1 2 3

A CRITICAL REVIEW ON DATE PITS ABSTRACT

God has not created anything useless in this world. Therefore use of such commodities

4considered waste can render unlimited benefits to mankind otherwise they will be creating 5unlimited episodes of problems . Take the example of only one such waste commodity and you 6will be astonished to know interesting facts about it. 7 This review article provides information on the nutritional and functional constituents of date

8pits from over 80 references. Date seeds contain higher protein (5.1 g/100 g) and fat (9.0 g/100 g) 9as compared to the date flesh. It is also high in dietary fiber (73.1 g/100 g), phenolics (3942 10mg/100 g) and antioxidants (80400 mol/100 g). 11 Date pit comprises 10-15% of the date fruit. Date pits were considered waste in the past and

12were a major waste produced during date syrup, pitted dates and other date products 13manufacturing. In the United States, pulverized ground date pits were used on a small scale, on dirt 14roads as a type of road base gravel. However, finding a way to make a profit on the pits would 15benefit date farmers substantially. 16 Some researchers used them in animal feed and outstanding results were obtained in respect

17of weight gain and less disease attack in animals. Diets, containing date pits, supported broiler 18weights and resulted in feed conversions comparable to, or better than, the control diet. Several 19investigations on the effects of pits in animal diets suggest the presence of substantial amounts of 20tannins, resistant starch and natural anabolic agents. 21 Further studies on date pits have exposed interesting and surprising facts about date pits.

22Date pits contain 22.50-80.20 % dietary fiber. The components of dietary fiber include cellulose, 23hemicellulose, pectins, hydrocolloids and lignin. Date pit dietary fiber consumption helps in 24preventing heart disease and cancer, normalization of blood lipids, regulation of glucose absorption 25and insulin secretion, prevention of constipation and diverticular disease. 26They contain 5.013.2% fat. It has been found that date seed oil (DSO) has better oxidative 27stability than most vegetables and also has a high antioxidant capacity owing to its richness in 28phenol and tocopherol compounds. Alpha-tocopherol constitutes 24.97% of the total tocopherols 29of DSO.

30

Date pit oil contains alpha-tocopherol 243.00 6.50 ppm and Polyphenols 319.00 7.30

31ppm. These phenolic components possess benefits, such as antioxidant, anti-carcinogenic, 32antimicrobial, anti-mutagenic and anti-inflammatory activities, as well as reduction of 33cardiovascular diseases. Date seed oil (DSO) can be used against male infertility by protecting 34spermatozoa against hydrogen peroxide (H2O2)-mediated damage and can improve sperm function, 35possibly owing to antioxidant properties. Date pits also contain significant higher contents of total 36phenolics (31024430 mg of gallic acid equivalents/100 g fresh weight) and antioxidant activity 37(580929 mol of Trolox equivalents/g fresh weight). 38 Date seed oil (DSO) has a protective effect against hydrogen peroxide (H2O2)-induced

39oxidative stress (OS) in human skin organ culture and suggested that the use of DSO as a dietary 40supplement may have beneficial effects in protecting against skin disorders in humans 41 The bioactive component(s), namely, protein and some derived polyphenolic compounds

42such as polysaccharides, lignans and bioflavonoids, are present in reasonable amounts in date pit 43which are reported to bind to the protein coat of virus and thus arrest absorption of the virus in the 44host cell. Therefore date pit extract has antiviral activity and has great potential to be used against 45viral infections like HIV. 46 Estrogens are a group of female sex steroid hormones that are synthesized in the ovary and

47placenta. They include estradiol, estriol, and estrone. The steroid sex hormone, estrone has been 48detected in the date pits as well as in the pollen grains of Phoenix dactylifera. The hormone estrone 49can be prepared commercially from date pits and can be used for treating different female estrogen 50related ailments. 51 This detailed information on nutritional and health promoting components of dates and their

52seeds will enhance our knowledge and appreciation for the use of dates in our daily diet and their 53seeds as a functional food ingredient. 54Keywords: Date pits, Antioxidants, Date pit oil. 55
56 57

58

59INTRODUCTION 60 61 BOTANICAL CLASSIFICATION The botanical name of the date palm, Phoenix dactylifera L., is presumably derived from a

62Phoenician name "phoenix", which means date palm, and "dactylifera" derived from a Greek word 63"daktulos" meaning a finger, illustrating the fruit's form (Linn, 1734). 64 Another source refers this botanical name to the legendary Egyptian bird, "Phoenix", which

65lived to be 500 years old, and cast itself into a fire from which it rose with renewed growth (Pliny, 661489; Van Zyl, 1983). This resemblance to the date palm, which can also re-grow after fire 67damage, makes the bird and the date palm share this name, while "dactylifera" originates from the 68Hebrew word "dachel" which describes the fruit's shape (Popenoe, 1938). 69 Belonging to the Angiosperms-Monocotyledones, Palmaceae is a family of about 200 genera

70and 1, 500 species (Dowson, 1982). Phoenix (Coryphoideae Phoeniceae) is one of the genera 71which contains a dozen species, all native to the tropical or subtropical regions of Africa or 72Southern Asia, including Phoenix dactylifera L. (Munier, 1973). According to Dransfield and Uhl, 73(1986) date palm is classified as follows: - Group: - Order: - Family: - Sub-family: - Tribe: - Genus: - Species: Spadiciflora Palmea Palmaceae Coryphyoideae Phoeniceae Phoenix Dactylifera L.

74

75TABLE 1 76Number of date varieties described per country

Egypt Egypt and Sudan Iran Iraq Morocco Tunisia USA Number of varieties described Author/Reference 26 Brown, 1924 22 Mason, 1925a,b 400 FAO, 1995 370 Dowson, 1923 244 Saaidi, 1979 250 Kearney, 1906 196 Nixon, 1950

77 78 79 PRODUCTION The date (Phoenix dactylifera L.) has been an important crop in arid and semiarid regions of

80the world. It has always played an important part in the economic and social lives of the people of 81these regions. The fruit of the date palm is well known as a staple food (Besbes et al., 2004a). The 82world production of dates has increased considerably during the last 30 years. Indeed, the 83production has tripled from 2,289,511 tonnes in 1974 to 6,772,068 tonnes in 2004 (FAOSTAT, 842005). 85 Dates are produced largely in the hot desert regions of Southwest Asia and North Africa, and

86are marketed worldwide as a high-value fruit crop. With the present uncertainty in the world food 87supply and an expected increase in demand, the date palm is likely to continue to provide a good 88source of low cost food. The world production of dates has increased from about 4.60 million tons 89in 1994 to 6.9 million tons in 2004 (FAO, 2007). Egypt, Saudi Arabia, Iran, and Iraq are the main 90producing countries (Table 2). Table 2 shows that Tunisia is leading the exporting countries in 91terms of gross exports (84.4 million US $), followed by Iran (36.4 million US $), Saudi Arabia 92(24.1 million US $), and Pakistan (22.5 million US $). However, the United States achieved the 93highest export price (3,339 US $/ton) among these countries, followed by Tunisia and Algeria, due 94to their strategy of growing top quality date varieties and targeting the high-value European 95markets (Zaid and Arias-Jimenez, 2002). 96Table 2 Cultivation area, production, export, and export value of dates for 97 2004. Countries Area Production Export Commercia Export 1000 1000 tons 1000 tons l value Hectares value 1000 US $ US $/ton Algeria 135 470 8 1,820 14,563 Egypt 35 1,166 3 457 1,370 Iran 185 880 95 383 36,430 Iraq 102 875 24 183 4,392 Oman 34 238 5 436 2,180 Pakistan 81 622 65 346 22,473

Saudi 145 901 Arabia Tunisia 45 122 UAE 186 760 USA 2 15 World 1,129 6,908 98 Source: (FAOSTAT). (FAO 2007). 99 100

44 40 60 4 377

548 2,110 219 3,339 786

24,090 84,382 13,127 13,357 296,248

The area under cultivation of dates in Pakistan and Sindh is 84,700 and 29,300 hectares

101respectively and its total production is 426,300 and 201,100 tones respectively. Only Khairpur 102district share is 34% of Pakistan and 83.33% of Sindh however the area is increasing day by day. 103Therefore, Khairpur district is known as Queen of Date palm in Pakistan. Khairpur, Kingri and Kot 104Diji are three major Date palm growing areas of the district. (Sayed, 2006-2007) 105 106 LIFE CYCLE AND FRUIT CHARACTERISTICS Dates pass through four stages of development known by their Arabic names; Kimri,

107Khalaal, Rutab, and Tamer. Many studies had discussed the physical and chemical development of 108dates as they pass through these stages (Sawaya et al., 1982;1983; Mustafa et al., 1986; Siddiqui 109and Gupta, 1994; El-Zoghbi,1994; Ahmed and Ahmed, 1995; Al-Hooti et al., 1997; Myhara et al., 1101999; Al-Shahib and Marshall, 2003). At the Kimri stages there is a rapid increase in size, weight, 111and sugar content. The moisture content at this stage is up to 85%. At the end of this stage the fruit 112starts to turn yellow or red depending on the variety. In the Khalaal stage weight gain is slow, the 113sucrose starts to be converted to glucose and fructose, the moisture content goes down, and tannins 114will start to precipitate and lose their astringency. In some varieties this latter process occurs 115rapidly, which makes the fruit palatable at the Khalaal stage. Normally the tips of the fruit start 116ripening by turning brown as they enter the Rutab stage which is characterized by a decrease in 117weight due to moisture loss, the conversion of sucrose into invert sugar (the degree depending on 118the variety) and a browning of the skin and softening of the tissues. The moisture content decreases 119to about 35% and the dates at this stage are sold as fresh fruit. Only when the dates are left to ripen 120further on the palm or sun dried they will develop into the Tamer (dried) stage. Therefore, dates 121distinguished from most other fruits in that they have a botanical maturity at least 3 commercial 122maturation levels, the sweet Khalaal, the Rutab, and the Tamer stage (Barreveld, 1993). There are 123over 2000 different date varieties (Amer, 1994). According to variety and growth conditions, date 124fruits (Tamer) vary in shape, size, and weight. Usually they are oblong in shape although certain 125varieties may be almost round. Length and width may vary from 18110mm to 832 mm,

126respectively and the average weight per fruit is about 2 to 60 grams (Zaid and Arias127Jimenez,2002). 128 Dates of date palm tree (Phoenix dactylifera L.) are popular among the population of the

129Middle East countries. A date is composed of a seed surrounded by a fleshy pericarp which 130constitutes about 8590% of date fruit weight (Elleuch et al., 2008). The seed constitutes between 13110% and 15% of date fruit weight (Hussein et al., 1998). 132 133 PROXIMATE COMPOSITION Chemical and nutritional constituents of date seeds were reported by numerous researchers.

134Al-Farsi et al., (2007) reported that the date seeds contain 3.17.1% moisture, 2.36.4% protein, 1355.013.2 fat, 0.91.8% ash and 22.580.2% dietary fiber. 136 Al-Farsi et al., (2007) evaluated the chemical composition of seeds of three native sun-dried

137date varieties from Oman (namely Mabseeli, Um-sellah, and Shahal). They found that 138carbohydrate was the predominant component in all varieties, followed by moisture, along with 139small amounts of protein, fat, and ash. Fat in seeds was ranged from 5.02 g/100 g in Mabseeli to 1405.90 g/100 g in Um-sellah. Seeds were, also, found to be good sources of dietary fiber, which 141varied between 77.75 and 80.15 g/100 g fresh weight. 142 The good nutritional value of date seeds is also based on their high dietary fiber content,

143which makes them suitable for the preparation of fiber-based foods and dietary supplements (Al144Farsi and Lee, 2008). 145 Table 3 Date seed composition.
Moistur e g/100 g 3.14 4.40 5.19 10.3 7.1 9.9 9.4 8.6 5.4 4.5 6.8 Protein g/100 g 3.92 5.40 2.29 5.7 6.0 5.2 5.04 4.73 6.43 5.92 5.1 Fat g/100 g 5.02 5.9 5.09 9.9 13.2 10.5 9.23 11.58 9.65 10.03 9.0 Ash g/100 g 1.03 1.16 0.89 1.4 1.8 1.0 1.0 1.02 1.04 1.05 1.1 Carbohydr ates g/100 g 86.89 83.14 86.54 72.7 71.9 73.4 75.3 74.07 77.48 78.5 78.0 Energy kcal/10 0g 387 386 379 385 412 391 386 401 403 408 394 Fiber g/100 g 79.84 80.15 77.75 67.8 64.5 68.8 73.1 Phenoli cs mg/100 g 4430 4293 3102 3942 Antioxida nts mol/100 g 58000 90300 92900 80400 References

Varieties

Mabseeli Umsellah Shahal Fard Khalas Lulu Deglet noor Allig Ruzeiz Sifri Average

(Al-Farsi et al., 2007) (Al-Farsi et al., 2007) (Al-Farsi et al., 2007) (Hamada et al., 2002) (Hamada et al., 2002) (Hamada et al., 2002) (Besbes et al., 2004) (Besbes et al., 2004) (Sawaya et al., 1984) (Sawaya et al., 1984)

146

Data are expressed on wet weight basis.

147Table 1 Proximate composition, water extract and water activity of roasted date pit powder Component Amount Wet basis, kg/100 kg sample 1.66 (0.31) 6.96 (0.24) 7.95 (0.43) 0.96 (0.02) 61.50 21.00 (0.007) 6.57 (0.31) 0.09 (0.005) Dry basis, kg/100 kg dry solids 7.08 8.08 0.98 62.31 21.35 6.68 -

Water Protein Fat Ash Carbohydrate * Crude fibre Others Water extract Water activity

152SUGAR CONTENTS 153Table 3 Sugar analysis of roasted date pits Sugar type Amount Wet basis, kg/100 kg sample Reducing sugars 1.09 Non-reducing 1.84 sugars Total sugars 2.93
154

148Note: Values in the parentheses are standard deviation. 149* Carbohydrate was estimated by difference. 150Source: (Rahman et al., 2006) 151

Dry basis, kg/100 kg dry solids 1.11 1.87 2.98

Source: (Rahman et al., 2006)

155 156 157

ANTIOXIDANT Reactive oxygen and nitrogen species, ROS/RNS are essential to energy supply,

158detoxification, chemical signaling and immune function. They are continuously produced in the 159human body and they are controlled by endogenous enzymes (superoxide dismutase, glutathione 160peroxidase, catalase). An exposure to external oxidant substances or a failure in the defense 161mechanisms results when there is an over-production of these species. The antioxidant hypothesis 162says that as antioxidants can prevent oxidative damages, increased intakes from the diet will also 163reduce the risks of chronic diseases (Stanner et al., 2004). 164 Chaira et al., 2007 reported about the composition of date pit from two varieties (Deglet nour

165and Alig ) as oil contents 10.13% in Deglet nour and 12.37% in Alig, In addition ethyl acetate 166extracts from both these varieties showed an important free radical scavenging activity towards 11671- diphenyl-2- picrylhydrazyl (DPPH) free radical. 168 169 PHENOLICS Phenolic components have been shown, also, to possess benefits, such as antioxidant, anti-

170carcinogenic, antimicrobial, anti-mutagenic and anti-inflammatory activities, as well as reduction 171of cardiovascular diseases. Thus, it is considered important to increase the antioxidant intake in the 172human diet and one way of achieving this is by enriching food with phenolics. As some synthetic

173antioxidants may exhibit toxicity, require high manufacturing costs and have lower efficiency than 174natural antioxidants, there is a need to identify natural and possibly to devise more economical 175ways to obtain effective antioxidants with potential to be incorporated into foods. At present, the 176natural antioxidants commercially produced include tocopherols, ascorbic acid and plant extracts 177(Al-Farsi and Lee, 2008). 178 Beyond compositional analysis, there is work of (Al-Farsi and Lee 2008) who researched the

179functional properties of date seeds. Their reported composition was 3.10-7.10% moisture, 2.301806.40 % protein, 5.00-13.20 fat, 0.90-1.80 % ash and 22.50-80.20 % dietary fiber. Also seeds 181contain high levels of phenolics (3102-4430 mg Gallic acid 1100 gm (Ardekani et al., 2010). 182 Phenolic compounds of date seeds mainly phenolic acids and flavonids, have been shown to

183possess such as antioxidant, anticarcinogenic, antimicrobial, antimutagenic and antiinflamatory 184activities (Halliwell, 1997 and Diplock et al., 1998), as well as reduction of cardiovascular diseases 185(Peterson and Dwyer, 1998). Thus it is considered important to increase the antioxidant intake in 186the human diet and one way achieving this is by enriching food with phenolics. 187 Table 7. Polyphenols, tocopherols and oxidative stability of date pit oil. Oils Parameters Date pits oil Palm kernel oil 44.40 2.30 198.00 4.50 187.00 4.01

188 189 190 191

Oxidative stability (hr) 41.60 2.00 -tocopherol ppm 243.00 6.50 Polyphenols ppm 319.00 7.30 Values are mean SE of three estimations. Source: Basunay and Al-Marzooq (2011)

Rahman et al., 2006 reported proximate analysis of roasted date pits to contain protein

1927.08%, Fat 8.08%, Ash 0.98%, Carbohydrates 62.31%, and Crude Fiber 21.35%. Sugar analysis of 193roasted date pits as reducing sugars 1.11%, Non-reducing sugars 1.87%, and Total sugars 2.98%. 194 Al-Farsi et al., (2007) reported date seeds contain significant higher contents of total

195phenolics (31024430 mg of gallic acid equivalents/100 g fresh weight) and antioxidant activity 196(580929 mol of Trolox equivalents/g fresh weight). Therefore date by-products (seeds) can be 197served as a good source of natural antioxidants and can potentially be considered as a functional 198food or functional food ingredient. 199 It has been found that date seed oil (DSO) has better oxidative stability than most vegetables

200and also has a high antioxidant capacity owing to its richness in phenol and tocopherol compounds 201(Besbes et al., 2004 b and c).

202

Ardekani et al (2010) reported that Iranian date palm seed has a relatively high antioxidant

203activity due to contribution of phenolics compounds. The Iranian date seeds are strong radical 204scavengers and can be considered as a good source of natural antioxidants for medicinal and 205commercial uses. 206 Table Antioxidant content and antioxidant capacity of date seeds Component Total phenolics (mg gallic acid equivalent kg1) Total flavonoids (mg rutin equivalent kg1) -Carotene (g kg1) Lutein (g kg1) -Cryptoxanthin (g kg1) Lycopene (g kg1) Zeaxanthin (g kg1) -Carotene (g kg1) FRAP (mol ferrous equivalent kg1) DPPH (% inhibition) Superoxide (% inhibition)
207 208

Value 24 600 63 674 3142 1599 20.4 19.5 10.8 ND 756 280 41.6 45.7

ND, not detected. Source: (Ibrahim, W H and Habib, H M 2011)

209 210

MINERAL CONTENTS OF DATE PITS Rahman et al., 2006 reported mineral Contents of roasted date pits as Sodium 165 mg/kg,

211Potassium 2540.96 mg/kg, Calcium 191.76 mg/kg, Iron 21.33 mg/kg, Copper 5.33 mg/kg, 212Magnesium 789.48 mg/kg, Manganese 6.19 mg/kg, Zinc 1.53 mg/kg, Phosphorous 1300 mg/kg, 213Lead < 0.05 mg/kg, Cadmium<0.05 mg/kg, Chromium <0.05 mg/kg. 214 Table 4. MINERAL CONTENTS OF DATE PITS Concentration (mg/kg samples) 160.00 10.50 189.35 11.02 2489.50 22.05 19.23 0.19 5.02 0.001 1256.23 16.05 811.30 13.09 1.67 0.001 7.12 0.02

Constituents Sodium (Na) Calcium (Ca) Potassium (k) Iron (Fe) Copper (Cu) Phosphorus (P) Magnesium (Mg) Zinc (Zn) Manganese (Mn) Values are mean SE of three estimations. Source: Basunay and Al-Marzooq (2011)

215 216 217

218Table 4 Mineral contents of roasted date pits Constituent Concentration (mg/kg sample) Sodium (Na) 165.00 Potassium (K) 2540.96 Calcium (Ca) 191.76 Iron (Fe 21.33 Copper (Cu) 5.33 Magnesium (Mg) 789.48 Manganese (Mn) 6.19 Zinc (Zn) 1.53 Phosphorus (P) 1300 Lead (Pb) <0.05 Cadmium (Cd) <0.05 Chromium (Cr) <0.05
219

Source: (Rahman et al., 2006)

220 221 222

FATTY ACID COMPOSITION OF DATE PITS Rahman et al., 2006 reported fatty acid composition of roasted date pits powder sample as

223C10 (capric) 0.0049%, C12 (lauric) 0.54%, C15 (pentadecanoic) 0.0006%, C16 (Palmitic) 0.21%, 224C18:1 (Oleic) 0.508%, C18:2 (Linoleic) 0.128%. Fatty acid composition of roasted date pits oil as 225C10 (capric) 0.062%, C12 (lauric) 6.793%, C15 (pentadecanoic) 0.0075%, C16 (Palmitic) 2.64%, 226C18:1 (Oleic) 6.39%, C18:2 (Linoleic) 1.61%. 227 Date seed oil has been reported to be composed of about 44% saturated fatty acids (SFA),

22841% monounsaturated fatty acids (MUFA) and 14% polyunsaturated fatty acids (PUFA), (Besbes 229et al., 2004a). 230 Table 5. Some physical and chemical properties of date pits oil compared with palm

231kernel oil. Oils Parameters Date pits oil Palm kernel oil 1.4500 0.001 1.50 0.10 0.74 0.003 1.42 0.13 19.30 1.05 254.00 5.01 1.70 0.15

232 233 234

Refractive index at 40C 1.4595 0.001 Color at yellow 35 Red 0.90 0.01 Free fatty acids (% as oleic acid) 0.05 0.002 Peroxide value meq. O2/ kg oil 1.73 0.15 Iodine number I2/100 g oil 46.50 2.13 Saponification number 213.00 4.50 Unsabonifiable matter (%) 1.65 0.13 Values are mean SE of three estimations. Source: Basunay and Al-Marzooq (2011)

235

Table 6. Fatty acids composition of date pits oil compared with palm kernel oil. Type of oils Date pits oil Palm kernel oil 2.87 0.15 3.11 0.19 45.88 1.90 16.27 0.85 15.66 0.71 2.16 0.19 26.25 1.31 2.57 0.18 0.89 0.11 0.00 0.00 70.29 6.30 29.71 1.50 100.00

Fatty acid (%)

236 237 238 239 Table 7. Unsaponifiable matter fraction (Hydrocarbons and sterols) of date pits oil 240compared with palm kernel oil. 241 Type of oils Compounds Date pits oil Palm kernel oil 0.05 0.001 0.04 0.001 1.11 0.10 1.83 0.80 0.42 0.01 3.09 1.02 0.40 0.01 7.85 2.80 15.85 4.03 11.79 3.01 0.45 0.01 11.61 3.05 7.50 2.72 62.70 5.62 0.00 0.00 2.10 019 10.67 2.93 4.00 1.15 21.33 3.21 38.10 4.01

C8:0 0.00 0.00 C10:0 0.25 0.001 C12:0 35.31 1.15 C14:0 0.04 0.001 C16:0 12.58 0.72 C18:0 3.30 0.10 C18:1 39.50 1.50 C18:2 8.20 0.61 C18:3 0.81 0.12 C20:0 0.02 0.001 Total saturated 51.00 3.50 Total unsaturated 49.00 3.10 Total fatty acids 100.00 Values are mean SE of three estimations. Source: Basunay and Al-Marzooq (2011)

242 243 244 245Table 2 Fatty acids composition of roasted date pits Fatty acid kg/100 kg kg/100 kg sample oil C10 (capric) 0.0049 0.062 C12 (lauric 0.54 6.793 C15 (pentadecanoic) 0.0006 0.0075 C16 (palmitic) 0.21 2.64 C18:1 (oleic 0.508 6.39 C18:2 (linoleic) 0.128 1.61
246Source: (Rahman et al., 2006)

C8 0.04 0.001 C10 0.02 0.001 C12 0.51 0.01 C14 1.12 0.10 C16 0.48 0.01 C18 3.15 1.01 C20 0.09 0.001 C22 6.79 2.15 C24 15.72 4.01 C26 10.79 2.91 C28 0.35 0.01 Squalene 11.23 3.03 C30 10.50 2.90 Total hydrocarbons 60.79 5.50 Cholesterol 0.00 0.00 Brassicaesterol 1.20 0.10 Campsterol 8.70 2.80 Stigmasterol 3.48 1.01 -sitosterol 25.83 3.59 Total sterols 39.26 4.03 Values are mean SE of three estimations. Source: Basunay and Al-Marzooq (2011)

kg/100 kg FA 0.35 38.81 0.043 15.09 36.51 9.20

247

248 249 250

USES OF DATE PITS AS ANIMAL FEED Vandepopuliere,Al-Yousef, and Lyons (1995) incorporated date pits in broiler starting diets

251at levels ranging from 5% to 27%. Diets, containing date pits, supported broiler weights and 252resulted in feed conversions comparable to, or better than, the control diet. Several investigations 253on the effects of pits in animal diets suggest the presence of substantial amounts of tannins, 254resistant starch (Hadarmi, 1999) and natural anabolic agents (Elgasim,Al-Yousef, & Humeida, 2551995). 256 The date seeds are considered as waste product of many date processing plants producing

257pitted dates (seed), date syrup and date confectionery. These date wastes are not consumed by 258humans and at present, are used mainly as animal feeds in the cattle, sheep, camel and poultry 259(Rahman et al., 2007). With world production of dates reaching 6.9 million tonnes in 2004, from 260this approximately 863 thousand tonnes of date seeds are year globally produced (FAO., 2007). 261Thus, utilization of such waste is very important to increase the income of this sector. 262 263 FUNCTIONAL FOODS Functional food components are health-promoting, nutritious materials from plant and

264microbial sources (Pszczola, 1998). They are components that have antimicrobial, anti265carcinogenic and other health-promoting activities, such as dietary fibre, vitamins, essential 266minerals, phytic acid, a-amylase inhibitors and tannins. 267 There is a particular lack of information on functional constituents of dates and their potential

268value as functional foods. Functional foods are defined as those foods that provide health benefits 269beyond basic nutrition (IFICF, 1998). Epidemiological studies have consistently shown that there 270are clear significant positive associations between intake of fruits and vegetables and reduced rate 271of heart diseases mortality, common cancers, and other degenerative diseases as well as ageing 272(Joseph et al., 1999; Dillard and German, 2000; Prior and Cao, 2000; Wargovich, 2000). This is 273attributed to the fact that these foods may provide an optimal mix of dietary fiber, natural 274antioxidants, and other biotic compounds. 275 276 USE IN BREAD Since a large quantity of date seeds are being produced as a waste material and the seeds

277contain a significant amount of bioactive phenolics and dietary fiber. Palm date seeds were 278evaluated by Almana & Mahmoud (Larrauri et al., 1995) as a source of dietary fiber. Finely milled

279date seed fiber had a total dietary fiber content of 71% while the coarsely milled fraction contained 28080% total dietary fiber. Total dietary fiber contents, rheological characteristics and sensory 281properties of Saudi Mafrood flat breads containing 0, 5, 10 and 15% date seed fibers were 282compared to control flat breads containing wheat bran. Rheological properties were similar for 283doughs containing coarse date seed fiber or wheat bran. Bread containing 10% coarse date seed 284fiber had a higher dietary fiber content and similar sensory properties to the wheat bran control. 285Breads containing the fine date seed fiber had higher dietary fiber contents than wheat bran 286controls, but lower color, flavor, odor, chewing, uniformity and overall acceptability sensory 287scores (Larrauri et al., 1995). 288 289 USE OF DATE PIT OIL IN BAKERY Briones et al., (2011) reported that little research has been undertaken on date seeds and this

290has focused particularly on their chemical composition for nutritional purposes. Hence, studies on 291product development from date seeds are limited. Some applications such as oil extraction from the 292seeds and the use of the seeds as a dietary-fiber provider in bakery formulations have been reported 293(Rahman et al., 2007). Others suggest some potential uses of the date seeds and their constituents 294in cosmetics, pharmaceuticals and to a lesser degree for food products (Devshony et al., 1992). 295 DATE PIT OIL MAYONNAISE 296 Date pit oil was used by Basunay and Al-Marzooq (2011) to replace conventional oil in

297producing mayonnaise, and sensory qualities were evaluated in comparison with commercial 298mayonnaise prepared from corn oil. The data demonstrated that mayonnaise containing date pit oil 299was superior in sensory characteristics as compared with control manufactured from corn oil. 300Results showed that the date pit oil could be used as non-traditional oil in some food processing 301such as mayonnaise products. 302USE IN MAKING DATE PIT JAM 303 Powdered date palm (Phoenix dactylifera) kernel (DPK) was used to produce edible jam.

304Saccharose, pectin, water, citric acid, ascorbic acid and Gum Arabic were added to the jam. Design 305Expert software was used to determine the optimum amount of the ingredients for the DPK jam. 306The design showed that there were ten variations of jams to be produced according to the content 307of different materials where the parameters such as amount of saccharose, pectin and dietary fiber 308from Gum Arabic were varied to give the best acceptability of the DPK jam samples. The jam 309samples produced were evaluated in terms of the sensory evaluation which includes taste, texture,

310aroma, appearance and overall acceptability by the panelist selected from students of Biochemical311Biotechnology Engineering, International Islamic University Malaysia. The samples were 312evaluated based on a five point hedonic scale, where zero (0.0) represented disliked extremely 313and four (4.0) represented liked extremely. All the data were analyzed by using Microsoft Excel 314spreadsheet. The results showed that 60% of panelists accept (> 3.1/4) the product while the others 31540 % moderately accept (2.55 2.64/4) the DPK jam samples. The average of overall acceptability 316is 2.98/4. This study is the outcome of the research done to find the acceptability of the production 317of DPK jams (Mirghani et al.,2012). 318 319 USE AS DATE PIT COFFEE Shafiur et al., 2006 suggested that caffeine-free date-pits-coffee could be developed when

320caffeine is a concern but a coffee-related flavor is desired. 321 A commercial product (Date Pits Powder Coffee Substitute) has also been introduced

322recently to the market (Rahman et al., 2007). 323 324 325 NUTRACEUTICAL VALUE USE AGAINST HYPERGLYCEMIA -Amylase and -glucosidase are key enzymes involved in carbohydrates breakdown and

326intestinal absorption, respectively. Inhibition of these enzymes hinders blood glucose level 327increase after a carbohydrate diet and can be an important strategy in the management of non328insulin-dependent diabetes mellitus (NIDDM). Dates-extract (DE) has inhibitory effect on 329amylase and -glucosidase.. The inhibition percentages on -amylase and -glucosidase are found 330to be in the range of 6-24% and 54%, respectively. The results clearly show the NIDDM treatment 331potential of date extract. Available anti-diabetic drugs used to treat non-insulin dependent diabetes 332mellitus NIDDM, such as Acarbose, strongly inhibits both enzymes. However, patients using 333Acarbose usually suffer from abdominal distention, flatulence, meteorism and possibly diarrhea 334(Bischoff, 1994). These side effects are caused by the excessive inhibition of pancreatic -amylase 335resulting in the abnormal bacterial fermentation of undigested carbohydrates in the colon 336(Bischoff, 1994; Horii, 1987). Therefore it is attractive to find a substance that has a strong 337inhibitory activity against -glucosidase, but minor effect on -amylase activity (Kwon et al., 3382006).

339 340

ANTIBACTERIAL ACTIVITY The crude methanol, acetone and water extracts of leaves and pits of three varieties (Barhee,

341Sukri and Rothana ) of Phoenix dactylifera were tested for antibacterial action against selected 342Gram positive and Gram negative pathogenic bacteria. Barring Enterococcus faecalis, the acetone 343and methanol extracts showed good antibacterial activity against Bacillus subtilis, Escherichia 344coli, Pseudomonas aeruginosa, Shigella flexeneri, Staphylococcus aureus, and Streptococcus 345pyogenes, whereas the water extract had very little effect on all test bacterial species and there was 346almost negligible effect on P. aeruginosa. Pits extracts of all three varieties of P. dactylifera were 347found to be more effective than leaves extracts. Results clearly showed that S. pyogenes was most 348sensitive pathogen to the crude extracts and had shown maximum zone of inhibition. Minimum 349inhibitory concentration (MIC) for S. pyogenes was found to be 1.3, 1.1, 1.6 and 1.4 mg/ml for 350methanol leaves and pits extracts and acetone leaves and pits extracts, respectively. ( Perveen, 351Bokhari and Soliman ,2012). 352ANTIVIRAL ACTIVITY 353 A crude acetone extract of the pit of date palm (Phoenix dactylifera L.) was prepared and

354its antiviral activity evaluated against lytic Pseudomonas phage ATCC 14209-B1, using 355Pseudomonas aeruginosa ATCC 25668 as the host cell. The antiviral activity of date pits was 356found to be mediated by binding to the phage, with minimum inhibitory concentration (MIC) of 357<10 mg /ml. The decimal reduction time (D-values), the concentration exponent (Z) and the phage 358inactivation kinetics were determined. The date pit extracts show a strong ability to inhibit the 359infectivity of Pseudomonas phage ATCC 14209-B1 and completely prevented bacterial lysis, 360which it is hoped will promote research into its potential as a novel antiviral agent against 361pathogenic human viruses.( Jassim and Naji,2007). 362 Date pit extract has promising antiviral activity, with minimum inhibitory concentration

363(MIC) concentrations found to be >10 mg/ml for Pseudomonas phage ATCC 14209-B1 (Fig. 1). It 364was also found that the 100 and 1000 mg/ml concentrations of date pit extract show a strong ability 365to inhibit the infectivity of Pseudomonas phage ATCC 14209- B1, as evidenced by the presence of 366higher numbers of Ps. aeruginosa cells surviving (Fig. 2). The results suggest that the cause of the 367inhibition could be due to interference with some aspect of the phages lytic cycle. The lytic cycle 368of the phage consists of three major phases (Stewart et al., 1998) : binding to a suitable host 369bacterium and injection of its genome;a period of intracellular production of new virions; and then 370lysis of the cell and release of progeny phage into the environment. To further understand the

371mechanism by which Ps. aeruginosa cells resisted phage lysis in the presence of date pit extract, 372the first of these stages,phage binding, was investigated. This effect was shown to be due to a 373direct effect of the extract on the phage itself rather than an effect on the host cell. It was found that 374the >100 mg/ml of date pit extract strongly inhibited the infectivity of Pseudomonas phage ATCC 37514209-B1 to Ps. aeruginosa (Fig. 3), and that this effect was abolished by heating. The phage 376infectivity inhibition may be attributable to date pit extracts heat-labile bioactive component(s) 377attaching to or modifying the surface of the phages.The bioactive component(s), namely, protein 378and some derived polyphenolic compounds such as polysaccharides, lignans and bioflavonoids, are 379present in reasonable amounts in date pit (Barreveld 2007), (Mutlak , Alywi and Maysara 1987), 380which were reported to act principally by binding to the protein coat and thus arrest absorption of 381the virus (Jassim and Naji ,2003). However, further studies are needed to investigate the direct 382effect of date pit protein and polyphenolic compounds on phage binding to host cells, and to 383determine whether the nucleic acid was damaged inside the phage capsid. In this study, date palm 384pit extract has demonstrated positive antiviral results (this effect was rapid, dose dependent, and 385appeared to be related to the adsorption stage of the phage replication cycle) Since HIV is an RNA 386virus it is not subjected to the proofreading that occurs during DNA transcription. As a 387consequence this kind of virus mutates very often. This enables HIV to grow resistant to 388antiretroviral pharmaceuticals quickly, and is one of the main reasons why an effective vaccine for 389HIV has not been developed yet (Jassim ,2005).In this regard, date palm pit extract could play an 390important role in a control replication of HIV-1 this hypothesis has been based from the results of 391date pit extract which has strongly inhibited the infectivity of the lytic Pseudomonas phage ATCC 39214209-B1 to Ps. Aeruginosa as a result of novel mechanism of interaction with binding of the 393phage to the host bacterium and injection of its genome. Therefore, it could conceivably play a role 394in combination therapy or even as a novel new class of anti-HIV phyto chemo-type therapy for 395drugs development fusion and attachment inhibitors similar to the Enfuvirtide (T-20) in which 396inhibit or blocks HIV from entering the CD4 cell are important drugs for the treatment of AIDS 397(Jassim ,2005). 398USE OF DATE PIT OIL FOR SKIN PROTECTION 399 Dammak et al., (2007) showed that date seed oil (DSO) has a protective effect against

400hydrogen peroxide (H2O2)-induced oxidative stress (OS) in human skin organ culture and

401suggested that the use of DSO as a dietary supplement may have beneficial effects in protecting 402against skin disorders in humans (Ben et al., 2009). 403 404 AMELIORATIVE ACTIVITY OF DATE PITS EXTRACT. The ameliorative activity of aqueous extracts of the flesh and pits of dates (Phoenix dactylifera

405L.) on carbon tetrachloride (CCl4)-induced hepatotoxicity was studied in rats. Sixty male Wistar 406rats were divided into six equal groups of 10. Four groups received extracts of flesh or pits of 407Phoenix dactylifera and intraperitoneal (IP) CCl4 (0.2 ml/100 g) either before or after 408administration of flesh or pits. Two groups were controls, one treated with CCl4 and one with only 409saline. Liver damage was assessed by liver morphology, histology, and estimation of plasma 410concentration of bilirubin and enzyme activities of aspartate aminotransferase, alanine

411aminotransferase, and alkaline phosphatase. Treatment with aqueous extract of date flesh or pits 412significantly reduced CCl4-induced elevation in plasma enzyme and bilirubin concentration and 413ameliorated morphological and histological liver damage in rats. This study suggests that CCl4414induced liver damage in rats can be ameliorated by treatment of extracts from date flesh or pits. 415( Qarawi et al 2004). 416Liver cirrhosis induced by CCl4 is perhaps the best-studied model of liver cirrhosis. 417(Cornelius,1993) Several mechanisms underlying this toxicity have been suggested.( Recknagel et 418al.,1989). The reduction of CCl4-induced elevated plasma activities of aspartate 419aminotransferase(AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and 420bilirubin level in animals pre- and post-treated with the aqueous extracts of date flesh or pits shows 421their ability to restore the normal functional status of the poisoned liver, and also to protect against 422subsequent CCl4 hepatotoxicity. The mechanism by which the date pits and flesh induces its 423hepatoprotective activity is not certain. However, it is possible that -sitosterol, a constituent of 424Phoenix dactylifera,( El-Mougy et al ., 1991) is at least partly responsible for the protective 425activity against CCl4 hepatotoxicity. (Nan-Lin and PinTome 1988) CCl4, the inactive metabolite, is 426transformed to a free radical through the microsomal cytochrome P-450-dependent enzyme, 427resulting in activation of CCl4 toxicity. An additional and important factor in the hepatoprotective 428activity of any drug is the ability of its constituents to inhibit the aromatase activity of cytochrome 429P-450, thereby favoring liver regeneration. On that basis, it is suggested that flavonoids in Phoenix 430dactylifera could be a factor contributing to its hepatoprotective ability through inhibition of 431cytochrome P-450 aromatase.( Kowalska, Brandt and Puett 1999)11 In addition, the recorded 432content of vitamin C in the date flesh and pits (0.179% and 0.137%, respectively) may also play a 433role in hepatoprotection. Extracts of date flesh and date pits are effective agents in the treatment 434and prevention of CCl4-induced hepatic cytotoxicity. The data suggest that the daily oral 435consumption of an aqueous extract of the flesh and pits of dates, and as a part of the daily diet ad 436libitum, was prophylactic to CCl4 poisoning, achieving about 80% protection with date palm flesh 437and 70% with pits. A similar percentage of protection was achieved when the aqueous extracts of 438the flesh and pits were used as a cure against CCl4 poisoning after toxicity was induced ( Qarawi 439et al 2004). 440 441 442 ESTROGEN LIKE ACTIVITY OF DATE PIT EXTRACT. Estrogens are a group of female sex steroid hormones that are synthesized in the ovary and

443placenta (Williams and Lemke 2002) (Lopez and Barecelo 2002). They include estradiol, estriol,

444and estrone. Estrone is a primary estrogenic component of several pharmaceutical preparations 445including those containing conjugated and esterified estrogens (Williams and Lemke 2002). 446However, estrogen therapy should be carefully prescribed due to its known side effects even in low 447amounts (Rang , Dale and Ritter 2002). Few studies have been cited about some parts of date palm 448trees. The steroid sex hormone, estrone has been detected in the date pits as well as in the pollen 449grains of Phoenix dactylifera.. These include the two studies by Heftmann and Bennett which 450reported the isolation of the hormone estrone from the seeds (date pits) and pollen grains of the 451date palm Phoenix dactylifera growing in California, U.S.A (Heftmann , KO and Bennett 1965) 452(Bennett, KO and Heftmann 1966). 453 454 USE OF DATE PIT OIL FOR MALE INFERTILITY Fatma et al (2009) reported that in vitro supplementation with date seed oil (DSO) can protect

455spermatozoa against hydrogen peroxide (H2O2)-mediated damage and can improve sperm function, 456possibly owing to antioxidant properties. The antioxidant effects of DSO were tested on human 457sperm motility, sperm viability, reacted acrosome and lipid peroxidation assessed in vitro after 458H2O2-mediated oxidative damage in spermatozoa. Sixteen patients (mean age: 35 years; range: 25 45945 years) referred to the HistologyEmbryology Laboratory of the Medicine Faculty of Sfax for 460semen analysis after 1224 months of sexual intercourse without conception were selected. After 461spermiogram, sperm selection by two interface discontinuous Sill Select gradient was performed, 462and selected spermatozoa were used in four experimental assays: control; incubation with 100 m 463H2O2; incubation with 0.1% date seed oil; and co-incubation with 0.1% DATE SEED OIL and 100 464m H2O2. Motility and viability were determined using World Health Organization criteria. 465Acrosome reaction and lipid peroxidation were assessed by staining with fluorescein 466isothiocyanate-Pisum sativum and spectrophotometric measurement of malondialdehyde,

467respectively. Results showed that incubation with H2O2 alone led to a significant increase in lipid 468peroxidation (57.83%, P < 0.05) associated with a significant decrease in sperm motility, sperm 469viability (after 30 min and 24 h) and percentage of reacted acrosome (P < 0.05). Date seed oil 470improved sperm motility after 24 h of incubation (P < 0.05) and protected spermatozoa against the 471deleterious effects of H2O2 on motility, viability, acrosome reaction and lipid peroxidation. 472Therefore supplementation with date seed oil may have a function in antioxidant protection against 473male infertility. 474EFFECT OF DATE PALM (PHOENIX DACTYLIFERA) SEED FIBERS ON PLASMA

475LIPID. 476 Five diets were investigated which include basal diet, cellulose diet and three defatted date

477seed fibers diets containing 1.5%, 2.5% and 5.2% date seed fiber. Wistar rats were randomly 478divided into five groups and the experiments proceeded for 27 days. The objective of study was to 479investigate the effect of date palm (Phoenix dactylifera) seeds crude fiber on triglycerides, 480cholesterol and high density and low density lipoproteins in plasma lipids in rats. The results 481revealed that diet based on date seed fiber had good potential as a source of dietary fiber in diet. 482Diet contained 1.5% date seed fibers is the most appropriate because of its action in reducing 483LDL cholesterol, total cholesterol and triglyceride. However, such level (1.5%) of date seeds fiber 484had no effect in HDL cholesterol level. (Salah and Maiman 2005). 485Effect of Date Pit Coffee on Total Homocysteine and Plasma Lipids in Rats 486 487 Al-Ghanem et al., (2010) in his studies used forty two male Wister albino rats (10010 g) 488to evaluate the effects of Arabian and date pit coffees administered orally at different 489concentrations (50, 100 and 150 mg/kg body weight/day) on total homocysteine and plasma lipids. 490The control group was administered water orally instead of coffee. All concentrations of Arabian 491and date pit coffees significantly (P<0.05) increased the concentration of homocysteine in the 492plasma of rats compared to the control group. The increase in this index was higher for the Arabian 493coffee compared to the date pit coffee, and increased with the increase of the concentration of the 494coffee. Total cholesterol (TC) significantly (P<0.05) increased in plasma of rats administered 495orally the Arabian and date pit coffees all concentrations except (50 mg/kg body weight/day) 496compared to the control group. However, the increase dose concentration of Arabian and date pit 497coffees had no further effect on the concentration of TC. There was no significant difference in this 498index between rats administered Arabian coffee and those administered date pit coffee. Low 499density lipoprotein-cholesterol (LDL-c) significantly (P<0.05) increased in plasma of rats 500administered orally Arabian coffee at all concentrations compared to the control group and it 501increased with the increase of the concentration of the coffee; however, there was no significant 502difference in this index between rats administered date pit coffee and the control group. Very low 503density lipoprotein- cholesterol (VLDL-c) significantly (P< 0.05) increased in plasma of rats 504administered orally date pit coffee all concentrations compared to the control group but the 505increase in dose concentration of date pit coffee had no further effect in the concentration of 506VLDL-c; however, there was no significant difference in this index between rats administered

507Arabian coffee and the control group. High density lipoprotein-cholesterol (HDL-c) significantly 508(P<0.05) increased in plasma of rats administered date pit coffee at the all concentrations 509compared to the control group and it increased with the increase of the concentration of the date pit 510coffee; however, there was no significant difference in this index between rats administered 511Arabian coffee and the control group. Triglyceride (TG) concentration in plasma of rats also 512increased significantly (P<0.05) in rats administered date pit coffee compared to those 513administered Arabian coffee but the increase in dose concentration of date pit coffee had no further 514effect in the concentration of TG; however, there was no significant difference in this index 515between rats administered Arabian coffee and the control group.In conclusion, the results suggest 516that Arabian and date pit coffees may increase homocysteine and TC and in serum of rats. Date pit 517coffee increased HDL-c and TG and VLDL-c in plasma of rats. Arabian coffee increased LDL-c in 518plasma of rats.
519

520 521

Effect of Date Pit Coffee on Antioxidant Enzymes in Rats Al-Ghanem et al., (2010) in his studies used forty two male Wister albino rats (10010 g) to

522evaluate the effects of Arabian and date pit coffees administered orally at different concentrations 523(50, 100 and 150 mg/kg body weight/day) on antioxidant enzymes. The control group was 524administered water orally instead of coffee. The activity of superoxide dismutase (SOD) 525significantly (P< 0.05) increased in serum of rats administered all doses of both Arabian and date 526pit coffees compared to the control group. This increase was higher in the rats administered the 527Arabian coffee compared to those administered the date pit coffee. However, the increase in dose 528concentration of Arabian coffee had no further effect on the activity of SOD. On the contrary, the 529increase in dose concentration of date pit coffee significantly (P< 0.05) decreased the activity of 530SOD, but its activity in its lowest concentration remained significantly higher compared to the 531control group. Results also showed a significant increase in the activity of catalase (CAT) in serum 532of rats administered all doses of Arabian coffee and the doses 50 and 100 mg/kg bw/day of date pit 533coffees compared to the control group. This increase was higher in the rats administered the 534Arabian coffee compared to those administered the date pit coffee, however, the increase in the 535concentration of the dose of Arabian coffee had no effect on the activity of CAT. On the contrary, 536the increase in dose concentration of date pit coffee significantly (P< 0.05) decreased the activity 537of CAT, and its activity at its highest concentration significantly decreased compared to the

538control group. Arabian and date pit coffees had no effect on the glutathione peroxidase (GPX) 539activity in the plasma of rats. In conclusion, the results suggest that Arabian and date pit coffees 540may increase the activities of SOD and CAT in serum and plasma of rats. However, they have no 541effect on the activity of GPX. 542CONCLUSION 543 In general, it is suggested that date by-products (seeds) could be served as a good source of 544natural antioxidants, dietary fiber and a wide range of components having nutraceutical importance 545and could potentially be considered as a functional food or functional food ingredient. It seems 546reasonable to conclude that a further characterization of the active ingredients may reveal useful 547compounds. 548

549

REVIEW OF LITERATURE

550Functional foods 551 For instance, phytic acid plays a major role in the treatment of cancer, hypercholesterolemia,

552hypercalcuria and kidney stones (Plaami, 1997). Work done recently has indicated that specific 553inositol polyphosphates have different functional potentials. Siren, Linne, and Persson (1991) 554outlined potential pharmacological applications for D-myo-inositol-1,2,6-trisphosphate, including 555anti-inflammatory effects, reduction in secondary complications of diabetes, interference with the 556vaso-constricting agent neuropeptide Y and, thus, reduction of hypertension. Date pits from the 557three varieties contained very little ash, which may indicate that the phytic acid concentration in 558date seeds is unusually small, i.e., in comparison to cereal grains and oil seeds. This conclusion is 559supported by the report of Attalla and Harraz (1996) who found that date pits of 11 cultivars grown 560in the Qassim region contained small amounts of phosphorus (0.190.26%). Pits contained a large 561quantity of fibre that may have health benefits. The difference between the quantities of neutral 562detergent fibre and acid detergent fibre gives the hemicellulose content of the pit sample. It should 563be noted, however, that the acid detergent fibre is very high, which may indicate the presence of 564substantial amounts of lignin and perhaps resistant starch in date pits. Furthermore, plant fibres can 565exhibit unique qualities, such as antioxidant capacity, inherent susceptibility to fermentation to 566release ferulic acid and glucose retardation indices. Due to their importance in developing 567functional foods from date pits, the carbohydrates should be characterized further, especially for 568the presence and quantities of resistant starches. Thorough investigation of these fibres, as well as 569other interesting and associated components in date pits, will determine their potential uses as 570functional foods. Date pits can be a source of dietary fibre without any negative impact on sensory 571quality of end-products if the pits are properly milled (Almana & Mahmoud, 1994). Other 572functional components that may be present in pits are selenium, which can be used as antioxidant 573(thus anticarcinogenic) in human therapeutics (Pszczola, 1998). Al-Showiman, Al-Tamrah, and 574BaOsman (1994) determined the selenium content in dates of some cultivars grown in Saudi 575Arabia. Selenium levels were found to be in the range 1.482.96 mg/g in the 10 varieties they 576studied, which were grown in different locations. Thus, further research is needed to characterize 577isolated components and to search for bioactive constituents with antimicrobial, antioxidant and 578other health promoting activities. 579

580

581MATERIALS 582

AND METHODS

Raw material will be procured from various date research institutes. Date pit powder will

583be produced by the method followed by Rehman et al.,(2007) with slight modification. Pits from 584the fully matured date fruit (tamar) will be cleaned by boiling them in water for an hour. The seeds 585will be separated by passing the water over a metal strain and they will be oven dried for 2 days at 58650 oC. Then dried whole pits will be roasted at 220 oC for 1520 min and cooled to ambient 587temperature. Roasted seeds will then be ground to a coarse setting size of 500 um. The ground 588material will be the roasted date-pit powder and it will be stored in airtight food-grade containers 589until used for experimentation. 590Chemical Analysis 591 The roasted date-pit powder will be analyzed for the moisture, ash, fat, protein and fiber

592content (AACC, 2000). 593Total Dietary Fiber (TDF) 594 The whole roasted date-pit powder samples will be analyzed for total dietary fiber

595according to AACC (2000). 596Soluble Dietary Fiber (SDF) 597 The whole roasted date-pit powder samples will be analyzed for soluble dietary fiber 598method as mentioned in AACC (2000), by employing Megazyme assay kit. 599Insoluble Dietary Fiber (IDF) 600 In soluble dietary fiber (IDF) in whole roasted date-pit powder will be determined by the

601procedure described in AACC (2000). 602Minerals Estimation 603 Minerals Na and K will be estimated through Flame photometer, while Mg, Ca, Zn, Cu,

604Mn and Fe will be measured through Atomic Absorption Spectrometer according to the method 605described in AOAC (1990). 606Sensory Evaluation of date pit coffee. 607 608(1998). 609 Statistical Analysis 610 611 612 The data of each parameter will be subjected to statistical analysis to determine the level of significance (Steel et al., 1997) and principle component analysis (Snedecor and Cochran, 1980). Sensory evaluation of bread will be performed by the procedure by Lawless and Heymann

613 614

Efficacy Studies. Induction of diabetes in rats

615 A freshly prepared solution of streptozotocin (45 mg/kg b.w.) in 0.1M citrate buffer, pH 6164.5 will be injected intraperitoneally to overnight fasted rats (Brosky and Logothelopoulos, 1969). 617FBG level will be estimated at the time of induction of diabetes and PPG will be checked regularly 618up to stable hyperglycemia, usually one week after streptozotocin injection. Depending on their 619fasting blood glucose level (BGL) the animals will be divided arbitrarily 620in to three groups (Gupta et al., 2005): 621(i) Sub diabetic animals, with nearly normal FBG of 80120 mg/dl but showing abnormal glucose 622tolerance. 623(ii) Mild diabetic animals with FBG of 120250 mg/dl. 624(iii) Severely diabetic animals showing FBG above 250 mg/dl.
625Assessment of hypoglycemic activity in normal healthy rats 626Initial testing was carried with the different doses of the seed extract in 24 normal healthy 627male rats fasted overnight. The animals were divided into four equal groups. Control rats 628(groupI) were given vehicle (distilled water) only while other groups IIIV received aqueous 629seed extract suspended in distilled water orally at doses 100, 250 and 500 mg/kg, 630respectively. Blood glucose levels were estimated before and after 2, 4, 6 and 8 h of seed 631extract administration. 632Assessment of activity of extract on glucose tolerance in sub and mild diabetic rats 633The hypoglycemic effect of aqueous extract of Roasted date-pit powder seeds in sub and mild 634diabetic rats was assessed by improvement of glucose tolerance. FBG was checked in 635overnight fasted rats and were divided into five groups of each. Control groups of sub and mild 636diabetic animals received vehicle (distilled water) only, whereas variable doses of 100, 250 637and 500 mg/kg of aqueous seed extract and a dose of 250 mg/kg of standard drug 638tolbutamide were administered orally to rest of the four groups of each, sub and mild diabetic 639animals. The rats of all the groups were given glucose (3 g/kg) after 90 min of the extract and 640drug administration. Blood samples were collected just prior to glucose administration (0 h) 641and 1, 2 and 3 h after glucose loading. 642Assessment of activity of extract in severely diabetic rats 643Study was carried on three groups (VVII) of six rats each. Group V served as normal healthy 644control, group VI as diabetic control and group VII was orally treated with a single dose of 250 645mg/kg b.w. of seed extract suspended in distilled water daily for 2 weeks. Control rats (group 646V and VI) were given vehicle (distilled water) only. Fasting blood glucose, total cholesterol, 647HDLcholesterol and triglyceride levels were estimated andLDL cholesterol was calculated at 648the beginning and after 7 and 14 days of experiment. Changes in body weight and urine sugar 649were also assessed. 650LD50 experiment 651Four groups of rats of both sex (six animals per group, three females and three males) and 652weighing about 180220 g were administered orally a single dose of either 2.5, 5, 10 or 15 653times of effective dose of water extract of seeds of Roasted date-pit powder. Then rats were 654observed for gross behavioural, neurologic, autonomic and toxic effects at short intervals of 655time for 24 h. Food consumption, feces and urine were also examined at 2 h and then at 6 h 656intervals for 24 h. 657Statstical analysis 658All the group data were statistically evaluated using Students t-test, expressed as the 659meanS.D. from six rats in each group.

660

661

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