Spectro Photometric Study of Lycopene Extract After Various Treatments of Fertilizers and Manure

 

Mr. Satish Y. Mane

Department of Chemistry,ShivneriMahavidyalayaShirurAnantpal,Dist-Latur, 413544. Maharashtra, India.

*CorrespondingAuthorE-mail:sssymane@gmail.com

 

ABSTRACT:

In present work extraction, isolation and spectrophotometric studyof lycopene was be done by various methods and effect of various fertilizer on lycopeneafter treatment on source plant was studied. The Several studies have been undertaken for the extraction of lycopene and for its quantification in various natural sources. Here we studied 13 types of treated tomato samples, using differentanalytical methods. The outcome of the study shows that the good analytical method for quantification of lycopene in tomatoes

 

KEYWORDS:Lycopene, extraction, hexane, acetone, benzene, fertilizers, farm manur, ethanol.

 

 


INTRODUCTION:

1The weakest singlet oxygen quencher of the antioxidants studied was α-tocopherol, which is found in much greater concentrations in many body metabolic reactions. Lycopene is a member of carotenoid family; is a lipid soluble antioxidant synthesized by many plants and microorganisms but not by animals and humans1 where it serves as an accessory light-gathering pigment and protect them against the toxic effects of oxygen and light. It is a red pigment without provitamin. Carotenoids are a group of vitamin A-like substances that give colour to fruits and vegetables and protect against various types of cancer, heart disease and blindness by destroying harmful free radicals in the body.2Lycopene, as an antioxidant, counteracts these detrimental effects by either reducing the rate the damage occurs, or by preventing it from occurring absolutely. Lycopene gives the red colour to tomatoes, papayas, pink grapefruits and watermelons.

 

 

Leafy green vegetables as well as deep orange fruits (apricots, cantaloupes) and vegetables (squash, sweet potatoes) are excellent sources of other diseasefighting carotenoids such as betacarotene and lutein. Lycopene is one of the most significant and useful carotenoids found in nature, and a potent antioxidant that has been shown to play critical role in cancer prevention.Lycopene is a carotenoid pigment and phytochemical found in tomatoes Lycopene extract from tomatoes is obtained by ethyl acetate extraction of the pulp of ripe red tomatoeswith subsequent removal of the solvent.3Chronic diseases, including cancer and cardiovascular disease, are the main causes of death in the Western world. Along with genetic factors and age, lifestyle and diet are also considered important risk factors,about 50% of all cancers have been ascribed to diet.Oxidative stress induced by reactive oxygen species is one of the main foci of recent research related to cancer and cardiovascular disease. Reactive oxygen species are highly reactive oxidant molecules that are generated endogenously through regular metabolic activity, lifestyle activity and diet.4The singlet quenching ability of lycopene is twice as high as that of β- carotene and 10 times higher than that of α-tocopherol and butylated hydroxyl toluene. All trans Lycopene as a result of the 11 conjugated carbon-carbon double bonds in its backbone, lycopene can theoretically assume 211 or 2048 geometrical configurations. However, it is now known that the biosynthesis in plants leads to the all-trans-form, and this is self-governing of its thermodynamic stability.5The structural distinctions of the predominant lycopene geometrical isomer is a member of the carotenoid family of pigments, lycopene is a potent antioxidant. The many conjugated double bonds of carotenoids make them potentially powerful antioxidants, and lycopene is no exception. Indeed, lycopene had the strongest singlet oxygen-quenching capacity of several carotenoids, with α-carotene, ß-carotene, and lutein next in capacity.6The product also contains oils, fats, waxes, and flavour components naturally occurring in tomatoes. Carotenoids, such as beta-caroten and lycopene, are important components of antioxidant defense against lipid peroxidation in living cells. Lycopene, an aliphatic hydrocarbon, has received particular attention as a result of studies indicating that it has highly proficient antioxidant and free radical scavenging capacity. This is the main reason that the development of tomato varieties with increased lycopene content requires resourceful selection and the ability to measure lycopene in thousands of samples.7Lycopene is a lipophelic, 40-carbon atom highly unsaturated, straight chain hydrocarbon containing 11 conjugated and 2 non-conjugated double bonds. The all-trans isomer of lycopene is the most predominant isomer in fresh tomatoes and is the most thermodynamically stable form. The many conjugated double bonds of lycopene make it a potentially powerful antioxidant, a characteristic assumed to be accountable for its beneficial effects. The antioxidant The Anti-Atherogenic Effects of Lycopene activity of lycopene is high light by its singlet oxygen-quenching property and its ability to trap peroxy1 radicals.8They react with cellular components, causing oxidative damage to such critical cellular biomolecules as lipids, proteins and DNA. There is strong evidence that this damage may play a significant role in the causation of several chronic diseases. Free radicals are unstable molecules produced during natural reactions in the body and in response to environmental triggers like smoking and sun exposure. They can be destructive and damage cells, contributing to the development of diseases such as cancer.9Sample (1.0-1.5 g powder) was extracted with 10mL acetone-petroleum ether (50% v/v). The upper lycopene-containing organic layer was removed by means of a pipette and collected in test tube. Extraction was repeatedin human plasma, lycopene is an isomeric mixture, containing at least 60% of the total lycopene as cis- isomers. All-trans, 5-cis, 9-cis, 13-cis, and 15-cis are the most commonly identified isomeric forms of lycopene with the stability sequence being 5-cis>all-trans>9-cis>13-cis>15- cis>7-cis>11-cis, so that the 5-cis-form is thermodynamically more stable than the all trans-isomer. Whereas a large number of geometrical isomers are theoretically possible for all-trans lycopene, according to only certain ethylenic groups of a lycopene molecule can participate in cis-trans isomerization because of steric hindrance. In fact, only about 72 lycopene cis isomers are structurally favorable.10The major colouring principle in tomato extract is lycopene; however, minor amounts of other carotenoid pigments may also be present.

 

MATERIAL AND METHODS:

1) Acetone-petroleum ether extraction method:-

1The extracts were combined, washed with 15mL saturated aqueous sodium chloride (NaCl) and removed the aqueous wash with a micropipette. The extract was washed with 10mL of 10% aqueous potassium carbonate (K2CO3) and removed the aqueous wash. The lycopene-containing organic layer was dried with a drying agent (calcium chloride). The excess solvent was allowed to evaporate at room temperature for a few minutes in the dark. The tubes containing lycopene extracts were covered with aluminium foil and stored in freezer until further analysis.

 

2) Acetone-Ethanol–Hexane Extraction Method:-

2Three solvents were used for comparison of extraction efficiency: (1) acetone, (2) ethanol, and (3) hexane. Tomato pastes were submitted to extraction with each solvent (1:10 v/v) by means of shaker for 30 minutes. The extraction was performed at room temperature with light protection by covering the vessels with aluminium foil. 3The lycopene contents were assayed for purity and concentration by UV–Vis spectroscopy at 472 nm (maximum of absorption for lycopene) .

 

Hexane extraction method:

4Sample (0.3 to 0.6 g powder) was weighed in a beaker, 5 mL BHT-acetone solution (0.05%, w/v), 5 mL ethanol and 10 mL hexane was added. The beaker was placed in a bowl of ice on a magnetic stirring plate, stirred for 15 min and added 3 mL distilled water. It was shaken for 5 min on ice and incubated at room temperature for 5 min to allow the separation of both phases. The upper layer containing lycopene was isolated by means of a pipette and collected in a test tube. The tubes containing lycopene extracts were covered with aluminium foil and stored in the freezer until further analysis.

 

Methanol Extraction Method:

5Fifty grams tomato paste was dehydrated by adding 65 ml methanol. This mixture was immediately shaken vigorously to prevent the formation of hard lumps. After 2 hr, the thick suspension was filtered; the dark red cake was shaken for another 15 min with 75 ml mixture of equal volume of methanol and carbon tetrachloride and separated by filtration. The carbon tetrachloride phase was transferred to a separatory funnel; added one volume of water and shaked well. After phase separation, the carbon tetrachloride phase was evaporated and the residue was diluted with about 2ml of benzene. Using a dropper, 1 ml of boiling methanol was added in portion, then crystals of crude lycopene were appeared immediately and the crystallization was completed by keeping the liquid at room temperature and ice bath, respectively. The crystals were washed 10 times using benzene and boiling methanol. Long, red lycopene prisms were observed under the microscope with some colorless impurity substances. For more purification, column chromatography on active acidic alumina using toluene as eluent was done. The deep red zone was collected. After complete evaporation of solvent, the residue was dissolved in 2 ml benzene. After recrystallization using boiling methanol, no colorless substances observed. Crystalline lycopene is not isomerized but has a tendency to autoxidation (or air oxidation), especially in light, so it was kept in dark evacuated glass tubes prior to use.

 

Spectrophotometry:

6Calibrate at zero absorptance using a blank of pure DMSO Measure absorbance of blank and samples at 645 and 663 nm no longer than 20 minutes after extraction procedure completed. A blank of pure DMSO will be included in each run. The absorbance of this blank will be subtracted from the absorbance readings of each sample before any calculations have been made.

 

RESULT AND DISCUSSION:

Sr. No.

Types of Treatment

Observed Optical density of Lycopene (mg/100g)

472nm

502nm

1

Rhizobium

11,48

10,30

2

Polutary manure

19,55

24,56

3

Cow dong

18,64

15,20

4

Trichoderma

13,20

14,32

5

Urea

3,41

5,30

6

Control

2,12

2,85

 

The extraction and isolation of lycopene has been carried out using different methods and the yield of lycopene showed to be high in natural sources of lycopene.A typical carotenoid such as lycopene displays maximum absorbance at 476nm. Spectrophotometry results revealed that lycopene showed maximum nabsorbance at 476nm, followed closely by absorbance at 503nm. The purified lycopene content was found to be the maximum in cherry tomato (88.87 mg/kg) as compared to Pakistani tomato (55.84 mg/kg) and watermelon (74.53 mg/kg). Lycopene content in tomato ranges from 55 to 181 mg/kg [41], 4.31 to 5.97 mg/100 g fw [4]

 

CONCLUSION:

Spectrophotometry showed the absorption peaks of carotenoids from these tomato samples in order to identify the sample with the highest lycopene concentration. Spectroscopy was used to quantify lycopene content in tomatoes varieties and showed a distinct vibration band at 846 cm-1 assigned to a trans CH deformation vibration of lycopene.

 

REFERENCES:

1         Pram YODJUN, Khantong and Chutima Separation of Lycopene/Solvent Mixture by Chitosan Membranes, Journal of Metals, Materials and Minerals, Vol.21 No.1 pp.107-113, (2011).

2         Tehniat Shahzad, Ijaz Ahmad, ShahnazChoudhry, Muhammad K Saeed, Muhammad N Khan, free radical scavenging activity of tomato, cherry tomato and watermelon: lycopene extraction, purification and quantification, International Journal of Pharmacyand Pharmaceutical Sciences, Vol 6, Suppl 2, ISSN-0975-1491, (2014).

3         A. Agarwal, SA Prahakaran and TM Said, J.Androl.653-660 (2005).

4         G. Turk, A. Atessahina, M. Sonmez, A. Yuce and Ceribasi, Theriogenology,778-785 (2006).

5         Wikepedia.

6         International Journal of Pharmaceutical Sciences Review and Research Volume 10, Issue 1, September October 2011; Article-018.

7         CMAJ SEPT. 19, 2000; 163 (6) Dr S. Agarwal and Rao are with the Department of Nutritional Sciences, Faculty of Medicine, University of Toronto, Toronto, Ont.

8         Journal of Pharmacognosy and Phytochemistry Volume 2 Issue 1.

9         IOSR Journal of Biotechnology and Biochemistry (IOSR-JBB) ISSN: 2455-264 X, Volume 1, Issue 5 (Jul–Aug.2015), PP 49-51.

10      Molecules 2010, 15, 959-987 doi 10.3390 molecules 15020959 ISSN 1420-3049 www.mdpi.com/journal/molecules

 

 

 

 

 

Received on 23.02.2018Modified on 09.04.2018

Accepted on 22.04.2018© AJRC All right reserved

Asian J. Research Chem. 2018; 11(3):607-609.

DOI:10.5958/0974-4150.2018.00109.8