506 Eur. J. Lipid Sci. Technol.

104 (2002) 506–512

M. Assunta Dessìa, Oxidative stability of polyunsaturated fatty acids:

Monica Deianaa,
Billy W. Dayb,
Antonella Rosaa,
Sebastiano Bannia,
Francesco P. Corongiua
a against temperature-dependent autoxidation and UVA (ultraviolet A, 320–380 nm)
Dipartimento di Biologia
Sperimentale, Sezione di
Patologia Sperimentale,
Università degli Studi di
Cagliari, Cittadella Univer-
sitaria, Monserrato (CA),
Italy
b Departments of Environ-
mental and Occupational
Health and Pharmaceuti-
cal Sciences, Graduate
School of Public Health
and School of Pharmacy,
University of Pittsburgh,
Pittsburgh, PA, USA
effect of squalene*
The propensity of polyunsaturated fatty acids (PUFAs) to undergo oxidation plays an
important role in the integrity of biological membrane and lipid containing foods. The
ability of squalene (SQ), a naturally occurring dehydrotriterpene present in animal and
plant tissues, to protect linoleic, linolenic, arachidonic and docosahexaenoic acids
mediated oxidation was assessed. The oxidation of PUFAs was protected in varying
degrees, with highest protection observed for linolenic, arachidonic and docosa-
hexaenoic acids. Linoleic acid was less protected. At a molar ratio of 7:1 (PUFA:SQ)
the inhibition of the oxidation process was 22% in the presence of linoleic acid and
about 50% in presence of the other PUFAs tested. The different protection exerted by
SQ against PUFAs with different degrees of unsaturation may be accounted for by the
higher stability of octadecadienoic acid hydroperoxide isomers compared with respec-
tive PUFA hydroperoxides. Observing mild UVA-mediated oxidation and the temper-
ature-dependent autoxidation reactions we found similarities in the oxidation pattern
and the protection exerted by SQ. These findings suggest that the reaction of autoxi-
dation is predominant and SQ acts mainly as peroxyl radical scavenger.
Keywords: Squalene, PUFA, autoxidation, hydroperoxide, antioxidant.

1 Introduction the understanding of the role and benefits of natural an-

Squalene (SQ), a dehydrotriterpene hydrocarbon with six
double bonds, is present in animal and plant tissues and
at high concentration in shark liver [1]. Nearly all tissues
synthesize SQ but it is generally converted to cholesterol.
In adult human skin SQ accumulates to levels approach-
ing 10% of the total lipids and represents one of the major
components of human skin surface lipids [2]. Skin is con-
Research Paper
tioxidants on the cutaneous pathophysiology of photoox-
idative stress can depend on the selection of endpoints in
the evaluation models of photoprotectants [12].
Squalene is a 30 carbon chain compound with 6 double
bonds that is structurally similar to β-carotene. Recent re-
ports have stipulated that olive oil contains 0.2%–0.7%
squalene, the average squalene intake in the United

stantly exposed to ultraviolet (UV) light and hence may be
under increased oxidative stress due to UVA-dependent
free radical generation. Solar UVA (320 nm – 380 nm) is a
significant factor in the induction of skin lipid peroxidation,
as the amount of UVA radiation reaching the earth’s sur-
face is about 20 times greater than that of UVB (280 nm –
320 nm) [3]. UVA radiation has been suggested to be an
important factor in carcinogenesis in animal exposed to
sunlight [4, 5]. The process leading to cancer is complex
and it has been suggested that singlet oxygen reacting di-
rectly with C=C double bonds to give peroxides might be
involved, thus triggering autoxidation reactions [3, 6, 7],
the products of which could act as second messengers
affecting cancer genes [8, 9]. This oxidative stress sug-
gests a protective role for antioxidants [10, 11]. However,
States of America being 30 mg/day, and that this intake
can reach 200–400 mg/day in Mediterranean countries
(reviewed in Smith [13] and Newmark [14]). SQ, scaveng-
ing both free radicals and oxygen reactive species, has
been shown to protect mice skin against γ-irradiation
[15, 16] and to be an important dietary cancer chemo-pre-
ventive agent [13, 14, 17, 18].
A model system for assessing the autoxidation of PUFAs
has been described [19]. This uses a reaction environ-
ment without solvents and involves a mild UVA exposure.
In this study we assessed the ability of SQ to protect
polyunsaturated fatty acids (PUFAs) against temperature
dependent autooxidation (to mimic exposure to varying
temperatures) and to mild UVA (to mimic singlet oxygen-

* Dedication: We dedicate this paper to the memory of Profes-

Correspondence: M. Assunta Dessì, Dipartimento di Biologia
Sperimentale, Sezione di Patologia Sperimentale, Università
degli Studi di Cagliari, Cittadella Universitaria, SS 554, Km 4.5,
09042 Monserrato (CA), Italy. Phone: +39-070-675-4125, Fax:
+39-070-675-4032; e-mail: dessima@unica.it
sor Francesco P. Corongiu who died on 17th August 2000.
Francesco’s vision and leadership played a pivotal role in es-
tablishing the Division of Experimental Pathology at the Univer-
sity of Cagliari. In addition to his great academic accomplish-
ments he was a great friend to all who knew him.

© 2002 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim 0931-5985/2002/0808-0506 $17.50+.50/0
Eur. J. Lipid Sci. Technol. 104 (2002) 506–512
dependent reaction) oxidation. We report that squalene
protected the oxidation of PUFAs at varying degrees and
suggest that these might be an important consideration
for the assessment of the photoprotective effects of natur-
al antioxidants.
2 Materials and methods
2.1 Autoxidation of PUFAs
All solvents used were high-performance liquid chro-
matography (HPLC) grade (Carlo Erba, Milano, Italy).
Autoxidation of PUFA was carried out as previously de-
scribed [19]. Briefly, 1 ml of a 3.6 mM solution in n-hexa-
ne of each PUFA (Sigma Chemical Co., St. Louis, MO,
USA), were dried under vacuum and incubated in a Dub-
noff water bath at 37 °C in a round bottom test tube. The
reaction was stopped, the mixture cooled to 0 °C and
0.5 ml of CH3CN/0.14% CH3COOH (v/v) added, when
about 70–80% of the PUFA were oxidized (linoleic acid,
24 h; linolenic acid, 14 h; arachidonic acid, 10 h; docosa-
hexaenoic acid, 5 h). Samples were then analyzed by
HPLC.
SQ (Sigma Chemical ) was mixed with PUFAs prepared
as above at the working final concentrations: 20, 40, 80,
160, 240, 320, 400, 480 nmoles achieved by adding ap-
propriate volumes of 2.43 mM SQ in n-hexane. Samples
were then dried down and incubated at 37 °C for different
lengths of time as described above. For comparative pur-
poses ancillary experiments were performed using
squalane (Sigma Chemical ), the saturated form of SQ.
2.2 UVA-mediated oxidation of PUFAs
UVA-mediated oxidation of PUFAs was carried out on
samples prepared as follows: 1 ml of 3.6 mM PUFA solu-
tions in n-hexane, was dried under vacuum in a glass
round bottom test tube. Unstoppered tubes, transparent
to UV radiation, were exposed to UVA monochromatic ra-
diations (366 nm) at a fluence rate of 250 kJ/m2, a physi-
ological dose of UVA as suggested by Vile and Tyrrell [6],
for various lengths of time. In all experiments, samples
were irradiated with the same lamp from a distance of
30 cm in a thermostatic system at 20 °C.
The reaction was stopped, adding 0.5 ml of CH3CN/
0.14% CH3COOH (v/v), when about 70–80% of the PUFA
were oxidized (linoleic acid, 56 h; linolenic acid, 20 h;
arachidonic acid, 15 h; docosahexaenoic acid, 8 h) and
samples were then analyzed by HPLC.
Samples containing SQ were prepared as described
above, and used at the concentrations of 240, 320, 400
and 480 nmol.
Prevention of PUFA oxidation by squalene 507
2.3 HPLC-diode array detector analyses
Separation of PUFAs was conducted essentially as de-
scribed in Banni et al. [19]. A Hewlett-Packard 1050 liquid
chromatograph equipped with a 1040M diode array de-
tector (Hewlett Packard, Palo Alto, CA, USA) was used. A
C18 Alltech Adsorbosphere column (Alltech, Deerfield, IL,
USA), 5 µm particle size, 250 × 4.6 mm, was used for
all separations, with a mobile phase of CH3CN/H2O/
CH3COOH (70:30:0.12, v/v/v) flowing at 1.5 ml/min. PU-
FA hydroperoxides were monitored at 234 nm and PUFAs
at 200 nm [19]. SQ was eluted with 100% methanol at a
flow rate of 1.5 ml/min and it was monitored at 203 nm.
Second derivative as well as conventional UV spectra
were generated, using the Phoenix 3D HP Chemstation
software, in order to confirm the peaks identification [20].
Further identification of components in the chromato-
graphic peaks was performed with the help of reference
compounds. Cis,trans-13-hydroperoxy-octadecadienoic
acid (c,t-13-HPODE) and cis,trans-9-hydroperoxy-oc-
tadecadienoic acid (c,t-9-HPODE) were purchased from
Cascade (Cascade Biochemical Ltd. London, UK).
2.4 Characterization of SQ oxidation
by-products
Characterization of SQ by-products formed during the
autoxidation or UVA-mediated oxidation of PUFA in the
presence of SQ was obtained by combining HPLC-UV
analyses with on-line atmospheric-pressure ionization
mass spectrometry. In these analyses, NH4O2CCH3
(2 mM) was added to the mobile phase to insure ioniza-
tion of the analytes. Pneumatically assisted electrospray
mass spectra were obtained on a Perkin Elmer/Sciex API
I mass spectrometer, equipped with an atmospheric-pres-
sure ionization source and an ionspray interface (Perkin
Elmer/Sciex, Toronto, Ontario, Canada). The interface
was maintained at 5 kV, and the orifice voltage was set at
70 V. High purity air was used as nebulizing gas at an
operating pressure of 276 kPa, and the curtain gas was
high purity N2 flowing at 0.6 l/min, heated to 55 °C. The
analytes in the HPLC mobile phase, after photodiode
array analysis, were introduced directly into the nebuliza-
tion ionization source with no splitting of the effluent. The
mass spectrometer was set to scan over the range of m/z
280 to 600 at a resolution of m/z 0.1, and a rate of one
scan per 8.67 s.
2.5 Statistical analysis
INSTAT software (GraphPad software, San Diego, CA,
USA) was used to calculate the means and standard de-
viations of three independent experiments, involving trip-
licate analyses for each sample/condition. One-way ANO-
VA was used to test whether the group means differed
significantly.
508 Dessì et al. Eur. J. Lipid Sci. Technol. 104 (2002) 506–512

3 Results
Oxidative patterns for all the PUFAs were similar, irre-
spective of their different oxidative stability due to the
number of double bonds. The oxidation parameters
(PUFA decrease and PUFA hydroperoxides formation)
were monitored at 1 h or 30 min intervals in this study.
In order to obtain an equivalent oxidative pattern for all
PUFAs, a preliminary set of experiments was performed
blocking the autoxidation, or UVA-mediated oxidation,
when PUFAs were oxidized to about 70–80%.
Fig. 1 shows the decrease of linoleic acid and the forma-
tion of its HPODE isomers during 24 h of autoxidation at
37 °C or 56 h of UVA-mediated oxidation at room temper-
ature (20 °C). Although the UVA-mediated oxidation at
20°C was slower than the autoxidation process at 37 °C,
the oxidation pattern was the same in the two experimen-
tal systems. The concentration of linoleic acid decreased
with time and the value of HPODE isomers increased
with a non-linear time course, in agreement with previous
reports [19, 21]. Linolenic, arachidonic and docosa-
hexaenoic acids had similar autoxidation pattern (Tab. 1 Fig. 1. Decrease of linoleic acid and formation of HPODE
and Tab. 2). The PUFAs showed the highest production of isomers during 56 h of UVA-mediated oxidation at 20 °C
PUFA hydroperoxides when about 40 % of their initial (A) or 24 h of autoxidation at 37 °C (B).
concentration was consumed (Fig. 1, Tab. 1, Tab. 2). This * = p <0.05; ** = p <0.01; *** = p <0.001 versus controls
point was reached at different times of incubation for the (samples at T = 0)

Tab. 1. Decrease of linoleic (A) and linolenic acid (B) and polyunsaturated fatty acids hydroperoxide (PUFA HP) formation
during autoxidation at 37 °C and UVA–mediated oxidation at 20 °C. * = p <0.05; *** = p <0.001 vs controls (samples at
T = 0).
A Autoxidation UVA-mediated oxidation
Incubation Residual PUFA PUFA HP Incubation Residual PUFA PUFA HP
time [h] [nmol] [nmol] time [h] [nmol] [nmol]
0 3528 ± 41 71 ± 12 0 3501 ± 41 80 ± 8
4 3413 ± 90* 220 ± 14*** 16 3281 ± 66*** 192 ± 38***
8 2969 ± 156*** 440 ± 39*** 24 2682 ± 7*** 322 ± 52***
12 2548 ± 55*** 711 ± 12*** 32 2346 ± 53*** 469 ± 6***
16 1906 ± 43*** 704 ± 5*** 40 1868 ± 62*** 516 ± 39***
20 1408 ± 36*** 575 ± 50*** 48 1299 ± 71*** 479 ± 63***
24 954 ± 33*** 423 ± 35*** 56 923 ± 19*** 342 ± 19***
B Autoxidation UVA-mediated oxidation
Incubation Residual PUFA PUFA HP Incubation Residual PUFA PUFA HP
time [h] [nmol] [nmol] time [h] [nmol] [nmol]
0 3508 ± 33 7± 3 0 3750 ± 96 34 ± 1
2 3295 ± 9*** 46 ± 2* 4 2934 ± 62*** 151 ± 2***
4 3242 ± 25*** 140 ± 28*** 8 1606 ± 105*** 311 ± 12***
6 3031 ± 15*** 268 ± 48*** 12 1288 ± 65*** 257 ± 10***
8 2571 ± 28*** 426 ± 19*** 16 718 ± 34*** 142 ± 8***
10 2195 ± 44*** 449 ± 27*** 20 427 ± 21*** 85 ± 2***
12 1789 ± 39*** 443 ± 26***
14 1154 ± 57*** 316 ± 5***
Eur. J. Lipid Sci. Technol. 104 (2002) 506–512 Prevention of PUFA oxidation by squalene 509

Tab. 2. Decrease of arachidonic (A) and docosahexaenoic acid (B) and hydroperoxide (PUFA HP) formation during
autoxidation at 37 °C and UVA–mediated oxidation at 20 °C. ** = p <0.01; *** = p <0.001 vs. controls (samples at T = 0).
A Autoxidation UVA-mediated oxidation
Incubation Residual PUFA PUFA HP Incubation Residual PUFA PUFA HP
time [h] [nmol] [nmol] time [h] [nmol] [nmol]
0 3637 ± 58 6± 1 0 3402 ± 10 7± 0
1 3544 ± 82 17 ± 2 3 3265 ± 11*** 81 ± 10**
2 3526 ± 40 34 ± 8 6 2765 ± 175*** 236 ± 2***
3 3377 ± 12** 92 ± 8*** 9 1534 ± 12*** 278 ± 13***
4 3335 ± 87*** 155 ± 17*** 12 774 ± 17*** 181 ± 2***
5 3119 ± 20*** 250 ± 37*** 15 476 ± 41*** 91 ± 3***
6 2803 ± 45*** 345 ± 8***
7 2381 ± 48*** 407 ± 17***
8 1775 ± 216*** 362 ± 4***
9 1344 ± 138*** 287 ± 1***
10 813 ± 87*** 186 ± 41***
B Autoxidation UVA-mediated oxidation
Incubation Residual PUFA PUFA HP Incubation Residual PUFA PUFA HP
time [h] [nmol] [nmol] time [h] [nmol] [nmol]
0 373 ± 83 32 ± 93 0 3459 ± 15 21 ± 2
0.5 3662 ± 20 62 ± 1** 2 2873 ± 1*** 165 ± 4***
1 3447 ± 7*** 129 ± 1*** 4 2099 ± 48*** 229 ± 6***
1.5 3376 ± 45*** 231 ± 1*** 6 1320 ± 125*** 228 ± 1***
2 3096 ± 11*** 363 ± 26*** 8 573 ± 25*** 176 ± 10***
2.5 2593 ± 113*** 419 ± 5***
3 2056 ± 53*** 405 ± 12***
3.5 1664 ± 91*** 350 ± 31***
4 1412 ± 38*** 230 ± 17***
4.5 700 ± 8*** 142 ± 23***
5 442 ± 36*** 75 ± 3***

different PUFAs, decreasing exponentially upon the num-
ber of double bonds (Fig. 2). This data would also concur
that the higher the number of double bonds, the greater
the oxidative propensity of the individual PUFA. HPODEs
formation was about 1.5-folds higher than hydroperoxides
formed during oxidation of the other PUFAs tested
(Tab. 1, Tab. 2). The antioxidant activity of SQ versus
PUFAs oxidation is shown in Fig. 3 and expressed as per-
centage of protection, calculated from the plot of concen-
tration-dependent residual fatty acid and hydroperoxides
formed. The protection exerted by SQ was the same in
the two experimental systems and it was linearly depen-
dent from its concentration. At the highest SQ concentra-
tion tested (480 nmol) linoleic acid was protected by 22%,
while the more unsaturated PUFAs were protected by
about 50% (Fig. 3). An ancillary experiment was per-
formed by substituting squalane for SQ. No protection
during linoleic acid autoxidation was detected at any
squalane concentration tested.

SQ consumption was higher in the presence of linoleic

Fig. 2. Correlation between the number of PUFA double
bonds and the time of incubation needed to reach the
maximum formation of PUFA hydroperoxides during aut-
oxidation at 37 °C.
acid than in the presence of the other PUFAs tested, dur-
ing both the autoxidation and the UVA-mediated oxidation
of PUFAs, and three major by-products derived from SQ
were formed (Fig. 4). The level of SQ hydroperoxide for-
mation varied upon the degree of unsaturation of PUFAs
either when taken at the end of the process (Fig. 5) or
when compared at the same time of incubation (Fig. 6).
The SQ oxidation by-products were detected exclusively
when SQ was incubated with PUFAs.
510 Dessì et al.
Fig. 3. Protection exerted by SQ at different concentra-
tions against PUFA UVA-mediated oxidation at 20 °C (A)
or autoxidation at 37 °C (B). Percentage of protection ver-
sus all the PUFA tested, calculated from the plot of con-
centration-dependent residual fatty acid and hydroperox-
ides formed, is extremely significant (p < 0.001) from
240 nmol of SQL (system A) and from 160 nmol of SQL
(system B).
4 Discussion
The decreasing concentration of linoleic acid and subse-
quent formation of HPODEs are important determinants
of autoxidizability of this fatty acid [19]. If this process is
slowed down, it becomes possible to use the system in
assessing antioxidant activity. Since the formation of
hydroperoxides during the time course of linoleic acid
autoxidation is not linear, misinterpretation of oxidative
propensity can result when both parameters are not mea-
Eur. J. Lipid Sci. Technol. 104 (2002) 506–512
Fig. 5. Relationship between SQ concentration and SQ
hydroperoxide formation during PUFA autoxidation at
37 °C: A – docosahexaenoic acid; B – linolenic acid; C –
arachidonic acid; D – linoleic acid.
Fig. 6. Relationship between SQ hydroperoxide forma-
tion and incubation time during PUFA autoxidation at
37 °C: A – docosahexaenoic acid; B – linolenic acid; C –
arachidonic acid; D – linoleic acid.
sured. In this paper the ability of SQ to act as scavenger
of peroxyl radicals and protect linolenic acid, arachidonic
acid and docosahexaenoic acid autoxidation was as-
sessed and compared with linoleic acid. Lipid peroxyl rad-
icals are the main reactive molecules during the temper-
ature-dependent PUFA autoxidation reactions in this
study. However, and as is evident from previous studies,
using physiological doses of UVA radiation, singlet oxy-
gen mediates UVA radiation-dependent peroxidation of
PUFA, a reaction that can be potentiated by the presence
Fig. 4. Typical chromatogram recorded at 203 nm
of SQ and its by-products formed during linoleic
acid oxidation. Peaks were identified by their UV
and mass spectra.
Eur. J. Lipid Sci. Technol. 104 (2002) 506–512
of iron and hydrogen peroxide [6]. In both the experimen-
tal systems, SQ exerted a significant antioxidant activity.
At a molar ratio of 7:1 (PUFA: SQ) the inhibition of the
oxidation process was 22% in the presence of linoleic
acid and about 50% in the presence of the other PUFAs
tested. The different protection exerted by SQ against
PUFAs that possess different degrees of unsaturation
may be accounted for by the higher stability of HPODE
isomers with respect to the other PUFA hydroperoxides.
PUFA hydroperoxides containing more than 2 double
bonds have the tendency to yield by-products not easily
converted to radicals capable of attacking other PUFA
molecules [21]. Therefore, substrates are exposed for a
longer period to HPODEs with respect to the other PUFA
hydroperoxides. As a consequence, SQ consumption
was higher and its antioxidant activity was lower in the
presence of linoleic acid than in the presence of linolenic,
arachidonic and docosahexanoic acids. The highest pro-
duction of hydroperoxides was attained when about 40%
of PUFA was oxidized. Concentrations of 700 or 500 nmol
for linoleic acid and about 400 or 300 nmol for the other
PUFAs tested were obtained depending on the experi-
mental system. Under our experimental conditions, SQ
oxidation by-products were detected exclusively when
SQ was incubated with PUFAs. Therefore SQ hydroper-
oxide formation was related to the capability of SQ to
scavenge the lipoperoxyl radicals. A similar reaction to
that proposed by Yeo and Shibamoto [22] under UV irra-
diation, may occur in the experimental condition reported
here, where a proton abstraction from SQ is followed by
the generation of SQ hydroperoxide and SQ hydroxide
Fig. 7. Proposed formation of SQ oxidation by-products,
SQ hydroperoxyde (SQOOH) and SQ hydroxide (SQOH)
during the reaction with lipoperoxyl radicals (LOO).
Prevention of PUFA oxidation by squalene 511
(Fig. 7). Yeo and Shibamoto [22] proposed that hydrogen
atom abstraction from SQ is likely to occur in the C-8 car-
bon. We confirm this hypothesis with the detection by
HPLC of the fragmentation byproduct, 6-methyl-5-hep-
ten-2-one (6-MHO), generated by the 6-hydroperoxide in-
termediate.
Using different experimental conditions, 37 °C in the
absence of solvent, it has been demonstrated that SQ is
more resistant to peroxyl radical attack than PUFAs [16].
This might explain why SQ, together with SQ hydroperox-
ide, is not able to propagate the chain reaction of lipid per-
oxidation, working as protective agent. The lack of proox-
idant activity suggests that SQ peroxyl radical is probably
stabilized by resonance in its isoprenic structure. SQ is an
efficient quencher of singlet oxygen [16], that mediates
UVA radiation-dependent peroxidation of PUFA under
physiological doses of UVA radiation [3, 6]. The similarity
of the oxidation pattern and the protection exerted by SQ,
between mild UVA-mediated oxidation and the temper-
ature-dependent autoxidation reaction, suggests that the
reaction of autoxidation is predominant and SQ acts
mainly as peroxyl radical scavenger, irrespective of the
initiator used to trigger the reaction. Under UVA radiation,
the formation of lipid peroxyl radical is time-dependent
and lacks the effect of temperature seen under the autox-
idation conditions.
SQ activity is linked to its double bond system since its
saturated form, squalane, does not exert any of the lipid
peroxyl radical scavenging activity. SQ is one of the major
components of human skin surface lipids [2]. Nearly all
tissues synthesize SQ, that is generally converted to cho-
lesterol; however, in adult human skin SQ accumulates to
levels approaching 10% of the total lipids. Even though
the ratio PUFA: SQ at which SQ exerts its maximum ac-
tivity seems very high (7:1), this ratio mirrors what is pre-
sent in the skin surface [2]. The surface of hairless human
skin is especially vulnerable to damaging free radicals
generated by a number of physiological and biological
processes and to exposure to UV radiation. In hairy pri-
mates, namely Pan and Gorilla, SQ represents 0.1% only
of the skin surface lipids [23]. SQ is an endogenous prod-
uct that constantly accumulates on the human skin sur-
face, where it is the most susceptible to UV irradiation [8].
SQ hydroperoxide did not act as prooxidant in our exper-
imental conditions. This contrasts with the reports that SQ
hydroperoxide stimulates the melanogenesis process in
human keratinocytes [8]. Thus the ratio of antioxidant/
substrate concentrations is critical when assessing an-
tioxidant efficacy. Nevertheless, we report the inhibition of
PUFA oxidation by SQ and suggest that this might provide
a defence against lipid peroxidation induced by autoxida-
tion reactions. Moreover, measurement of thiobarbituric
acid reactive materials (TBARs) has been used to assess
512 Dessì et al.
the index of UV induced oxidative stress [3, 6, 24, 25], the
profile of PUFA stability offers useful markers for the as-
sessment of the redox modifying agents at sites of
UVA/UVB radiation generated damage.
The current interest in the cancer preventive properties of
squalene is noteworthy primarily from the standpoint that
experimental studies have show that squalene can effec-
tively inhibit chemically-induced colon, lung and skin tu-
morigenesis in rodents [9, 17, 18]. Both Smith [13] and
Newmark [14] in their reviews of the current status of
chemopreventive potential of squalene refer to the reduc-
tion of mutated ras oncogene activation by squalene
which is primarily a consequence of the inhibition of the
activity of HMG-CoA (3-hydroxy-3-methylglutaryl coen-
zyme A) reductase catalytic activity in vivo, thus reducing
the availability of farnesyl pyrophosphate which is re-
quired for the prenylation of the ras oncogene [26]. Al-
though the protective effects from increased levels of
squalene in animal studies are not known we report that
increased concentrations of squalene over the concentra-
tion of PUFAs in vitro renders protection to the oxidation
of the PUFAs.
Acknowledgements
This work was supported by Consiglio Nazionale delle
Ricerche (CNR-Roma, Italy) Contract N° 96.0498.ST74
and by Regione Autonoma della Sardegna (RAS-PIC-
INTERREG II).
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[Received: March 1, 2002; accepted: May 23, 2002]