Asian Journal of Dairy and Food Research, volume 42 issue 4 (december 2023) : 557-563

Effects of Binders on Physico-chemical Characteristics of Egg Albumen Paneer (EAP)

Kushal J. Thakuria1, Dilip R. Nath1, Ankur Das1, Mineswar Hazarika1, Saurabh Kumar Laskar1, Rashmi R. Saikia1, Sadhana Choudhury1
1Department of Livestock Products Technology, College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati-781 022, Assam, India.
Cite article:- Thakuria J. Kushal, Nath R. Dilip, Das Ankur, Hazarika Mineswar, Laskar Kumar Saurabh, Saikia R. Rashmi, Choudhury Sadhana (2023). Effects of Binders on Physico-chemical Characteristics of Egg Albumen Paneer (EAP) . Asian Journal of Dairy and Food Research. 42(4): 557-563. doi: 10.18805/ajdfr.DR-1835.
Background: An innovative food product from egg albumen analogous to the conventional paneer was developed by inclusion of different types of binders and their effects on the physico-chemical qualities of the product were studied.

Methods: Different cereal flours were used as binders viz., juoha rice flour (JRF), glutinous rice flour (GRF), wheat flour (WF) and oat flour (OF) in different ratios, with the egg albumen as the base material for the production of egg albumen paneer (EAP). EAP cubes were packed in food grade high density polyethylene bags and stored at 5-7oC for physico-chemical evaluation upto 90 days. Rehydration properties of the dried products were also studied.

Result: Studies of EAP revealed that pH and water activity differ significant (P<0.01) among the treatments groups and decreased gradually during storage irrespective of the treatment groups and days of storage. The Thiobarbituric acid and Tyrosine values also showed significant (P<0.01) differences among the treatment formulations as well as during the storage days. The rehydration percentage, increase in volume after rehydration and coefficient of rehydration recorded highest in T1 than the other samples. Both the rehydration ratio and rehydration percentage were recorded highest in T2 group.
Eggs are an important source of high quality protein with only some calories and have been recognized as a food of high nutritional value, with the egg albumen concentrating the bulk of egg proteins with almost no fat. The quest for newer and variety of egg products are continuously on rise in the form of number of egg based products viz. egg coated potato (Muller,1994), premixed flavoured egg product (Wu et al.,1995), egg flakes containing monosodium glutamate and onion/garlic extracts (Lee et al. 1998), egg white chips containing stabilizers and flavouring (Yang et al., 2000), formulated fried egg (Merkle et al., 2003), other than the basics products like boiled eggs, egg in curry, omelette and egg pakoda. Inclusion of the cereal flours in the food formulations may have many health related advantages (Bazzano et al., 2003; Liu et al. 2003; Schatzkin et al. 2007), while the dietary fibers in cereal flours act as volume enhancer, binder, bulking agent, stabilizer and also improve product’s texture (Cofrades et al. 2000; Sanchez-Zapata et  al., 2010). Oat bran contains higher amount of soluble dietary fibre as compared to wheat flour which contains higher amount of total dietary fiber and insoluble dietary fibre (Talukdar and Sharma, 2010). With this background knowledge, it was planned to develop an egg albumen paneer with different binders as an alternative to traditional milk paneer with added benefits.
The study was conducted in the Department of Livestock Products Technology, College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati in the year 2016.
Product preparation
Hen eggs were carefully collected from the nearby retail market and checked for quality. Binders and spices were procured from the departmental super market. The formulations were made after a trial study with different percentage of the binders to obtain the desired paneer like shape and texture. Dry binder mix was prepared by mixing the dry ingredients manually in a bowl as per the different formulations like Control (wheat flour-15%, rice flour- 5%), T1 (glutinous rice flour-10%, joha rice flour-10%), T2 (glutinous rice flour-8%, oat flour-12%), T3 (joha rice flour-8%, oat flour-12%) and T4 (glutinous rice flour-4%, joha rice flour-4% and oat flour-12%). Eggs white was separated from the yolk manually. The batter for egg albumen paneer was prepared by mixing the liquid albumen at 77%, mallic acid, citric acid and spice mix 0.5% each as preservatives and  salt 1.5% for taste with the dry binder mix in a mechanical hand blender (Bajaj, India) for 3-5 mins to obtain a homogenous consistency of the batter. It was then poured to a stainless steel mould lined with Aluminium foil and cooked at a temperature range of 80-85oC for 40-45 mins in steam, followed by cooling to ambient temperature. The loaf obtained was then cut into uniform cubes (1 x 1 x 1) inch and vacuum packaged in food grade high density polyethylene bags and stored at 5-7oC temperature for quality studies at regular interval upto 90 days. For studying the rehydration properties of the product, cubes were dried in a hot air oven at 80-85oC for 5-6 hours to get the dehydrated EAP cubes and thereafter evaluated for various parameters.
Physico-chemical properties
Physico-chemical properties of the samples were recorded on the day of production and subsequently on on 5th, 10th, 15th, 20th and upto 90th day for the following parameters.
The pH (Hydrogen Ion Concentration) was determined according to the method of Pippen et al. (1965), using a Digital pH Meter (Make: Metrohm, Model: 801 Stirrer).
Thiobarbituric acid (TBA) value
The Thiobarbituric acid (TBA) Value was determined by employing the method as described by Witte et al. (1970).
Tyrosine value
Tyrosine value was determined by following the method of Strange et al. (1977).
Water activity (aw)
Water activity (aw) was determined with the help of a Water Activity Meter (Make: AQUA LAB, Model: 4TE) by taking 10g of dehydrated EAP sample in a sample cup and introduced in to the machine for calculation of the water activity (aw). The aw was recorded on the day of production as well as on the subsequent storage days for all the product batches.
Volume increase after rehydration
Rehydration of the cubes was done by soaking the dried cubes in cold water in a beaker for 10 minutes followed by allowing the drainage of the excess water. For 5 dried cubes 20ml of water was used. The per cent increase in volume after rehydration was determined by measuring the volume (length x breadth x height) of rehydrated cubes.

Coefficient of rehydration
Coefficient of rehydration, rehydration ratio and per cent rehydration in terms of per cent water in rehydrated EAP were calculated as described by Pawar et al. (2012).
Coefficient of rehydration =

Rehydration ratio
Wt of EAP:Wt. of rehydrated EAP
Per cent water in the rehydrated EAP (Rehydration%)
(Rehydration,%) =
*(Weight of EAP taken for rehydration - Moisture (g) in EAP before rehydration)
Statistical analysis
Experimental data obtained from the experiment was analyzed following the standard statistical method given by Snedecor and Cochran (1994). Five batches of EAP of each formulation including the control ones were prepared to obtain the data required for statistical analysis. Analysis of variance (ANOVA) with honest significant difference (HSD) test for mean comparison was used to highlight significant differences among the samples. Statistical tests were performed with a 5% or 1% significance level using the SPSS program version 20 (IBM Corp, 2011).
Physico-chemical properties
The pH values differed significantly (P<0.01) (Table 1) among the treatment groups and also in the storage days with a slow but gradual decreasing trend irrespective of different formulations and days of storage. The lowest pH values were recorded in the C group as compared to others in all the storage days. However, the interaction between the treatment groups and storage days was non-significant. The relatively higher pH values recorded initially in the treatment groups as compared to the C formulation might be due to the fact that all the treated formulations had high content of rice flours, a very rich source of carbohydrate and also oat flours added in fresh during EAP preparation. But during the storage days these carbohydrate were slowly acted by different lipolytic enzymes (Buege et al. 1978) (Saccharolytic/Glycolytic) resulting in the production of different acids, thus lowering the pH values (high acidity) slowly and gradually till the end of storage period. A slightly lower pH value reported by Deepthi et al. (2011a) might be related to the difference in varieties of RF and oat flour incorporated in EAP production.

Table 1: pH of EAP (Mean ± SE).

Thiobarbituric acid (TBA) values
The TBA values showed a gradual increasing trend irrespective of treatment groups and days of storage, however itwas within the permissible limit of 2 mg malonaldehyde/kg of food product. On day-1, the highest TBARS value was recorded in T4 while it was lowest in T1 group (Table 2). However, on 90th day the highest TBARS value was recorded in C and the lowest in T2 formulation. The gradual increase in TBARS values of EAP might be related to the lipid oxidation caused by different lipolytic enzymes (Buege et al. 1978) present in EAP for which there were gradual increase in malonaldehyde values during the storage days. Moreover, albumen has high foaming property that entraps oxygen within it and this might have also contributed in oxidation of lipid Deepthi et al. (2011a). Considering the fact that the lipid content of egg albumen is very less i.e.,1.09% (Bashir et al.2015) as compared to the flours incorporated in EAP and this could be the possible reasons for very slow but gradual rise in TBARS values during the storage days. The TBARS values recorded for C formulation in the study was nearer to the ones reported by Deepthi et al. (2011a) who also reported TBARS values that varies from 0.66±0.04 to 1.24±0.07 in EAP during the storage period of 6 months. Several other investigators also reported an increase in TBARS values along with the increase in storage period although most of them had worked on certain meat products (Jebin 2011; Sebranek et al. 2005).

Table 2: TBARS values (mg malonaldehyde/kg) of EAP (Mean±SE).

Tyrosine value
Tyrosine values of EAP increased gradually irrespective of the treatment groups and days of storage. The lowest Tyrosine values were recorded on day-1, having lowest value in T3 and highest in C formulation (Table 3). A very similar trend was also observed on 90th day of storage with highest value in C and lowest in T3 formulations. The gradual increase in Tyrosine values of EAP might be related to the continuous protein denaturation by the different proteolytic enzymes i.e. Proteases/Cathepsins and others present in EAP. Moreover, egg albumen is very rich in protein content i.e., 3.48% (Bashir et al. 2015) which might have facilitated the proteolysis process during the storage days resulting in a gradual rise of tyrosine values of EAP. Several other workers also reported increase in Tyrosine values of their meat food products during the storage period (Mahmmod et al. 2014).

Table 3: Tyrosine values of EAP(Mean ± SE).

Water activity (aw)
Water activity (aw) of EAP revealed a gradual decrease in values irrespective of treatment groups and the days of storage and varies between 0.5795±0.005 to 0.7357±0.004 during storage (Table 4). On day-1, the T3 formulation recorded highest and T1 formulation recorded lowest aw. However, on 90th day of storage, the T3 and C formulation registered highest and lowest aw values respectively. The gradual fall in aw values of EAP might be a corollary to the continuous drop in pH values during the storage days and vacuum packaging of EAPs with high density polyethylene (HDPE) packaging material. This gradual fall in aw values of EAP during their storage time might have resulted in the absence of microbial count as observed in the present study. The aw values recorded for the C formulation were at variance with the ones reported by Deepthi et al. (2011a) for EAPs prepared with same incorporation levels of WF and RFs. This discrepancy could be attributed to the vacuum packaging of EAP with HDPE packaging material, besides, storing of the product (EAP) at refrigeration temperature (5 -7oC).

Table 4: Water activity of EAP (Mean±SE).

Increase in volume on rehydration
The increase in volume on rehydration values of EAP ranges between 120.69±1.57 to 153.07±7.00. There was no significant differences in increase in volume on rehydration among the T3 and T4 groups whereas the other formulations recorded a significant difference (P<0.05) (Table 5). The highest increase in volume on rehydration value recorded in T1 formulation might be due to incorporation of only rice flours (10% GRF and 10% JRF) in EAP preparation since rice flours has higher water binding capacity as compared to wheat and oat flours. The lowest increase in volume value recorded in the control group might be a reflection of incorporation of high percentages of wheat flour (15%) which has poor water absorption capacity. The results obtained in the study for increase in volume on rehydration were in contrast to that of Deepthi et al. (2011b). The higher increase in volume on rehydration values recorded in the present studies might be due to the differences in the product formulation, i.e. incorporation of GRF and JRF, besides oat flour.

Table 5: Increase in volume on rehydration, co-efficient of rehydration, rehydration ratio and rehydration percentage values of EAP (Mean±SE).

Coefficient of rehydration
No significant differences in coefficient of rehydration were found among the T1, T2, T3 and T4 groups, however, the control group showed a significant difference (P<0.05) (Table 5) with the other formulations. The highest coefficient of rehydration recorded in T1 formulation might be due to incorporation of only rice flours having higher water binding capacity.  A much lower coefficient of rehydration registered in the control group might be due to the same explanation as has been given for rehydration of EAP. Lower coefficient of rehydration value as reported by Deepthi et al. (2011a) might be due to their evaluation of EAP at monthly intervals and also on storage of the product at 27±2oC which was much higher than the one followed in the present study i.e. 5-7oC.
Rehydration ratio
There was no significant differences in rehydration ratios among the control and T4 groups whereas, the T1, T2, T3 groups recorded a significant difference (P<0.05) (Table 5) with the other formulations. The highest rehydration ratio recorded in T2 formulation might be a reflection of much higher fiber content in oat flour (3.53-5.87% fiber, Youssef et al., 2016) that was added at 12% along with 8% level GRF (0.9%, Itthivadhanapong and Sangnark, 2016). The study further revealed almost equal rehydration ratio of EAP in Control and T4 formulations. This could be attributed to much lower fiber content in JRF (0.25-0.75%, Roy et al., 2010) and slightly higher fiber content in oat and wheat flours incorporated in Tand Control formulations of EAP. 
Rehydration percentage
The rehydration percentage of different formulations of EAP varies between 87.06±2.31 to 104.92±5.38 per cent. No significant differences in rehydration percentages were observed between the T3 and T4 groups. On the contrary, the C, T1 and T2 groups revealed significant differences (P<0.05) (Table 5) with the other two formulations. The highest rehydration percentage recorded in T2 formulation might again be a reflection of high fiber content in GRF while the lowest in control group might be ascribed to incorporation of 15% wheat flour (low in fiber content) and only 5% rice flour (RF).
The results for Rehydration percentage recorded in the study for the control group was almost in conformity with the findings of Deepthi et al. (2011a) who also recorded (84.10±5.35) rehydration percentage on ‘0’ day of storage of EAP prepared by incorporation of same levels of WF and RF as followed in the control formulation of EAP in the present study.
An ease and ready-to-use product was prepared from egg albumen by incorporating GRF, JRF and Oat flour other than WF and RF. The physico-chemical characteristics of the product obtained in the study were reported. Treatments groups have a better quality aspects in terms of pH, TBA, Tyrosine value and water activity, than the control samples. Rehydration properties were also found to be better for the treatment groups than the control ones, with special mentioning of the T1 and T2 groups. The product alike to the traditional milk based paneer and has a better acceptability in terms of its physico-chemical properties and can be used in our day to day diet in the form of curry, snacks, etc.
The authors express sincere gratitude and thankfulness to the Dean, Faculty of Veterinary Science, AAU, Khanapara, Guwahati, Dr B.N. Saikia, for providing the necessary facilities and financial aid to carry out the research programme successfully.
All authors declare that they have no conflicts of interest.

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