SQUALENE

Wini Trilaksani
Bambang Riyanto

DEPARTEMEN TEKNOLOGI HASIL PERAIRAN

FAKULTAS PERIKANAN DAN ILMU KELAUTAN
INSTITUT PERTANIAN BOGOR
2017
Squalene Extraction
Squalene Extraction
SQUALENE
• An unsaturated aliphatic hydrocarbon, C30H50, that is the
biosynthetic acyclic precursor to tetracyclic steroids such
as cholesterol.
• It is widely found in animals, plants, and fungi and is
present in high concentrations in shark liver oil and
human sebum.
• Except squalene, the liver oil of the deep see shark
contains vitamin A, D3 and E, Jodium and alcohol, as well
as alcoxyglycerols and other immunity enhancing
nutritional substances.
• As it turns out now the synergetic effect of squalene
together with the other components improve not only
the sharks immunity against cancer and other diseases,
but supports the animal with an increased endurance.
SQUALENE
• Squalene from shark liver oil is named in Latin
‘squalene exogene oleum’. The medical name is
Spinacene or Supraene.
• The local people on the Japanese peninsula of Izu
called the substance already for centuries
"Samedawa" or a cure-all. They used it for a wide
range of diseases from constipation to cancer. In
ancient China the miraculous effects of shark liver oil
are recorded for posterity in the pharmaceutical book
of "Honzukomuko".
 PHYSICAL DESCRIPTION: Clear, slightly yellow liquid with a faint
Squalene odor. Density 0.858 g / cm3.; 0.8584 at 68° F (NTP, 1992); Faint agreeable odor
Boiling Point
421.3 deg C 545° F at 25 mm Hg (NTP, 1992)
Melting Point
-4.8 deg C -103° F (NTP, 1992)

Molecular Weight Flash Point

410.73 g/mol 110 deg C - closed cup greater than 235° F (NTP, 1992)

Solubility
Synonym(s): (E,E,E,E) -SQUALENE; SPINACEN; SPINACENE;
Practically insoluble in water.
SQUALEN; TRANS-SQUALENE
Freely soluble in ether, petroleum ether,
Squalene; Trans-Squalene; Spinacene; 111-02-4; Supraene; carbon tetrachloride, acetone, other fat
2,6,10,15,19,23-Hexamethyltetracosa-2,6,10,14,18,22- solvents; sparingly soluble in alcohol, glacial
hexaene acetic acid
Slightly soluble in alcohol; soluble in
About SQUALENE: Squalene is a naturally occuring lipid in both lipids and organic solvents
plants and animals. This ingredient is listed in the PETA's Caring less than 1 mg/mL at 66° F (NTP, 1992)
Consumer guide as primarily derived from animals, especially
shark liver oil. Vapor Pressure
6.3X10-6 mm Hg at 25 deg C (est)
Squalene
Stability
Stable under recommended storage conditions.
Decomposition
When heated to decomposition it emits acrid smoke and
irritating vapors.
Viscosity Spectral Properties
12 cP at 25 deg C Index of Refraction: 1.4990 at 20 deg C
Surface Tension
33.9 mN/m at 22 deg C (100 g/L)

Kovats Retention Index

Standard non-polar 2808

Semi-standard non-polar 2835.8, 2847.1, 2814, 2819, 2832, 2833, 2809.1
Standard polar 2865
SQUALENE
• Squalene is the biochemical precursor to the whole family
of steroids. Oxidation (via squalene monooxygenase) of
one of the terminal double bonds of squalene yields 2,3-
squalene oxide, which undergoes enzyme-catalyzed
cyclization to afford lanosterol, which is then elaborated
into cholesterol and other steroids.
• Squalene is an ancient molecule. In plants, squalene is the
precursor to stigmasterol. In certain fungi, it is the
precursor to ergosterol. However, blue-green algae and
some bacteria do not manufacture squalene, and must
acquire it from the environment if they need it.
SQUALENE
• Squalene is a natural 30-carbon organic compound
originally obtained for commercial purposes
primarily from shark liver oil (hence its name, as
Squalus is a genus of sharks), although plant
sources (primarily vegetable oils) are now used as
well, including amaranth seed, rice bran, wheat
germ, and olives. Yeast cells have been genetically
engineered to produce commercially useful
quantities of "synthetic" squalene.
 SQUALENE
• Squalene, an isoprenoid compound structurally similar to
beta-carotene, is an intermediate metabolite in the synthesis
of cholesterol.
• Squalene is a poly-unsaturated hydrocarbon liquid (C30 H50)
which occurs naturally, ubiquitously in human tissue in small
amounts.
• In humans, about 60 percent of dietary squalene is absorbed.
It is transported in serum generally in association with very
low density lipoproteins and is distributed ubiquitously in
human tissues, with the greatest concentration in the skin,
where it is one of the major components of skin surface
lipids.
Squalene
• Squalene is not very susceptible to peroxidation and
appears to function in the skin as a quencher of singlet
oxygen, protecting human skin surface from lipid
peroxidation due to exposure to UV and other sources of
ionizing radiation.
• Supplementation of squalene to mice has resulted in
marked increases in cellular and non-specific immune
functions in a dose-dependent manner.
• Squalene may also act as a "sink" for highly lipophilic
xenobiotics. Since it is a nonpolar substance, it has a
higher affinity for un-ionized drugs.
Squalene
• In animals, supplementation of the diet with squalene
can reduce cholesterol and triglyceride levels.
• In humans, squalene might be a useful addition to
potentiate the effects of some cholesterol-lowering
drugs.
• The primary therapeutic use of squalene currently is as
an adjunctive therapy in a variety of cancers.
• Although epidemiological, experimental and animal
evidence suggests anti-cancer properties, to date no
human trials have been conducted to verify the role this
nutrient might have in cancer therapy regimens.
Squalene
• The non-saturated hydrocarbon C30H50, in order to
stabilize, attaches hydrogen ions from water and acids in
the body, and in the process, frees oxygen to the body.
It was established that the non-mythical Squalene
stimulates the metabolism of the body by supplying an
abundance of oxygen to the tissues.
• Squalene is unique oil high in Vitamins A and D that is
found in high concentrations within shark liver. It also
occurs naturally within the human body where it
supports natural lubrication for our skin and supports
oxygen delivery and toxin removal within all our body's
cells.
Historical Importation-Shark
squalene
• It is generally assumed that squalene was an
essential substance in the very beginning of life.
After millions of years of evolutionary development
the liver of the deep sea shark forms the richest
source of natural squalene.
• Shark liver oil consists of about 60-85 % of this
organic matter. Olive oil has a squalene
concentration of only 0,4 –0,8 %. During the
production process from olive oil squalene can
easily be distilled to a concentration of about 99%.
Historical Importation-Shark
squalene
• Squalene is a low density compound often stored in the
bodies of cartilaginous fish such as sharks, which lack a
swim bladder and must therefore reduce their body
density with fats and oils.
• Squalene, which is stored mainly in the shark's liver, is
lighter than water with a specific gravity of 0.855.
Recently it has become a trend for sharks to be hunted to
process their livers for the purpose of making squalene
health capsules.
• Environmental and other concerns over shark hunting
have motivated its extraction from vegetable sources or
biosynthetic processes instead
Historical Importation-Shark
squalene
• Since time immemorial Japan uses seafood as a staple diet.
It’s not surprising then that in 1906 a Japanese chemist Dr.
Mitsumaro Tsjujimoto, specialised in oils and fats, published
his scientific research about the secrets of shark liver oil. He
discovered that a specific substance in the oil was
responsible for the healing properties. He called it squalene
because the people of Japan and neighboured islands
extract the oil only from sharks livers of the Squalidae family.
• According to fishery authorities this order represents the
second largest order and includes 7 families and about 113
species and their most distinguishing feature is the missing
anal fin. Most species also have a spine in front of the first
dorsal fin. The majority are deep water species, and some
have luminescent organs. Some species can reach a length
for instance the Greenland shark, Somniosus microcephalus,
of 7m.
Historical Importation-Shark
squalene
• 6 kinds are used for the production of shark liver oil: the Aizame
shark or Dog Shark, the nurse shark, de Kasumizame or Fog shark,
the Gulper Shark, the Thompson’s Shark or bird beak shark and the
Cat Shark.
• The squalidae sharks live 500 to 1000 m below sea level. They have
no fish maw but carry a conspicuous big liver. This organ makes up
25 % of their body weight and is of vital importance for their
existence in greater depth. Because of a unique metabolism these
sharks can survive in great depth, where is lack on oxygen.
• Unlike whales, which are a viviparous and an often endangered
species, most of the squalidae sharks show an successful
ovoviviparous or in some cases also an oviparous reproduction, with
up to twelve eggs at a time. The squalidae family feed mainly on
plankton, algae en small fishes and reach a maximum body length of
1,5 m.
Squalene sources
• The most abundant source of squalene is from the livers of
deep sea sharks to be found at depths of as much as 1,500
metres. It occurs in Shark liver oils as the major
component, comprising up to 85% of the oil. Squalene is
not found in sharks living at shallow sea levels.
• Several type of sharks live 3,000 feet under the sea. At this
depth, sunlight and oxygen are almost nil. The
Atmospheric pressure is intense and conditions are harsh.
The ability of these sharks to survive under a hostile
environment is due largely to the size and content of their
liver which contains pure Squalene. It is Squalene which
supplies much of the volume of oxygen needed in their
bodies, providing strength and stamina.
• All plants and animals produce squalene as a
biochemical intermediate, including humans. It
occurs in high concentrations in the stomach oil of
birds in the order Procellariiformes.
• Squalene is a hydrocarbon and a triterpene, and 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.
Health Benefits
• Adaptogen ( Adaptation Syndrome )
Squalene is non-toxic, 100% natural and has no
side effects. Its beneficial action is not limited to a
particular organ. If any, initial reactions result from
toxins being released from the body.
• Cell-Invigorating
Squalene revitalizes weakened body cells and helps
revive cell generation. Its chief attribute is the
protection it affords cells from oxidation reactions.
The human body has about 6 billion oxygen reliant
cells. Oxygenation promotes good health to the
most basic level of life - the cell.
• Purifying
Squalene helps clean, purify, and detoxify the blood from
toxins, facilitating blood circulation. It cleanses the
gastrointestinal tract and kidneys, causing better bowel
movement and urination. Many diseases are cured if the
blood is purified, a condition Squalene contributes to.
• Curbing of Bad Cholesterol
Squalene is good cholesterol that is made up of high
density lipid (HDL) which prevents the hardening of
blood vessels and deposition of cholesterol plaques.
To test the authenticity of pure Squalene, you can place
the deep-sea shark liver oil extract in the freezer. Even at
low temperatures, it should not freeze, allowing the
substance to flow freely into your system, pumping
oxygen into the cells.
• Anti-Aging
Since Squalene prevents cell damage, beautiful, healthy cells
are produced. Linked together with lipoproteins, it later
forms lipofuscin, an anti-aging substance. It also prevents
the formation of harmful lipid peroxide which destroys
various kinds of vitamins.

• Oxygen Control
Squalene carries oxygen in the cellular level, causing further
improvement in organ function through cellular metabolism,
preventing the acidotic cell syndrome where cells become
acidic, deteriorate and die due to lack of oxygen.

There is a significant relationship between the degree of one's

health and the amount of oxygen consumption by body cells.
In today's polluted environment, lack of exercise, and poor
lifestyle, Squalene gives your body a beneficial source of
oxygen.
• Sterilizing
Squalene's terpene gives a sterilizing effect,
combating the growth of various germs such as
coliform bacilli, dysentery bacilli, micrococcus
pyocynanel, staphylococcus, hemolytic
streptococcus, and candida albicans.
• Generation of Body Hormones
Squalene naturally increases male potency and
vitality through a better body. It also helps
regulate the female menstrual cycle and improves
irregular and abnormal cycles.
• Other Actions:
Squalene - reduces various aches and pains - helps
body organs such as the kidneys, liver and gallbladder
function properly - helps digestive system function
properly by reducing gas troptosis conditions, helping
to shrink hemorrhoids and curbing obesity –
• Acts as a relaxant, giving added vigor and vitality
without the hyper-activity associated with other food
supplements - generates hair and smoothens skin -
exhibits a penetrating action with immediate effects
on topical applications - helps prevent various kinds
of diseases and speeds up the healing process in
most conditions of ill health.
Main properties
• Health as well as dysbalance of our body has its
origin on the cellular level. A wide range of
squalene’s core properties influence this level.
Therefore it find a wide application:
• Anti-oxidation - powerful Anti-oxidant
• Oxygen generator - vast recuperation
• Immune stimulator - good immune response
• Fat regulator/energy modulator - natural vitality
Squalene - A strong natural anti-oxidant

• The biochemical structure of squalene is C30 H50

(C30: 6n-omega 2) all trans isoprenoid, that means a
C30 polyprenyl compound hold 6 prenyl (better
known as isoprenoid or isoprene).
• Because of a double bound structure this isoprenoid
has the function of a strong anti-oxidant and a
natural antibiotic.
• The molecular weight of squalene is heavier than
that of EPA or DHA, through which all active bonding
positions of the receptor can be maximal belayed.
As consequence squalene permeates not only
deeper in cells but also faster and more effective.
Squalene- A unique oxygen generator
• The unsaturated hydrocarbons of squalene (C30 H50)
bind hydrogen ions from water and saturate with that
squalene (C30 H62). This chemical process release 3
unbound oxygen molecules (O2):

• C30H50 (squalene) + 6 H20 (water) > C3) H62 + 3 O2

(oxygen).
Squalene: a unique oxygen generator
• Squalene has shown efficiency in improving the quantity and quality of the oxygen
saturation in cells, leading in a impressive number of cases to an enhanced vitality.
• As an omega 2 fatty acid squalene has a redox function which releases oxygen
from water, resulting in this above mentioned increased oxygen level in the cells.
• Squalene is in fact an oxygen generator which provides our organism with a
sufficient oxygen level and distributes it into the remotest cells of our body.
• Squalene will mainly lay up in our skin and the adipose tissue, the natural stock of
this substance. The increased oxygen level stimulates also the natural metabolism.
• There is strong empiric evidence that the combination of squalene with regular
medication produces good and sometimes even better results in the recovery
process. In comparison to adults the body of babies and pre-schoolers show the
highest levels of squalene. This fact could explain the energetic behaviour and the
vivid eyes of young and healthy children.
• Starting with an age of 30 the squalene level in our organism drops significantly.
Therefore it is not surprising that round an age of 30 the sport performances of
most humans decline. Squalene can intercept this lack of energy and improve the
power output.
 Powerful immune stimulator
• An important substance in shark liver oil are the alkoxyglycerols
(AKG’s), also known as glycerol ether lipids, G-E’s), which are
stimulating the immune system and protect us against
uncontrolled and malignant cell growth.
• Swedish biochemist Sven Brohult discovered that AKG’s support
the production of white corpuscles, which are responsible for an
optimal function of our immune system.
• Alkoxyglycerols are also present in organs being essential for our
immune defence like spleen, lymph nodes as well as in bone
marrow and mother’s milk. Because of an hostile environment a
baby need to strengthen his immune system in no time.
• The main substance of mother’s milk, alkoxyglycerol, stimulates
and boost this process. Shark liver oil contain much more of this
substance than other fish oils.
Sustainable properties (strong
points) of Squalene
Shark liver oil has proofed to work out sustainable and effectively
and can be seen as nutrition supplement with the following
properties and applications:
•Anti-oxidant (intercepting free radicals)
•Anti-biotic (anti bacterial, anti-infectious)
•Anti-coagulant ( reducing thrombocyte aggregation, disintegrate
blood clots)
•Anti-histaminic (suppressing histamine)
•Anti-phlogiston (combat inflammation)
•Anti-allergic (preventing allergic reactions)
•Anti-carcinogen (anti-cancer through differentiation of cancer
cells to adipocyt [fat cell] apoptosis [cell death] and necrosis
[cell decomposition])
•Digestive (stimulating digestion)
•Immune stimulant (improving resistance, supporting
development and activity of phagocytes and lymphocytes)
Derivative used as a skin moisturizer
in cosmetics
• Squalene is one of the most common lipids produced by
human skin cells. It is a natural moisturizer, and occurs as
a major component of nasal sebum.
• Squalane is a saturated form of squalene in which the
double bonds have been eliminated by hydrogenation.
Squalane is less susceptible to oxidation than squalene.
Squalane is thus more commonly used than squalene in
personal care products, such as moisturizers.
• Toxicology studies have determined that in the
concentrations used in cosmetics, both squalene and
squalane have low acute toxicity, and are not significant
human skin irritants or sensitizers.
Use as an adjuvant in vaccines
• Immunologic adjuvants are substances, administered
in conjunction with a vaccine, that stimulate the
immune system and increase the response to the
vaccine. Squalene is not itself an adjuvant, but it has
been used in conjunction with surfactants in certain
adjuvant formulations.
• An adjuvant using squalene is Novartis' proprietary
adjuvant MF59, which is added to influenza vaccines to
help stimulate the human body's immune response
through production of CD4 memory cells. It is the first
oil-in-water influenza vaccine adjuvant to be
commercialized in combination with a seasonal
influenza virus vaccine.
• A later study reported that about one in ten people
have squalene antibodies in their blood, regardless
of whether or not they received squalene from a
vaccination. A later study confirmed this result,
and also showed that vaccination with squalene-
containing vaccines do not alter the levels of these
naturally-occurring antibodies.
• Oil-water suspensions, including MF59, were
associated with the ability to induce lupus
autoantibodies in non-autoimmune mice.
• In one study, endogenous squalene was linked to
autoimmune arthritis in rats. An epidemiologic
analysis of safety data on MF59 seasonal and
pandemic influenza vaccines showed no evidence
of increased risk of vaccine adverse events of
potential autoimmune origin.
• The World Health Organization and the US
Department of Defense have both published
extensive reports that emphasize that squalene is
a chemical naturally occurring in the human body,
present even in oils of human fingerprints.
• WHO goes further to explain that squalene has
been present in over 22 million flu vaccines given
to patients in Europe since 1997 and there have
never been significant vaccine-related adverse
events.
Drug Warning
Squalene supplementation
Squalene supplementation should be avoided in infants,
children, pregnant women and nursing mothers.PDR for
Nutritional Supplements 2nd ed. Thomson Reuters,
Montvale, NJ 2008, p. 595
• Those taking squalene supplements may have mild
gastrointestinal symptoms such as diarrhea.PDR for
Nutritional Supplements 2nd ed. Thomson Reuters,
Montvale, NJ 2008, p. 595
• Squalene should not be confused with squalamine,
which is an unusual steroid found in the dogfish shark
and which has antibiotic properties.PDR for
Nutritional Supplements 2nd ed. Thomson Reuters,
Montvale, NJ 2008, p. 595
Metabolism/Metabolites
• A review of the oxidization of squalene, a specific human
compound produced by the sebaceous gland, is proposed.
Such chemical transformation induces important
consequences at various levels. Squalene by-products,
mostly under peroxidized forms, lead to comedogenesis,
contribute to the development of inflammatory acne and
possibly modify the skin relief (wrinkling).
• Experimental conditions of oxidation and/or photo-
oxidation mechanisms are exposed, suggesting that they
could possibly be bio-markers of atmospheric pollution
upon skin. Ozone, long UVA rays, cigarette smoke ... are
shown powerful oxidizing agents of squalene.
• Some in vitro, ex vivo and in vivo testings are proposed as
examples, aiming at studying ingredients or products
capable of boosting or counteracting such chemical
changes that, globally, bring adverse effects to various
cutaneous compartments.[Pham DM et al; Int J Cosmet
Sci 2015 Feb 6. doi: 10.1111/ics.12208. Abstract: PubMed
REFERENCES
• Ernst, Josef; Sheldrick, William S.; Fuhrhop, Juergen H. (1976).
"Crystal structure of squalene". Angewandte Chemie. 88 (24): 851.
• Bloch, Konrad E. (1983). "Sterol, Structure and Membrane Functio".
Critical Reviews in Biochemistry and Molecular Biology. 14: 47–92
• Wolosik K1, Knas M, Zalewska A, Niczyporuk M, Przystupa AW. The
importance and perspective of plant-based squalene in cosmetology.
J Cosmet Sci. 2013 Jan-Feb;64(1):59-66.
• Spanova, Miroslava & Daum, Guenther. Squalene - biochemistry,
molecular biology, process biotechnology, and applications. Eur. J.
Lipid Sci. Technol. 2011
REFERENCES
• Final Report on the Safety Assessment of Squalane and Squalene".
International Journal of Toxicology. 1 (2): 37–56. 1982.
• Mosca, F.; Tritto, E.; Muzzi, A.; Monaci, E.; Bagnoli, F.; Iavarone, C.;
O'Hagan, D.; Rappuoli, R.; De Gregorio, E. (2008). "Molecular and
cellular signatures of human vaccine adjuvants". Proceedings of the
National Academy of Sciences. 105 (30): 10501–6.
• Kalvodova, Lucie (2010). "Squalene-based oil-in-water emulsion
adjuvants perturb metabolism of neutral lipids and enhance lipid
droplet formation". Biochemical and Biophysical Research
Communications. 393 (3): 350–5.
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