Extraction of Beta Carotene from Selected Dried and Fresh Samples of Vegetables

 

Amita Gupta,  Ajita Dixit*, Ajay Sinha and Kamlesh Mittal

Rungta College of Engineering and Technology, Raipur (C.G.)

*Corresponding Author E-mail: ajita.dixit@gmail.com

Fresh  and dried vegetable samples were chosen for the extraction of beta carotene laboratory method was chosen to perform experiment . In present investigation dried vegetable were used for the extraction of beta carotene .Data of samples were compared with fresh sample of Beta carotene. It is a phytochemical helping people in reducing many diseases using it along with their daily diet.

 

 

INTRODUCTION:

Beta Carotene is a strongly red orange  colored pigment abundant in various vegetables and fruits β-Carotene is composed of two retinal groups, and is broken down in the mucosa of the human small intestine by 15,15'-monooxygenase to retinal, a form of vitamin A. β-Carotene can be stored in the liver  and converted to retinal as needed. More Out of 600 only 50 components liver can convert into active vitamin A.(1,2,3,4) organic molecule with eleven double bonds act as an antioxidants.(5,6) The two ends of the β-carotene molecule are structurally identical, and are called β-rings, Once inside the intestinal cell, cartotenoids can be converted into Vitamin A Stored in the liver, carotenoids get back into circulation via other lipoproteins, the same once that carry cholesterol in the blood  beta carotene is converted into two molecule of vitamin A (7,8)In 1831 β-carotene was first isolated  by Wackro Deand its structure in 1931  deduced by  Karrer  about 100 years  later (9)The group of nine carbon atoms at each end form a β-ring. Carotenoids are tetraterpenoid   organic pigments that are naturally occurring in the chloroplasts and chloroplasts of plants and some other photosynthetic organisms like algae, some types of fungus some bacteria  Carotenoids are generally not manufactured by species in the animal kingdom, although one species of aphid is known to have acquired the genes for synthesis of the carotenoid toluene from fungi, by the known phenomenon of horizontal gene transfer.

 

Carotenes contribute to photosynthesis by transmitting the light energy they absorb from chlorophyll. They also protect plant tissues by helping to absorb the energy from singlet oxygen, an excited form of the oxygen molecule O₂ which is formed during photosynthesis.

 

Material and Method

Material

Dried sample, Carbon tetra chloride, Anhydrous sodium sulphate, Benzene, Methanol Chloroform, Concentrated Nitric Acid, Antimony (Chemicals were purchased from Merck Company) Distilled water was used.

 

Sampling

Thin Orange Carrots 1Kg were purchased from Chhattisgarh (Raipur) from Telebandha vegetable market, Madhya Pradesh (Jabalpur) Gwari Ghat vegetable market, Maharashtra (Nagpur) local vender of vegetable   market.  One kilogram carrots were taken and crushed into small pieces. Crushed pieces were dried for three nights and four days in dark room under air.

 

Procedure

Dried, crushed and mashed carrots were filled in two liters of brown bottle. About 1200ml of Carbon Tetra Chloride was added into the above brown bottle. The bottle was shaken for ten minutes and kept in a water bath for two to three minutes which is later filtered.   The three times processed filtrate was kept in five liters brown bottle. 1000ml double distilled water was added to it then it was well shaken for twenty to thirty minutes and about one liter of this shaken solution was shifted to separating funnel which was allowed to stand for two to three minutes. Yellow colour liquid as lower layer was separated into a three liter conical flask by separating funnel which was covered with foil. Ten grams anhydrous sodium sulfate was added to it to remove the water content then filtered into the steam distillation flask then subjected to steam distillation till yellow coloured oily liquid was left behind, this liquid was transferred into a 100ml beaker. Crystalline liquid was obtained by steam distillation method. Now 25ml of benzene was added into it and then drop wise methanol was added into it, oily mass was obtained after keeping it for some time. Oily mass was separated and weighed and subjected to test. Every sample was processed three times to get optimum result.

 

Test for β-Carotene

1.     0.12 grams of β-Carotene was dissolved in 2ml chloroform and concentrated H₂SO₄ was added to it. The acid layer turned blue which confirms the presence of β- Carotene.

2.     0.12grams of β-Carotene was dissolved chloroform, and a drop of fuming HNO₃ was added to it. An immediate blue colorization was produced which then turned green and finally yellow.

3.     0.12grams of β-Carotene was added to chloroform, add a solution of antimony tetrachloride was added to it. A dark blue colorization was observed.

 

A result from tests proves that the obtained oily crystalline mass was of β-Carotene.

 

Natural beta-carotene consists of two isomers; that is, two different structures of the same (5,6)molecule. One is the all-trans and the other is the 9-cis; each contains the same atoms but in different arrangements. However, synthetic beta-carotene consists of only the all-trans form. In experiments, Ben-Amotz found that the natural 9-cis is a potent antioxidant that rapidly quenches free radicals. If that's the case, it would give natural beta-carotene a powerful advantage over the synthetic. Ben-Amotz believes that the all-trans form found in synthetic beta-carotene is not an antioxidant at all and is in fact a "pro-oxidant," which is why it damages cells and leads to cancer. Thus it is advisable people should intake this useful phytochemical  using fruits and vegetables in their daily diet(10).

 

Precaution           

Since these carotenoids are very sensitive to photo chemical air oxidation hence the solution as well solid were protected from undue exposure of light. Thus in every process sample should be protected  from light to get optimum result(11)

 

RESULT AND DISCUSSION:

According to USDA (12,13) fresh Carrots contains nearly 10 mg/100 gms Fresh carrots contain 9-10 mg/100 gm  where as 100 gm dried sample contain 5.7 mg/100 gm to   6mg/100gm.  It had been observed if it is dried in sun quantity further reduced  drastically C1  sample consider for Chhattisgarh has lesser value than Madhya Pradesh Maharashtra it has least quantity as compare to MP and Chhattisgarh .

 

Sample 1

C1               100                                                     5372

P1               100                                                     6002

 

 

Sample 2

C1               100                                                     5872

P1               100                                                     6012

 

 

 

Sample 3  

C1               100             5317           100             10.10

P1               100             5908           100             11.03

1/2 kg dried sample was reported between 80 to 65 gm.

 

 

 

Concentration of Beta Carotene Micro gm/ml	Absorbance at 450 nm
0.1000 0.2000 0.3000 0.4000 0.5000 0.6000 0.7000 0.8000	0.0100 0.0300 0.0500 0.0700 0.0900 0.1100 0.1300 0.1400

 

At 0.031 concentration value of carrot is 0.053  at 450nm. Value comes nearly 10983 Micro grams  using statical data .

 

Concentration of Beta Carotene Micro gm/ml	Absorbance at 450 nm
0.1000 0.2000 0.3000 0.4000 0.5000 0.6000 0.7000 0.8000	0.0142 0.0328 0.0514 0.0699 0.0886 0.1071 0.1257 0.1443

 

CONCLUSION:

Central part of India has almost same quantity of beta carotene between 6 mg to 5 but has least in Maharashtra. Fresh colored vegetables should be used and at home it should be kept in dark to reduce its nutrition value. Dried or crushed packed vegetables should be avoided as it had been observed it has lower value of beta carotene. As dried form of vegetable has 50% to 60% lesser quantity in beta carotene than fresh vegetable.

 

REFERENCE:

1.       Organic Chemistry ‘Solomons FryHle’ 8^th edition.

2.       Organic Chemistry ‘Stanley H pine’  5^th Ed. ( 2009)page no. 873.

3.       Organic Chemistry ‘Francis and Corey’ Page no. 1093.

4.       Chemistry and Natural Products ‘S.V. Bhat, M. Sivakumar’  page no. 200-203.

5.       Tarwadi, V.V.K.U, Mangale, S and Chiplonkar, S.A, Journal of Chromatography, 200,13: 885-891.

6.       http/// www.Winkipidia.org.

7.       Kramer, T.R. and Burri, B. Journal of American Clinical Nutrition, (1997).  65:871-75.

8.       Sun, J., Giraud, D.W., Moxley, R.A., et al. International Journal of Vitamin and Mineral Research ( 1997) 67: 155-163.

9.       Bio-chemistry ‘ Zoary’ 8^th Edition

10.     Levy, J., Bosin, E., et al. Nutrition and Cancer, (1996). 24: 257-66.

11.     Chemistry of Natural Products   Sujata V. Bhat, Bhimsen A. Nagasampagi, Meenakshi Sivakumar (2005).   

12.     http///USDA Agriculture Research service  –Nci carotenoid database/carrot/index.html

13.     http///USDA-NCI Carotenoid Database (www.nal.usda.gov/ fnlc / foodcomp /data /carot/index.html).

 

 

Received on 07.01.2013         Modified on 19.01.2013

Accepted on 06.02.2013         © AJRC All right reserved