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Vol 2 | Issue 3 | Jul – Sep 2023                                                                           Indian J Pharm Drug Studies | 118 

Original Article 

Spectrophotometric Characterization of lycopene Phytosomes from Fruit 

Peels of Lycopersicum esculentum 

Ramakrishna Shabaraya A1, Viresh K Chandur2 

From, 1Prof. and Principal, 2Asso. Prof.Department of Pharmaceutics, Srinivas College of Pharmacy Mangalore. 

 ABSTRACT  

Among all colored fruits tomatoes are one of the rich source of lycopene, which is essentially known for its anti-oxidant 

activity and effective in treating major diseases like cancer, heart diseases. It is insoluble in water and sensitive to light and 

temperature but, it is soluble with various proportions in aprotic solvents like petroleum ether, diethyl ether, and acetone. Best 

combination of these solvents was made used to extract lycopene and characterized for its standardization by means of 

phytochemical testing, melting point, λ max, FTIR and DSC studies showing the effective way of extraction with good yield 

up to 8%, phytochemical investigation showed presence of flavonoids, turpinoids and steroids. UV Spectrophotometric 

analysis shows λ max at 471nm. FTIR studies show the presence of conjugated carbon double bonds essential to get red color 

to the product and open ring structure on both the ends. DSC endotherms, the melting peak obtained for the phytosomal 

complex was less than the melting point obtained for the lycopene and is due to the less cohesive force energy of crystal lattice 

of the complex. 

Key words: Lycopene, FTIR, UV Spectrophotometer, DSC, Endotherms. 

ecent intriguing possibilities for the treatment of 

chronic diseases stand out as phytophar-

maceuticals. New therapy options for a variety of 

diseases are made possible by phytochemicals that have 

fewer side effects and cost less. Natural pigments called 

carotenoids are produced by microbes and plants to protect 

cells from photosensitization and act as light-blocking 

substances during photosynthesis [1]. About 60 of the 

more than 700 carotenoids that have been classified can be 

present in the human diet [2]. Carotenes and xanthophylls, 

including lutein and beta-cryptoxanthin, are important 

dietary groups [3]. Up to 95% of the total plasma 

carotenoid concentration is made up of six different 

compounds: -carotene, -carotene, -cryptoxanthin, lutein, 

zeaxanthin, and lycopene. Food is one factor that affects 

plasma concentrations of carotenoids [4]. In human plasma 

and tissue, lycopene is a very significant carotenoid and 

accounts for up to 50% of the body's total carotenoids. 

Access this article online 

Received – 24th March 2023 

Initial Review – 04th April 2023 

Accepted – 14th April 2023 Quick Response Code 

Lycopene is a carotenoid pigment that is primarily 

present in foods that are red in hue. Due to its conjugated 

double bonds, lycopene is a potent antioxidant. Lycopene 

prevents oxidation of vascular cells and lipoproteins 

through its antioxidant action. In vitro tests on human 

lymphoid cells, low-density lipoprotein, and plasma have 

shown that lycopene is an efficient singlet oxygen 

quencher [5]. Lycopene-rich meals have been 

demonstrated to reduce the risk of cancer and 

cardiovascular disease. 

The primary red colour in fruits, lycopene is a linear, 

unsaturated hydrocarbon carotenoid. Lycopene, which is 

present in tomatoes, belongs to a class of carotenoids with 

a broad polyene chain that contains 35–40 carbon atoms. 

Some of these polyene chains are ended by two 6-carbon 

rings.Antioxidant capabilities of carotenoids have the 

potential to slow down the ageing process and many 

degenerative diseasbes. Lycopene is a necessary nutrient 

that must be consumed daily. Lycopene's potential health 

_______________________________________ 

Correspondence to: Viresh K Chandur, Asso. Prof. 

Department of Pharmaceutics, Srinivas College of 

Pharmacy, Mangalore. Karnataka, India. Tel: +91 

9742565141, Email: viresh.chandur2009@gmail.com 

R 

mailto:viresh.chandur2009@gmail.com


Shabaraya A & Chandur K                                                     Spectrophotometric Characterization of lycopene 

Vol 2 | Issue 3 | Jul – Sep 2023                                                                           Indian J Pharm Drug Studies | 119 

consequences have also recently been researched. 

Lycopene is believed to have antioxidant characteristics 

that are primarily responsible for these positive effects [6]. 

MATERIALS & METHODS 

Collection of Tomato fruits: Tomatoes were collected 

from local market at Mangalore of Karnataka in the month 

of January 2018. The fruit was authenticated by Botanical 

department Regional Science Center Pilikula Mangalore. 

Morphological study of Tomato fruit 

Tomato fruit: The tomato is the edible, often red, 

fruit/berry of the plant Solanumlycopersicum, Syn: Lycope-

rsicumesculentum, commonly known as a tomato plant. The 

plant belongs to the nightshade family, Solanaceae. 

Tomato is consumed in diverse ways, including raw, as an 

ingredient in many dishes, sauces, salads, and drinks. 

While tomatoes are botanically berry-type fruits, they are 

considered culinary vegetables as an ingredient or side 

dish for savory meals. Numerous varieties of tomato are 

widely grown in temperate climates across the world, 

with greenhouses allowing its production throughout the 

year. The plants typically grow to 1–3 meters (3–10 ft.) in 

height and have a weak stem that sprawls. It is 

a perennial in its native habitat, and cultivated as 

an annual. Fruit size varies according to cultivator, with a 

width range of 0.5–4 inches (1.3–10.2 cm) 

Preliminary preparation for the peel of tomato: The 

ripe tomatoes stored at 4ºC were used within 48 h for 

isolation of lycopene. The peels of tomato were separated 

using blanching method. The removed peels were dried at 

room temperature (24-25°C) for 2-3 h and then packed in 

zip-lock polyethylene bags.  

Method 1: Extraction of Lycopene from Tomato Peel: The 

dried peels were stored at 4°C before grinding with a small 

amount of solvent system (n-hexane: acetone: ethanol in 

2:1:1 v/v) in the presence of carbon dioxide. It was then 

sonicated using ultrasonic crusher followed by magnetic 

stirring for 4 h in an inert environment (carbon dioxide) 

under dark conditions. The resulting extract was collected 

in an amber color glass container with the presence of 

carbon dioxide environment and stored at 4ºC for further 

processing. The extract was then dried using rotary flash 

evaporator by heating at 60°C at 50 rpm in dark condition 

[1-2]. 

Method 2: Raw tomato slices were blended with water 

(1:1) and boiled for 10 Minutes. The aqueous suspension 

was vacuum filtered to reduce water to 15% and treated 

with Acetone (1:1) under high speedy blender and filtered, 

filtrate containing β- carotenes and other carotenoids was 

separated. Residue was treated with diethyl ether (1:1) 

under high speedy blender and filtered. Filtrate containing 

lycopene was separated and lyophilized. 

Pre-Formulation Investigation of Lycopene Extract 

1. Shinoda test: a little quantity of extract was dissolved 

in alcohol + few fragments of Mg turnings + conc. HCl 

drop wise. 

2. Lead acetate test: lead acetate solution was added to 

small amount of extract. 

3. Alkaline reagent test: Increasing amount of sodium 

hydroxide was added to the sample of extract. 

4. Ferric chloride test: Extract + Ferric chloride solution 

Detection of Steroids  

1. Lieberman Burchard’s test: 2mg of dry extract was 

dissolved in acetic anhydride, heated to boiling, cooled 

and then 1 ml of conc. H2SO4 added. 

2. Salkowski reaction: 2mg of dry extract was shaken 

with CHCl3. To the CHCl3 layer, H2SO4 was added 

slowly along the sides of test tube [4]. 

UV-VIS Spectrophotometric Determination of 

Lycopene in Tomato Peel extract. 

Determination of λ max: Solution of Lycopene in Diethyl 

ether was prepared and scanned using UV–VIS spectroph-

otometer to read λ max in the wavelength of 200–600 nm. 

Determination of Standard Graph: 100 mg of tomato 

extract (Lycopene) was dissolved in diethyl ether and the 

obtained solution is serially diluted to obtain descending 

concentrations and the final volume was made up to 10 ml 

using diethyl ether. Finally the absorbance values were 

plotted on a graph with variable concentration on x-axis 

and absorbance at 471 nm on Y-axis. The regression 

coefficient value and the equation for the graph were also 

derived so as to ease the calculations of concentration of 

lycopene in tomato samples [8]. 

FTIR spectrum of lycopene was taken by FTIR 

Spectrophotometer: Lycopene was scanned between 

wave number ranges of 4000 cm-1 to 650 cm-1. Major 

peaks of the spectra were interpreted to determine the 

respective functional groups present. To authenticate the 

spectrum, FTIR spectrum of the extracted lycopene was 

compared with spectrum of imported lycopene, spectrum 

given by HisarPhytoextracts [9]. 

Differential Scanning Calorimetric (DSC) studies of 

Lycopene Phytosomes: Lycopene Phytosomeswere 



Shabaraya A & Chandur K                                                     Spectrophotometric Characterization of lycopene 

Vol 2 | Issue 3 | Jul – Sep 2023                                                                           Indian J Pharm Drug Studies | 120 

subjected to Differential Scanning Calorimetric using SDT 

Q600 V20.9 Build 20 calorimeter. The instrument 

comprised of calorimeter, flow controller, thermal analyser 

and operating software. The instrument was designed to 

supply heat to the sample, so that its temperature was 

raised precisely. The sample was heated in sealed 

aluminium pans under nitrogen flow (30 ml/min) at a 

scanning range of 30 to 800oC. Empty aluminium pan was 

used as a reference. The heat flow as afunction of 

temperature was measured for the Lycopene Phytosomes. 

The DSC of Lycopene Phytosomeswas obtained from 

which melting point was recorded [10]. 

RESULTS 

Phytoconstiuents investigation found to possess flavonoids 

and Terpenoids in tomato peel extract (Table 1 – 3). 

Qualitative determination of λmax and FTIR studies. 

UV-VIS Spectrophotometric Detection of Lycopene in 

Tomato Peel Extract. 

Identification of Lycopene: -  

i) Determination of λmax 

Figure 1: Determination of λmax of Lycopene form Tomato Peel extract. 

As reported λ max for solution of lycopene in diethyl ether 

is 472 nm and during investigation it was found to be 471 

and 502 nm (Figure 1 & Figure 2). 

Quantitative determination –The Percentage yield from 

marketed powder and Tomato peels was found to be 8.5% 

and 8.0 % respectively. 

From fruit authentication and from the qualitative & 

quantitative determination it was concluded that selected 

component from tomato was Lycopene. 

The phytochemical studies confirms the presence of the 

above said constituents and further the spectrophotometric 

determination was compared with that of the marketed 

tomato extract containing Lycopene, confirming the 

presence of lycopene which showed same λ Max as that of 

the standard Figure 2. FTIR spectral studies confirms 

about the presence of the same functional groups as that of 

the standard (Figure 3, 4, 5). 

Drug Excipients Compatability Study                        

FTIR Studies of Tomato Extract

Figure 2: Standard Graph of Lycopene. 

y = 0.4419x

R² = 0.9961

0

0.2

0.4

0.6

0.8

1

1.2

1.4

0 0.5 1 1.5 2 2.5 3

A
b

s 
a

t 
4

7
1

 n
m

Conc in mg/ml

Standard Graph of Lycopene

Series1

Linear (Series1)



Shabaraya A & Chandur K                                                     Spectrophotometric Characterization of lycopene 

Vol 2 | Issue 3 | Jul – Sep 2023                                                                           Indian J Pharm Drug Studies | 121 

Figure 3: FT-IR spectrum of Lycopene 

Figure 4: FT-IR spectrum of Phosphatidylcholine 

Table 1: Peak picking of Lycopene 

Frequency Amide Lipid 
C-H bending 

C-C and C-C-H stretching 
Water 

Reported 1650 and 1540 cm-1 1730-1765 cm-1 

And 3000-2800 cm-1 

1100-1400 cm-1 3700-3000&1600-1700 

Observed 1610 1735 1049 2922 & 1610 

Table 2: Peak picking of Phosphatidylcholine 

Frequency Amide Lipid 
C-H bending 

C-C and C-C-H stretching 
Water 

Reported 1650 and 1540 cm-1 
1730-1765 cm-1 

And 3000-2800 cm-1 
1100-1400 cm-1 3700-3000&1600-1700 

Observed 1610 1735 1049 2922 & 1610 

Table 3: Peak picking of Formulation C1 

Origin 
Frequency (Cm−1) 

Observed Given 

C−C−H stretch, 3338.78 3072−3055, 3033−3007 m 

methylene/methyl 

H stretch, alkene 

2924.09 

2852.72 

2981−2962, 2926−2894 

2894 m to s 

C=C stretch 1602.85 1636, 1558 m 

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PC



Shabaraya A & Chandur K                                                     Spectrophotometric Characterization of lycopene 

Vol 2 | Issue 3 | Jul – Sep 2023                                                                           Indian J Pharm Drug Studies | 122 

C=O Stretch 1734.01 1735 

C−H deformation, methyl 1373.32 1399−1375, 1364−1350 m 

C−H out-of-plane, (E) 

disubstituted double bond 
1004.91 976−946 s 

Figure 5: FT-IR spectrum of Formulation C1 

Differential scanning calorimetric studies 

 
Figure 6: DCS of Lycopene phytosomes. 

DISCUSSION 

The maxima are decidedly red-shifted from the maxima at 

505, 475, and 445 that are observed for lycopene in 

organic solvent such as hexane as observed by Davis AR 

et al. Sanchit S and Oishee C observed the absorbance 

maxima of lycopene at 502 nm using hexane. The FTIR 

spectra showed typical bands same as by Aghel N et al., 

observed arising from amide (1650 and 1540 [cm-1]) and 

lipid (1730-1765 [cm-1] and 3000-2800 [cm-1]) groups. 

Other bands occur at 1477-1400 [cm-1] (C-H bending), 

1100-1400 [cm-1] (C-C and C-C-H stretching), and 1170-

1115 [cm-1] (C-O stretching). Strong and broad absorption 

bands of water are shown in the 3700-3000 [cm-1] and 

1600-1700 [cm-1] range. The frequency region between 

1200 and 900 [cm-1] shows intense bands attributed to (C-

O-C) 1352 vibrational modes of various carbohydrates and 

acids, which are abundant groups in tomatoes. Frequencies 

of all types of deformations are found below 1000 [cm-1]. 

The spectral signal obtained at a frequency of 957 [cm-1] 

can be attributed to the presence of Trans CH out-of-plane 

deformation vibration of lycopene. 

The DSC thermogram obtained for the phytosome is 

shown in Figures 6. The thermogram of the lycopene 

phytosomes showed endothermic peaks at 66.43oC. These 

melting peaks obtained were less than the melting point 

obtained for the Lycopene (173.28°C). The decrease in the 

melting point of the lycopene might be due to the less 

cohesive force energy of crystal lattice of the complex 

formed as showed by Haixiang W et al one peak around 44 

◦C, probably because of loss of water, another peak at 

about 162.5 ◦C, likely due to its melting point. However, 

the DSC curve of the inclusion complexes showed 

different features of free molecules and the physical 

mixtures, indicating that there was probable interaction 

between the lycopene and β-CD. These results evidenced 

that the lycopene was embedded into the cavity of the β-

CD. Phytochemical screening showed the presence of 

flavonoids, Terpenoids & Steroids as seen by Sravanthi J 

et al. Better yield was seen as compared with marketed 

spray dried tomato powder of 8%. 

CONCLUSION 

Hence, Tomato can be a good natural source of lycopene. 

Methods have been developed for extraction and analytical 

process for quantification and qualification. An absorption 

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Shabaraya A & Chandur K                                                     Spectrophotometric Characterization of lycopene 

Vol 2 | Issue 3 | Jul – Sep 2023                                                                           Indian J Pharm Drug Studies | 123 

maximum was found to be at 471 nm. FTIR, a better tool 

for understand drug excipient interactions and identifying 

the functional groups which complies with screening tests. 

In the DSC endotherms, the melting peak obtained for the 

phytosomal complex was less than the melting point 

obtained for the lycopene and is due to the less cohesive 

force energy of crystal lattice of the complex.  

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How to cite this article: Ramakrishna Shabaraya A, 

Viresh K Chandur.Spectrophotometric Characterization 

of lycopene Phytosomes from Fruit Peels of 

Lycopersicum esculentum. Indian J Pharm Drug Studies. 

2023: 2(3) 118-123. 

Funding: None              Conflict of Interest: None Stated 

 


