research obtained that the value of squalene was 75.83 1.14 percent. To discover the purification and safety to consume squalene which had been isolated from oil of the liver’s shark cucut botol was used a method which available in AOAC, therefore that could be done elucidation of the structure of isolate with any instruments such as KLT, FTIR, GC, GC-MS, and NMR. Structure elucidation of isolate compare with standard of squalene had result as follows: a. KLT: Rf isolate = 0.457, Rf standard = 0,457, b. FTIR = 2980– 2850 CH of methyl and methylene; 1668.58 &amp; 1446.74 a double chain of carbon (C=C); 1381.16 and 835.25 = branch chain that all for isolate was the same with standard; GC tR = 9.065 minute of isolate and standard was 9.124 minutes; GC-MS the main fraction with mass 69 was one unit of isoprene (C5H9); the value of proton NMR (δ = ppm) were 1.601 and 1.674, 2.016, 5.150 and 5.155 for isolate; the standard were 1.599 and 1.670, 2.016, 5.155. The result of elucidation structure could be concluded that the isolate was the same with the squalene standard. Therefore, isolate could be able to consume safety as natural health food for all consumers because it had already pure and free from other compound such as cholesterol.

Amaranth squalene reduces serum and liver lipid levels in rats fed a cholesterol diet

In this study, the hypocholesterolaemic effect of amaranth grain, oil and squalene are examined. In experiment 1, rats are given a semi-purified diet containing 1% (w/w) cholesterol for four weeks and either amaranth grain (AG; 300 g/kg) or amaranth oil (AO; 90 g/kg) substituted in experimental groups. Both AG and AO lowered serum and hepatic cholesterol and triglyceride levels. Faecal excretion of cholesterol and bile acid in the AO group increased, while AG ANected only bile acid excretion. In experiment 2, rats were fed the cholesterol diet for four weeks and injected (i.p.) with saline (control), amaranth squalene (AS) or shark liver squalene (SS, 200 mg/kg) for seven days. The hypolipidaemic effects of AS were evident in both serum and liver. In addition, AS markedly increased faecal excretions of cholesterol and bile acid, and slightly inhibited 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity. In contrast, none of these effects were observed in the SS group. This preliminary study suggests that the cholesterol-lowering effect of AS may be mediated by increased faecal elimination of steroids through interference with cholesterol absorption, and that different sources of squalene (plant versus animal) may ANect cholesterol metabolism differently.

Development and application of an analytical method for the determination of squalene in formulations of anthrax vaccine adsorbed

Specific lots of Anthrax Vaccine Adsorbed, administered to members of the US Armed Forces, have been described on various Internet sites and in news articles as a source of squalene, a chemical purported by these media to be associated with the Gulf War Syndrome. We have developed and validated a method using high-performance liquid chromatography with ultraviolet detection for the determination of squalene in anthrax vaccine preparations. The method has a limit of detection of 140 parts per billion and has been successfully applied to a commercial vaccine known to contain squalene. We have applied this method to 17 lots of Anthrax Vaccine Adsorbed administered to members of the US Armed Forces. No squalene has been detected in any lot. The results of these analyses provide direct evidence for the absence of squalene as an ingredient or a manufacturing contaminant in Anthrax Vaccine Adsorbed.