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Enhancement of the nutritional value of whey drink by supplementing with leaves of Moringa oleifera.


Malnutrition results in retarded physical and mental growth as well as lack of resistance against diseases, whereas, protein deficiency results in poor health and reduced capacity for physical work (NNS, 2004). Plants have natural antioxidants and polyphenolic compounds that have anticarinogenic, cardiac and hepatic protective activity (Jeong et al., 2004; Kris-Etherton et al, 2002; Middleton et al., 2000). M. oleifera is fast growing tree and its flowers, leaves and pods are consumed as vegetable and tender roots are preserved in the form of tasty pickles (Anwar et al., 2007). It has high contents of protein and vitamins (Juliani et al., 2009; Quarcoo, 2008; Fuglie, 1999).

Whey is the by-product of cheese, containing nearly half Of the total solids present in milk (Walzem et al., 2002). Cheese manufacturing units of Pakistan throw thousands of liters of untreated whey into the drain on daily basis which must be treated before discharge into the environment (Marwaha and Kennedy, 1998). Soft drinks may also be manufactured from whey by making use of different types of stabilizers and fruit concentrates of orange, apple, banana, mango and lemon (Niketic and Marinkovic, 1984). Keeping in view the utilization of whey, the present investigation was aimed to prepare nutritious whey drink by exploiting the massive nutritional potential of M. oleifera.

Materials and Methods

Materials. Dried M. oleifera leaves were ground in a laboratory scale grinder (Moulinex), sieved through mesh size 50, stored in amber colour glass bottles, sealed and stored at -40[degrees]C till further usage in this study. All the chemicals used in this study were HPLC grade and obtained from Sigma Chemical Co. (St. Louis MO).

Treatments. Moringa oleifera leaf powder (MOLP) was incorporated into whey drink at four different concentrations i.e., 1% MOLP ([T.sub.1]), 2% MOLP ([T.sub.2]), 3% MOLP ([T.sub.3]), 4% MOLP ([T.sub.4]), and compared with a control ([T.sub.0]). The formulation of control comprised of banana pulp 5%, sugar 5%, citric acid (20% solution) 0.5%, carageenan 0.25% and whey to make volume up to the mark, colour and flavour 1 mL/L. All the ingredients were dissolved in the whey, pasteurised in the beakers at 65[degrees]C for 30 min, immediately cooled down, filled in clean sanitised pet bottles and stored at 4[degrees]C for further analysis. Each treatment was replicated three times.

Analysis. Fat, pH, acidity, protein, lactose, ash content and total solids were determined by following the respective methods AOAC 7.093, AOAC 7.094 and AOAC 7.095 (AOAC, 2000). Ca, K and Mg were analysed on atomic absorption spectrophotometer (Model: AA240, Varian, Australia) briefly, 5 g sample was taken into a conical flask, 10 mL of concentrated nitric acid was added and contents of the flask were heated for 20 min. After cooling, 5 mL perchloric acid was added and again heated vigorously till the volume was reduced to 2-3 mL followed by cooling. The contents were then diluted in 50 mL volumetric flask by using distilled water. Absorbance of each mineral was determined on the respective wave length, also the unknown concentration determined by constructing calibration curve using 8 standards of each mineral ([R.sup.2] = 0.9891 to 0.9961). Concentration of iron was determined by using the standard method AOAC 999.11 of AOAC (2000). Vitamin assay was performed on HPLC by following the respective methods AOAC 971.30, AOAC 948.26, and AOAC 975.42 (AOAC, 2000). Free fatty acids and peroxide values were determined by using standard methods cd 8-53 and cd 1-25 of Firestone (1997). For the determination of total phenolic contents of MOLP, 1 mL (methanolic extract of leaf powder) was poured into 11 mL screw capped test tube, 2 mL (0.2-molar) folin ciocalteu, 2 mL sodium carbonate (7.5%) added, mixed and stored in dark for 20 min at ambient temperature and absorbance measured at 765-nm in visible region on a double beam spectrophotometer (Shidmadzu, Japan). The concentration of phenolic compounds was measured on a standard curve constructed by using gallic acid as standard (R = 0.9943) by following the method of Anwar et al. (2007). The sensory evaluation of whey drink was conducted by a panel of 10 trained judges performed on a 9 point hedonic scale (1--the worst; 9--the best) as prescribed by Larmond (1987). The samples were evaluated for texture, taste, smell and overall acceptability. Samples were presented in glasses, coded with three digit random numbers and all servings were completely randomized. The data of triplicate experiments designed completely randomized (CRD). All data expressed as Mean [+ or -] SD, the significant difference among the treatments calculated by using Duncan's Multiple Range Test SAS 9.1 Statistical Software.

Results and Discussion

Composition of whey drink supplemented with M. oleifera leaves. The results of chemical composition of whey drink show that fat, protein, lactose, ash and total solids content were 0.42%, 0.65%, 4.38%, 0.58% and 6.11%. Addition of MOLP at all the four levels did not have any negative effect on pH and acidity of supplemented whey drink (Table 1).

As the concentration of MOLP increased to 2%, fat content increased significantly (P<0.05). The highest fat content 0.64% observed in [T.sub.4] which was 52% over the control. Protein content of the supplemented whey drinks increased in a concentration dependent manner. Increase in protein content was 164% in [T.sub.4] as compared to the control. The increase in fat and protein content was due to the presence of higher concentration of these constituents in MOLP. Addition of MOLP at all levels increased the ash content Of whey drinks significantly (P<0.05), [T.sub.1] and [T.sub.2] comparison with each other (P>0.05), increased to 52 and 65% in [T.sub.3] and [T.sub.4], respectively, as compared to the control. Increasing trend was observed in total solids with increased level of supplementation. The highest total solids recorded in [T.sub.4] (16.93%) followed by [T.sub.3] (15.84%) and [T.sub.2] (14.89%).

Total phenolic contents: Total phenolic content of MOLP was 7.4g/100gon dry matter basis (GAE). The total phenolic contents of Sesamum indicum cake extract was 1.94% on dry matter basis (Mohdaly et al., 2011). InM olifera concerentration of phenolic compounds was considerably higher than S. indicum (Mohdaly et al., 2011) and canola hull (Naczk and Shahidi, 1998).

Minerals content of whey drink supplemented with M. oleifera leaves. Calcium content of all the treatments increased in a dose dependent manner. The results showed the increasing level of calcium content of fortified whey drink by 10%, 19%, 31% and 47%, respectively, as compared to the control, while the effect on potassium content of the drinks was non-significant (Table 2). Potassium content ranged from 1226 to 1245 mg/L among different treatments and control. The elevation in calcium content of the experimental samples was due to the higher concentration of calcium in MOLP. The addition of MOLP at 4% level increased the iron content of the whey drink by 675% over the control.

The addition of MOLP at 1% level ([T.sub.1]) increased the iron content from 0.17 mg/L to 44 mg/L which was 257% higher than the control (Aney et al., 2009) while, studying the nutritional value of M. oleifera reported that the calcium, magnesium and iron content of 100g dried MOLP was 2000 mg, 1328 mg and 28.2 mg, respectively. As recommendations of FAO, WHO 400 mg and 1200 mg calcium is required on daily basis for the children of 1-3 years age and nursing women. Half of the total calcium requirement for nursing women may be easily and economically fulfilled by consuming just two glasses of 250 mL of whey drink supplemented with 4% MOLP, almost 75% of the calcium requirements of the children can be met by 250 mL of the fortified drink. M. oleifera leaves demonstrated higher vitamin A than carrots, more calcium and potassium than milk and banana with superior amino acid profile resembling to egg proteins (Juliani et al., 2009). Milk is not a complete food because of its lower iron and vitamin C contents, fortification of milk with iron is usually practiced in poor and malnourished countries. Most of the strategies involve the usage of sulphates of iron, which are unnatural and accelerate the oxidative breakdown (Fox and McSweeney, 2003). The iron requirement of 10 and 15 mg per day for children and nursing women can be efficiently fulfilled by 250 and 300 mL of supplemented ([T.sub.4]) whey drink. Feeding M. oleifera supplemented diet significantly improved the health status of school going babies and pregnant women in Senegal who gave birth to healthy babies (Juliani et al., 2009).

Feeding 10 g leaves to pregnant women may provide adequate concentration of essential micro nutrients to save them from becoming anemic, physically and mentally weak (Fahey, 2005). This could be the simple, economical and efficient way of preventing and correcting the anemic conditions of millions of children and women of the poor nations who cannot afford medication and expensive nutritional supplements.

Vitamins content of whey drink supplemented with M. oleifera leaves. The addition of MOLP at all levels significantly (P<0.05) increased the vitamin [B.sub.5], [B.sub.2], [B.sub.6] and vitamin C content (Table 3). The increase in vitamins [B.sub.5], [B.sub.2], [B.sub.6] and C was 158%, 372%, 193% and 50%, respectively, in [T.sub.4]. The increase in vitamin content was due to the chemical composition of the MOLP which resulted in higher concentrations of these vitamins in the whey drinks. The addition of MOLP did not have any effect on free fatty acids and peroxide value of whey drink at all levels. The recommended daily allowance of vitamin [B.sub.2] is 0.8 and 1.8 mg for children and nursing women, 300 mL of supplemented whey drink at 2% MOLP can meet 100% body's requirement with respect to this vitamin. 423 and 17 mg/100 g, vitamin [B.sub.2] and C were present in Ethiopian Moringa stenopetala (Price, 1985). Anwar et al. (2007) studied the effect of M.oleifera addition on oxidative stability of some vegetable oils and found that blend of soybean and sunflower containing 80% (MOO) has induction time from 1.12 to 5.99 h, 1.47 to 6.22 h, recorded the increase in oxidative stability 435% and 323%, respectively.

Sensory evaluation. The results of sensory evaluation (Table 4) indicated that supplementation of MOLP up to [T.sub.2] level did not have any negative effect on texture and taste score. Texture and taste score of [T.sub.1] and [T.sub.2] were at par with the control (P>0.05). With the increasing increments of MOLP, the score for these two parameters decreased. Some panelists criticised [T.sub.3] and [T.sub.4] for relatively higher coarse texture. Level of criticism for [T.sub.4] was not adverse as the people are already habitual of adding green leaves of coriander and other herbs in the butter milk. Quarcoo (2008) studied the development of M. oleifera based beverage and reported that addition of increasing concentration of M.oleifera leaf powder tended to make the colour greener and decreased the colour score. The low colour score was attributed to the uncommon green colour of the beverage; the lower concentrations were rated higher for this parameter. This problem could have been resolved by homogenising the supplemented whey drinks at high pressure. Homogenisation is commercially carried out to make the nectars and high fruit preparations homogenous and smooth (Spreer, 2005). Score for smell of all the treatments and control was non-significantly influenced from each other. The overall acceptability score of [T.sub.4] was 6.9 out of 9, which was more than 76% of the total score.


The addition of M. oleifera leaf powder at all concentrations improved the nutritional value of the whey drink. Protein and iron content significantly increased in the supplemented drinks. 300 mL of supplemented whey drink can fulfill 100% requirement of vitamin [B.sub.2] of nursing women. Highly nutritious whey drink can be successfully prepared by the addition of MOLP up to 4% level; whey drink supplemented with MOLP should be homogenised for better sensory characteristics.


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Kris-Etherton, P.M., Hecker, K.D., Bonanome, A., Coval, S.M., Binkoski, A.E., Hilpert, K. F., Griel, A.E., Etherton, T.D. 2002. Bioactive compounds in foods: Their role in prevention of cardiovascular disease and cancer. The American Journal of Medicine, 113 Suppl9 B: 71S-88S.

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Muhammad Nadeem *, Imtiaz Hussain, Muhammad Abdullah and Fazal Rehman

Department of Dairy Technology, University of Veterinary and Animal Science, Lahore, Pakistan

(received April 16, 2012; revised April 10, 2013; accepted April 11, 2013)

* Author for correspondence;
Table 1. Effect of MOLP supplementation on composition of
whey drink

Treatments   pH                       Acidity


[T.sub.0]    6.43 [+ or -] 0.99 (a)   0.23 [+ or -] 0.10 (a)
[T.sub.1]    6.37 [+ or -] 0.83 (a)   0.24 [+ or -] 0.06 (a)
[T.sub.2]    6.38 [+ or -] 0.66 (a)   0.24 [+ or -] 0.09 (a)
[T.sub.3]    6.35 [+ or -] 0.77 (a)   0.25 [+ or -] 0.14 (a)
[T.sub.4]    6.38 [+ or -] 1.02 (a)   0.24 [+ or -] 0.12 (a)

Treatments   Fat                      Protein


[T.sub.0]    0.42 [+ or -] 0.08 (c)   0.67 [+ or -] 0.13 (e)
[T.sub.1]    0.45 [+ or -] 0.16 (c)   0.94 [+ or -] 0.11 (d)
[T.sub.2]    0.51 [+ or -] 0.14 (b)   1.19 [+ or -] 0.34 (c)
[T.sub.3]    0.55 [+ or -] 0.11 (b)   1.46 [+ or -] 0.29 (b)
[T.sub.4]    0.64 [+ or -] 0.19 (a)   1.73 [+ or -] 0.43 (a)

Treatments   Lactose                  Ash


[T.sub.0]    4.65 [+ or -] 0.21 (a)   0.60 [+ or -] 0.14 (c)
[T.sub.1]    4.56 [+ or -] 0.15 (a)   0.71 [+ or -] 0.22 (b)
[T.sub.2]    4.39 [+ or -] 0.11 (b)   0.85 [+ or -] 0.12 (b)
[T.sub.3]    4.31 [+ or -] 0.08 (b)   0.91 [+ or -] 0.18 (a)
[T.sub.4]    4.25 [+ or -] 0.14 (b)   0.99 [+ or -] 0.24 (a)

Treatments   TS


[T.sub.0]    12.75 [+ or -] 0.21 (e)
[T.sub.1]    13.80 [+ or -] 0.18 (d)
[T.sub.2]    14.89 [+ or -] 0.35 (c)
[T.sub.3]    15.84 [+ or -] 0.42 (b)
[T.sub.4]    16.93 [+ or -] 0.29 (a)

Means of triplicate experiment; means with same letters in
same column are statistically non-significant (P>0.05); TS
= total solids; [T.sub.0] = control without any addition
of MOLP; [T.sub.1] = 1% MOLP; [T.sub.2] 2% MOLP; [T.sub.3]
3% MOLP; [T.sub.4] 4% MOLP

Table 2. Effect of MOLP supplementation on mineral
content of whey drink

Treatments   Calcium                 Potassium

             (mg/100 g)

[T.sub.0]    268 [+ or -] 0.93 (d)   1245 [+ or -] 6.13 (a)
[T.sub.1]    297 [+ or -] 0.81 (d)   1231 [+ or -] 2.48 (a)
[T.sub.2]    320 [+ or -] 0.54 (c)   1226 [+ or -] 1.05 (a)
[T.sub.3]    353 [+ or -] 0.77 (b)   1238 [+ or -] 0.78 (a)
[T.sub.4]    394 [+ or -] 1.06 (a)   1241 [+ or -] 3.16 (a)

Treatments   Magnesium              Iron

             (mg/100 g)

[T.sub.0]    35 [+ or -] 0.43 (b)   0.17 [+ or -] 0.03 (e)
[T.sub.1]    38 [+ or -] 1.24 (b)   44 [+ or -] 0.09 (d)
[T.sub.2]    42 [+ or -] 0.06 (b)   75 [+ or -] 0.34 (c)
[T.sub.3]    45 [+ or -] 0.31 (a)   99 [+ or -] 0.52 (b)
[T.sub.4]    48 [+ or -] 0.19 (a)   115 [+ or -] 0.05 (a)

Means of triplicate experiment; means with same letters in
a column are statistically non-significant; refer Table 1 for
the detail of treatments.

Table 3. Effect of MOLP supplementation on vitamin
content and stability of whey drink

Treatments   Vitamin                   Vitamin
             [B.sub.5]                 [B.sub.2]


[T.sub.0]    3.94 [+ or -] 0.03 (e)    2.14 [+ or -] 0.16 (e)
[T.sub.1]    5.59 [+ or -] 0.14 (d)    4.16 [+ or -] 0.24 (d)
[T.sub.2]    7.12 [+ or -] 0.11 (c)    6.06 [+ or -] 0.42 (c)
[T.sub.3]    8.68 [+ or -] 0.31 (b)    9.03 [+ or -] 0.34 (b)
[T.sub.4]    10.20 [+ or -] 0.43 (a)   10.12 [+ or -] 0.66 (a)

Treatments   Vitamin E                Vitamin C


[T.sub.0]    2.85 [+ or -] 0.05 (e)   1.46 [+ or -] 0.21 (e)
[T.sub.1]    3.91 [+ or -] 0.18 (d)   1.69 [+ or -] 0.19 (d)
[T.sub.2]    5.02 [+ or -] 0.45 (c)   1.83 [+ or -] 0.41 (c)
[T.sub.3]    6.15 [+ or -] 0.29 (b)   2.01 [+ or -] 0.12 (b)
[T.sub.4]    8.36 [+ or -] 0.45 (a)   2.20 [+ or -] 0.67 (a)

Treatments   FFA%                     PV (meq/kg)

[T.sub.0]    0.10 [+ or -] 0.01 (a)   0.23 [+ or -] 0.03 (a)
[T.sub.1]    0.10 [+ or -] 0.01 (a)   0.23 [+ or -] 0.01 (a)
[T.sub.2]    0.11 [+ or -] 0.03 (a)   0.25 [+ or -] 0.05 (a)
[T.sub.3]    0.11 [+ or -] 0.01 (a)   0.26 [+ or -] 0.02 (a)
[T.sub.4]    0.11 [+ or -] 0.02 (a)   0.26 [+ or -] 0.09 (a)

Means of triplicate experiment; means with same letters
in same column are statistically non-significant; refer
Table-1 for the detail of treatments; FFA = free fatty
acids; PV = peroxide value.

Table 4. Effect of MOLP supplementation on sensory
characteristics of whey drink

Treatments   Texture                 Taste

[T.sub.0]    7.6 [+ or -] 0.11 (a)   7.9 [+ or -] 0.23 (a)
[T.sub.1]    7.3 [+ or -] 0.08 (a)   7.6 [+ or -] 0.09 (a)
[T.sub.2]    7.4 [+ or -] 0.14 (a)   7.5 [+ or -] 0.11 (a)
[T.sub.3]    7.0 [+ or -] 0.03 (b)   7.0 [+ or -] 0.17 (b)
[T.sub.4]    6.8 [+ or -] 0.06 (b)   6.6 [+ or -] 0.14 (b)

Treatments   Smell                   Overall

[T.sub.0]    7.5 [+ or -] 0.08 (a)   7.5 [+ or -] 0.03 (a)
[T.sub.1]    7.4 [+ or -] 0.02 (a)   7.2 [+ or -] 0.14 (a)
[T.sub.2]    7.5 [+ or -] 0.04 (a)   7.3 [+ or -] 0.05 (a)
[T.sub.3]    7.2 [+ or -] 0.19 (a)   7.0 [+ or -] 0.09 (b)
[T.sub.4]    7.2 [+ or -] 0.12 (a)   6.9 [+ or -] 0.04 (b)

Means of triplicate experiment; means with same letters in
same column are statistically non-significant; refer Table 1
for the detail of treatments.
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Author:Nadeem, Muhammad; Hussain, Imtiaz; Abdullah, Muhammad; Rehman, Fazal
Publication:Pakistan Journal of Scientific and Industrial Research Series B: Biological Sciences
Article Type:Report
Geographic Code:9PAKI
Date:Nov 1, 2013
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