CHAPTER 7 CHAPTER 7 CHAPTER 7 CHAPTER 7

DEVELOPMENT OF FUNCTIONAL FOODS

USING LACTUCA SATIVA

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7.1. INTRODUCTION

"Functional food is a natural or processed food that contains known

biologically-active compounds which when defined in quantitative and qualitative

amounts provides a clinically proven and documented health benefit, and thus, an

important source in the prevention, management and treatment of chronic diseases of

the modern age". Currently, dozens of functional food components from plants

(phytochemicals) as well as animals are under investigation for their potential role in

disease prevention and health promotion. The general category of functional foods

includes processed food or foods fortified with health-promoting additives, like

vitamin-enriched products. Products considered functional generally do not include

products where fortification has been done to meet government regulations and the

change is not recorded on the label as a significant addition. An example of this type

of fortification would be the historic addition of iodine to table salt, or Vitamin D to

milk, done to resolve public health problems such as ricketts. Fermented foods with

live cultures are considered functional foods with probiotic benefits. Functional foods

are an emerging field in food science due to their increasing popularity with health-

conscious consumers and the ability of marketers to create new interest in existing

products. Functional foods, as a form of medicalised food, open opportunities to

market food through its therapeutic potential. Functional foods obscure the boundaries

between food and drugs and in doing so they raise questions about a fundamental

public health concern, namely, whether a balanced diet is sufficient for health or

whether a normal diet still requires additives such as those provided by functional

foods (Williams and Ghosh, 2008; Lawrence and Germov, 2012).

The term “Nutraceutical” combined from “Nutrition” and “Pharmaceutical” in

1989 by Dr. Stephen De Felice, defined nutraceutical as food or part of a food that

provides medical or health benefits (De Felice, 2002). Nutraceuticals also refer to

natural functional/medical foods or bioactive phytochemicals that have health

promoting, disease preventing or medicinal properties. These nutraceuticals normally

contain the required amount of vitamins, lipids, proteins, carbohydrates, minerals, or

other necessary nutrients, depending on their emphases (Zeisel, 1999; Whitman,

2001; Rajasekaran et al., 2008).

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7.2. Development of Lactuca sativa based compressed bar

The new trend for consumption of healthy, innovative and practical food,

which has occurred recently, has leaded the market of cereal-bars to a gradual growth.

Cereal-bars are considered healthy type of food, because they are rich in fiber,

however, poor in fat (Bower and Whitten, 2000; Palazzolo, 2003). Compressed food

bars made from dehydrated food items provide for the Armed Forces, the advantages

of greater convenience and stability and a higher concentration of energy and

nutrients than conventional canned foods. The nutritious energy bars have gained

more importance and popularity in the global market during recent years and today

the market is offering wide variety of bars under different names. Indigenously made

bars like Horlicks multicereal nutri bar, Rite bite choco delite bars, low fat bar,

sugarless bar, fruit choco bars, etc. are getting popular among Indian consumers. The

various varieties of bars available in the global market with good organoleptic

properties and consumer appeal are referred by names such as chewy cereal granola

bars, organic bar, choco bar, muffin bar, fruit filled bars, etc. These bars are generally

packed in metallised polyester films and have a limited shelf life of 3 to 4 months.

7.3. Development of Lactuca sativa based fruit spread

Fruit is a food group which is receiving more attention among the population

due to its interesting and healthy properties such as high functional and nutritional

value, being rich in fibre, minerals, vitamins and terpens antioxidant compounds

(Cavanah et al., 2003; Randhir et al., 2005). Fruits contain many different dietary

phytonutrients which contribute to the prevention of degenerative diseases caused by

oxidative stress (Kaur & Kapoor, 2002). The intake of food rich in phenolic acids,

polyphenols and flavonoids scavenge free radicals such as peroxide, hydroperoxide or

lipid peroxyl, thus inhibiting the oxidative mechanism that leads to degenerative

diseases (Halvorsen et al., 2006; Ho et al.,2007; Pellegrini et al., 2003).

The basic principle of jam and marmalade manufacture is the boiling together

of fruit, sugars and water. For marmalade, the peel is cooked separately prior to

mixing in. Jelly manufacture involves boiling the fruit with water and then the extract

after filtration is boiled with the sugars (Kirk and Sawyer, 1991). Fruit and vegetable

spreads means all of the recognized fruits and those vegetables recognized as suitable

in making jams, including chestnuts, ginger, melon, rhubarb, tomato.

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7.4. Development of Lactuca sativa based squash

The major source of biologically active substances, such as vitamins and

secondary metabolites (polyphenols, carotenoids, sterols, glucosinolates, and

saponins) is present in herbs, fruits and vegetables (Alothman et al., 2009; Cassileth,

2008; Yang et al., 2007). The consumption of fruits and vegetables is globally

insufficient and should be encouraged, and it may be useful to enhance fruit

concentrations of vitamins and secondary metabolites by genetic and/or

environmental approaches (Poiroux-Gonord et al., 2010).

Squash is non-alcoholic concentrated syrup that is usually fruit-flavoured and

usually made from fruit juice, water, and sugar or a sugar substitute. Modern squashes

may also contain food colouring and additional flavouring. Some traditional squashes

contain herbal extracts.

7.5. Materials and methods

7.5.a. Raw materials

All the ingredients for preparation of Lactuca sativa based compressed bar

were procured from local market.

7.5.b. Preparation of Lactuca sativa based compressed bar

For the preparation of 2.0 kg of Lactuca sativa based compressed bar, 640.0 g

flaked rice, 320.0 g dalia, 200.0 g bengal gram and 200.0 g green gram were soaked

in wet muslin cloth for 2 h and dried in the hot air oven at 50° C for 2 hrs, after

drying, all the four samples were grounded and sieved to get fine powder. Later, all

the samples were packed in sealed containers. 240.0 g oats was roasted in an

aluminium pan at low flame, grinded, sieved and packed in sealed container. All the

above ingredients were mixed thoroughly, 640.0 g skim milk powder, 2.0 g amchur

powder, 480.0 g desiccated coconut and 640.0 g powdered sugar 2.5 % Lactuca sativa

lyophilized powder was also added and mixed well. Finally, 200.0 g each honey and

date syrup was mixed together and added to the above mixed sample, they act as

binding agents. 40.0 g cardamom powder was also added for providing aroma. 80.0 g

each cashew nut and raisins were added as taste buds. All the ingredients were mixed

well. The mixtures were conditioned at 20-30° C for 1 hr. A 50 g of each mixture was

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fed into a rectangular mould (2½" square 55) and pressed with a load of 5000 kg

using Carver hydraulic press (Carver 2625 model 12- 102T hydraulic laboratory press

12 ton capacity manually operated full threaded column 10" x 7" flatten size, 5 – 1/8"

press stroke) to obtain bars. The applied pressure 2½" square mould (Load/Area) was

standardized as 124 kg/cm2 to obtain a compression ratio of 2.066). The bar samples

were packed in PP (polypropylene, 75µ) and heat sealed. Alternatively, the above bar

samples were stored under 5° C, ambient (15-35°C) and 37° C temperature

conditions.

7.5.c. Preparation of Lactuca sativa based fruit spread

Aloe vera gel devoid of green skin (20g)

Blend with apple pulp (20g)

Filter through four layered muslin cloth

Add 10.0 g of partially purified extract

pH adjust to 3.25 by adding citric acid

Add sugar 75g

Pectin 1g

Boil (68-70oBrix)

Transfer to glass bottles for setting

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7.5.d.Preparation of Lactuca sativa based squash

7.6. Sensory evaluation

Sensory evaluation was carried out in Lactuca sativa based compressed bar for

their colour, aroma, taste, texture and overall acceptability (OAA) on 9-point hedonic

scale by semi-trained panel of 15 members. The 9-point Hedonic scale grading was as

follows: 9=Excellent, 8=Very good, 7=Good, 6= Good above fair, 5= Fair, 4= Fair

above poor, 3= Poor, 2= Very poor, 1= extremely poor.

7.7. Phytochemical studies

• Total polyphenols (Test was performed as described previously in Chapter

No. 2)

• Flavonoids (Test was performed as described previously in Chapter No.2)

• Antioxidant activity by DPPH (Test was performed as described previously

in Chapter No. 2)

7.8. Anti-anxiety activity

7.8.a. Experimental design

The Lactuca sativa (LS) based functional foods were treated to animals for 15

days and it was analysed for anti anxiety property by behavioral parameters. The

animals were divided into four groups with six animals per each group.

Group I: Control

Pineapple pulp (30g) and 10 ml of L sativa extract

Pasteurize for 30 min at 80o C

Add Sugar (40.72 g) and water (21.37 ml), boil to syrup consistency

Cool and filter through muslin cloth

observe through refractometer to get 40o Brix

Add 0.7g cirtic acid and 0.06g sodium benzoate

Store the squash in sealed bottles

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Group II: LS based compressed bar

Group III: LS based fruit spread

Group IV: LS based squash

Elevated plus maze test (Test was performed as described previously in Chapter No.

3)

Light dark box test (Test was performed as described previously in Chapter No. 3)

7.9. Results and discussion

Lactuca sativa based compressed bar was developed (Figure 40), fruit spread

(Figure 41), and squash (Figure 42).

Lactuca sativa has bitter taste, in order to mask its bitter taste, compressed bar

was chosen because it is sweet to taste. The results revealed that the contents of total

polyphenols and flavonoids respectively, also observed in radical scavenging activity

of Lactuca sativa based compressed bar, fruit spread, and squash (Table 56). The

sensory studies (Table 57) of the compressed bar indicates that the acceptability of the

product was ‛good’ initially with a score of 7.9 on 9 point hedonic scale.

The behavioral parameters were carried out. EPM is considered one of the

most widely validated tests for assaying new benzodiazepine-like anxiolytic agents

(Bhattacharya and Satyan 1997). It is well known that the anxiolytic agents increase

the motor activity which is measured by time spent by the animal in the open arms

(Hui et al., 2001; de Melo et al., 2006). Products that increase open arm exploration

are considered as anxiolytic and reverse holds true for anxiogenics. In our study, we

observed that Lactuca sativa based compressed bar, fruit spread, and squash fed group

(p < 0.01) significantly increases time spent in the open arms with associated decrease

in closed arms when compared to control group (Table 58).

The light/dark box is a characteristic tool used in the assessment of anxiety.

The basic measure is the animal’s preference for dark, enclosed places over bright,

exposed places. Time spent in the lit (light) half of the arena, and the related

exploratory behaviours, are reliable parameters for assessing anxiolytic effects that

may be useful in identifying and/or screening of anxiolytic and anxiogenic agents.

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Significant increase in the number of entries into light zone and time spent in

the light compartment (p < 0.05) was seen with Lactuca sativa based compressed bar,

fruit spread, and squash fed group compared to control group (Table 58).

Lactuca sativa based

functional foods

Flavonoids

(mg/100g)

Total

polyphenols

(mg/100g)

DPPH

(%)

Compressed bar 62.20 140.04 79.45

Fruit spread 70.50 150.20 81.55

Squash 72.25 120.15 80.50

Table 56. Total polyphenols, flavonoids and DPPH scavenging activity of Lactuca

sativa based functional foods.

Sensory attributes

Compressed bar Fruit spread Squash

Colour 7.7±0.22 7.5±0.22 7.9±0.12

Aroma 7.6±0.10 7.9±0.10 7.6±0.20

Taste 7.9±0.14 7.6±0.14 7.9±0.22

Texture 7.8±0.20 7.8±0.20 7.6±0.10

Overall acceptance 7.9±0.14 7.6±0.14 7.8±0.22

Table 57. Sensory attributes of Lactuca sativa based functional foods, values

expressed as Mean ± SD of 3 individual samples.

Groups

Elevated Plus

Maze Test Light Dark box Test

Time spent in

open arm

(sec/5min)

No of entries

to light zone

Time in light

zone (sec/5min)

Latency to first

entry into light

zone

Control 69.6±14.36 9.5±5.55 85.51±20.1 48.9±18.2

Compressed

bar 113.7±11.60* 13.2±1.75* 110.00±08.5* 18.7±04.2*

Fruit spread 105.7±08.50* 13.0±3.50* 135.46±11.1* 14.5±06.4*

Squash 115.7±10.60* 14.0±3.10* 136.46±11.0* 15.5±04.5*

Table 58. Effect of Lactuca sativa based functional foods on the behavior. Time

spent in open arm by mice behaviour in elevated plus maze and the time spent in light

zone by mice behaviour in light and dark test was measured. * indicates p <0.05

compared with control group.

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Figure 40. Lactuca sativa based anti-anxiety compressed bar

Figure 41. Lactuca sativa based antianxiety fruit spread

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Figure 42. Lactuca sativa based anti-anxiety drink (squash)

7.10. SUMMARY AND CONCLUSION

Functional food which claims health boosting affect play a significant role in

the reduction of health care costs associated with diet related illness. These foods are

being marketed for disease prevention and treatment as consumers takes an

increasingly proactive approach to health. Although, the functional food products are

assessed for safety before going to market yet these are reported to interact with

conventional drugs. Therefore proper regulation and consultation with health care

providers is key for the success of these products. The regulatory issues regarding

these categories is different among different countries but still their future seems to be

big, better and brighter in coming few years. Functional foods and nutraceuticals were

developed using Lactuca sativa lyophilized powder.

The study concluded that Lactuca sativa incorporated nutraceuticles viz.,

compressed bar, fruit spread and squash possesses a marked anti-antianxiety property

with antioxidant activity.