GC-MS Chemical profile, Antioxidant Activity, and Sun Protection Factor of Essential Oil of Tea Tree (Melaleuca alternifolia) and Rosemary (Rosmarinus officinalis L.)

Abstract

The present investigation aimed to determine the oxidative potential, sun protection factor value, and half-maximal inhibitory concentration of rosemary essential oil and tea tree essential oil. These two essential oils have gained popularity as active ingredients in many cosmetic preparations due to their antioxidant activity, whether used individually or in combination. Gas chromatography-mass spectroscopy is used to identify the presence of different phytochemical constituents in essential oils. The GC-MS (Gas chromatography-mass spectroscopy) chemical analysis of tea tree oil revealed 34 and rosemary oil revealed 35 volatile chemical components with sesquiterpene hydrocarbon, monoterpenes (42.77%), and alcohols (41.01%) as major detected classes. The 2, 2-diphenylpicrylhydrazyl (DPPH) and nitric oxide-free scavenging activity techniques were used to investigate the antioxidant capacity of these oils. It was found that both tea tree and rosemary oil have the potential to slow down skin aging through their anti-oxidative properties using the approach of free radical scavenging activity. The UV spectroscopy method was used to determine the sun protection factor, and the sun protection values of rosemary and tea tree oil were found to be 8.45 and 6.85, respectively. Rosemary oil was an extremely promising tea tree essential oil for anti-aging and sunburn prevention. The study's findings indicated that rosemary and tea tree essential oil can both offer a synergistic sun protection factor effect, antioxidant property, and anti-aging or extra activity of cosmetic preparations.

Introduction

Essential oils have been around for centuries in many cultures for their medicinal and therapeutic uses and have enhanced lives for thousands of years ¹. Essential oils (EOs) are aromatic oily liquidscomposed of a complex mixture of volatile compounds and are produced byaromatic plants as secondary metabolites ². Theuse of essential oils as a natural antioxidant is of great interest in cosmeticformulation since most commonly used synthetic antioxidants such asbutylhydroxytoluene (BHT) are suspected to be harmful side effects on humanhealth ⁵. The antioxidants arebeneficial in two ways: on the one hand, they aid in the declination of activeconstituents in cosmeceuticals products; on the other hand, antioxidantsprotect skin damage and slow the aging process by improving skin gleam andminimizing age spots, sun spots, fine lines, and wrinkles ⁴. It would seem that vitamins and nutritionalfactors have an effect on the skin’s antioxidative protection and thatcombining multiple antioxidants at the same time has a synergistic effect ⁶⁷⁸. Sunburn, photodermatoses, hyperpigmentation, photoaging of theskin, and precancerous lesions and cancers are all side effects of UV exposure.The mechanisms discussed in this paper are involved in the formation of theseclinical changes in the skin. ⁹. UV exposure causes skin sideeffects such as sunburn, photodermatoses, hyperpigmentation, photoaging of theskin, and precancerous lesions and cancers. The mechanisms discussed in thispaper are involved in the formation of these clinical differences in the skin ¹⁰.

Rosemary (Rosmarinusofficinalis L.), from abotanical perspective, this plant is expressive because it contains over 240active pharmacological and nutritional composites. Antifungal properties of theRosemary plant have been discovered through pharmacological research. Gas chromatography-massspectrometry was used to find out the chemical constituents of essential oil (EOs)ofrosemary upstanding region collected in Kerman province. Kerman province’srosemary essential yield was calculated to be 3.2 percent. The essential oilincluded 35 composites that accounted for 98.74% of the total oils ¹¹. The principal constituents were α-pinene(14.62%), camphor (12.67%), verbenone (10.19%), and 1, 8-cineole (10.63 %) ¹². Carnosic acid, carnosol, rosmarinic acid, and hesperidindominate these plants’ polyphenolic histories. The antioxidant components ofrosemary have been identified as cyclic diterpene diphenols, carnosolic acid,and carnosol ². The antioxidant activity of essential oil is one of thebiological properties of highly considerable interest in cosmeceuticalformulation.

Tea tree oil (Melaleucaalternifolia) is used principally to supplement endeavors to validate itsuse, and its reputed therapeutic antioxidant properties have been studied invitro and, in some cases, in vivo. It is a volatile essential oil derivedprimarily from the Australian native plant Melaleuca alternifolia ¹³. TTO is used as the main ingredient in many topicalpreparations used to treat cutaneous infections due to its anti-agingproperties ¹⁴¹⁵. The essential oil is high in triglycerides, free fatty acids, tocopherols, sterols, phospholipids, waxes, squalene, and phenolic compounds. According to top dermatologists, facial oils are the missing piece in traditional beauty routines. Skin moisture levels decline with age, causing dehydration and making fine lines and wrinkles more visible. Oils not only hydrate the skin, but their high antioxidant content protects cells from free radical damage, preventing further ageing¹⁶. Oils are excellent for delivering desired skin benefits inspot applications ¹⁷.

The antioxidant capacity and sun protection factor of tea tree and rosemary oil are primarily attributed to terpinen-4-ol, a major constituent of the EOs (essential oils). As a result, the terpinen-4-ol content was limited to 30% with no upper limit to maximize anti-aging activity. In contrast, a 15% upper limit and no less significant boundary was established for 1,8-cineole, though the reasoning behind this may not have been entirely sound. ¹⁸. For many years, cineole was mistakenly thought to be a skinand mucous membrane irritant, which fueled efforts to reduce its concentrationin TTO (tea tree oil). This reputation will be built on anecdotal evidence fromthe past and unsubstantiated statements ⁸.

Material and Methods

Sample collection andprocessing

The samples were collected for RMO (Rosemary oil) and TTO (Tea tree oil) testing, and their active chemical ingredients piqued the experimenters’ interest due to their antioxidative activity. The essential oils of the tea tree and rosemary plant (leaves) were purchased from Natural aroma product private limited New Delhi. As a free experimental sample from Shanghai, China, DPPH (2,2-diphenylpicrylhydrazyl) was obtained. Sigma-Aldrich Chemicals Pvt. Ltd. The naphthyl ethylenediamine dihydrochloride and L- ascorbic acid was given by Limited, USA. All chemicals and solvents were of the highest quality.

Characterizationof RMO(Rosemary oil)/TTO(Tea tree oil):

The physical properties of an oil, such as appearance(color), aroma(odor), specific gravity, saponification number, and acid number, were used to classify it ⁵.

Saponification Number

To determine the saponification value, add one ml of essential oil to 50 ml of ethanol-mixed KOH(potassium hydroxide) solution (0.5N) and reflux for thirty minutes to ensure complete dissolution of all chemicals. After adding one mL of phenolphthalein indicator, the sample solution was titrated with 0.5 N Hydrochloric acid solutions until the solution’s color changed. A similar procedure was followed with a blank sample solution. The saponification number was then calculated using the following formula:

Formula for Saponification number = 56.1 x Normality/ Weight _(sample)(Vs-Vb)

Where,

Molecular weight of potassium hydroxide = 56.1, Vs = Sample titer value, and Vb = blank titer value.

Acidvalue

One gm of EOs (essential oils) was accurately measured and added to a solution mixture of 25 milliliters of diethyl ether and 25 milliliters of C2H5OH (ethyl alcohol) to calculate the acid value. After adding 1 ml of the indicator phenolphthalein, the solution was titrated with 0.1 N potassium hydroxide solution and filled in the burette until the color was obtained ¹⁹. The acidnumber was determined as the following formula:

Formula for Acid value =56.1(molecular weight of KOH) x Normality x Volume/Mass

GC-MSanalysis of TTO/RMO

The chemical constituents present in the EOs (essential oils) samples were identified using GCMS apparatus (Agilent-5973, Aligent Technologies Inc., USA); an ABINNO Wax capillary Colum (30 m length, Inner diameter 0.25 mm & 0.25 m film consistency) was used under the following conditions column roaster temperature was 70.0 ^oC &sample injection heat was 250.0 ^oC The gas chromatography operatingconditions were as follows: gas carrier flow rate 2.0 mL/min, columntemperature programming from 70 ^oC to 270^oC at 4^oC/min,injector temperatures of 216 and 275^oC, respectively, and massspectroscopy operating process was scanned over the 40.850 m/z range. Todifferentiate individual oil constituents, the chromatogram data of both EOselement peaks were compared to the standard database from (Wiley 8 databaselibrary).

Estimationof oxidative activities for TTO (tea tree oil)/RMO (rosemary oil)

TTO/RMO antioxidant activity was examined using the following methods:

Nitric oxide (NO) assay

2,2-diphenyl -1-picryl-hydrazyl-hydrate (DPPH) assay

Nitric oxide free radical scavenging activity

The ability of antioxidant-containing samples to scavenge nitric oxide free radicals is measured by this assay. At physiological pH, sodium nitroprusside (SNP) spontaneously initiates the formation of nitric oxide, which then reacts with oxygen to form nitrite ions, as measured by the Griess Illosvoy reaction. As a positive control, standard ascorbic acid was dissolved in methanol, just like essential oils. Oil samples and standard concentrations ranging from 10 to 1000 g/ml were mixed with 2 ml of a 10 mM SNP solution prepared in 0.5 mM PBS at pH 7.4. The final solution was incubated at 25°C for 120 minutes. Following incubation, 0.5 mL of oil sample and 0.5 mL of Griess reagent were mixed. In a UV-visible double beam spectrophotometer, the abs of the colored solution were measured at 546 nm in comparison to the control (10 mmol/L SNP in phosphate buffer solution without essential oil or standard sample). The rate of inhibition of nitric oxide free radical scavenging activity by essential oil was calculated using the equation below ²⁰.

(ABS- Absorbance)

The percentage inhibition was calculated for each test solution dilution, and a chart with concentration (gm/mL) and free radical inhibit percentage was generated to obtain a linear equation. The half-maximal inhibitory concentration value was calculated using the linear equation. The IC50 value is defined as the sample concentration required for scavenging 50% of the nitric oxide free radical. The experiment was carried out three times.

2,2-diphenylpicrylhydrazyl (DPPH) free radical scavenging activity:

After a few minor adjustments, the DPPH(2, 2-diphenylpicrylhydrazyl) free radical scavenging assay was used to assess DPPH activities in a 96-well microtiter plate. A sample stock solution volume of 100L produced final concentrations of 500, 250, 125, 63, 16, 8, 4, and 2 g/mL after being diluted twice. The positive control was ascorbic acid. Following that, each well received 100 L of DPPH (2, 2-diphenylpicrylhydrazyl) at a concentration of 0.04 percent (w/v). The mixture was blended and incubated for 30 minutes at room temperature in the dark. A microplate reader was used to calculate the abs value at 515 nm. The methanol sample serves as a blank for the test sample. The control well contained a DPPH solution made of methanol. Each well’s total volume is 200 L. On each sample, three copies of each test were run. The percentage of inhibition was calculated ²¹.

The rate of inhibition was calculated for each test solution dilution, and a chart was built to achieve a linear equation, with concentration and rate of inhibition, and the half maximal inhibitory concentration was measured using the equation. The IC50 (half maximal inhibitory concentration) value is the sample concentration needed to scavenge 50% of the NO (nitric oxide) assay.

Estimation of SPF (sun protection factor) value

There are two types of in vitro SPF techniques. A very simple mathematical equation was developed to replace the in vitro method used in methods that measure UV absorption or transmission through hydrogel product films in quartz plates or biomembranes, as well as methods that rely on spectrophotometric analysis of diluted solutions to determine the absorption properties of essential oil agents ²².

The COPILA standard was used to calculate in vitro SPF, which involves calculating the percentage of a product’s transmittance over the UV spectra, subjective by the EE weighting factor at a range of absorbance ²³.

Where,

EE =Erythemogenic Effect of radiation at a wavelength (λ), I (λ) =Intensity of solar light at wavelength, abs =Absorbanc by a test sample of the standard solution. It was calculated that a standard microemulsion preparation containing 8% homosalate presented an SPF value of 4, calculated by a UV spectrophotometer.

The values for the term (I) λ x (EE) λ are constant, which were calculated, and are shown in Table 1

Table 1: Value of Intensity (λ) and EE (λ) used SPF calculation.

Wavelength	(I)λ x (EE)λ
290	0.0165
295	0.0819
300	0.2912
305	0.3217
310	0.1867
315	0.0778
320	0.0197
Total 1

(I) λis the sunlight spectra intensity, (EE) λis the erythemal action spectrum the intensity spectrum & erythemal spectrum are constant

The sample (1g) was weighed, transferred to a volumetric flask of 100 mL, filled to volume with ethanol, mixed for 15 minutes, and then filtered through Whatman filters. A 5.0 mL sample was transferred to a 25 mL volumetric flask after being diluted to volume with methanol. The absorption values were measured between 290 and 320 nm (every 5 nm). Each measurement was taken three times, and the final number represents the average of those three measurements. The Mansur equation was then used to calculate the sun protection factor values for the formulations. The in vitro method is replaced by a combination of two relatively simple mathematical equations ²⁴.

Results

TTO& RMO physicochemical characterization

TTO yield was discovered to be between one and two percent. Tea tree oil, also known as Melaleuca alternifoliaoil, is a water-soluble essential oil with a distinct camphoraceous aroma and a yellowish to nearly clear color. At 25 °C, the acid and saponification numbers of the EOs were 1.01-1.22 (mg KOH/g), 177.56-200.45 (mg KOH/g), and 0.8950-0.9050 g/mL, respectively.

RMO yield was found to be between 0.5 and 2%. Rosmarinus officinalis Linn, a member of the Lamiaceae family, flowering tops and leaves were used to create rosemary essential oil. Colorless to pale yellow transparent liquid of herbal camphoraceous minty aromatic woody balsamic medicinal phenolic essential oil. At 25°C, the acid values of the oil were 0.5-1.12 (mg KOH/g), 185-195 (mg KOH/g), and 0.908g/mL, respectively.

GCMS (Gas chromatography-mass spectroscopy) Analysis: A complete GCMS analysis of tea tree and rosemary essential oils was performed on the obtained essential oil.

GCMSanalysis of RMO

The gas chromatogram of rosemary oil is shown in Figure 1(A), and its chemical composition is shown in Table 2. RMO’s gas chromatography-mass spectroscopy analysis revealed 35 chemical constituents. Carnosol (28%), terpinen-4-ol (20.6%), β-pinene (10.55%), caryophyllene (2.54%), epi-camphor (9.97%), camphor (7.26%), α-pinene (1.9%), and 1,8-cineole were the most prevalent components of RMO oil. Because of their age-defying and antioxidant properties, these chemicals can reduce or prevent oxidative stress, and they can be used in skincare routines to slow skin aging. As a result, monoterpenes became prevalent, which is why they are important in cosmeceuticals and perfumes ²⁵.

GCMSanalysis of TTO

Figure 1(B) depicts the gas chromatogram of rosemary oil, and table 3 lists its chemical components. TTO’s gas chromatogram revealed a total of 34 chemical constituents. The majority of the oil (44.5%) was composed of Cineole-1, 8 (4.51%) Cyclohexanol, 8.78% Terpineol, 2.65% Terpineol-4-ol, 12.11% (10.5%) α-pinene, β-pinene (5.81%) Terpinolene. Because of their anti-aging and antioxidant properties, these substances can be used in skincare regimens to delay skin aging and possibly reduce or prevent oxidative stress. Other elements were present in lesser amounts. The primary goal of the GCMS study was to detect the presence of monoterpenes, oxygenated monoterpenes, alcohols, sesquiterpene hydrocarbons, and diterpenes in essential oils because they are primarily responsible for antioxidant activity, sun protection factors, and skin aging ²⁶.

Table 2: Rosemary oil chemical constituent’s details by GC-MS analysis.

S. no	RT (Retention Time)	Content percentage (%)	Chemical constituents
1	8.81	0.9	α-thujene
2	9.092	1.9	α-pinene
3	9.684	1.26	Camphor
4	9.882	10.55	β-pinene
5	11.221	1.44	Myrcene
6	12.700	1.888	p-cymene
7	13.082	0.344	Limonene
8	14.805	0.444	Linalool
9	16.983	0.324	Citronellol acetate
10	17.578	1.999	0-cymene
11	18.065	20.6	Terpinen-4-ol
12	20.795	1.688	α-terpineol
13	22.892	1.99	Citronellol
14	23.053	0.139	Z-linalol oxide
15	23.645	1.54	Caryophyllene
16	24.686	0.12	ᵧ-terpinene
17	26.777	0.999	Citronellyl Acetate
18	27.139	2.97	Epi- camphor
19	28.001	18.76	1,8 cineole
20	28.575	2.7	rosmanol,
21	29.604	1.77	epirosmanol,
22	30.616	1.44	hesperidin,
23	31.802	0.567	Camphene hydrate
24	33.329	1.345	E-citronellyl tiglat
25	34.045	0.9999	Aromatic oxygenated Monoterpenes
26	34.587	0.67	rosmarinic acid
27	35.297	0.522	Sesquiterpene Hydrocarbons
28	36.165	1.777	(E)- caryophyllene
29	36.255	2.098	isorosmanol.
30	37.113	1.339	Carophyllnol
31	39.291	1.666	Oxygenated Sesquiterpenes
32	40.579	1.700	Citronellyl butyrate
33	47.994	28.0	Carnosol
34	49.073	1.44	Oxygenated Monoterpenes
35	49.165	1.111	Carsonic acid

Table 3: Chemical constituent composition of Tea tree oil by GCMS analysis.

S. no	RT (Retention Time)	Content percentage (%)	Chemical constituents
1	8.81	0.9	α-thujene
2	9.092	1.9	α-pinene
3	9.684	1.26	Camphor
4	9.882	10.55	β-pinene
5	11.221	1.44	Myrcene
6	12.700	1.888	p-cymene
7	13.082	0.344	Limonene
8	14.805	0.444	Linalool
9	16.983	0.324	Citronellol acetate
10	17.578	1.999	0-cymene
11	18.065	20.6	Terpinen-4-ol
12	20.795	1.688	α-terpineol
13	22.892	1.99	Citronellol
14	23.053	0.139	Z-linalol oxide
15	23.645	1.54	Caryophyllene
16	24.686	0.12	ᵧ-terpinene
17	26.777	0.999	Citronellyl Acetate
18	27.139	2.97	Epi- camphor
19	28.001	18.76	1,8 cineole
20	28.575	2.7	rosmanol,
21	29.604	1.77	epirosmanol,
22	30.616	1.44	hesperidin,
23	31.802	0.567	Camphene hydrate
24	33.329	1.345	E-citronellyl tiglat
25	34.045	0.9999	Aromatic oxygenated Monoterpenes
26	34.587	0.67	rosmarinic acid
27	35.297	0.522	Sesquiterpene Hydrocarbons
28	36.165	1.777	(E)- caryophyllene
29	36.255	2.098	isorosmanol.
30	37.113	1.339	Carophyllnol
31	39.291	1.666	Oxygenated Sesquiterpenes
32	40.579	1.700	Citronellyl butyrate
33	47.994	28.0	Carnosol
34	49.073	1.44	Oxygenated Monoterpenes
35	49.165	1.111	Carsonic acid

Figure 1: Gas chromatogram of RMO / TTO by GCMS analysis.

Discussion

Determinationof In- vitro antioxidant capacity:

Significant quantities of essential oils are naturally responsible for antioxidant capacity. We used two techniques in this investigation: To ascertain the antioxidative potency of rosemary oil and tea tree oil

Nitric oxide assay

2, 2-diphenyl-1-picryl-hydrazyl-hydrate assay

Nitricoxide free radical scavenging activity:

Table 4 and Figure 2 (A) show that the nitric oxide radical scavenging of oils ranges between 23.13 and 81.57% and 36.55 and 90.04%, respectively. Tea tree essential oil (23.13%) and rosemary essential oil (36.55%) had the lowest activity at 0.10 g/ml, while at 1.0 mg/ml, tea tree essential oil (81.57%) and rosemary essential oil (90.04%) had the highest activity. At this dose, the EOs have greater than 50% scavenge efficacy, with an IC50 value of 0.50 mg/ml (62.76%) ². The essentialoils TTO and RMO have oxidation properties, as evidenced by the decreasedabsorbance of the reaction mixture.

Table 4: Nitric oxide free radical scavenging activity for TTO and RMO.

Concentration (mg/ml)	Absorbance of Blank	Absorbance of standard	Absorbance of (Tea tree oil)	Absorbance of (Rosemary oil	% FRSA of Standard	% FRSA of TTO	% FRSA of RMO
0	0	0	0	0	0	0	0
0.1	1.677	0.489	1.289	1.064	70.84	23.13	36.55
0.25	1.677	0.390	0.755	0.607	76.74	54.97	63.80
0.5	1.677	0.286	0.672	0.396	82.94	59.92	76.38
0.75	1.677	0.192	0.377	0.190	88.55	77.51	88.67
1.00	1.677	0.123	0.309	0.167	92.66	81.57	90.04

Table 1:

2, 2-diphenyl-1-picryl-hydrazyl-hydrate free radical scavenging activity:

The free radical scavenging activity (FRSA) of essential oils ranges from 28.42 to 83.34% and 30.96 to 88.44%, according to TTO and RMO measurements. TTO has the lowest free radical scavenging activity of 28.42% at 0.10 mg/ml and the highest at 83.34% at 1.00 mg/ml when compared to regular ascorbic acid. RMO has the lowest (30.96% at 0.10 mg/ml) and highest (88.44% at 1.00 mg/ml) free radical scavenging activity when compared to regular ascorbic acid. The IC50 value was 0.25 mg/ml, and the free radical scavenging activity was 63.23%. The results shown in Table 5 and Figure 2(B) show that the antioxidants’ free radical scavenging abilities cause the constant and violet-blue color core to change to a yellowish color. When the results are compared, we can conclude that TTO and RMO essential oils both have beneficial antioxidant properties, and their concentration increases their ability to scavenge free radicals ²⁵.

Table: 5 DPPH assays of Tea tree oil & Rosemary oil.

Concentration (mg/mL)	Abs of Blank	Abs of standard	Abs of Sample oil(Rosemary oil	Abs of Sample oil(Tea tree oil)	% FRSA of Standard	% FRSA of RMO	% FRSA of TTO
0	0	0	0	0	0	0	0
0.1	4.024	0.199	2.778	2.880	95.05	30.96	28.42
0.25	4.024	0.190	2.315	2.567	95.27	42.47	36.20
0.5	4.024	0.180	1.470	1.988	95.52	63.49	50.59
0.75	4.024	0.163	0.698	0.876	95.94	82.65	78.23
1.00	4.024	0.155	0.465	0.670	96.14	88.44	83.34

FRSA- Free radical scavenging Activity

Using the DPPH assay technique, the IC 50 values for Tea tree essential oil and Rosemary essential oil are 0.25g/ml and 0.25g/ml, respectively. Tea tree essential oil has half maximal concentrations of 0.25 g/ml and 0.50 g/ml in the Nitric Oxide Scavenging technique. Both essential oils were 50% effective against free radicals at these concentrations ¹⁷. When theresults were compared, it was discovered that both rosemary and tea treeessential oils had potent antioxidant properties that increased withconcentration.

Figure 2: (B) DPPH free radical scavenging assay of TTO and RMO.

Table 6: IC50 (half maximal inhibitory concentration) Value of Tea tree Oil.

IC50 (mg/ml)	Standard	Tea tree essential oil
Nitric Oxide Scavenging Method	<0.1	0.25
DPPH Scavenging Method	<0.1	0.25

Table 7: IC50 (half maximal inhibitory concentration) Value of Rosemary Oil.

IC50 (mg/ml)	Standard	Rosemary essential oil
Nitric Oxide Scavenging Method	<0.1	0.50
DPPH Scavenging Method	<0.1	0.25

Figure 3: IC50 (half maximal inhibitory concentration) value of TTO/RMO by DPPH/Nitric oxide activity:

Determination of in vitro sun protection factor for TTO/ RMO

SPF determination is a useful test for vetting substances when developing cosmeceutical products. The SPF rating demonstrates the efficacy of the microemulsion. The higher the sun protection factor, the better the formulation’s UV protection against light. To be effective in preventing photoaging, sunburn, skin wrinkles, and other skin damage, essential oils must absorb a significant amount of UV – radiation (290-400 nm). An examination of the literature revealed a scarcity of studies on how to calculate the SPF of essential oils. This is due to two factors. The first is that essential oils are highly flammable and have very different properties than thick, fatty vegetable oils; as a result, they do not provide adequate sun protection when used alone. Money is another consideration. ²³. Plant-derivedessential oils contain a wide range of chemical constituents. The percentage ofthese chemical components varies by harvest and batch depending on the growingconditions of the plants. Due to the conflicting information on essential oils,any SPF study would cost millions of dollars. This is one of the primaryreasons why the scientific community has decided against funding essential oilSPF research. The TTO and RMO SPF values were 6.85 and 8.45, respectively,according to the current investigation (Table 4). It was discovered that theevaluated essential oils have low SPF ratings for the antioxidant agentprimarily used for producing defense against oxidative stress and combating thebothersome free radicals that frequently accompany prolonged sun exposure. Eventhrough this, they always provide numerous skin benefits ²⁶.

Table 8: SPF (Sun Protection Factor) Value of TTO/RMO.

Wavelength ( nm)	Absorbance ofTTO	Absorbance of RMO
290	0.905± 0.08	0.955± 0.14
295	00867± 0.11	0.939± 0.08
300	0.785± 0.07	0.899± 0.16
305	0.630± 0.16	0.830± 0.19
310	0.523± 0.09	0.799± 0.18
315	0.445± 0.07	0.732± 0.12
320	0.433± 0.11	0.675± 0.14
Calculated Sun Protection Factor	6.85	8.45

n = 3

Conclusions

According to the present research work, tea tree and rosemary oil can prevent or reduce oxidative stress and can be used for anti-aging purposes due to their anti-oxidative properties. The essential oil of TTO and its majority chemical constituent 1, 8 cineole, terpineol-4-ol, β- pinene α-pinene, and above all rosemary essential oil and its chemical constituent’s carnosol, camphor, terpinen-4-ol, are active against in-vitro free radical scavenging activity & skin aging. The sun protection factor results show that tea tree oil/rosemary oil can be used to make microemulsions that protect the skin from sunburn. From the result, it can be concluded that Tea tree oil had a lower sun protection factor than rosemary essential oils. TTO and RMO essential oils have good antioxidant properties and their free radical scavenging activities (in DPPH and Nitric oxide scavenging methods) increase with concentration and have a synergistic photo-protective effect on exposure to UV light.

Acknowledgment

The authors would like to thank the Deanship and Vice-Presidency for Scientific Research at IFTM University in Uttar Pradesh for providing ongoing financial support and lab space for this project.

Conflict of Interests

There are no declared conflicts of interest.

List of Abbreviations

TTO: Tea tree oil; RMO: Rosemary oil; IC50: Half-maximal inhibitory concentration; SPF: Sun protection factor; UV: Ultraviolet; GCMS: Gas chromatography Mass spectroscopy; Abs: Absorbance

References

1. Matts, P. J.; Alard, V.; Brown, M.W.; Ferrero, L.; Barlag, Gers. H.; Issachar, N.; Moyal, D.; Wolber, R.; The COLIPA in vitro UVA method: a standard and reproducible measure of sunscreen UVA protection. Int. J. Cosmet. Sci. 2010, 32, 35–46.

2. Rašković, A.; Milanović, Isidora.; Pavlović, N.; Ćebović, T.; Vukmirović, S.; Mikov, M.; Antioxidant activity of rosemary (Rosmarinus officinalis L.) essential oil and its hepatoprotective potential. BMC Complement. Altern. Med. 2014, 14, 225.

3. Khotimah, H.; Ismail, D. D. L.; Widasmara, D.; Riawan, W.; Retnaningtyas, E.; Nugraheni, R.W.; Puspita, O. E.; Adianingsih, O. R.; Mardiyah, M.; Setiawan, A.; Ameliorative effect of gel combination of Centella asiatica extract transfersomes and rosemary essential oil nanoemulsion against UVB-induced skin aging in Balb/c mice. F1000Research 2022, 11, 288.

4. Lohani, A.; Verma, A.; Hema, G.; Pathak, K.; Topical Delivery of Geranium/Calendula Essential Oil-Entrapped Ethanolic Lipid Vesicular Cream to Combat Skin Aging. BioMed Res. Int. 2021, 1–13 .

5. Pazyar, N.; Yaghoobi, R.; Bagherani, N.; Kazerouni, A.; A review of applications of tea tree oil in dermatology. Int. J. Dermatol. 2013, 52, 784–790.

6. rJiang, Y.; Wu, Nan.; Fu, Y. J.; Wang, W.; Luo, M.; Zhao, C. J.; Zu, Gang. Y.; Liu, X. L.; Chemical composition and antimicrobial activity of the essential oil of Rosemary. Environ. Toxicol. Pharmacol. 2011, 32, 63–68.

7. Gavaric, N.; Mozina, S. S.; Kladar, N.; Bozin, B.; Chemical Profile, Antioxidant and Antibacterial Activity of Thyme and Oregano Essential Oils, Thymol and Carvacrol and Their Possible Synergism. J. Essent. Oil Bear. Plants 2015, 18, 1013–1021.

8. Pazyar, N.; Yaghoobi, R.; Bagherani, N.; Kazerouni, A.; A review of applications of tea tree oil in dermatology. Int. J. Dermatol. 2013, 52, 784–790.

9. Forni, C; Francesco, F.; Bartoli, M.; Pieretti, S.; Facchiano, A.; D’ Arcangelo, D.; Norelli, S.; Valle, G.; Nisini, R.; Beninati, S.; Tabolacci, C.; Jadeja, R..N.; Beneficial Role of Phytochemicals on Oxidative Stress and Age-Related Diseases. BioMed Res. Int. 2019, 1–16 .

10. Pandel, R.; Poljšak, B.; Godic, A.; Dahmane, R.; Skin Photoaging and the Role of Antioxidants in Its Prevention. ISRN Dermatol. 2013, 1–11 .

11. Thomsen, N. A.; Hammer, K. A.; Riley, T. V.; Van Belkum, A.; Carson, C. F.; Effect of habituation to tea tree (Melaleuca alternifolia) oil on the subsequent susceptibility of Staphylococcus spp. to antimicrobials, triclosan, tea tree oil, terpinen-4-ol and carvacrol. Int. J. Antimicrob. Agents 2013, 41, 343–351.

12. Yousuf, S.; Singh, A.; Yousuf, T.; Evaluation of Free Radical Scavenging activities of Essential Oil Extracted from Valeriana hardwickii. 2021, 5, 2724–2731.

13. Hendra, R.; Ahmad, S.; Oskoueian, E.; Sukari, A.; Shukor, M. Y.; Antioxidant, Anti-inflammatory and Cytotoxicity of Phaleria macrocarpa (Boerl.) Scheff Fruit. BMC Complement. Altern. Med. 2011, 11, 110.

14. Sameer Ali, Z.; Muhammad, D.; Zrieki, A.; In vitro Assessment of sun protection factor (SPF) and Antioxidant activity of Viola odorata extracts. Res. J. Pharm. Technol. 2022, 655–660 doi:10.52711/0974-360X.2022.00108.

15. Garzoli, S.; Masci, V. L.; Franceschi, S.; Tiezzi, A.; Giacomello, P.; ovidi, E.; Headspace/GC–MS Analysis and Investigation of Antibacterial, Antioxidant and Cytotoxic Activity of Essential Oils and Hydrolates from Rosmarinus officinalis L. and Lavandula angustifolia Miller. Foods 2021, 10, 1768.

16. Adewinogo, S. O.; Sharma, R.; Africa, C.; W. J.; Marnewick, J. L.; & Hussein, A. A.; Chemical Composition and Cosmeceutical Potential of the Essential Oil of Oncosiphon suffruticosum (L.) Källersjö. Plants 2021, 10, 1315.

17. Lohani, A.; Mishra, A. K.; Verma, A.; Cosmeceutical potential of geranium and calendula essential oil: Determination of antioxidant activity and in vitro sun protection factor. J. Cosmet. Dermatol. 2019, 18, 550–557.