THE BIOACTIVE OF PINUS MERKUSII NEEDLE AND BARK EXTRACT AS ANTIOXIDANT AND ANTIAGING

Pinus merkusii is a native pine species to Southeast Asia and has used as an oleoresins source and raw material for pulp and paper industries. This plant also possesses several biological activities, such as anti-inflammatory and larvicidal activity. This study aims to evaluate the antioxidant and antiaging activity of P. merkusii needle and bark. The qualitative phytochemical screening was used to evaluate the presence of secondary metabolites compounds. DPPH (2,2-diphenyl-1-picryl-hydrazyl) methods evaluated the antioxidant activity, and an anti-tyrosinase assay was used to evaluate the antiaging activity. Phytochemical analysis showed flavonoids, phenols, alkaloids, tannins, and terpenoids in both extracts. Bark extract showed the presence of saponins and triterpenoids, while needle extract possesses steroids. The antioxidant activity (IC50) of P. merkusii bark extract was 59.32 ± 1.74 µg/mL, stronger than needle extract (68.67 ± 1.47 µg/mL). Also, the bark extract showed higher inhibitory activity of tyrosinase (IC50) 74.97 ± 1.54 µg/mL than needle extract (96.08 ± 1.77 µg/mL). From this investigation, P. merkusii bark extracts appeared to have more potential as a natural source of antioxidants and antiaging and might be beneficial in these subjects.


INTRODUCTION
. The degradation of ECM connected with skin aging and collated with the increasing activity of certain enzymes involved in skin aging [15], [16]. The role of antioxidants in preventing skin aging is essential since these compounds can neutralize free radicals by donating or accepting an electron to complete the unpaired molecules [17]. Moreover, antioxidant to prevent skin aging is also supported by its anti-inflammatory properties [17].
Pinus merkusii is the native species to Southeast Asia known as Sumatra pine, located in Indonesia, mainly in northern Sumatra; Aceh, Tapanuli, and Kerinci [18], [19]. P. merkusii mostly found in acidic and poor soils with an elevation between 800 and 200 m above sea level [18]. It is known that Indonesia is among the top three countries for the production of P. merkusii resin [20].
This pine has been used as a natural source for oleoresins to produce gum rosin and turpentine, while the wood is used as raw material for pulp and paper industries [21]. In the pulp industry, pine bark is mostly left discarded or used for fuel. The discarded pine bark left in large amount since pine bark accounts for 10-15% of the whole pine tree [22].
Several studies revealed that pine has high therapeutic value and has potential as a drug in the future due to its bioactivity, such as antioxidant, antimicrobial, antifungal, and anti-inflammatory [23]. P. pinaster, as the common pine that could be found in several places since it was easy to grow was reported to have anti-inflammatory, antioxidant, and wound healing activity in its essential oil [24].
Previous studies demonstrated that phenolic compounds such as matairesinol and nortrachelogenin and flavonoid compound such as pinocembrin are found in P. merkusii bark extract [19]. This is also confirmed by the findings [25] who identified flavonoid compounds from P. merkusii bark extract. The antioxidant, anti-inflammatory, and antifungal properties of flavonoid are considered essential in pharmaceutical and cosmetic applications [26]. For example, Pinocembrin, a flavonoid, has been reported to have the capability of absorbing UV rays, which enhance the possibility of its usage as a sunscreen in photoprotection [27].
Pinocembrin was also used as an antifungal since it could inhibit the mycelial growth of Penicillium italicum on the skin (peng).
Matairesinol, a lignan from the phenolic compound, were also reported to have potential as antiaging agents [28].
Although the information about P.

a. Preparation and Sample Extraction
The preparation and extraction of samples were determined by the following previously method with modification [29].

b. Phytochemical Screening
The filtrated of P. merkusii needle and bark was evaluated by qualitative assay of common plant secondary metabolites. The screening was carried out for flavonoids, alkaloids, tannins [30], saponins [31], phenols [32], steroid, terpenoids [33], and triterpenoid [34]. The changing of color, frothing, or precipitate formation was used for the test response.

c. Antioxidant Capacity Analysis
The antioxidant capacity of extracts was investigated using a DPPH assay. This assay was followed the previous method [35]. Scavenging activity (%) = (Ac -As) Ac × 100% Ac = negative control absorbance (without sample) As = sample absorbance The IC50 value was also determined as the sample concentration (µg/mL) required to inhibit 50% of the activity (IC50) calculated from a dose-response curve using GraphPad software (San Diego, CA, USA).

d. Anti-tyrosinase Activity Assay
The crude extracts were dissolved into   [42]. In previous research, [25] found that ethyl acetate extract of bark of P.merkusii contained flavonoid compound while tannins were identified in ethanolic-extract of P.

Antioxidant Activity
Since  µg/mL [44]. The antioxidant capacity of pine needle extract was also studied previously by [45]