Chapter 2 REVIEW OF LITERATURE

The use of flaxseed as a functional food is gaining more popularity in the recent years. It is grown from the ancient times for food and other uses. Flaxseed is an excellent source of nutrients including protein, soluble and insoluble dietary fiber as well as omega-3 fatty acids. The flaxseed can be used as a whole or it can be incorporated in different other foods including bakery products. A lot of work has been carried out on the composite flour technology with special reference to improve the wheat flour quality through blending it with other flours. In this section some of the recent literature is reviewed under the following headings. 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 2.1 Wheat flour Potential of flaxseed use in different food products Storage and cooking stability of flaxseed Rheological characteristics of composite flours Chapatti baking quality Bread baking quality Biological assay of protein quality Health benefits of flaxseed Wheat flour Wheat is considered as the principal cereal grain produced, consumed and traded in the world. Wheat, rice, barley, corn and sorghum provide around 68% of the total world food supplies. The wheat is acknowledged as a staple food and the economical source of protein and calories for the people all over the world (Health and welfare, 1990). Wheat and wheat products besides providing

protein and carbohydrates also are major sources of minerals and dietary fiber (Sidhu et al., 1999). Wheat is the leading cereal grains produced, consumed and processed in Pakistan and it contributes as the cheapest and principal source of protein and calories (Zahoor, 2003). The wheat grains were cultivated on an area of 8414 thousand hectares with a provisionally estimated production of 21.75 million tons during the 2007-08 seasons (GOP, 2008). In Pakistan, wheat is used for the preparation of several products such as: chapattis, rotis, paratha, nan, bread, buns, biscuits, cakes, pastries, patties, pan cakes and many others (Pyler, 1988). The 70% of the total wheat produced in Pakistan is consumed in the form of chapatti (unleavened flat breads) and its variants like tandoori roti, nans, prathas and poories, while the rest of 30% is used for other bakery products such as breads, cookies, cakes and pastries etc (Butt et al, 1997). The chemical composition of whole wheat flour for moisture, ash, crude protein, crude fat, crude fiber, nitrogen free extract, wet gluten and dry gluten range from 9.38 to 10.43%, 1.32 to 1.85%, 10.13 to 14.74%, 1.96 to 2.52%, 2.31 to 2.99%, 78.71 to 85.37%, 23.53 to 38.71% and 7.51 to 13.52%, respectively among different Pakistani wheat varieties (Ahmad, 2001). The poor segments of populations meet their dietary needs from foods based on tubers and cereals and limiting their intake of fruits, vegetables and costly animal foodstuffs (Allen, 1991). The main intervention strategies against micronutrient malnutrition include supplementation, dietary improvement and fortification of foods. The food based approaches namely food fortification and dietary improvement appeared to be the most sustainable, potentially long lasting strategic measures against micronutrient malnutrition (Hurrell, 1997). The technology of composite flour refers to the process of amalgamation of wheat flour with the flours of other edible plant materials. It makes the use of local and potentially health beneficial raw materials for the production of high quality, healthy and economical food products for the masses.

2.2

Potential of flaxseed use in different food products The flax (Linum Usitatissimum) is a blue flowering rabi crop and is a

member of family linaceae, commonly known as Alsi (Gujrati, Hindi and Punjabi).The flaxseed and linseed are the other names which are often used interchangeably. The flax in North America when eaten by humans is named as flaxseed and named as linseed when it used for linoleum flooring. However, Europeons use the term flaxseed to explain about the varieties grown for manufacturing of linen (BeMiller et al., 1993). The history of flaxseed cultivation has been found as early as 3,000 B.C. The flaxseed grain is oval and flat, slightly larger than a sesame seed possessing about 2.5x5.0x1.5 mm size. The colour of flaxseed ranges from reddish brown to a light yellow (Freeman, 1995). The texture of flaxseed is crisp and chewy possessing a pleasant nutty taste (Carter, 1996). The proximate composition of flaxseed indicates that it contains 30% protein, 35 % lipids and 35 % fiber; though it varies depending upon the seed variety, cropping year, cropping location and environmental conditions. The protein content in flaxseed has been reported to range from 10.5% to 31% depending on environment and growing conditions (Baijpai et al., 1985). The flaxseed is not a good source of starch. The soluble fiber is approximately a quarter of the total fiber present in flaxseed. Mucilaginous gum is the major component of the soluble fiber in flaxseed and its composition varies from 7 10 g per 100 g (Carter, 1993). The alpha linolenic acid (ALA) which is an omega-3 fatty acid forms the mass composition of polyunsaturated fatty acids in flaxseed. It is present about 50% of the total fatty acids (Daun and Przybyliski, 2000). ALA can not be synthesized by the human body from any other substance therefore it is considered as an essential fatty acid. The essential fatty acids requirements for the human body can be fulfilled by intake of flaxseed products (Morris, 2004). 100g of flaxseed provides 100% of the recommended daily allowance (RDA) for

manganese and potassium, 57-65% of the RDA of phosphorus and iron, and 1335% for zinc, calcium and copper while its recommended daily intake is 25 to 50 grams (Anon, 1994). Specialty grains (bulgar wheat, barley, flax seed, sprouted grains and rice extract) are used in breakfast cereals to add novel textures, flavours and colours in these products (Kuntz, 1998). The chapatti prepared from the composite flours containing 15 g roasted flaxseed powder, 0.05 g oil and 80 g of whole wheat flour was evaluated for its chemical composition. That single serving of chapatti contained 29.9 grams carbohydrates, 6.3 grams protein, 5.9 grams fat and 198 K calories (Soniya et al., 2004). Similarly chapattis containing 20% or lower levels of the full fat flaxseed flour were found acceptable by the panelists with respect to their sensory attributes. The mineral contents of the chapattis increased significantly as level of the replacement of the full fat flaxseed flour was increased in the wheat flour (Hussain, 2004). The breads containing 10 and 13% levels of linseeds were characterized by higher amounts of protein, fat, dietary fiber, macro and microelements in comparison to standard bread. There was a significant increase in Fe, Zn and Mn contents of the bread with the increasing level of flax seed flour in the wheat flour (Gambus et el., 2004 and Naz, 2000). The ground flaxseed used @ 30% to 50% has been found to be acceptable in muffins and quick breads (Alpers and Morse, 1996). The cookies prepared from wheat flour containing 20% whole flax grains, partially defatted flaxseed flour and full fat flaxseed flour were found acceptable with respect to their sensory attributes (Hussain, 2006). Gambus et al. (2003) investigated the quality of 2 bakery products (cookies and muffins) containing brown and yellow linseeds (flaxseed) flour at 11 and 9%, respectively. These levels of replacement did not affect the sensory

properties of flax hermit cookies and flax muffins. The increased linseed content improved the dietary and nutritional values of both types of bakery products due to the increased total protein and dietary fiber contents. The study

conducted by Zaib-un-Nisa (2000) showed that protein content of biscuits made from composite flour containing 15% ground flaxseed increased from 6.50% to 8.52%, fat content increased from 26.13% to 31.45%, fiber content increased from 0.15% to 3.78% and ash content increased from 0.26% to 1.00%. The supplementation of flaxseed flour up to 15% showed no deleterious affect on the sensory attributes of biscuits. The minerals like Fe, Zn and Mn contents increased significantly with 15% supplementation of flaxseed flour. According to Gambus et el. (2004), 3% increase in linseed in the recipe for flax hermit cookies and 5% rise of linseed in flax muffins resulted in an increase in the amounts of proteins, dietary fiber, micro and macro minerals. Frank and Sarah (2006) also studied the effect of combining flaxseed meal (15%) and soy flour (5% and 10%) on the product quality in production of yeast bread. Addition of flax and soy flour resulted in the reduction of bread volume. The sensory attributes like crust and crumb colour were also affected due to the addition of soy/flaxseed flours and darker colour was observed in both products. No significant differences in moisture in the flax and soy/flax breads were recorded as compared to control breads prepared form wheat flour alone. Kocka and Anil (2007) reported that addition of flaxseed in wheat flour at 15 and 20% levels resulted in decrease of dough stability, lower crust and crumb scores as compared to the control breads prepared form 100% wheat flour. In another study conducted by Pohjanheimo et al. (2006), the effect of addition of flaxseed in bread rolls and cinnamon rolls on their sensory profile and chemical composition was studied. At 0 and 6 days of storage, no significant changes were observed in their sensory attributes. The higher contents of saturated fatty acids were observed in cinnamon rolls. Both the bakery products were found to be higher in alpha linolenic acid contents. The fiber contents of bread rolls were also increased.

2.3

Storage and cooking stability of flaxseed Now a days interest has led to produce a wide variety of foods referred to

as functional foods and nutraceuticals because these type of foods offer either therapeutic or preventive medicinal health benefits in addition to provision of their basic nutrition. Addis and Warner (1991) found that in higher oil containing functional foods, lipid per oxidation results in the deterioration of their nutritive value and stability. The flavor and taste of such foods or the foods in which these foods are added are also affected (Barlow, 1990). The oxidative rancidity is a major cause of development of bad odour and flavour in the foods containing higher oil contents. Free radical formation is responsible for that process. The byproduct of this process is the development of peroxides. The measurement of peroxide value helps to determine the level of amount of rancidity present (Martin, 2004). The flaxseed meal or oil can easily be incorporated into common food products such as breads, rolls, cereals, muffins, margarines, and salad dressings (Fitzpatrick, 2006). The flaxseed oil is a source of the essential fatty acids, such as linoleic acid (LA, C18:2 n 6) and linolenic acid (LNA, C18:3 n 3), required in the diet for normal health (Dobson, 2002; Lee and Lip, 2003). The level of LNA in flaxseed oil is generally higher than 50% of the total fatty acids (FAs). LNA has been reported to play an important role in the avoidance of immune disorders, cardiovascular diseases and certain type of cancers (KrisEtherton et al., 2002). Although there is a great concern in the exploitation of flaxseed oil as a nutraceutical supplement, owing to its high content of LNA which is prone to rapid oxidation, hence resulting in quality deterioration. The study of Hauman (1998) indicated that flaxseed oil is high in unsaturated fatty acids mainly in alpha-linolenic acid (ALA) which is about 52 % of the total fatty acid contents of flaxseed oil. The instability of flaxseed oil to oxidation is the restraining factor for its incorporation in food products. (Kolodziejczyk and Fedec, 1995) reported that the stability of oil can be extended by suitable storage which is vital for improving the oxidative stability of flaxseed oils. ALA in

remote form or as a part of an extracted oil is generally considered susceptible to oxidation as it is highly unsaturated and oxidation in oils is encouraged by both heat (auto-oxidation) and light (photo-oxidation). However, ALA in the intact seed of flax has proven particularly resistant to oxidation. The storage stability of flaxseed is of major importance before using it in any food product. Due to its high content of oil, proper cooking and storage assures the better stability of high oil bearing flaxseeds in different products. Several researchers have worked on the storage and cooking stability of flaxseed. The maccroni was fortified with ground flaxseed at level of 10 and 20% were dried at 40 oC, 70 oC and 90 oC temperatures. The dried maccroni after storage for 32 weeks, conjugated diene values indicated no significant increase while ALA levels in the fortified maccroni remained unchanged during the whole storage period (Hall et al., 2005). The study conducted by Lee et al. (2000) concluded that maccroni containing 20% ground flaxseed flour showed significant storage stability. ALA content and storage stability of the spaghetti were not affected by the processing method. Fatty acids profile of the pasta remained consistent during the storage. Conjugated dienes and headspace analysis also indicated non significant variations. Both the whole and coarsely ground flaxseed showed long-term storage stability at room temperature. No changes in the peroxide values were observed after 308 days of storage at temperatures of 22C. The percentage of ALA in fat extracted from the stored flaxseed samples also remained intact i.e unchanged (Ratnayake et al., 1992). The roasting of flaxseed significantly affected the peroxide value (POV). Roasted samples showed less peroxide value as compared to the non roasted flaxseed samples. The storage time significantly affected the peroxide value as it is increased with the 4 week storage period, which ultimately resulted in the decrease in the stability of flaxseed. Storage temperatures also affected the

peroxide value. The samples stored at 30 oC gave higher POV than at 25 oC storage samples which yielded lower POV. Non roasted samples showed statistically (P<0.05) higher free fatty acids (FFA) contents than roasted samples. The fatty acid profile showed low variation, indicating insignificant changes in terms of linolenic acid content (Scott et al., 2004). Flaxseed can be stored in different materials that affect its stability. The peroxide values were relatively unchanged and the ALA content was not affected in flaxseed packaged in loosely packed plastic bags and protected from light when stored at room temperatures for 20 months in warehouse conditions (Daun, 2001). Flaxseed spread out in trays showed ample deterioration (high peroxide levels) after 14 weeks of storage. Flaxseed stored in bags showed very little increase in peroxide levels and fatty acids remained unaffected during storage period (Wiesenborn et al., 2004). Flaxseed samples milled and stored at 73 oF for 4 months in paper bags with plastic liners were tested after 0, 33, 66, 96 and 128 days of storage intervals. Slight increase in peroxide values or changes in alpha linolenic acid content was noticed. No difference in odor or taste of fresh and stored samples of flaxseed by a sensory panel (Malcolmson et al., 2000). Negligible changes in chemical parameters of oxidation like peroxide value and free fatty acids were observed. No difference was observed in the aroma intensity of the water slurries of fresh and stored samples (Malcolmson et al., 1998). Similarly flaxseed samples stored in glass tube remained unchanged regarding their ALA content and showed stability to heat and light for 280 days during storage at room temperature. Little changes in head space oxygen were recorded during the said period (Chen et al., 1994). Research conducted by Cunnane et al. (1995a) showed no significant reduction during baking process in the thiobarbituric acid-reactive substances and alpha-linolenate contents of muffins supplemented with 25 g of flaxseed.

Malcolmson (1997) reported that 36 consumers could not tell the difference between the tastes of yeast bread baked with the either fresh or stored milled flaxseed @ 11% of the flour weight in the recipe. In another study, Manthey et al. (2002b) concluded that processing and cooking of spaghetti did not significantly affect the alpha linolenic acid contents. During baking, minimal loss of alpha-linolenic acid from flaxseed was observed that is helpful for the incorporation of flaxseed in different food products (Chen et al., 1994). No changes in peroxide values and fatty acids composition were observed when whole and milled flaxseed was heated for 60 minutes at either 212 oF or 662 oF (Ratnayake et al., 1992). Maccaroni was fortified with 20% (by weight) ground flaxseed, stored for 20 weeks and cooked in boiling water for 12 minutes. ALA content remained the same during the whole storage process and after cooking (Manthey et al., 2002 a). 2.4 Rheological characteristics of composite flours The mechanical and rheological properties of the dough exert promising effect on the overall quality of baked products (Blokshma and Bushuk, 1988). The arrangement and interaction of constituents (especially proteins) and the structure of materials are the responsible factors affecting the rheological properties (Bushuk, 1985). The final product quality depends upon the dough rheology taking place during the processing of the constituents (Lindahl, 1990). The nature of ingredients, their proportions, mixing time and beating conditions are responsible for the quality of batter which finally determines the baked product quality (Baixauli et al., 2007). The dough rheological properties are influenced by the structure of the aggregates and their tendency to interact with each other. Quality and quantity of the proteins affect the water absorption capacity of the dough (Finney, 1984). The farinograph or mixograph are two most common methods used for measuring the rheological properties of dough during mixing (Mani et al., 1992). The mixing of flours results in the hydration

which leads to the formation of gluten matrix. During mixing process, air is also incorporated into the dough system (Bushuk, 1985). The mixograph provides an indication of the mixing requirements of flour (Hoseney and Finney, 1974). The peak height depends on the gluten content of the wheat flour. The wheat variety Barani-83 of Pakistan showed higher gluten contents which resulted in the increase of peak height (Anjum and Walker, 2000). The rheological properties of the composite flours are significantly affected as the level of replacement of cottonseed flour in the wheat flour was enhanced (Bajwa, 1997). Similarly 10% replacement of the defatted peanut flour in the wheat flour altered the water absorption capacity and extensibility of the dough mix (Rao and Vakil, 1980), while no significant affect on the peak height and mixing time was observed with the 10% addition of cotton seed flours in the two different types of wheat flours (Rasool, 2004). More water absorption with sticky dough is observed, as the fiber content of chapattis was increased and the mixing time of the dough was decreased (Ahluwalia and Kuar, 2001). Addition of cottonseed, peanut, safflower and soy flour in wheat reduced the mixing tolerance while increase in the water absorption is observed (Mathews, 1972). The water absorption of composite flour containing 10% chickpea flour decreased significantly from 68.40% at (0 day) to 66.5 % (60days) as a function of storage time (Shahzadi, 2004). Flaxseed mucilage is composed of mainly polymeric carbohydrates while galacturonic acid, rhammose, galactose, fructose, glucose are also present in small quantities. It can help to improve the water absorption characteristics of the dough (Fedeniuk and Biliaderis, 1989). The flaxseed may function as natural additive that retards the staling of the bread (Pohjanheimo et al., 2006) as its gum possesses excellent water binding capacity and technological benefits (Konecsni et al., 2005). The farinographic studies of bread prepared from wheat flour replaced with flaxseed flour @ 5, 10, 15, and 20% showed that water absorption, dough development time and mixing tolerance index increased as the amount of flaxseed flour increased, while dough

stability decreased at 10, 15 and 20% of flaxseed flour supplementation. The addition of 15 and 20% levels of flaxseed flour showed a decrease in the dough extensibility. The doughs containing 10, 15 and 20 % flaxseed flour exhibited resistance to extension comparable to that of control dough containing no flaxseed flour (Koca and Anil, 2007). 2.5 Chapatti baking quality The wheats grown in Pakistan are mainly used for the preparation of unleavened flat breads, which is locally known as chapatti or roti. The chapattis are unleavened and are prepared from whole-wheat flour or atta (95% extracted flour). The chapattis are the staple diet of the Indian subcontinent especially Pakistan and some parts of the Middle East (Anjum et al., 2002; Nurul-Islam and Johansen, 1987 and Gujral and Pathak, 2002). Wheat is a basic raw material for the preparation of chapattis and its other similar variants like rotis and naans. The quality of chapattis is influenced by the wheat quality (Haridas et al., 1986 and Prabhasankar, 2002). Among the different Triticum species of wheat, Triticum aestivum is more suitable for the production of a fully puffed chapatti with soft and pliable texture, whereas dry and hard textured chapattis are prepared from Triticum durum and Triticum diccocum wheats (Haridas, 1993). Medium hard wheat, having a protein content of 9.510.5% has been found to be suitable for making chapattis (Schofield, 1992). The gliadin contents of wheat flour effects the chapatti quality and wheat varieties containing higher amounts of gliadin protein results in poor quality chapattis while albumin and globulin did not impart any significant effect on the chapatti-making quality (Prabhasankar, 2002). Like any other baked product, dough forms an important transitional product between flour and chapatti. The dough consistency is dependent upon the amount of water added which ultimately influences the chapatti quality. The chapattis are baked from both sides on an iron plate, and finally puffed on a coal or gas fire (Austin and Ram, 1971). Chapattis are

generally prepared twice a day for lunch and dinner. The chapattis are normally consumed fresh immediately after baking because storage of chapttis results in staling and the product become hard to masticate (Shaikh et al., 2007). The most important parameters of chapatti quality are flavour and texture and are evaluated as soft texture, greater pliability, light creamish brown colour with small brown spots, slight chewiness, fully puffed, and baked wheatish aroma (Shaikh et al., 2007; Haridas et al., 1986). The chapatti should be easily torn and flexible so that it can be folded by the forefinger and thumb (Dhaliwal et al., 1996). This product is made by mixing the flour and water to develop the dough, after relaxing; balls are sheeted (Gujral and Pathak, 2002). The size and shape of the chapattis vary from region to region, culture to culture and family to family but normally these are circular or round disc in shape and size of 150-200 mm in diameter and 1-4 mm thickness. After rolling, chapattis are baked on a pan by contact heating and puffed directly on gas fired burner and consists mostly of crust with little crumb (Sridhar and Rao, 1993; Gupta, 1990). The composite flours can be prepared and used to improve the nutritional and technological properties of the chapattis. Gandhi et al. (1983) demonstrated that chapattis made from wheat flour containing 10-15% soy flour were of satisfactory flavour, texture, appearance and overall acceptability by trained sensory panel, although flavour and texture were significantly different from the all-wheat-flour chapattis. Similarly Lindell and Walker (1984) also found that soy supplemented wheat flour chapattis would fulfill a nutritional need as they are higher in fat and protein with a well balanced profile of amino acids. Khan et al. (2005) and Anjum et al. (2006) observed significant improvement in the mineral contents (Fe, Zn, Ca, Mg and Cu) of chapattis supplemented with soy flour /hulls and found that soy flour and hulls can be replaced up to 24 and 4.5 % levels, respectively to produce organoleptically acceptable chapattis.

Chapattis made from defatted soy bean flour supplemented wheat flour were found acceptable but those made from rape seed and sunflower flours were unacceptable (Jain et al., 2000). The protein content was increased from 11.9% to 19.8% at 20% blending level of defatted soy flour. The textural parameters like cohesiveness, springiness, hardness and chewiness were also affected by the incorporation of defatted soy flour (Gandhi et al., 2000). In another study conducted by Gujral and Pathak (2002), it is observed that whole wheat flour replaced with flours from millets, rice, corn, barley and black gram showed chapattis with higher extensibility even after 24 hrs of storage. Some of the additives like sodium caseinate also considerably enhanced the texture of chapattis. Sekhon et al. (1980) concluded that triticale flour can be blended with wheat at a level of 50% to prepare acceptable chapattis. Arya et al. (1978) concluded that wheat flours with more fat contents produced chapattis with a softer and smooth texture and better folding ability. Barley can be blended up to 40% with wheat flour to yield acceptable quality of chapattis (Sood et al., 1992). The chapattis prepared from the composite flour containing 15g roasted flaxseed powders, 0.05g oil and 80g of whole wheat flour provided 29.9g carbohydrates, 6.3g protein, 5.9g fat and 198 K calories (Soniya et al., 2004). The texture of chapattis became progressively harder with storage at both room and refrigerated temperatures. A decrease in sensory quality and acceptability of the chapattis was observed with storage. The rate of staling was lower at refrigerated temperature than that of room temperature storage of chapattis (Sheikh et al., 2007). The texture of chapattis can be affected by the hardness of wheat. The flour with higher moisture retention and gelatinization of starch in hard wheats results in the chapattis with soft and pliable texture (Srivastava et al., 2000). 2.6 Bread baking quality The bread is an ideal product that can serve as a functional food since it is an important part of daily part of a large segment of population through out the

world (McKechnie, 1983). Bread consumption cannot be replaced by any other single food product. It is consumed in various forms in many countries (Faridi et al., 1982). Wheat flour is the major ingredient in pan breads (Klopfenstein and Hoseney, 1995) while flour, water and yeast are reported as key ingredients for breads by Osuji, (2006). It is reported by Pomeranz (1990) that quality and quantity of wheat protein plays an important role in baking technology. Horefall et al. (2007) reported that breads can be prepared from composite flours containing 5, 10, 15, 20 and 30% plantain flour in wheat flour. The quality of breads based on sensoric attributes is described by appearance, aroma, texture and flavour (Lawless and Heyman, 1999; Meilgard et al., 2007). Wang et al. (2002) reported that addition or increase of dietary fiber in breads results in the decrease in loaf volume, increase in crumb firmness and darker crumb appearance of bread. Filipovic et al., (2007) also found that incorporation of fibers (modified and unmodified) showed decrease in scores assigned to volume and crumb quality of bread. The sensory scores for appearance, texture and flavour of breads have been reported to be decrease by the incorporation of non wheat flours in wheat flours (Shittu et al., 2007). Frank and Sarah (2006) found that addition of 15% flaxseed meal in bread, negatively affected the volume, crust colour and crumb colour of breads. Koca and Anil (2007) showed that crumb darkness increased by increasing the level of flaxseed flours levels. A significant decrease in assigning scores to all the sensory attributes has also been supported by the study conducted by Naz, (2000) who observed that breads exceeding 15% flaxseed supplementation in wheat flour resulted in lower scores for texture, crumb colour, grain, and volume and crust colours. 2.7 Biological assay of protein quality The maintenance of good health is primarily dependent upon the utilization of good quality proteins. The proteins from animal sources are

thought to be superior in quality than the proteins obtained from vegetable sources (Salcedo-Chavez et al., 2002). The nutritive value of plant proteins is low due to their imbalanced amino acid profile while animal proteins are more expensive (Shils et al., 2002; Bodwell, 1979). The plant proteins are the major contributor to the dietary protein requirements in the developing countries (Bodwell, 1979). Due to this reason, protein malnutrition is a common problem in the major population segments of these countries (Rangel et al., 2003). Cereal based diets are responsible to divest the indispensable amino acids (Seena et al., 2006). A survey of the world wheat collection has revealed that total lysine variation in common wheat protein was no more than 0.5 %, which was less than needed for nutritional balance ( Johonson et al., 1985 ). Among different amino acids, lysine is considered to be more heat susceptible amino acid (Carpenter, 1960) and it is the first limiting amino acid in cereal products (Meredith and caster, 1984). The gap between the protein requirement and supply can be abridge by amino acid fortification, uses of protein mixtures (protein supplementation and complementation), genetic modification of food crops and identification and evaluation of underexploited sources (Egbe and Akinyele, 1990; Bodwell, 1979). Several factors affect the definitive quality of protein in vivo. The quality of proteins is best judged by their biological evaluation (Sogi et al., 2004). The amino acid content and profile is a critical determinant of protein quality and most of the methods which measure protein quality are directly or indirectly related to the efficacy with which they can satisfy amino acid requirements (Srikantia, 1981). The technology of composite flour preparation is initially referred to the process of blending of wheat flour with legume and cereal flours for making biscuits and bread. However, the declaration can also be used with respect to blending of other non wheat flours like roots, tubers or

other raw materials (Dendy, 1992). The legume proteins have been used widely for many years in the food industry as a food product constituent to improve the nutritional value of foods because of their better amino acids profile (Horax et

al., 2004). The bakery products can be improved in their nutritional profile by using the composite flour technology. Chapattis (unleavened flat breads) can be nutritionally improved by the composite flour technologies (Rasool, 2004). In a biological study conducted by Shehata and Fryer, (1970) concluded that rats fed on diets with 10 and 20 percent chickpea flour showed protein efficiency ratios of 1.46 and 1.56, which were better than that of diets with 0 percent chickpea flour. Maqbool et al., (1987) observed that protein efficiency ratio, net protein utilization and biological value of supplemented rotis (unleavened flat breads baked in earthen oven) were improved by the addition of soybean flour. Similarly, Rasool (2004) reported that biological value and net protein utilization of chapattis increased due to the addition of cotton seed flour in the wheat flour. As the first limiting amino acid in wheat flour is lysine, but its quantity in flaxseed flour is 6.8 mg/100g of protein (Dev and Quensel, 1986) which is higher than soy flour i.e 5.8 mg/100g of protein (Friedman and Levin, 1989) and wheat flour. Thus addition of flaxseed flour in chapattis and bread can be helpful for improving the biological parameters of these products. 2.8 Health benefits of flaxseed Flaxseed is a complex plant material in which a number of components including dietary fiber, flaxseed oil, protein and phenolic compounds are mainly responsible for a number of health benefits. Some of studies showing the health benefits of whole flaxseed, defatted flaxseed meal, flaxseed oil and individual flaxseed components are reviewed under this section. There are two groups of omega fats; omega-3 and omega-6 fatty acids. Linolenic acid, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) are the three types of omega-3 fatty acids and are nutritionally important. All three fatty acids have been shown to reduce the risk of cardiovascular disease. Linolenic acid is naturally present in canola, flaxseed, and walnuts whereas the other two types of omega fats are mainly present in fish like salmon, mackerel

and herring (Hurteau, 2004). Ground flaxseed is high in omega3-fatty acids, which have been shown to reduce hypertension, cholesterol and triglyceride level (Oomah and Mazza, 1998). Flaxseed acts as good source for lowering cholesterol and improving heart functions due to the eicosanoides derived from omega-3-fatty acids (Simopoulos, 1999). It is revealed in many studies that monounsaturated and polyunsaturated fatty acids are responsible for lowering the total cholesterol in diet (Zambon et al., 2000). The studies on lipid profile of male Wister rats fed high fat diets like flaxseed and trout (sources of polyunsaturated fatty acids), peanut (source of monounsaturated fatty acids) and chicken skin (source of saturated fatty acids) indicated that total cholesterol levels in rats fed flaxseed diet were lower than in rats fed on the other fats (Cintra et al., 2006). Alpha Linolenic acid (ALA) from flaxseed exerts positive effects on blood lipids. The dietary alpha linolenic acid was found be as effective as oleic (18:2n-6) and linoleic acid (18:2n-6) in the reduction of plasma total cholesterol, low density lipoprotein cholesterol and very low density lipoprotein cholesterol in 20-34 years old healthy men (Chan et al., 1991). In another study, 12 g alpha linolenic acid was taken three times a day by a group of healthy young women in the form of flaxseed oil capsules and compared with other group given the flaxseed flour supplemented products. Impressive reduction in blood lipids was observed in both cases (Cunnane et al., 1993). Incorporating full fat flaxseed meal into the diet eliminated the adverse effect of hydrogenated soybean oil on serum cholesterol levels in hypercholestrolemic rats. Full fat flaxseed meal enhanced the cholesterol-lowering effect of diets containing flaxseed oil (Ranhotra et al., 1993). The flaxseed protein was also found to be effective in lowering plasma cholesterol and triacylglycerides (Bhathena et al., 2002). Dietary fiber is a communal word used to describe a variety of plant substances that are not easily digested by the enzymes responsible for digestion

in humans (Eastwood and Passmore, 1983). There are two types of dietary fiber; soluble and insoluble. Soluble fibers like psyllium, guar gum and oats are responsible for reduction in the total cholesterol and LDL cholesterol (Glore et al., 1994). The flaxseed fiber is also considered to reduce the blood glucose and cholesterol levels by delaying and reducing their absorption in the body (Shen et al., 1998). Increasing the level of soluble fiber of guar gum, chick pea, and lentil in wheat flour contributed towards the reduction of blood glucose, total cholesterol and increase in the HDL cholesterol of Sprague Dawley rats fed on these fibers (Butt et al., 2007). The studies of Jenkins et al. (1999) showed a 5 and 8% reduction, respectively, in serum total and LDL cholesterol levels in subjects fed partially defatted flaxseed. These researchers attributed the LDL reduction to soluble fiber component of the flaxseed. Cunnane et al. (1995b) reported that blood glucose level reduced by about 27% when bread containing flaxseed was consumed, while Bhathena et al. (2003) observed 26% and 41% reductions in the plasma total cholesterol of leans and obese rats, respectively due to the effect of inclusion of flaxseed meal in their diets. A significant reduction in total serum cholesterol of weanling rats fed on 20-40% flaxseed was also observed by Ratnayake et al. (1992). Gambus et al. (2001) conducted an experiment on Wister rats which were fed standard bread or bread containing 10 or 13% milled flax seeds for 19 days, and observed 47.0 and 48.5% reduction in average blood cholesterol concentration and low-density lipoprotein fraction, respectively as compared with the group of rats fed on standard bread with no added flaxseed. In a similar study done by Kritchevsky (1995), it was observed that rats fed a 20% flaxseed diet showed a 25% additional reduction in serum and liver cholesterol than rats fed a diet containing 10% flaxseed. Cunnane et al. (1995b) stated that 8% reduction in LDL protein and 30% increase in bowel movement observed in adults consuming 50g flaxseed per day for 4 weeks. Gambus et al. (2004) in an experiment on rats fed flaxseed bread achieved impressive hypocholesteric

effects. Hasler et al. (2000) observed 6.9% reduction in total cholesterol of women fed on bread and muffins supplemented with 38 g of flaxseed for a period of six weeks. Lemay et al. (2002) noticed small changes in cholesterol levels of 25 hypercholestrolemic women fed on diet containing 40 g flaxseed/day. Arjmandi et al., (1998) reported that women (38) with mild, moderate or severe hypercholesterolemia were provided with 38 g of flaxseed incorporated into bread or muffins as part of their daily diet for six weeks. The total cholesterol, LDL-cholesterol and lipoprotein (a) concentration reduced

significantly, possibly due to the activity of linolenic or linoleic acids, fiber and non-protein constituents present in this seed. In a clinical study by Lucas et al. (2002, 40 g of either ground flaxseed or a wheat-based comparative control regimen was fed to postmenopausal women on daily basis for three months. The flaxseed supplementation resulted in 6% reduction in both serum total and HDLcholesterol, whereas no effect was observed due to use of control regimen. The flaxseed regimen reduced the serum levels of both LDL and HDL cholesterol by 4.7% and triglyceride by 12.8%. Prasad (1999) studied the effect of secoisolariciresinol diglucoside (SDG) isolated from flaxseed in rabbits and concluded that SDG helped to decrease hypercholestrolemic atherosclerosis due to reduction in serum total and LDL cholesterol. Increase in HDL-cholesterol and antioxidant reserves was also observed. Dodin et al., (2005) reported that the intake of flaxseed reduced serum total (-0.200.51) mmol/liter and high- density lipoprotein (-0.080.24) mmol/liter cholesterol concentrations of menopausal women who consumed 40g flaxseed/day. The wheat is staple food for the inhabitants of Pakistan. It is used to meet their energy and protein requirements. The quality of wheat proteins is not as good as needed to meet the body requirements. Therefore most of the population could not meet their requirements for these nutrients from other sources (e.g. animals) due to their poor economic status and are facing nutrient deficiencies.

Their deficiencies can be ameliorated by improving the wheat nutrients through dietary interventions like fortification, enrichment and supplementation. The flaxseed grains are good source of nutrients and also provide health benefits. The flaxseed can be included in the bakery products like chapattis and bread. The addition of flaxseed in wheat flour can be achieved keeping in view the sensory parameters of chapattis and breads. The storage stability of flaxseed for about 2 months makes it suitable for its blending with wheat flour. The flaxseed proteins can enhance the quality parameters of bread and chapattis. The addition of flaxseed in wheat flour can also contribute towards the several health benefits and can offer a cheap treatment for many diseases e.g diabetes,

hypercholesterolemia and cardiovascular complications which are more prevalent in Pakistan.