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Journal of Food Research; Vol. 1, No.

3; 2012 ISSN 1927-0887 E-ISSN 1927-0895 Published by Canadian Center of Science and Education

Development of Fermented Beverage Using RSM and Nutrients Evaluation – I. Fermented Ashgourd Beverage
C. S. Devaki1 & K. S. Premavalli1
1

Food Preservation Discipline, Defence Food Research Laboratory, Mysore, India

Correspondence: K. S. Premavalli, Food Preservation Discipline, Defence Food Research Laboratory, Siddarthanagar, Mysore 570011, Karnataka, India. Tel: 91-821–247-3828. E-mail: dfrlmysore@sancharnet.in Received: January 6, 2012 doi:10.5539/jfr.v1n3p138 Abstract Ash gourd (Benincasa hispida) is valued for its nutritive and medicinal properties and further value addition is being attempted by fermentation process. In the present study, the optimization of the fermentation process with reference to yeast concentration and the period has been attempted by using RSM. The statistical design gave 13 formulations, where yeast concentration was from 1 to 5.8% and the fermentation process period varied from 0.5 to 17.5 days. The product varied formulations had total phenols ranged from 24.8 to 43.6mg%, antioxidant 12.3 to 27%, acidity 0.067 to 0.6%, total volatiles 0.0025 to 0.078%, alcohol 4.6-12.4%, overall acceptability 6.9 to 8.4, thiamine 115.7 to 367 µg%, riboflavin 32.4 to 159 µg%, niacin 6.7 to 139.4 µg%, pyridoxine 71.68 to 174.1 µg% and vitamin C from 0.02 to 0.21mg%. Yeast concentration of 3% and fermentation time for 3 days was the optimized process conditions achieved with the best fit of desirability 0.92. Keywords: ashgourd, dry yeast, fermentation, rSM, vitamins 1. Introduction Ash gourd (Benincasa hispida), a member of the family Cucurbitaceae is one of the familiar crops that are grown primarily for its use as a vegetable and usually recognized for its nutritional and medicinal properties especially in Asian countries. As a rich source of functionally important bioactives and therapeutics such as triterpenes, phenolics, sterols, glycosides and soluble dietary fiber the vegetable has been widely used for therapeutic treatments. (Gupta & Premavalli, 2010; Sivarajan & Balachandran, 1994; Aslokar et al., 1992) Ashgourd is widely cultivated throughout the world and stored in the whole form that it is available all the year round. In India, ashgourd is used in curries or is coated with sugar or syrup and eaten as a sweet and recently Majumdar et al. (2009; 2010) have reported on the stabilization of ashgourd juice and ashgourd blended juice. However the fermentation process improves the quality in terms of nutrients and flavour. Therefore, an attempt has been made to develop fermented ashgourd beverage by optimization of the process with commercially available dry yeast and the study is oriented towards the analysis of nutritional profile of the product. Fermented foods provide several health benefits. Though vegetables are often used for fermentation in other countries (Yoon et al., 2006; Filomena et al., 2008) it is less commonly used in India. Vegetables being good source of vitamins and minerals, further availability can be improved by fermentation process (Campbell, 1994; Marica et al., 2007). Fermentation process is known for improving digestibility and flavour. As well Yoon, et al. (2006) showed that Fermented cabbage juice serve as a healthy beverage for vegetarians and lactose – allergic consumers. Beetroot and carrot juices were fermented with the use of brewer’s yeast autolysate by Marica et al. (2007); while 3-5% yeast with 10-12days was followed by Joshi and Attri (2005) for fermentation of apple, plum and pear juices. However, the studies on nutrient profile has not received much attention thus the fermentation conditions followed for vegetable juices and the changes in their nutrients still needs a radical approach to achieve the best quality product. Therefore in the present study, the strengthful raw material ashgourd, statistical design using RSM, an effective tool for optimization have been used with an attempt to bring out the nutritional strength of the fermentation process. Accepted: January 19, 2012 Online Published: July 2, 2012

URL: http://dx.doi.org/10.5539/jfr.v1n3p138

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and n denoted the number of independent variables i.83 17.www. The statistical software package design expert 8. dry yeast and sugar were procured from local market. Process variables their levels and experimental design Process variables -1. 0.3 Experimental Design The percentage of yeast and time of fermentation was as per the runs obtained by design expert statistical software. antioxidants (%). blanched at 90°C for 3 mins and extracted the juice using a juice extractor (RayLons Metal Works.5 to 17. The parameters that influence the product quality.8%) and fermented at room temperature for 0. satisfactory experimental results. HPLC certified solvents and M/s Sigma Aldrich. and xi. vitamin-C(mg%). B2.51 X2 ±1 0 ± 1. B3. A total of 13 level combinations (design points) were generated for the two independent variables and the alpha values in the design outside the ranges were selected for rotatability of the design.e.414 0 0 0. USA. 1.49 The regression analysis of the responses was conducted by fitting to the suitable model represented by the following equation Y  0   i X i  i 1 n  i 1 n ii X i2  i  j 1  n  ii X i X ij Where. The factorial design considered 4 factorial points. removed the rind portion. The extracted juice was processed further using sugar syrup. 2010).ccsenet. Louis. Stat . For HPLC analyses HPLC grade solvents and standards were procured from M/s SK Chemicals.17 0. All the chemical and reagents used for the analysis were AR grade. Table 1. India).5. Burdick and Jackson. 4 axial points and 5 central points leading to 13 sets of experiments (Wadikar et al. Minneapolis. 3.414 (augmented form) 5. Each independent variable investigated in this experiment had five levels which were -1.e. Variables Dry Yeast concentration and fermentation period were selected as independent variables and total phenols (mg%). at least.org/jfr Journal of Food Research Vol..1 Materials Good quality raw materials i. Bombay. Optimized ranges of the variables are shown in Table 1. was used to construct the experimental design as well as analyze the data. The center point (the level combination in which the value of each coded variable was 0) was repeated five times for the two-variable design and was selected keeping the ingredients at levels expected to yield.. acceptability and functionality were taken as responses. 2012 2. B6. No. St. -1. 2.0.total volatiles (%). and ßij were the linear. ßo was the value of the fitted response at the center point of the design. while ßi. ßii. The number of design points was obtained on basis of the number of independent variables decided. alcohol (%) and Organoleptic evaluation (score) were selected as the responses.Water soluble vitamins (µg%)-B1.414 0 -1 (factorial point) 1 3 0 (centre point) 3 9 Runs 4 2 2 5 +1 (factorial point) 5 15 +1. A Central Composite Rotatable Design was used without blocking. MN.2. Yeast was added (1. respectively.4142. acidity (%). +1 and +1. in this case n is 2. 2. and cut into pieces.5 days period.414 (augmented form) Dry yeast (%) Fermentation (Days) X1 ±1 ± 1.4142. Materials and Methods 2.Ease Inc. quadratic and interactive-effect regression terms. xij are independent variables in coded values represented by X1 and X2 in 139 . ash gourd (Benincasa hispida).2 Raw Material Processing Ash gourd was de-skinned.

The analysis for all the parameters was carried out in duplicate and the average value has been reported.4 150. Table 2.org/jfr Journal of Food Research Vol.98±0.9 5.1 222.8ml/min.8 3 3 3 3 3 3 3 3 3 3 15 15 9 9 0.2 2 7. 09 8.4 107.17 0.9 8.27±0.47 0.70 113.6 114.9 Score Y6 7.2 9 7.18 0.7 142.7 12.8 43.0 19.0 149. pyridoxine and vitamin C analyses were performed using HPLC on reverse phase C18 analytical column (Knauer.0039 0.1 12.0 18.73±0.5 71.8 29.0 31.60 mg% Y11 0.20 Note: * Over All Acceptability (OAA) scored on nine point Hedonic scale.2 4.57 0. 49 9 9 9 9 9 24.0056 0.4 129. total volatiles (AOAC. 21 7.60 129.0 4.1 139.0 17.02±0..1 43.21 0.3 24. 3.16 0. 07 7.28 0.3 22.5 Antioxi dants Acidity Total Volatile s % Y3 0. 1986).0420 0.8 150.89 37.47 0.4 0 µg% Y7 36. 4 16.19±0. 01 6. 2009).6 24.1±0.6 96.9 4.e.31 0. riboflavin. thiamine.8 32.7 137.04 0.5 32.2 20.7 46.7 210. 54 7.37 0. 1. 56 7.07 0. 2. 1970) and alcohol by HPLC on reverse phase C18 column with isocratic elution of solvent orthophosphoric acid and water at 0.70 66.14 0.1 251.8 4.91±0.06 0. No X1 X2 Total Pheno ls mg% Y1 1 5 1 5 0.0 µg% Y9 6.19 0.0025 0.0 223.00 76.1 32. 21 7.8 17.48 0.47 % Y4 0.1 87.7 159.4 132.7 35.50 166. The samples were analysed immediately after the completion of the fermentation process.0190 0.6 9.8 37.3 29.07±0.1±0.ccsenet.5 79.0029 0.2 130.5 1 17.2 2 7. Water Soluble Vitamins i.7 133.2±0.70 132.7 15.3 6 7.2 17.38±0.18 0.0 218.9 4. After fermentation for varied periods the respective samples were filtered in muslin cloth immediately and centrifuged to get a clear juice.1 174. 140 .50 32. Actual experimental combinations and response values Exp.61 0.0 281. No.48 0.1 7 5.3 27.5 212.4 Analytical Evaluation The fermented ashgourd juice was analysed for total phenols and free radical scavenging capacity (DPPH activity) (Sandra et al.www.19 0.6 4.61 18.0420 0.40 66.9 µg% Y8 153. 2010).10 0.2 30.2±0.5 166.8 27.1±0.6 5. 1 15. Then the bottles were sealed and pasteurized for 15mins at 90ºC.1 216.5 64. 11 7..40 140.5 24.0780 0.0053 0.6 % Y5 8. niacin.46 0.0045 0.7 31. acidity (Ranganna. with UV detector (215nm).04 0.7 115.0051 0.0089 0.2 µg% Y10 95.02 0.0059 Alcoh ol OAA* Thiami ne Riboflav in Niaci n Pyridoxi ne VitaminC % Y2 14.40 92.46 20. 2012 Table 2.4 7.4 14.

03*** 0.39**** 27.34 -22.024 17.766 101.72*** 5.048** 0.6%.32 <0.38 33. Samples were randomly drawn for each experimental block. Results and Discussions The experimental central composite rotatable design with independent variables and responses is given in Table 2.8 to 43.2**** 8.92 0.042* 0.022**** -0.ccsenet.91 for B2 respectively indicating that all the values were more than 90%.1325*** 99.02 to 0.034 12767.7* 98.0008 0.93 6.99 <0.org/jfr Journal of Food Research Vol.85**** 1.50**** 2. coded and served to the panelists.01 110.4.067 to 0. The regression coefficient. riboflavin 32. Since overall acceptability is an important criterion for the product acceptance.001.www.013 -0.69 -8.95 -8.47 0. 1.936*** 0. Coefficient of second order polynomial regression models Coefficient Total Phenols mg% Y1 β0 β1 β2 β11 β22 β12 R2 % p% Mean SD F-value 31.09**** 0.84 <0. it has been taken as one of the responses.41 <0. 2010).63 0. All main. taste.017**** -0.73 µg% Y9 22.6 0.069*** 0.0001 19.0066 27. Over the 13 combinations total phenols ranged from 24.139**** -0. 3. but because of the subjective nature of the response the model is considered significant. alcohol 4.22 -3.48**** -29.6-12.67 % Y3 0. Table 3.07**** 99. B3 and 0.7 -3.9**** 2.18 0. linear. Negative sign of a coefficient at linear level indicates decrease in response with an increase in level of the variable where as at interactive level. overall acceptability. The adequacy was calculated by F-ratio.11 µg% Y10 101.275 99.0015 0.0046 2868. ** p<0. antioxidants.02 0. aroma.49**** 1.95 6.283 681. Overall acceptability 6. acidity 0.40 0. 0.9 to 8. acidity.188 577.16 <0.7* 10. consistency and overall acceptability using a nine-point hedonic scale (Majumdar et al. 2012 2.515** 99.4 µg%.0001 213. Quadratic response surface models were selected for all the responses.7 0.016*** 0.93 for vitamin C.58**** 99.033** -0.79**** 1.0001 44.95 for total volatiles.0001 0.53 75.18 0. 141 .54 % Y5 4.13 -26.7**** 1.62 <0.02 -44.008 -0.392 14.012 -0.7 to 139. quadratic and interactive effects were calculated for each model.89 µg% Y7 216. level of one variable could be increased while that of the other decreased to obtain the same response. alcohol.39**** -0.8**** -0.001 0. No.33 0.7 to 367 µg%. The R2 value for B6 was 0.0001 6.025**** 99.0025 to 0. pyridoxine 71.6mg%.57 Alcohol OAA* Thiamine Riboflavin Niacin Pyridoxine Vitamin-C Note 1: Significance: **** p>0.01.34**** 0.078%.76.01 23.13 0.31**** 1.0001 33.7** -6. Note 2: * Over All Acceptability (OAA) scored on nine point Hedonic scale. B1. R2 value was more than 90% and lack of fit was highly non-significant.69 83.21mg%.7*** 91.0025 92. mean.8 42.63 <0.76 <0.95 % Y2 17. thiamine 115. 3. The effects of the independent variables yeast and fermentation time on the response at linear.68 to 174.20 15. 0. antioxidant 12.95 <0.48 Antioxidants Acidity Total Volatiles % Y4 0.043 238.1 10.001 0.8** 53. * p<0. total volatiles 0.47 0.99 for total phenols.3 to 27%.054 4.05.1**** 0.14 0.87**** 0.. niacin 6.384 563.05 118.75 <0.45 µg% Y8 131. *** p<0.0015 95.0001. quadratic and interactive levels are presented in Table 3. correlation coefficient for the responses were 0.3**** 19.1 µg% and vitamin C from 0.61 4. The sign and magnitude of the coefficients indicate the effect of the variable on the responses.4%. coefficient correlation and lack of fit test. standard deviation.5 Organoleptic Evaluation A Semi trained panel evaluated the samples for color.9 Score Y6 7.105**** 0.39 mg% Y11 0.0001 7.75 <0.4 to 159 µg%.

500 1. 3.500 1. Abbas (2006) reported that yeasts can also synthesize a number of bioactive compounds which can serve as antioxidants.113 X22 + 0.000 -0.735 X12 + 1. Yeast and fermentation time has positive significant effect (p<0. No.00 45 Perturbation 40 A B 35 Total Phenols 30 B A 25 20 -1.001) positive effect on total phenols and antioxidants.275 X1 X2 Antioxidants = 17. As yeast concentration increased total phenols increased and as fermentation increased there was increase in antioxidant level of beverage.000 -0. 1. Table 3 reveals that at linear level. Total Phenols Design-Expert® Software Factor Coding: Actual Antioxidants Actual Factors A: Yeast = 3. Design-Expert® Software Factor Coding: Actual Total Phenols Actual Factors A: Yeast = 3.00 B: Fermentation = 9.000 Deviation from Reference Point (Coded Units) 1a.506 X1 + 2.99 The response surface for total phenols and antioxidants in relation to yeast and fermentation time is shown in Figures 1a and 1b respectively.96 + 6. Antioxidant Figure 1.719+ 0.org/jfr Journal of Food Research Vol. 2012 3.01) on antioxidant level at interactive level.500 0.487 X2 + 1. glutathione and tocopherols with interesting antioxidant properties while Cruz et al. (1999) reported that the formation of these substances can be induced in yeasts grown under stressing conditions or in response to fermentation medium ingredients such us phenolics or additives that are known to be toxic to cells grown aerobically.515 X1 X2 R2 = 0.000 0. Perturbation graphs depicting effect of independent variables on total phenols and antioxidant levels of the fermented beverage 142 . The multiple coded equations in terms of coded factors generated for their responses are shown below: Total Phenols = 31. Several Candida and Saccharomyces species produce compounds such as caroteniods.www.859 X2 + 1. citric acid.936 X22 + 0.725 X1 + 5.0001 for antioxidants and p<0.00 B: Fermentation = 9.038 X12 + 0.99 R2 = 0.000 0.001 on total phenols.500 0.00 30 Perturbation 25 B Antioxidants 20 A A 15 B 10 -1.ccsenet.000 Deviation from Reference Point (Coded Units) 1b. yeast and fermentation time had a significant (p<0.1 Effect of Variables on Functional Parameters Total phenols and antioxidants were considered as functional parameters for the independent variables. At quadratic level both yeast and fermentation time a positive highly significant level at p<0.

1 8 8 8.0001) positive effect on acidity and alcohol.14 + 0. Querol and Fleet (2006) reported that in wines.0001 and on total volatiles at p<0.878 X12 + 0.99 Figures 2a to 2d represents the change in quality parameters of fermented beverage with respect to dry yeast concentration and fermentation time.9 7.9 7.9 7.00 Colour 8.793 X1 + 1.9 7.77 0.0934 X1 + 0. At linear level yeast and fermentation time had a significant (p<0. consistency and colour is given in Table 4.org/jfr Journal of Food Research Vol.9 7.81 0. alcohol and OAA at p<0.99 R2 = 0.8 7. Lopez et al.6 7.99 R2 = 0.00 0.8 7. Table 4. n=10 Sl.80 0.139 X2 – 0.0001).9 + 0. The multiple coded equations in terms of coded factors generated for their responses are shown below: Acidity = 0.6 7. taste.8 7. Yeast had positive effect on alcohol. 1. on overall acceptability at p<0.8 7.3465 X22 + 0.14 Taste 7. However.0.001.012 X1 – 0.1 8 8 8.474 + 0. It is clear from table that fermentation time influenced all quality parameters.8 7. (2008) also showed a positive effect of fermentation process by confirming that yeast are present throughout the fermentative process and it is generally accepted that they can produce compounds with important organoleptic attributes determining the quality and flavour of the final product.6 7. Increase in yeast concentration increased acidity and alcohol.9 7.9 7.8 7.0015 X12 + 0.12 Note: * Overall acceptability.8 7.0001) positive effect on acidity.58 X1 X2 Overall Acceptability = 7.001 on OAA. Total volatiles. Variables had significant (p<0. while total volatiles was not significant.9 7. Sensory score of optimized composition.025 X1 X2 Total Volatiles = 0.8 7.0082 .05 0. while yeast had negative effect on total volatiles.0224 X2 – 0.0.2 Effect of Variables on Quality parameters Acidity.2 8.319 X2 + 1. 143 .9 7.0175 X12 .9 7. No 1 2 3 4 5 6 7 8 9 10 Mean SD ± Flavour 7.1 7.www.1056 X22 + 0.0001.6 7.08 OAA* 7.397 X2 – 0. overall acceptability. alcohol and p<0. alcohol and OAA had positive effect while acidity had negative effect. No.ccsenet. 3.9 7. among the 1.0001) positive effect on total volatiles and overall acceptability was negatively affected (p<0. while acidity and OAA had negative effect and total volatiles was not significant. Fermentation time affected acidity.0165 X22 + 0. more than 400 are produced by yeasts. Effect of independent variable on sensory attributes such as flavour.0015 X1 X2 Alcohol = 4.5 7.1325 X1 X2 R2 = 0. 2012 3.6 7. alcohol and total volatiles were selected as quality parameters. fermentation time had a significant (p<0. alcohol were significant at p<0.6 7. As fermentation time increased there was increase in alcohol and total volatiles where as OAA decreased.8 7. no significant effect of yeast had seen on total volatiles and overall acceptability.907 X22 + 2.9 7.05.95 R2 = 0.0429 X12 + 0.1 8.9 8. Esters produced by yeasts can contribute to both aroma and flavour and several factors can contribute to aroma production by yeasts. At quadratic level yeast effect on acidity.0008 X1 + 0.9 7.13 Consistency 8 8 8 8 8 8 8 8 8 8 8.8 7.000 volatile compounds identified.

01) at quadratic level. pyridoxine (vitamin B6) and pantothenic acid.00 1. 1.6 10 Acceptability 7.500 0.000 B: Fermentation A: Yeast Deviation from Reference Point (Coded Units) 2a.03 0.00 3.00 2. No.000 -0.2065 X12 + 8.9514 X1 – 8.3945 X2 + 27.99 R2 = 0.0331 X12 – 0.6992 X22 + 83.6925 X2 – 8.141 + 10.500 1.3 Effect of variables on nutritional parameters Viljoen (2006) reported that yeasts appear to be active microorganisms synthesising substances such as vitamins.500 1.www.0480 X22 + 0.001) positive effect on vitamin C at linear level. Yeast concentration significantly (p<0.000 -0.7271 X22 + 10.180 + 0. while riboflavin was insignificant at linear level and fermentation time affected significantly (p<0.2205 X12 – 3.00 9. Niacin was highly significant (p<0.99 R2 = 0.0025 X1 X2 R2 = 0.00 4.0001) negative effect on linear model and p<0.91 R2 = 0.0001) at linear level and was insignificant at quadratic level. 2012 Design-Expert® Software Factor Coding: Actual Acidity Design points above predicted value Design points below predicted value 0.00 6.7525 X1 X2 Niacin = 22.00 -0.6 0.2 A B 0 A 0. The model obtained from the regression can be written as follows for nutritional parameters.00 4.00 6.ccsenet.00 9.00 12.000 B: Fermentation A: Yeast Deviation from Reference Point (Coded Units) 2c.00 B: Fermentation = 9.000 0.500 0. Alcohol 2d.76 R2 = 0.8 -1.00 2.05 on quadratic model. Yeast concentration had a significant (p<0.80 + 42.075 X1 X2 Riboflavin = 131. Thiamine = 216. Total Volatiles Design-Expert® Software Factor Coding: Actual Acceptability Actual Factors A: Yeast = 3.73 – 3. it can be concluded that thiamine had significant (p<0.01 on B6 at 144 .4 4.00 3.6 14 2b. Yeast concentration and fermentation time had negative effect on thiamine and riboflavin at linear and quadratic level.00 12.00 6.0001) affected niacin at quadratic level and fermentation time significantly affected at p<0. Overall Acceptability Figure 2. or breakdown complex carbohydrates which will have an impact on product.4 0.8 X1 = A: Yeast X2 = B: Fermentation 12 7.5 Total Volatiles 0.00 5.01 -1.000 0.7 Design-Expert® Software Factor Coding: Actual Total Volatiles Actual Factors A: Yeast = 3.04 X1 = A: Yeast X2 = B: Fermentation 0.4811 X1 – 29.6268 X1 + 0.3 0.3157 X1 + 19.03 – 44.4 Alcohol 8 6 7.00 8 Perturbation B 7.8010 X2 – 0. amino acids and purines.00 3. 3.00 B: Fermentation = 9.067 0.93 From the above equation.0609 X1 + 0. p<0.8461 X22 + 53.2 4 7 A A B 15.6575 X1 X2 Pyridoxine = 101.05.3855 X2 + 33.02 Acidity 0.0133 X2 – 0.00 0.00 3. Thiamine affected significantly (p<0. nicotinic acid.1 15.53 X1 X2 Vitamin C = 0.00 1.org/jfr Journal of Food Research Vol.7317 X12 – 6. Abbas (2006) also reported that among the vitamins and other enzyme cofactors that are accumulated and/or synthesized by yeast are thiamine (vitamin B1).1393 X12 – 26.01 0. Perturbation graph and 3D plots depicting effect of independent variables on Quality Parameters of the fermented beverage 3.00 5.05 Perturbation B 0.3412 X22 – 22. Acidity Design-Expert® Software Factor Coding: Actual Alcohol Design points above predicted value Design points below predicted value 12.0001) at linear level.61 0.

000 -0.000 -1.00 6.ccsenet. It is clear from figures 3c to 3e.00 5.00 2. niacin at p<0.500 0.00 2. Riboflavin Design-Expert® Software Factor Coding: Actual Niacin Actual Factors A: Yeast = 3. there was increase in niacin.7 400 Design-Expert® Software Factor Coding: Actual Riboflavin Design points above predicted value Design points below predicted value 159 32.00 12.05 while time had no significant effect on pyridoxine and Vitamin C retention.001.00 B: Fermentation A: Yeast B: Fermentation A: Yeast 3a.00 B: Fermentation = 9.05 0 15.00 15. Design-Expert® Software Factor Coding: Actual Thiamine Design points above predicted value Design points below predicted value 367 115.00 12.00 4.00 2.00 B: Fermentation = 9.00 3.15 Vitamin C 0.00 1.000 Deviation from Reference Point (Coded Units) Deviation from Reference Point (Coded Units) 3c.00 140 Perturbation Design-Expert® Software Factor Coding: Actual Pyridoxine Actual Factors A: Yeast = 3.21 0.00 1. riboflavin at p<0.00 1. At interactive level.00 5.000 -0.00 5. 1.00 6.00 6.1 0.00 B: Fermentation A: Yeast 3e.000 0. as there is increase in yeast concentration. Thiamine 3b.org/jfr Journal of Food Research Vol. pyridoxine and vitamin C where thiamine and riboflavin were not influenced much.00 3.4 180 X1 = A: Yeast X2 = B: Fermentation 350 X1 = A: Yeast X2 = B: Fermentation 160 140 300 120 100 250 Riboflavin Thiamine 200 80 60 40 20 150 100 15. Niacin Design-Expert® Software Factor Coding: Actual Vitamin C Design points above predicted value Design points below predicted value 0.00 9.00 3.25 3d.00 9.500 0. thus the fermentation time has great influence on vitamin profile.00 3.2 0.500 1.0001.00 3. Perturbation graph and 3D plots depicting effect of independent variables on Nutritional Parameters of the fermented beverage 145 .00 12.00 9.01) negatively on vitamin C at quadratic level.000 0. 3.02 0.00 180 Perturbation 120 160 100 A A 140 80 Pyridoxine Niacin A 120 60 B 100 40 B B 20 80 B A 0 60 -1. No.500 1. 2012 quadratic level while it was significant (p<0. Vitamin C Figure 3. yeast and fermentation time affected the thiamine significantly at p<0. With increase in fermentation time there was decrease in B vitamins.00 4.www.00 4.00 3. Pyridoxine X1 = A: Yeast X2 = B: Fermentation 0.

DC: AOAC International. (1994).1 149 0. Food Research International. Using the optimized condition the prepared beverage from fermentation process contained more total phenolics.36 12. antioxidants. In Querol. K. Predicted and actual response values Responses Total Phenols Antioxidants Acidity Total Volatiles Alcohol OAA Thiamine Riboflavin Niacin Pyridoxine Vitamin C 4. P.org/jfr Journal of Food Research Vol. F values were significant in all selected responses and a high R2 value of >90% showed fitness of the polynomial regression models for describing the effect of variables.22 0. Conclusion RSM showed that quadratic response surface models were fitted. aromas. it meets the PFA specifications under the food category.1 and overall acceptability score 7. The results of the study indicated that the effect of yeast concentration and fermentation time were significant to all the selected responses and RSM could be useful in optimizing the yeast concentration and fermentation time with maximum retention of vitamins and minimum amount of alcohol formation. Sensory score of optimized composition is given Table 4.).11mg%.8. L. H.doi. B2. K.0. OAA 7. http://dx. acidity 0. The predicted response value as against actual value for responses as shown in Table 5 were in concurrence with each other. hence the similar fitted models are suitable for predicting the responses. 2012 3.1016/0963-9969(94)90093-0 146 Predicted 29. 253-257. Production of antioxidants.92. A S Bawa.6 0.11 Actual 28. consistency 8.0025 4. New Delhi: CSIR Part I. pp.4 Optimization of Independent Variables Numerical optimization of independent variables.6 7. B6 and Vitamin C when compared with fresh ashgourd juice.22. colour 8. A. The aim of the experiment was alcohol content was minimized and OAA was increased.). Yeasts in Food and Beverages. B6 149 µg% and vitamin-C 0. Table 5.9 255. B3 51. (Eds.6 50. Acknowledgement The authors sincerely thank the Director Dr.7 7. V. taste 7. and vitamins by yeasts. Since the alcohol content is within 5%.doi. B2 119. O. Campbell.34%. (1st ed. flavours.9 140 0.17 . & Chakre.www.. Aslokar. DFRL. The criteria used along with the predicted and actual values of the response are given in Table 5.0025%.org/10. Kakkar. J.34 0.1 µg%.1007/978-3-540-28398-0_10 AOAC (1970).org/10.7 119. (1992). B3.21 0. Mysore for his support and encouragement. (2006).8 244. C.9. 1. http://dx. 3. Fermented foods – a world perspective. alcohol 4..6 µg%.0020 4. No.4 mg%.7µg%. The optimized results were total phenols 29. References Abbas. total volatiles 0. colours.8. antioxidants and total phenols with 3% dry yeast concentration and 3 days fermentation time.ccsenet. 285-334.6 78. Vitamin B1. Glossary of Indian medicinal plants with active principles. & Fleet.38 13. G. 27. Overall the fermentation time and yeast concentration is optimized with maximum retention of vitamins. antioxidants 13. A.6 51. The optimized fermented beverage had flavour 7. yeast and fermentation time were optimized using Design Expert Software.8.7%.. vitamin B1 255. The solution which was obtained was 3% yeast and 3 days fermentation time with best fit desirability of 0. Official methods of analysis. Washington.

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