MOLECULAR SCREENING OF Ocimum basilicum FOR

ANTI INFLAMMATORY REACTION AND ORTHOPEDIC

APPLICATION

Dr. D. KALYANI BY
Teaching Fellow, Nithiya M (2019116029)
Division of Biomedical Engineering, Sangamithra B (2019116038)
College of Engineering, Guindy. Surya T.G. (2019116047)
 ABSTRACT
Ocimum basilicum commonly known as sweet basil, is a culinary plant from Lamiaceae family
(mint), Asian in origin. Ocimum basilicum plays a key role in many ailments due to its varied
therapeutic properties. This project focus on the phytochemical profiling by GC-MS and FT-IR
analysis of Ocimum basilicum solvent extract. The objective of the project is to find the main
constituent in Ocimum basilicum and its implementation in biomedical applications. Estragole
was found to be the main constituent in Ocimum leaf extract and its application in orthopedic
application in done by 3D slicer and Fusion 360 software. The analysis of Ocimum basilicum
leaf extract in reducing the inflammatory action is studied by Autodock analysis. Further
extensive research has to be done to find its biocompatibility and feasibility of its
implementation in healthcare.
 LITERATURE SURVEY
S. AUTHOR NAME OF THE VOLUME NO.,ISSUE INFERENCE
NO JOURNAL NO., ISSN, YEAR OF
PUBLICATION
1. Aswathy Valsan, Preliminary Res. Jr. of Agril. Sci. (Jul- The phytochemical such as alkaloids,
Athulya Bose, Phytochemical Aug 2022) 13(4): 925– steroids, flavonoids, tannins, phenols
Gopika K. N, Screening of 930 and several other aromatic compounds
Amrutha C. S, Indigenous Medicinal of plants serve a defense mechanism.
Anil Kumar A. K, Plants Ocimum
Jayamol K. V and tenuiflorum, Ocimum
Shyam Kumar basilicum, and Ocimum
gratissimum
2. James Redfern, Using Soxhlet Ethanol Journal of microbiology & This article presents information about
Malcolm Extraction to Produce biology education, May the soxhlet extraction method in the
Kinninmonth and and Test Plant 2014, p. 45-46 teaching laboratory.
Dariel Burdass Material (Essential
Oils) for Their
Antimicrobial
Properties
3. Dina Salah Eldin GC-MS analysis, ISSN 2455- Essential oil of O. basilicum leaves showed
Mohammad and antioxidant, antimicrobial 3301 wjpmr, different peaks nineteen Compounds identified.
Hassan S. Khalid cytotoxicity, activities of 2016, 2(6), 08- The Compound with high percentage area was
leaves essential oil Ocimum 13. Eugenol (54.78%).
basilicum and Ocimum
gratissimum l. (lamiaceae)
4. Mohanad Jawad Evaluation of anti-bacterial ISSN 2141- Fourier transform infrared (FT-IR) analyses of O.
Kadhim, Azhar activity and bioactive 250; Vol. 8(6), basilicum revealed the existence of aliphatic
Abdulameer chemical analysis of pp. 127-146, fluoro compounds, alcohols, ethers, carboxlic
Sosa and Imad Ocimum basilicum using June 2016 acids, esters, nitro compounds, alkanes, and
Hadi Hameed Fourier transform infrared phenols.
(FT-IR) and gas
chromatography mass
spectrometry (GC-MS)
techniques
5. Min Sook Jeong, Methodological Scientific Electron spin resonance (ESR) is a useful tool for
Kyeong-Nam considerations of electron Reports , detecting ROS formation.
Yu, Hyun Hoon spin resonance spin trapping 6:26347, DOI:
Chung and Soo techniques for measuring 10.1038/srep26
Jin Park reactive oxygen species 347
generated from metal oxide
nanomaterials
 WORK FLOW
Ocimum basilicum (leaf) is collected and dried.

Leaf is crushed to small pieces after 20 days.

Preparation of leaf extract using ethanol as a solvent by Soxhlet extraction method.

FTIR (ATR mode) and GC-MS

Detection of ROS

Molecular docking analysis using AutoDock

Application of leaf extract in orthopedic application using 3D Slicer and Fusion 360
 Why Ocimum basilicum?
• Basil possesses strong anti-inflammatory properties which help to fight against various
diseases and disorders.
• The essential oil of Ocimum basilicum contains methyl eugenol and linalool has
bactericidal properties. Essential oils present in leaf to fight inflammation and help to
relieve inflammatory signs and symptoms like swelling, pain and redness of affected part.
• It helps to restore the digestive health of a person. Basil helps to treat indigestion and
other digestive troubles. It restores the pH of the stomach by balancing the acid within.
 Ocimum basilicum

AFTER 20 DAYS CRUSHED LEAF

Sample collected at Pamban Swamigal Temple, Thiruvanmiyur
 SOXHLET EXTRACTION

Experimental setup is made for Soxhlet extraction.

Magnetic Stirrer (300-500 rpm) - gradually heat the solvent

Solvent begin to evaporate, moving through the apparatus to the condenser.

Condensate drips into the reservoir

Once the level of the solvent reaches the siphon it pours back into the air blow
Experimental setup After 16 hours
Experimental setup After 16 hours
Ethanol separation using water bath Leaf extract
• Qualitative analysis I

It was performed by color’s test which shows the presence of alkaloids, flavonoids, terpenoids, steroids.

• Qualitative analysis II

It was performed by UV-visible spectroscopic analysis, supports the results obtained by Qualitative analysis I.

Table 1: UV-VISIBLE Spectroscopy

S.NO WAVELENGTH ABSORBANCE PHYTOCHEMICAL REFERENCE

(nm) (L/mol cm) PAPER
1. 404 1.2882 Flavonoids [6]
2. 344 0.9173 Coumarin, Alkaloids [4], [1]
3. 666 0.3994 Chlorophyll [6]
4. 502 0.1325 Carotenoids, Flavonoids [5]
5. 534 0.1066 Tannins [5]
 ATR-FTIR Spectroscopy

• FTIR stands for ‘Fourier-transform infrared’ spectroscopy: a technique that can be used to procure an
infrared spectrum of either the emission or absorption of a liquid, solid, or gas sample.

• Principle: When infrared (IR) radiation passes through a sample, some of the radiation is absorbed. The
radiation that passes through the sample is recorded.

SPECIFICATION:

• Spectra Range: 500-4000 cm-1

• Sample Required: 1 ml

• Crystal: Diamond brazed into Tungsten

Carbide

ATR – FTIR Spectrometer

ATR-FTIR spectra of Ocimum bascilicum leaf extract
Table 2: Functional groups obtained from Ocimum leaf extract in ATR- FTIR mode
S.NO WAVELENGTH (cm-1) FUNCTIONAL GROUP

1. 3339.14 =C–H(methene)
2. 2973.18 CH3(methyl)
3. 2889.29 -C–H (CH3, CH2)
4. 2124.11 -C≡C-(ethyne)
5. 1650.19 -C=O(carbonyl alkene)
6. 1381.67 -C–H (CH3)(methyl alkane)
7. 1325.60 C-O-C(aromatic ether)
8. 1274.61 -C–O; -CH2-(carbonyl)
9. 1085.64 -C–O(carbonyl alkane)
10. 1044.09 -HC=CH-(ethene)
11. 879.42 =CH2(alkene)
12. 801.74 -C–H; -HC=CH-(ethene , methyl)
13. 635.13 unknown
 Gas chromatography–mass spectrometry
•Gas chromatography–mass spectrometry (GC-MS) is an analytical method that
combines the features of gas-chromatography and mass spectrometry to identify
different substances within a test sample.

•Gas chromatography mass spectrometry (GC-MS) is an instrumental technique,

comprising a gas chromatograph (GC) coupled to a mass spectrometer (MS), by
which complex mixtures of chemicals may be separated, identified and quantified.
This makes it ideal for the analysis of the hundreds of relatively low molecular
weight compounds found in organic materials. In order for a compound to be
analyzed by GC-MS it must be sufficiently volatile and thermally stable.
 PROCEDURE
• 1ml of leaf extract was injected into gas chromatography mass spectroscopy automatic
system. The maximum number of hit was counted as 1 and the result was compared with
MS library.

• The following result was obtained, the retention time of the peak 8.288mins which was
maximum single peak obtained under GC-MS.

GC-MS LAB IN SAIF-IITM DILUTED SAMPLE

 GC-MS spectra of Ocimum basilicum leaf extract

GC-MS analysis of Ocimum leaf ethanolic extract reveals single strong compound
estragole was found to be maximum contributor by giving a single peak.
Table.3: GC-MS analysis of Ocimum basilicum leaf extract
S.NO RETENTION TIME (mins) COMPOUNDS [9]
1. 6.12 1,1-dichloroethane
2. 7.2 1,2-dichloroethane
3. 8.28 ESTRAGOLE
4. 9.6 ethylene dichloride
5. 10.19 1,2-dichloropropane
6. 10.9 sec-butyl acetate
7. 11.3 iso-butyl acetate
8. 12.8 3-hexanone
9. 13.12 2-ethoxyethanol
10. 13.8 cis-2-hexen-1-ol
11. 14.9 1,4-dichloro-2-butene
12. 15.99 iso-propylbenzene
13. 18.7 methyl benzoate
A

Fig A reveals the standard spectral library whereas B is spectral library for Ocimum basilicum leaf extract.
Table 4: Ionisation table

S.no Ions(m/z ratio) Abundance(%) Compounds [10]

1. 148 100 Estragole

2. 147 65 1,2-dimethylpropanal
3. 77 46 1,1-ethylbutanal
4. 121 45 cis-3-hexen-1-ol
5. 117 45 cis-2-hexen-1-ol
6. 91 30 methyl benzoate

The ion with the highest intensity (m/z) 148 for estragole is stable and
predominant those ionic peak is taken as 100% and considered as base peak.
CONCLUSION FROM THE GC-MS
• Formula- C10H12O
• Compound- Estragole
• Chemical structure-

• CAS NO- 140-67-0

• Retention time(mins)- 8.288
• Area- 3533143
• Area(%)- 100
• Library Id- 149572
• Relevant score/matching index- 961
ELECTRON SPIN RESONANCE
• ESR spectroscopy is an absorption spectroscopy which involves the absorption of
radiation in the microwave region (104 –10 6 MHz) containing one or more unpaired
electrons.
• It takes place under the influence of an applied magnetic field.

JEOL Model JES FA200

ESR spectra of spectra of Ocimum bascilicum leaf extract
ESR results of Ocimum bascilicum leaf extract

Magnetic field - 0 to 800mT

Intensity - 95.691 to -42.905
Nanoparticle - Manganese (Mn)
Maximum intensity - 128.701

• ROS are usually very reactive, but present in low concentration in healthy state.
• ROS elevated during inflammatory reactions.
• Hence drug can be designed to suppress ROS generation by inhibiting anti-
inflammatory protein.
INFLAMMATORY ACTION

• An inflammatory response occurs immediately following bone fracture.

• Cytokines modulate the immune response to infection or inflammation and regulate inflammation
itself via a complex network of interactions. However, excessive inflammatory cytokine production
can lead to tissue damage, hemodynamic changes, organ failure.

• Pro-inflammatory proteins :

• Interferon-gamma

• Interleukin-33

• Interleukin-12

• Interferon alpha
DOCKING USING AUTODOCK

• Docking is a molecular modeling technique that is used to predict how

a protein (enzyme) interacts with small molecules (ligands).

• Autodock (software) is a suite of automated docking tools. It is

designed to predict how small molecules, such as substrates or drug
candidates, bind to a receptor of known 3D structure.
Docking ligand- Estragole
Docking proteins- interferon-gamma, interleukin-33, interleukin-12, interferon alpha2
 FLOWCHART

INF- γ, INF- α2, IL-12 and IL-

33 are downloaded from Protein Ligand structure estragole is
data bank. downloaded from Pubchem.

Imported into molegro (molecular viewer) and removing

other substrates.

Protein and ligand are loaded into autodock

SOFTWARE USED:
software.
1. AutoDock
3D grid generation around receptor and ligand
2. Molegro
3. Discovery studio
Defining the docking parameters and running the
docking simulation

Analysis of docked complex

DOCKING PROCEDURE

1. Preparation of receptor & ligand files.

The preparation step starts with pdb files of receptor (protein.pdb) and ligand
(ligand.pdb), which are added hydrogens and then saved as RH.pdb & LH.pdb.

Prepreparation in molegro Selection of ligand

2.Calculation of affinity maps by using a 3D grid around the receptor &
ligand.

3D grid generation
3. Defining the docking parameters and running the docking simulation.
Using commands:

Execution of docking
The protocol is followed for all the four proteins are docked with estragole.
Analysis is done using molecular viewer and discovery studio software.

Analysis using molegro

Molecular viewer helps to find type of bonding, bond length
and bond strength.
Conformation information

Each conformation has their own information. The information is tabulated as

below.
Table 5: Conformation information
Protein: interferon gamma Ligand: estragole
TOTAL
BINDING INHIB VANDERWALLS TORSIONAL UNBOUND
LIGAND INTERMOLECULAR ELECTROSTATIC INTERNAL
CONFORMATION ENERGY CONSTANT ENERGY ENERGY ENERGY
EFFICIENCY ENERGY(kcal/mol) ENERGY (kcal/mol) ENERGY
(kcal/mol) (pM) (kcal/mol) (kcal/mol) (kcal/mol)
(kcal/mol)
1 -5.0 -0.45 217.88 -5.89 -5.89 0.0 -0.26 0.89 -0.26
2 -4.93 -0.45 241.86 -5.83 -5.83 0.0 -0.26 0.89 -0.26
3 -4.93 -0.45 242.7 -5.83 -5.83 0.0 -0.27 0.89 -0.27
4 -4.89 -0.44 261.06 -5.78 -5.78 0.0 -0.26 0.89 -0.26
5 -4.88 -0.44 262.69 -5.78 -5.78 0.0 -0.25 0.89 -0.25
6 -4.85 -0.44 278.41 -5.75 -5.75 0.0 -0.27 0.89 -0.27
7 -4.27 -0.39 746.77 -5.16 -5.16 0.0 -0.27 0.89 -0.27
8 -4.19 -0.38 954.46 -5.08 -5.08 0.0 -0.23 0.89 -0.23
9 -4.10 -0.37 980.2 -5.0 -5.0 0.0 -0.25 0.89 -0.25
10 -4.09 -0.37 1.01 -4.98 -4.98 0.0 -0.26 0.89 -0.26
Table 6: Bonding information

Bond length and strength

BONDLENGTH
ATOM ID BONDING TYPE BONDED TO AMINOACID
(Å)

1 Alkyl Proline 223 4.53

2 Pi-cation Lysine 200 3.24

10 Alkyl Valine 199 4.67

Discovery studio analyzed image: 2D structured protein-ligand interaction

Hydrogen bond Hydrophobicity

Table 7: Conformation information
Protein: interferon alpha-2 Ligand: estragole

TOTAL
BINDING INHIB TORSIONAL UNBOUND
LIGAND INTERMOLECULAR VANDERWALLS ELECTROSTATIC INTERNAL
CONFORMATION ENERGY CONSTANT ENERGY(kcal/ ENERGY
EFFICIENCY ENERGY (kcal/mol) ENERGY(kcal/mol) ENERGY(kcal/mol) ENERGY
(kcal/mol) (pM) mol) (kcal/mol)
(kcal/mol)
1 -4.26 -0.39 758.51 -5.15 -5.15 0.0 -0.27 0.89 -0.27

2 -4.17 -0.38 875.96 -5.07 -5.07 0.0 -0.29 0.89 -0.29

3 -4.07 -0.37 1.04 -4.96 -4.96 0.0 -0.27 0.89 -0.27
4 -4.06 -0.37 1.05 -4.96 -4.96 0.0 -0.24 0.89 -0.24

5 -4.04 -0.37 1.1 -4.93 -4.93 0.0 -0.29 0.89 -0.29

6 -3.78 -0.34 1.71 -4.67 -4.67 0.0 -0.25 0.89 -0.25

7 -3.67 -0.33 2.05 -4.56 -4.56 0.0 -0.28 0.89 -0.28

8 -3.53 -0.32 2.57 -4.43 -4.43 0.0 -0.29 0.89 -0.29

9 -3.5 -0.32 2.7 -4.4 -4.4 0.0 -0.23 0.89 -0.23

10 -3.23 -0.29 4.31 -4.12 -4.12 0.0 -0.28 0.89 -0.28
Table 8: Bonding information

Bond length and strength

BONDLENGTH
ATOM ID BONDING TYPE BONDED TO AMINOACID
(Å)

0 Carbon-Hydrogen Serotonin 306 2.52

1 Alkyl Cysteine 278 4.62

2 Pi-alkyl Cysteine 278, valine 230 4.14,5.37

10 Alkyl Proline 223 4.98

Discovery studio analyzed image: 2D structured protein-ligand interaction

Hydrogen bond Hydrophobicity

Table 9: Conformation information
Protein: interleukin-33 Ligand: estragole
TOTAL
BINDING INHIB TORSIONAL UNBOUND
LIGAND INTERMOLECULAR VANDERWALLS ELECTROSTATIC INTERNAL
CONFORMATION ENERGY CONSTANT ENERGY ENERGY
EFFICIENCY ENERGY(kcal/mol) ENERGY(kcal/mol) ENERGY(kcal/mol) ENERGY
(kcal/mol) (pM) (kcal/mol) (kcal/mol)
(kcal/mol)
1 -4.86 -0.44 274.92 -5.75 -5.75 0.0 -0.24 0.89 -0.24

2 -4.79 -0.44 307.3 -5.69 -5.69 0.0 -0.26 0.89 -0.26

3 -4.75 -0.43 327.39 -5.65 -5.65 0.0 -0.26 0.89 -0.26

4 -4.73 -0.43 343.67 -5.62 -5.62 0.0 -0.27 0.89 -0.27

5 -4.73 -0.43 343.6 -5.62 -5.62 0.0 -0.25 0.89 -0.25

6 -4.73 -0.43 341.2 -5.62 -5.62 0.0 -0.27 0.89 -0.27

7 -4.72 -0.43 344.67 -5.62 -5.62 0.0 -0.28 0.89 -0.28

8 -4.69 -0.43 363.94 -5.59 -5.59 0.0 -0.28 0.89 -0.28

9 -4.67 -0.42 376.0 -5.57 -5.57 0.0 -0.27 0.89 -0.27

10 -4.66 -0.42 384.22 -5.55 -5.55 0.0 -0.27 0.89 -0.27
Table 10: Bonding information
Bond length and strength
BONDLENGTH
ATOM ID BONDING TYPE BONDED TO AMINOACID
(Å)

1 Pi-alkyl Tyrosine 54 5.01

Alkyl Leucine 102 4.22

5 Pi-pi t-shaped Phenylalanine 107 5.24

Pi-alkyl Leucine 102 4.94

Pi-alkyl Isoleucine 6 5.44

10 Alkyl Isoleucine 3 5.07

Discovery studio analyzed image: 2D structured protein-ligand interaction

Hydrogen bond Hydrophobicity

Table 11: Conformation information
Protein: interleukin-12 Ligand: estragole
TOTAL
BINDING INHIB TORSIONAL UNBOUND
LIGAND INTERMOLECULAR VANDERWALLS ELECTROSTATIC INTERNAL
CONFORMATION ENERGY(kcal CONSTANT(p ENERGY(kcal/ ENERGY(kca
EFFICIENCY ENERGY(kcal/mol) ENERGY(kcal/mol) ENERGY(kcal/mol) ENERGY(kcal
/mol) M) mol) l/mol)
/mol)
1 -3.7 -0.34 1.93 -4.6 -4.63 0.04 -0.23 0.89 -0.23

2 -3.69 -0.34 1.96 -4.59 -4.62 0.03 -0.25 0.89 -0.25

3 -3.54 -0.32 2.53 -4.44 -4.43 -0.01 -0.19 0.89 -0.19

4 -3.52 -0.32 2.63 -4.41 -4.33 -0.08 -0.21 0.89 -0.21

5 -3.49 -0.32 2.76 -4.39 -4.4 0.01 -0.2 0.89 -0.2

6 -3.27 -0.3 4.02 -4.16 -4.11 -0.05 -0.25 0.89 -0.25

7 -3.24 -0.29 4.19 -4.14 -4.15 0.01 -0.21 0.89 -0.21

8 -2.78 -0.25 9.21 -3.67 -3.59 -0.09 -0.22 0.89 -0.22

9 -2.73 -0.25 10.03 -3.62 -3.61 -0.01 -0.24 0.89 -0.24

10 -2.67 -0.24 11.06 -3.56 -3.5 -0.07 -0.23 0.89 -0.23
Table 12: Bonding information
Bond length and strength

BONDLENGTH
ATOM ID BONDING TYPE BONDED TO AMINOACID
(Å)

0 Hydrogen Phenylalanine 132 2.11

1 Alkyl Alanine 151 4.10

7 Alkyl Alanine 151 5.17

10 Alkyl Leucine 130 4.57

Discovery studio analyzed image: 2D structured protein-ligand interaction

Hydrogen bond Hydrophobicity

 Table 13: Conformation table for all the proteins

INTERMOLEC TOTAL
BINDING INHIB VANDERWAL ELECTROSTAT TORSIONAL
PROTEINS DOCKED LIGAND ULAR INTERNAL
ENERGY CONSTANT LS ENERGY IC ENERGY ENERGY
WITH ESTRAGOLE EFFICIENCY ENERGY(kcal ENERGY
(kcal/mol) (pM) (kcal/mol) (kcal/mol) (kcal/mol)
/mol) (kcal/mol)

INTERFERON-γ -5.0 -0.45 217.88 -5.89 -5.89 0.0 -0.26 0.89

INTERFERON-α2 -4.26 -0.39 758.51 -5.15 -5.15 0.0 -0.27 0.89

INTERLEUKIN-33 -4.86 -0.44 274.92 -5.75 -5.75 0.0 -0.24 0.89

INTERLEUKIN-12 -3.7 -0.34 1.93 -4.6 -4.63 0.04 -0.23 0.89

CONCLUSION FROM GC-MS:

• Estragole with Interferon gamma, interferon alpha-2, interleukin-33 and interleukin-12 has binding energy of -4.09, -
3.23, -4.66 and -2.67 respectively. Interleukin-12 with estragole has the lowest binding energy of -2.67 of other.
Hence interleukin-12 is tightly bounded with estragole which is more reactive to suppress the activity of pro-
inflammatory property of interleukin-12.

• Highly efficient fragment hit makes it easier to optimize the fragment into a drug-like compound. Interleukin-12 has
the highest ligand efficiency and others have sufficiently high ligand efficiency.

• The more hydrogen bonds between protein-ligand, the more stable the protein-ligand interactions. All four proteins
have hydrogen bonding distributed evenly over the surface.

• Hydrophobicity is thought to be one of the primary forces driving the folding of proteins. On average, hydrophobic
residues occur preferentially in the core, whereas polar residues tend to occur at the surface of a folded protein.
Interferon-gamma and interleukin-33 have higher hydrophobicity while other two proteins have sufficiently small
hydrophobicity.
Application of leaf extract in orthopedic application
• 3D Slicer is a free and open source software package for image
analysis and scientific visualization. Slicer's capabilities include: SOFTWARE USED:
Handling DICOM images, Volume renderings, Image segmentation. 3D SLICER
• Fusion 360 is a cloud-based 3D modeling CAD software platform FUSION 360
to create 3D designs, collaborate, manage data, create toolpaths, and
run simulations to validate designs. Fusion 360 is also the tool of
choice for manufacturing, machining, engineering and industrial
design experts.

• CT scan DICOM files - https://www.embodi3d.com/files/file/8130-

ct-scan-dicom-files-for-instructables-tutorial/
3D SLICER PROCEDURE
Step 1 :View DICOM image as 3D image using Grayscale Model Maker
Step 2 : Remove the CT table using Segment editor
Step 3 : Extract only the Femur bone using segment editor
Step 4 :Extract the bone as .stl file and import this file to fusion 360.
FUSION 360 PROCEDURE
Step 1 : Import .stl file and convert the mesh to solid

WITH MESH WITHOUT MESH

Step 2: Create a crack in the bone

DIMENSION:
AREA 18354.315 mm^2
LOOP 785.61 mm
LENGTH
Step 3: Add the patch to the Cracked surface
Step 4: Add the material to the patch and modify the properties of Estragole and
hydroxyapatite[11] to it.
PROPERTIES VALUE
Thermal 5.800 E-01
conductivity W/(m.K)
Specific heat 4.1183 J/(g° C)
Thermal expansion 12 µm/ (m.°C)
coefficient
Young’s modulus 6 GPa
Poisson’s ratio 0.27
Shear modulus 5120MPa
Density 0.965 g/cm3
Yield strength 18 MPa
Tensile strength 1.5 MPa

Note: Temperature at 25°C

CONCLUSION
• The presence of phytochemicals is revealed by quanlitative and UV- visible spectroscopy
analysis.
• The presence of high quantity of functional groups are revealed in FTIR spectroscopy.
• The main constituents of Ocimum basilicum estragole is identified in GC-MS.
• In ESR, the ROS level is found to be low, so it can be used in medical field.
• In autodock, the main constituent of Ocimum basilicum estragole is docked with pro-
inflammatory proteins and it is found that the estragole bind with the pro- inflammatory
protein and reduce the inflammatory effect.
• As the first phase of bone fracture is inflammation, it is concluded that estragole can be
used in orthopaedic application.
• The prototyping model is designed using fusion 360, where a crack is created and a patch
of estragole and hydroxyapatite is implemented.
 FUTURE WORKS

• Screening of biocompatibility of the patch has to be carried to check for its health
care applications.

• Preparation of a prototype using electrospinning or 3D printing.

• Pressure analysis of bone fracture and suitable software prototyping.

 REFERENCE
1. Azeana Zahari, Abdulwali Ablat and Noridayu Omer (2016), ‘Ultraviolet-visible study on acidbase equilibria of aporphine alkaloids
with antiplasmodial and antioxidant activities from Alseodaphne corneri and Dehaasia longipedicellata’, Scientific Reports | 6:21517 |
DOI: 10.1038, pp. 1-10.
2. Ioana-Raluca, Suica-Bunghez and Sofia Teodorescu (2017), ‘Characterization of antioxidant activity and phytochemical compounds,
metal nanoparticles obtained by Sideritis scardica extracts’, Academia Română, 62(6-7), pp. 545-552.
3. Praveena Ch, Swaroopa Rani S and Veeresham C (2013), ‘Phytochemical investigation of Calophyllum inophyllum Linn.’, Natural
Products Chemistry & Research Article, ISSN: 2329-6836 NPCR, Volume 1, pp.1-4.
4. Rajat Buragohain (2015), ‘Screening and quantification of phytochemicals and evaluation of antioxidant activity of Albizia chinensis’,
International Journal of Current Microbiology and Applied Sciences, ISSN: 2319-7706, Volume 4 Number 9, pp. 305-313.
5. Shazia Kanwal Malik, Maqsood Ahmed and Farah Khan (2018), ‘Identification of novel anticancer terpenoids from Prosopis juliflora
(Sw) DC (Leguminosae) pods’, Tropical Journal of Pharmaceutical Research, ISSN: 1596-5996, pp. 1-8.
6. Ummah Hafsa Mukta, Roni Roy and A. F. M. Shahid Ud Daula (2020), ‘Phytochemical analysis, antioxidant and antidiarrhoeal
activities of methanol extract of Fimbristylis miliacea (L.) Vahl’, Journal of Pharmacognosy and Phytotherapy, ISSN 2141-2502, Vol.
12(1), pp. 10-18.
7. Carmen Socaciu ,Florinela Fetea , Floricuta Ranga (2020), ‘Attenuated Total Reflectance -Fourier Transform Infrared Spectoscopy
with chemometrics to control the botanical authenticity and Quality of cold pressed’, Mdpi, 10, 8695; doi:10.3390, pp.4-15.
8. Bolaji Lilian Ilesanmi, Linda Schollum, Barbara Kuhn, Michelle McConnell and Sonya Mros (2019), ‘Inflammatory markers and bone
health in postmenopausal women: a cross-sectional overview’, Multidiscipline Digital Publication Inst Proc. 2019;8(1):28.
9. Solvent retention data, Dean rood (2015), agilent technologies.
10. Gas Chromatography – Mass Spectrometry (GC−MS) ,Tara M. Lovestead
11. ESTRAGOLE PROPERTIES: Estragole | C10H12O – PubChem, HYDROXYAPATITE: Pubmed, NCBI
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