221 ENDOLICHENIC FUNGI EXTRACTS
phenolic, and flavonoid contents of lichens have been proven by various studies.33,34 However,
the small amount of lichens in nature limits the use of these ingredients in many sectors,
including cosmetics. Therefore, it is important to detect and prove the antioxidant and
bioactivities of ELF species isolated from lichens, especially in order to prevent skin damage
caused by UV. The reason for this is the slow growth of lichens in nature and their limited
availability, which limits their use. However, endolichenic fungi isolated from lichens are
thought to be important for their use in the cosmetic industry as they grow faster and their
extracts are obtained in a standardized manner.35 For these reasons, total antioxidant, phenolic
and flavonoid contents of lichens, and isolated ELF extracts were determined in this study.
The total antioxidant activity of extracts from Usnea sp., L. pulmonaria, and B. capillaris
lichens and their ELF extracts was determined using the DPPH method in vitro. In this
study, the total antioxidant activities of lichen extracts at 1 mg/mL concentration were
found to be 34.86% for B. Capillaris, 21.74% for L. Pulmonaria, and 23.94% for Usnea
sp. extracts. ELF extracts isolated from L. pulmonaria lichen exhibited similar antioxidant
activities as their host lichen extracts as shown in Table II (18.82–24.01). Among all
extracts, the highest DPPH activity was determined for T20-P26 (91.77%) and T20-B02
(87.82%) isolated from B. capillaris lichen. T22-P07 ELF extract isolated from Usnea sp.
exhibited 70.23%, and T22-B07 extract exhibited 74.26% total antioxidant activity, which
was considerably higher than that of the Usnea sp. from which they were isolated. It was
observed that T20-P26, T20-B02 isolated from Bryoria and T22-P07, T22-B07 isolated
from Usnea sp. were comparable to and even higher than the positive control L-ascorbic
acid at a concentration of 1 mg/mL. As a result, when the total antioxidant activities were
examined, it was observed that especially T20-B02, T20-P26 isolated from B.capillaris and
T22-P07, T22-B07 ELF extracts isolated from Usnea sp. had higher antioxidant activities
than their host lichen species. It is known that antioxidants play a role in preventing damage
Table II
Antioxidant Activity of Lichen and ELF Extracts
Name of the lichen and ELF DPPH
TPC
(mg GAE/g extract)
TFC
(mg Rutin/g extract)
Positive control L-Ascorbic Acid
(1 mg/ml)
74.20
Gallic Acid
(1 mg/ml)
444.67
Rutin
(1 mg/mL)
1008.96
Lobaria pulmonaria 21.74 ± 0.002 86.53 ± 13.39 ND
T04-P01 18.82 ± 0.003 8.85 ± 0.33 399.33 ± 2.31
T04-P03 22.57 ± 0.003 16.10 ± 9.31 212.37 ± 1.28
T04-P13 24.01 ± 0.02 23.42 ± 0.71 204.75 ± 1.15
Bryoria capillaris 34,86 ± 0.037 35.57 ± 2.433 ND
T20-B02 87.82 ± 0.01 128.87 ± 6.59 ND
T20-P07 11,89 ± 0.008 20.73 ± 5.90 274.27 ± 2.02
T20-P10 18.82 ± 0.01 16.81 ± 6.52 373.93 ± 3.89
T20-P26 91.77 ± 0.001 129.37 ± 10.58 51.45 ± 0.85
T20-P27 27.21 ± 0.003 33.03 ± 17.45 ND
Usnea sp. 23.94 ± 0.003 83.14 ± 5.57 263.632 ± 1.63
T22-P07 70.23 ± 0.002 77.68 ± 12.02 80.163 ± 1.02
T22-B07 74.26 ± 0.01 96.08 ± 6.04 ND
ND: No data

222 JOURNAL OF COSMETIC SCIENCE
such as sun-induced pigmentation and anti-aging.36,37 Although lichens are known to be a
potential source of antioxidants, their slow growth and limited numbers restrict their use
in the cosmetics industry. From this point of view, the determination of high antioxidant
content in ELF extracts in our study shows that it can be evaluated as a potential antioxidant
source in cosmetic products and can be used as a sustainable and environmentally friendly
source with economic benefits.
The TPC was calculated in terms of gallic acid equivalent. Although L. pulmonaria had
the highest phenolic content among the lichen species (86.53 mg GAE/g), the phenolic
contents of ELF extracts isolated from this lichen were found to be quite low. On the other
hand, T20-B02 and T20-P26 isolated from B. capillaris had the highest phenolic content,
with 128.87 mg GAE/g and 129.37 mg GAE/g, respectively, similarly to these extract total
antioxidant activities. The TPC of Usnea sp. mg GAE/gTPC activities of the ELF samples
showed similar results to Usnea sp. As a result, it was found that the majority of ELF
extracts had lower TPC activities than their host lichen species. The reason for this finding
may be due to exposure of lichens to various environmental factors such as temperature,
pressure, light, and wind in their surrounding environments. These different environmental
conditions provide the production of secondary metabolites of lichens and increase their
bioactivity. Due to the high phenolic contents of T20-B02 and T20-P26 isolated from
B.capillaris and T22-B07 ELF samples isolated from Usnea sp., we may suggest that they
can be used as potential sources in cosmetic industry.
When the total flavonoid content (TFC) was examined, L. pulmonaria did not show any TFC
activity, but the TFC activities of ELF extracts isolated from this species were found to be
in the range of 204.75–399.33 mg rutin/g. Similarly, no results were found for B. capillaris,
but the amount of flavonoids in the ELF extracts isolated from these lichens ranged from
51.45–373.93 mg rutin/g. Although the TFC value of Usnea sp. was found to be 263.32 mg
rutin/g, the TFC values of ELF extracts isolated from this species were lower than those
isolated from the lichen.
Lichen samples are a natural source of antioxidants.38,39 However, their slow growth in
nature limits their active use in industries such as cosmetics, despite their high antioxidant
activity. For this reason, it is thought that it is important to determine the bioactivities,
especially the antioxidant activities, of ELF samples isolated from lichens. In particular,
it has been reported that the antioxidant activity of the (3R)-5-hydroxymellein (150)
compound of ELF000039 species isolated from Parmotrema austrosinense lichen was found to
be high with an IC50 value of 1,170.8 µg/mL.40 Also, two new polyketides from Curvularia
trifolii isolated from Usnea sp. exhibited antioxidant activities with IC50 values of 4 and
1.3 mg/ml, respectively.41 However, studies on the bioactivities of ELF samples are limited,
and further detailed researches are needed.
Poornima et al. investigated the total antioxidant and phenolic contents of ELF extracts
isolated from different lichen species. They reported varying results in the total antioxidant
activities (10–90%) of ELF extracts isolated from different lichen species.42 They also
reported similar results in the total phenolic contents to those of the total antioxidant
activities. Similarly, the total antioxidant, phenolic, and flavonoid contents of ELF extracts
isolated from three different Usnea lichens were determined. They reported that the ELF
extracts had strong antioxidant activity, while the total phenolic and flavonoid content
results showed differences.43 According to the literature, there is a positive correlation
between antioxidant activity and phenolic content.44,45 Similar to this data, we also observed
a positive correlation between TPC of T20-B02 and T20-P26 isolated from the species B.