squalene.pptx in Sharks Byproducts and Uses

Squalene Shark

1. SQUALENE Presented by Arya Kumari FPT-MB3-01

2. History From ancient times, fishermen all over the world benefited from the wonderful properties of the oil extracted from the liver of sharks living beneath 1,000 m. The shoguns from ancient Japan recognized the benefits of the deep-sea sharks liver oil, as a source of power, force, energy, and vitality, calling it “Tokubetsu no Miyage,” meaning “precious gift.” This oil was also known and used by coastal residents and fishermen in Micronesia, who referred to it as “miraculous oil.” Locals from Japanese peninsula Izu called this shark liver oil “ Samedawa ,” meaning “cure-all.” They accustomed themselves to use it to cure a wide range of conditions in rough sea. Its usage is also recorded in ancient C hinese texts Honzokomuko .

3. History Squalene was discovered in 1906 by the Japanese researcher Dr. Mitsumaru Tsujimoto , an expert in oils and fats at Tokyo Industrial Testing Station. He separated the unsaponifiable fraction from the shark liver oil “ kuroko-zame ” and discovered the existence of a highly unsaturated hydrocarbon . Ten years later, Tsujimoto succeeded to obtain by fractional vacuum of the liver oil from two deep-sea shark species an unsaturated hydrocarbon, with the chemical formula C 30 H 50 , which he named “squalene” . The name came from the denomination of the sharks' family: Squalidae . After this successful innovation Dr. Keijiro Kogami reported the most beneficial effects of squalene on human health in 1930 (Fatma, , 2013).

4. Squalene Squalene is a highly unsaturated isoprenoid hydrocarbon (C30H50) organic compound. Source - shark liver oil. Tropical Deep Sea Shark ( Centrophorous artomarginatus ) contains high quantity 50% Squalene in its liver oil. In India- Centrophous scalpraus Plant sources - Amaranth seed, rice bran, wheat germ and olives. Micro organisms - yeast strain (Pseudozyma sp) Microalgae (Schizochytrium mangrovei)

5. Squalene All plants and animals produce squalene as a biochemical intermediate, including humans. It is a natural and vital part of the synthesis of all plant and animal sterols, including cholesterol, steroid hormones, and vitamin D in the human body. Nobel Prize winner , Paul Karrer , revealed that squalene was found in human body (Heller et al., 1963). Squalene stimulates the activity of immune cells (macrophages) that founds in the inner and outer layer of human bodies (Heller et al., 1963).

6. Chemical structure of squalene Structurally squalene (2,6,10,15,19,23- hexamethyl-6,6,10,14,18,20tetracosahexane) is a 30 carbon nonpolar compound with an extremely unsaturated linear chain . It is essential for the steroids and an isoprenoid with six isoprene units , so it acts as a naturally occurring isoprenoid compound . The long carbon chain structure of squalene is molded into three interrelated closed rings with six carbon atoms in each ring and attached to a five carbon ring with a prenyl side-chain in the presence of enzymes.

7. Why squalene is present in high amount in liver of seep sea sharks? In the case of deep-sea sharks, the liver is the main organ for lipids' storage, being in the same time an energy source and means for adjusting the buoyancy. For example,   Centrophorus artomarginatus   deep-sea sharks which live in waters at 600 to 1000 m depth, without sunlight, manage to survive where pressure is consistently high and the oxygen supply is very poor, due to this compound from their liver, which accounts for 25% to 30% of their total body weight . The liver of the shark   Centrophorus squamosus  represents 18.1% from the body mass; 77.2% by weight of the liver composition is oil and the squalene concentration in this oil is 79.6%.

8. Physico-chemical properties Squalene is glassy-white in colour , nonpolar in nature and demonstrate low viscosity . The oil has a pleasant bland taste with almost no odour . Squalene is not stable as it is highly unsaturated and easily get oxidised on exposure to atmosphere oxygen. Oxidized squalene has very little biological properties and nutritional value. The most common factors that promote the oxidation process of squalene include temperature, metals, water, oxygen and light.

9. Physical Properties of squalene Chemical formula C 30 H 50 Molar mass 410.73 g·mol −1 Appearance Pale yellow, translucent liquid Density 0.858 g cm −3 Melting point −5 °C (23 °F; 268 K) Boiling point 285 °C (545 °F; 558 K) at 3.3  kPa Viscosity 12 cP (at 20 °C)

10. Market

11. Squalene importance Squalene is a molecule with a long carbon chain it tends to have hydrophobic properties that is of particular interest in industry because it can be used to transport liposoluble compounds in an effective and economic ways. Squalene participates in the formation of steroid hormones, bile acids, steroids, and sterols synthesized though mevalonic acid pathway . Human epidermal sebum is composed by triacylglycerides, free fatty acids (57%), wax esters (26%), and squalene (12%), the use of compounds present in human sebum as squalene in cosmetics reduces the possibility of allergies and is highly appreciated in cosmetic industry due its emollient and antioxidant properties. Squalene reduces serum cholesterol due this triterpenoid may act as a substrate for HMG-CoA reductase (3-hydroxy-3-methylglutaryl Co-A).

12. Weight and Cholesterol Control Oral administration of squalene can produce benefits when ingested in the body. Between 60% and 80% of the exogenous Squalene is absorbed and distributed to various tissues, while the endogenous Squalene is first synthesized in the liver and then transported to the skin or organs through blood. It has been established that it is absorbed faster in the circulatory system than cholesterol, where it is deposited as excess in adipose tissue or muscle tissue.(F. E.sra G¨unes et al,2013).

13. Antioxidant It has six double bonds ; however, it is very stable to peroxidation and other reactions related to unsaturated compounds. Evidence indicates that the antioxidant activity comes from its capacity to trap oxygen singlets during the auto hydrolytic reaction processes and oxidation products (peroxides, SQ-OOH, SQ-OH, and isomers of Squalene oxides). (D. M. Pham et al,2015). Squalene double bonds react with oxygen on the surface of the skin and prevent photooxidation by the rays (S. Ekanayake Mudiyanselage et al.,2003). The entrapment velocity of O- radicals by SQ, and this was superior compared to other lipids even compared to additive hydroxytoluene(HBT) and food antioxidant frequently used.(Kohno et al.) The antioxidant capacity it is not only a function of concentration but also of temperature conditions (<62°C) and interaction with other acids fatty acids, which reduce the oxidation rate due to competitive oxidation ( E. Psomiadou et al,1999 ).

14. Emollient and Moisturizer Squalene relieves skin irritations, through topical applications which prevent photoaging and cancer induced by UV light. Treatments of it with offer protection to burns, and topical applications of creams added with antioxidants (vitamin E) can increase the barrier of the skin and reduce the attack of bacteria and fungi (V. Kostyuk et al.,2012).

15. Detoxifying Squalene, by its nonpolar nature, has an affinity to nonionized drugs that allows it to function as a purifier of xenobiotic substances in the human body. It has been found that SQ improves the elimination of hexachlorobenzene(HCB, organochlorine xenobiotic) through feces when it is supplied in 8 % concentrations in the diet. Other xenobiotic substances such as theophylline and strychnine can also be eliminated in feces when the intake of it is greater(G. S. Kelly,1999) Other studies in pediatric patients have determined that it helps stimulate liver detoxification enzymes, such as the P450 enzyme or for detoxification with lead and other toxic substances; consumption of it in infants also showed greater growth in height and better neuromotor development .(A. L. Ronco ,2013).

16. Anticancer. A hypothesis on the mechanism of inhibition in anticancer activity indicates that Squalene reduces the activity of the HMG-CoA reductase enzyme by limiting the steps towards normal cholesterol synthesis and intermediate stages where geranyl diphosphate (GDP) and farnesyl diphosphate (FDP) produce important substrates for the biosynthesis of ubiquinones and for the prenylation (farnesylation) of proteins. Inhibition of these proteins inactivates and reduces signal transduction in the proliferation and differentiation of active cells such as oncogenes and GTP-binding proteins.(T. J. Smith,2000). Properties of Squalene on cell proliferation, apoptosis, the level of reactive oxygen species (ROS), and oxidative damage to DNA in human breast cells. It was found that it did not have significant activity in cell proliferation rates; however, it exerted effects on epithelial cells in a dose-dependent manner of Squalene. It was observed that it reduced levels of intracellular ROS and oxidative damage induced by H 2 O 2 .(Warleta et al. )

17. Cosmetics It has been reported that Squalene is used as a fixing for perfumes and the elaboration of lipsticks because it accelerates the dispersion of the dye and produces greater brightness. When Squalene is applied to hair and skin exposed to the sun, it helps restore lost oils and easily forms emulsions with other lipophilic substances which allows not to oxidize quickly.(K. Wołosik et al,2013).

18. EXTRACTION METHODS Soxhlet extraction (organic solvent extraction) The most common method used as standard and extract is considered to be 100 % of extractable matter. Hexane is typically the solvent used for large scale extractions due to its relatively low cost and high extraction efficiency. Ultrasonic extraction Combined with organic extraction can achieve higher yield

19. Deep-sea sharks Livers Chopped or ground into smaller pieces to increase the surface area for extraction Homogenized with hexane, ethanol Soxhlet extraction, or ultrasound-assisted extraction Filtration solvent is removed by rotary evaporator Storage

20. Super Critical Fluid Extraction Supercritical fluid extraction (SCFE) can be used to extract polar compounds. Supercritical fluids have diffusivity as gas so can penetrate solid materials, high density and solvation power as liquids, these fluids are compressible and little pressure changes its properties. SCFE have been studied due its advantages against conventional extraction and extract have better quality, biostability and easy to remove from extracted matrix. CO 2 is used to extract oil due its convenience characteristics as non-toxic, non-flammable, easy to remove and economic solvent and also reduces Thermolabile degradation in extracted compounds. Squalene SCFE have been performed by several researchers even when is considered as an expensive technology it extract with high purity.

21. Supercritical fluid Extraction Temperature above the critical point of CO2 (31.1°C) and the pressure above its critical pressure (73.8 atm).

22. Demerits The use of marine animal oil as a source of Squalene has been limited by animal protection regulations and the presence of organic pollutants (POPs) as organochlorine pesticides, polycyclic aromatic hydrocarbons, dioxins, or heavy metals that cause cancer. This has generated interest in finding new natural sources, especially of plant origin. Stability and proper storage. Sustainability concerns

23. A way forward Shift of source Gene transfer technology Biotechnological production Cosmetics Squalene supplements Vaccine adjuvants Toxicological investigations of Squalene for metals and toxic substances. As food additives Extraction procedure Antibacterial drug

24. References M. Azalia Lozano-Grande et al.(2018); Review Article on Plant Sources, Extraction Methods, and Uses of Squalene. K. Gopakumar ;A textbook of Fish Processing Technology Ovidiu Popa et al.(2015); Methods for Obtaining and Determination of Squalene from Natural Sources