Meena Vankudre, Vina D. L. Putra and Vaibhav Zamare
disease and reduced frequency of several types of can- cers, most notably prostate (Giovannucci et al., 2002). Lycopene can prevent carcinogenesis by protecting vital bio molecules including DNA, proteins, enzymes and lipids (Breemen et al., 2008 and Salvadori, 2007). It has been shown to have the highest antioxidant activ- ity among the carotenoids in cell protection against free radicals (Salvadori et al., 2007). Supplementation with tomato based products also increase lycopene level in human serum and protect against UV-light induced erythema (Aust et al., 2003).
Lycopene content in tomatoes can vary signifi- cantly, depending on the type of tomato and ripening. In the reddest strain, lycopene levels are close to 50mg kg-1, compared with only 5mg kg-1 in the yellow strain (Veazie et al., 2006). The colour was generally an accurate indicator of lycopene content, with a yellow cultivar containing less lycopene than red cultivars, and the red cultivars containing more than an orange culti- var. However, black cultivars as a group did not contain more lycopene than red (Cox et al., 2003).
Processing of food helps to release the lycopene from food matrix, thus improving accessibility of the lipophilic compound for the formation of lipid micelles together with dietary lipid and bile acid. Lycopene uptake is higher after ingestion of processed tomatoes (e.g tomato paste, tomato juice) (Stahl and Sies, 2006).
The antioxidant and anticancer properties of Lyco- pene make it an ideal component for daily food supple- ments. It is one of the popular pigments highly accepted by food industry as food additives and also for its health benefits (Rao and Agarwal, 1999). At the sixty-seventh meeting of The Joint FAO/WHO Expert Committee on Food Additives (JECFA), it was agreed that both syn- thetic lycopene and lycopene extracted from any organ- isms were acceptable as food colours and established a group ADI of 0-0.5 mg/kg body weight/day for both preparations (FAO/WHO 2006).
Nowadays, Lycopene supplements are available in the form of tablets or gel beads which are relatively high of cost. For this reason, improved form of Lycopene sup- plement is developed in the form of widely consumed cookies. This provides the consumer the easiest way to include Lycopene in the diet by consuming one package, to fulfil the daily requirement of lycopene.
MATERIALS AND METHODS
EXTRACTION AND PURIFICATION OF LYCOPENE
Tomatoes, obtained from the local market, were ground with water in a mortar and pestle to obtain a paste. The
paste was mixed with acetone and stirred for 3-4 min- utes (Aghel et al., 2011). The extra liquid was removed and the residue is collected by squeezing in between fil- ter paper. The red dehydrated paste was the mixed with mixture of n-hexane and ethyl acetate (1:1). Anhydrous MgSO4 was added to remove the extra water content. The solution was filtered by using filter paper to sepa- rate chunks of tomato paste. The final solution was sub- jected into column chromatography packed with silica gel by using solvent mixture of n-hexane and acetone (1:1) to elute ß-carotene and pure Lycopene (Lehman, 2009). Lycopene extract from tomato was a dark-red viscous liquid. It was freely soluble in ethyl acetate and n-hexane, partially soluble in ethanol and acetone, and insoluble in water.
EVAPORATION OF SOLVENT
The collected compound was subjected to rotary evapo- rator to evaporate and recollect the solvent. The water bath was maintained at temperature 80o . The solvent, such as ethyl acetate and n-hexane were recollected and reused for another round of extraction.
COLOR TEST FOR CAROTENOID
To the small amount of extracted sample, concentrated sulphuric acid was added drop by drop. Blue colour was obtained, indicating the presence of carotenoid (Maheswari et al., 2012).
QUALITATIVE AND QUANTITATIVE ANALYSIS OF LYCOPENE
The pure lycopene sample was analyzed by thin layer chromatography by using Lycopene standard. Other qualitative analysis was done by using Double beam UV-vis spectrophotometer at wavelength range 400- 500nm. The concentration was also determined by this method and by comparing with standard. A lycopene solution in n-hexane showed an absorption maximum at approximately 472 nm (Alda et al., 2009).
COOKIES AND LYCOPENE EMBEDDED CREAM PRODUCTION
The cookies are made by following the simple butter cookies recipe (Lamke et al., 2011). The whole batter pro- duces 24 cookies. The cookies are coated with lycopene embedded chocolate cream. 1 cookie weighs about 27.8 grams and contains 1 mg of lycopene. The product is formulated in such a way that they will provide about 3 mg lycopene per serving.
