For Official Use Only

RPF-III Final Report

Studies on alternate solvent for extraction of oil from soybean and its effect on the nutritional quality of the meal

Project No: Institute Code No: ICAR Code No: Period: Investigators(s):

Dr.A.P.Gandhi
Principal Scientist (Biochemistry)

Dr.Krishna Jha
Principal Scientist (Microbiology)

Dr.K.C.Joshi
Senior Scientist (Biochemistry)

Soybean Processing and Utilization Center Central Institute of Agricultural Engineering Bhopal-462038.

RPF-III
1. 2. 3. 4. 5. 6. 7. Institute Code No: ICAR Code No: Name and address of the Research Institute: Central Institute of Agricultural Engineering, Bhopal. Title of the project: Studies on alternate solvent for extraction of soy oil and its effect on nutritional quality of the meal Title of the sub project: Nil Name and designation of the leader: Dr. A.P.Gandhi Principal Scientist (Biochemistry) Name(s) and designation(s) Project Leader and associate(s) together with time proposed to be spent and work to be done Sl.No 1 Name Dr.A.P.Gandhi Designation Principal Scientist (Biochemistry) Principal Scientist (Microbiology) Senior Scientist (Biochemistry) Time to be spent 25% Work to be done Implementation of the project, data collection, analysis and report writing. Experimentation, data analysis and preparation of reports. Experimentation, data analysis and preparation of reports.

Dr.Krishna Jha

25%

Dr.K.C.Joshi

25%

8.Location of the research project with complete address: Soybean Processing and Utilization Center, Central Institute of Agricultural Engineering, Bhopal. 9.Period: 3 Years, October 1998 to September 2001. 10.Objectives: i. To optimize the parameters for extraction of oil from soybean using different solvents like n- heptane, iso-propanol, n-propanol, ethanol and mixtures there of along with presently used n-hexane. ii. To study the quality of oil before and after refining. iii. To assess the nutritional quality of solvent extracted meal and its value addition.

iv. To analyze the techno-economic feasibility of the alternate solvent system. 11. Detailed Progress Report: n-hexane is normally used as a solvent for extraction of oil from oil seeds. But due to its concern on the health, toxicological and the environmental, a search is on for identifying the alternate solvent, which may be as effective as n-hexane and cause minimum health hazards. n-hexane may enter the body both by inhalation and skin absorption, even though the absorption is low both ways. Acute exposure to vapors leads to various symptoms like heaviness, nausea, headache, numbness, muscular weakness, irritation of eyes and nose, dermatitis, chemical pneumonia, giddiness etc. Death may occur at a concentration of 5000 ppm. The threshold limit for the hexane vapor as recommended by ISO is 50 ppm where as BIS prescribes 170 ppm. The operating losses of n-hexane ranges between one liter and ten liters per ton of seed processed. The current search was prompted by the 1990 Amendments to the Clean Air Act, which listed nhexane as a hazardous pollutant. This legislation caused a great concern in the oil seed extraction industry. Hence there is a need for a better solvent, which gives better oil extraction and simultaneous removal of undesirable compounds. Johnson and Lusas (1983) reviewed the alternative solvents for oil extraction. Based on the information available the following solvents were selected for study: n-heptane, iso-propanol, n-propanol, ethanol and their azeotropic mixtures. These are based on the following characters: Plentiful. Non-toxic to workers. Ability to use in the existing equipment. Besides extraction of triglycerides they should remove toxins, beany flavours, etc. hence the quality of the meal may be upgraded. Less ability to extract phosphotides. Easily recovered. Stability of oil due to extraction of more tocopherols. Non-reactive to equipment. High purity. Relative high boiling points hence few losses.

The chemical and physical properties of the selected solvents are given as under:
Solvent Boiling point,oC Latent heat of vaporization, cal/g Sp.heatcal/g/oC Sp.gravity at 20oC Viscosity, cp at 20oC Surface tension, dyne/cm Dielectric constant Flash point.0C Explosive limits, %vol in air Water solubility/l

Water nhexane nheptane Ethanol Isopropanol npropanol

100 69 98.4 78.3 82.5 97.2

540.1 79.9 75.5 204 159.3 162.6

1.018 0.533 0.528 0.61 0.596 0.586

1.000 0.659 0.684 0.785 0.786 0.805

1.005 0.312 0.417 1.22 2.40 2.256

72.75 18.40 20.14 22.10 20.80 23.8

80.36 1.89 1.92 25.7 18.60 20.1

NF -23 -4.0 12 11.70 22.00

NF 1.2-7.7 1.0-7.0 3.319.0 2.512.0 2.613.5

0.05 0.05 NF NF NF

Solubility Curves: The miscibility data for soybean oil in different solvents is required to obtain the critical solution temperature. This data will be useful in the design of an extraction process. Dynamic method was used. The details are as follows: Methodology: Known volumes of oil and solvent in the ratio of 30:20 were taken so as to provide excess oil at fixed temperatures and stirred for 30 minutes. The stirring was stopped and allowed the mixture to settle for 30 minutes. About 10 ml of the clear supernatant solution was drawn out into already weighed conical flasks and weighed again to get the weights of solution drawn. The solvent in the solution was evaporated on a vigorously boiling water bath and after its evaporation; the flask was dried and weighed to a constant weight. From the above data the percentage solubility was calculated. The commercially available refined soybean oil was tested for its solubility in different solvents like n-hexane, heptane, n-propanol, iso-propanol and ethanol. The temperature tested was 0oC, 10oC, 20oC, 30oC, 35oC and 40oC for all solvents and in case of ethanol up to 70oC was tried. From the data the percentage solubility was calculated. The results are presented as under.

Solubility of soybean oil in different solvents: The data are given in Table 1. Table.1 Solubility of soybean oil in solvents Temperature.oC 0 10 20 30 35 40 50 70 n-hexane Miscible Solubility % in Solvents n-heptane Iso-propanol Miscible 7.43 21.81 45.18 66.62 82.62 Miscible Ethanol Immiscible Immiscible Immiscible 4.2 11.2 20.4 40.7 Miscible

The results indicate that the soybean oil is soluble even at low temperatures in n-hexane and n-heptane. In case of isopropanol the solubility was 7.43% at 0oC and increased with increase in temperature. It was completely soluble at 40oC.The oil was immiscible in ethanol up to 20oC and later increased with increase in temperature. It was completely soluble at 70oC.The results thus indicate that the oil extraction can be selectively done with different solvents. The optimum temperature in the case of ethanol was 70oC, nearer to its boiling point. In other solvents like hexane and heptane the oil is soluble even at low temperatures inferring there by that the cold extraction can also serve the purpose. However the time taken may be longer in this case. In all the cases the solubility of oil increases steadily until the critical temperature is reached. At critical temperature any amount of oil can be dissolved. Oil and solvent are completely miscible in all proportions at or above that temperature. Solvent Extraction Studies: The soybean oil was extracted using different solvent systems. The flake thickness namely 0.3mm, 0.6mm and 1.2mm were chosen for studies. The period of extraction was up to 8 hours in traditional soxhlet apparatus. The percentage of oil extraction was calculated at 1h, 2h, 4h, 6h and 8h. n-hexane: The data is presented in Table 2. Table2 Oil extraction with n-hexane Hours of extraction 1 2 4 6 8 0.3mm 77.0 85.0 90.0 98.7 99.5 % of oil extracted 0.6mm 75.0 84.0 89.5 98.0 99.1 1.2mm 73.0 78.0 86.0 96.5 99.0

The data indicated that the flake thickness effect the extraction of oil. With increase in flake thickness the percentage of oil extraction decreased. It is more with 0.3mm. The oil extracted also increased with the period of extraction. In about 6-8 hours on lab scale experiments approximately 99% oil was extracted. n-hexane had the capacity to extract 77% of oil in first one hour of extraction with 0.33mm thick flakes. Iso-propylalcohol: The results are given in Table 3. Table 3 Oil extractions with iso-propylalcohol Hours of extraction 1 2 4 6 8 0.3mm 73.0 89.0 94.0 98.0 99.5 % of oil extracted 0.6mm 69.0 87.0 92.5 95.0 99.2 1.2mm 47.0 63.0 87.5 93.1 98.9

The data indicate that isopropyl alcohol extracts 73% oil in the first hour of extraction with 0.33 mm flakes. In about 8 hours 99.5% oil was extracted. The thickness of the flakes effected the oil extraction in this case also. n-heptane: The data are presented in Table 4. Table 4 Oil extractions with n-heptane Hours of extraction 1 2 4 6 8 0.3mm 70.0 82.0 92.0 98.0 99.0 % of oil extracted 0.6mm 66.0 74.0 84.0 96.0 98.2 1.2mm 64.0 72.5 81.0 95.0 99.0

The results show that in the first one hour of extraction 70% of oil was obtained. It increased with the time. In 8 hours 99% was extracted.

Ethanol: The results are shown in Table 5. Table 5 Oil extractions with ethanol % of oil extracted 0.3mm 0.6mm 1.2mm 1 71.0 68.0 67.0 2 84.0 76.0 75.0 4 92.0 86.0 84.0 6 98.0 98.0 94.0 8 99.0 99.5 99.2 The results reveal that the ethyl alcohol also effectively extracts the oil from soybean. It increases with the time. The thickness of the flakes also influences the system. n-propanol: The results are given in Table 6. Table 6 Oil extractions with n-propanol Hours of extraction 1 2 4 6 8 0.3mm 70.0 81.0 86.0 90.0 98.0 % of oil extracted 0.6mm 68.0 78.0 80.0 89.5 97.0 1.2mm 65.0 71.0 78.0 88.0 96.8 Hours of extraction

The data indicate the possibility of the oil extraction with n-propanol. Absolute solvent system: Experiments were conducted with absolute n-hexane and n-heptane at different boiling points till their respective optimum boiling points and the oil yields were measured during 8 hours continuous operation. The results are presented as under. n-hexane: The extraction temperatures were 50oC, 60oC and 70oC. The results are shown in Table 7. Table 7. Extraction of oil with absolute n-hexane Temperature,oC 50 60 70 2 55.0 77.0 83.0 % oil extracted Hours of extraction 4 6 60.0 68.0 84.0 92.0 90.0 97.8

8 82.0 94.7 99.5

The data indicate that the maximum extraction of 99.5% was obtained in 8 hours of extraction at 70oC and the extraction rate increased with the temperature and also the duration of extraction n-heptane: The data on the extraction at different temperatures are given in Table 8. Table 8 Extraction of oil from soy flakes with absolute n-heptane Temperature,oC 60 70 80 90 100 2 61.0 63.0 65.0 70.0 78.0 % oil extracted Hours of extraction 4 6 68.0 72.0 70.0 78.0 73.0 80.0 78.0 85.0 84.0 91.0

8 81.0 88.0 90.0 96.0 99.0

The results clearly indicate that the maximum extraction of 99 % was obtained at 100oC in 8 hours. Aqueous solvent system: Different solvent mixtures in combination with water were prepared at 10% and 20% level. The extractions were carried out at different temperatures and the oil recovery was estimated. Ethanol: The results with ethanol are presented in Table 9. Table 9. Oil recovery with aqueous ethanol Temp,oC 50 60 70 80 % oil recovered 90:10(Solvent:Water) 80:20(Solvent: Water) 2h 4h 6h 8h 2h 4h 6h 56.0 58.0 70.0 74.0 53.0 60.0 65.0 60.0 68.0 80.0 84.0 57.0 70.0 77.0 72.0 76.0 87.0 92.0 64.0 75.0 83.0 76.0 81.0 90.0 97.0 71.0 78.0 86.0

8h 71.0 82.0 91.0 95.0

The data indicated that the extraction of oil increased with the increase in temperature till the boiling point was attained. With more water incorporation the rate of extraction was slowed down and also affected the oil recovery at the termination of run. About 95-97% of extraction was achieved at 8 hours run.

Iso-propanol: The results are shown in Table 10. Table 10 Oil recoveries with aqueous iso-propanol Temp, oC 50 60 70 80 2h 57.0 62.0 68.0 80.0 % Oil recovered 90:10(Solvent: Water) 80:20(Solvent: Water) 4h 6h 8h 2h 4h 6h 60.0 71.0 74.0 53.0 58.0 61.0 78.0 82.0 86.0 56.0 72.0 78.0 80.0 87.0 93.0 67.0 77.0 84.0 86.0 92.0 97.8 72.0 82.0 90.0

8h 68.0 85.0 89.0 94.0

The results also agree with the earlier observations with ethanol. With 10% water incorporation the maximum oil recovery was 97.8% as compared to 94% with 20% water in the solvent. With the increase in temperature the oil recovery was also increased.

n-propanol: The experiments were also conducted with n-propanol and the results are presented in Table 11. Table 11 Oil recovery with aqueous n-propanol Temp,oC 60 70 80 90 100 2h 54.0 57.0 61.0 69.0 78.0 % Oil recovered 90:10(Solvent: Water) 80:20(Solvent: Water) 4h 6h 8h 2h 4h 6h 58.0 61.0 78.0 50.0 55.0 57.0 65.0 69.0 85.0 55.0 60.0 64.0 69.0 76.0 88.0 58.0 62.0 69.0 74.0 80.0 91.0 65.0 69.0 72.0 82.0 85.0 96.0 72.0 77.0 80.0

8h 67.0 77.0 82.0 89.0 92.0

The data clearly indicate that the rate of extraction increases with the increase in time of extraction and also temperature till its boiling point. The data further indicate that the boiling points of water solvent mixtures vary as compared to their absolute solvent counter parts. Azeotropic mixtures: Another set of experiments was conducted with azeotropes of these solvents. Azeotrope is a constant boiling mixture that distills with out change in composition. These azeotropes thus prepared have boiling points below that of either component. The details are given in Table 12.

Table 12 Azeotropes prepared from the solvents under study Components n-heptane Ethanol n-hexane Ethanol n-hexane IPA n-hexane n-propanol IPA Water Ethanol Water Water n-propanol Boiling point,oC 98.4 78.5 69.0 78.5 69.0 82.3 69.0 97.2 82.3 100.0 78.5 100.0 100.0 97.2 Azeotropic b.p. oC 71.8(72.0) 58.4(58.7) 60.8(61.0) 65.1(65.7) 79.9(80.4) 75.7(76.2) 87.8(88.1) Composition, wt% 52.0 48.0 79.0 21.0 78.0 22.0 96.0 4.0 87.8 12.2 95.6 4.4 28.2 71.8

The boiling points of so prepared azeotropic mixtures were determined in triplicate and the average values were reported here. The figures in parentheses indicate the standard reported values in the literature. Slight differences observed in the b.p of these azeotropes determined at this place may probably due to the difference in the atmospheric pressure. The soy oil was extracted with the above azeotropes and the data are presented in Table 13. The extractions were conducted up to 8 hours in most cases and in some up to 10 hours till the maximum oil was recovered. The oil yields were recorded at 2h, 4h, 6h, 8h and 10 h . The data indicate that all azeotropes were efficient in extracting oil. Table 13 Soy oil recovery with different azeotropes Azeotrope n-hexane+IPA nhexane+ethanol nheptane+ethanol n-hexane+npropanol Water+ethanol Water+npropanol IPA+water Boiling point,oC 61.0 58.7 72.0 65.7 78.2 88.0 80.0 2 64.0 52.0 64.0 68.0 54.0 57.0 56.0 % Oil recovery Period of extraction, hours 4 6 8 72.0 89.0 99.5 65.0 82.0 96.0 72.0 71.0 65.0 65.0 68.0 84.0 85.0 81.0 84.0 85.0 94.0 97.0 95.0 96.0 94.0

10 99.0 99.0 99.6 99.0 99.0 99.8

The above data clearly indicate that the azeotropes were efficient in extracting oil but took longer period (10 hours in most cases). The main advantage with them was lower boiling points as compared to their respective solvents hence less energy requirement in the overall process. The amount of solvents required were also less so that the residual solvent levels in the meal might be reduced to a considerable level. Water with IPA was the best, which yielded 99.8% oil recovery in 10 hours. Quality of soy meal and oil from different solvent extractions: Absolute solvent systems: Meal: The quality of the meal obtained from different solvent extraction studies was analyzed and presented in Table 14. Table 14 Quality of soy meal Parameters Moisture,%(w.b) Protein, % Fat, % Ash, % Carbohydrates, % Total bacterial counts,/gx103 Total fungal counts, /gx103 Salmonella Coliforms Residual solvent, ppm TI, mg/g Urease, change in pH units NSI, % Fiber, % n-hexane 6.25 52.2 0.25 5.1 36.25 1.2 0.7 100 2.0 0 19 3.5 n-heptane 6.81 50.2 0.45 5.24 37.50 0.77 0.77 78 1.4 0 6 3.3 Solvent n-propanol 6.7 51.0 0.32 5.30 36.68 0.67 0.67 69 1.3 0.19 8 3.8 IPA 6.5 52.81 0.20 5.40 34.84 0.45 0.45 120 1.2 0.13 11 3.6 Ethanol 6.6 50.12 0.50 5.20 37.58 0.2 0.2 20 1.3 0 13 3.5

The data show that the soy meal has about 50.12 to 52.81% protein. It has less than 0.5% oil in most of the cases indicating that all the solvents extracted the oil effectively. The microbial counts were with in the permissible limits of BIS. The anti nutritional factors were at the minimum. Almost 90% destruction of TI was achieved. The NSI values of the meal before desolventization were in the range of 40-58% and after desolventization they were in between 6.0-19.0 indicating the severity of the heat treatment. The higher the NSI values the lower was the denaturation of proteins.

Oil: The oil obtained from these solvent extractions were analyzed for various characteristics before and after refining. The results are given in Table 15. Table 15 Characteristics of soybean oil from absolute solvents Characteristics n-hexane Crude Phosphorus, ppm Peroxide value, meq/kg FFA, % Neutral oil, % Total volatile matter, % Water, % Refined Phosphorus, ppm Peroxide value, meq/kg FFA, % Color Copper, ppm Iron, ppm 413 2.0 0.18 97.7 0.51 0.19 <1 0 0.05 Colorless 0.003 0.04 n-heptane 300 2.0 0.18 99.4 0.52 0.1 <1 0 0.03 Colorless 0.002 0.04 Solvent n-propanol 10 2.1 0.17 99.6 0.48 0.2 <1 0 0.02 Colorless 0.003 0.03 IPA 5 2.3 0.19 99.8 0.47 0.1 <1 0 0.03 Colorless 0.003 0.02 Ethanol 4 2.1 0.18 99.5 0.50 0.1 <1 0 0.04 Colorless 0.003 0.04

The results indicate that the oils from different solvent extractions have identical quality except in phosphorus content in crude oil. The crude oils contain almost negligible quantities of undesirable factors like FFA, peroxides etc. The crude oils were refined using the standard technique. After refining all the oils were colorless and the FFA and peroxides were completely removed. Higher neutral oils in alcohol extractions indicate the highest degree of purity.

Aqueous solvent systems: Solvent: Water, 90:10:

Meal: The soy meal from aqueous solvent systems 90:10 at normal b.p were also analyzed for different characteristics and shown in Table 16. Table 16 Characteristics of soy meal from aqueous solvent mixture (90:10) Parameters Moisture,%(w.b) Protein, % Fat, % Ash, % Total bacterial counts,/gx103 Total fungal counts, /gx103 Salmonella Coliforms Residual solvent, ppm TI, mg/g Urease, change in pH units NSI, % Fiber, % n-propanol 6.0 52.7 0.6 5.8 1.5 0.7 76 1.1 0 7 3.7 IPA 6.3 51.9 0.5 5.60 1.6 0.51 63 0.9 0 9 3.6 Ethanol 6.1 53.0 0.7 5.4 0.9 0.28 114 1.0 0 10 3.5

The data revel that the residual solvents in the meal varied from 63-114 ppm. The TI was in the range of 0.90 to 1.1mg/g. The NSI values were in between 6 and 16%. The total fungal and bacterial counts were with in the permissible limits.

Oil:

The oil obtained from different solvent water mixture, 90:10 at normal b.p before and after refining were analyzed for various characters and presented in Table 17. Table 17 Characteristics of soybean oil from aqueous solvent mixture (90:10) Characteristics n-propanol Crude Phosphorus, ppm Peroxide value, meq/kg FFA, % Neutral oil, % Total volatile matter, % Water, % Refined Phosphorus, ppm Peroxide value, meq/kg FFA, % Color Copper, ppm Iron, ppm 8 1.7 0.13 99.9 0.42 0.20 Solvent IPA 4 2.0 0.12 99.8 0.47 0.16 <1 0 0.01 Colorless 0.002 0.02 <1 0 0.02 Colorless 0.002 0.03 Ethanol 2 2.0 0.11 99.6 0.50 0.18 <1 0 0.02 Colorless 0.002 0.04

The results indicate that the phosphorus in the crude oils were very low in the alcohol extracted oils. After refining the values were reduced to <1. in all cases. The highest degree of purity of oil was obtained with n-propanol, IPA and ethanol extractions.

Solvent: Water, 80:20:

The desolventized soy meal from 80:20 aqueous solvent systems were analyzed for various quality characters and given in Table 18. Table 18 Characteristics of soy meal from aqueous solvent mixture (80:20) Parameters Moisture,%(w.b) Protein, % Fat, % Ash, % Total bacterial counts,/gx103 Total fungal counts, /gx103 Salmonella Coliforms Residual solvent, ppm TI, mg/g Urease, change in pH units NSI, % Fiber, % n-propanol 6.5 52.0 0.6 5.7 1.7 0.73 73 0.82 0 5 3.7 IPA 6.2 51.7 0.5 5.5 1.9 0.60 59 1.0 0 7 3.6 Ethanol 6.0 53.1 0.7 5.4 1.1 0.33 110 1.0 0 9 3.5

The data indicated that the meal samples contained residual solvents ranging from 59 to 110 ppm and lowest in IPA. The TI activities were also at minimum. The NSI values were also lower. The total microbial load was in the prescribed limits.

Oil: The oil was also analyzed before and after refining and the data are shown in Table 19.

Table 19 Characteristics of soybean oil from aqueous solvent mixture (80:20) Characteristics n-propanol Crude Phosphorus, ppm Peroxide value, meq/kg FFA, % Neutral oil, % Total volatile matter, % Water, % Refined Phosphorus, ppm Peroxide value, meq/kg FFA, % Color Copper, ppm Iron, ppm 9 1.8 0.15 99.9 0.44 0.23 <1 0 0.004 Colorless 0.001 0.02 Solvent IPA 6 2.2 0.14 99.8 0.46 0.18 <1 0 0.002 Colorless 0.002 0.03 Ethanol 4 1.9 0.12 99.4 0.52 0.20 <1 0 0.002 Colorless 0.003 0.04

The results indicated that the phosphorus in the crude oils were very low in the alcohol extracted meals. After refining the values were reduced to<1 in all cases. The peroxides were in between 1.8 and 2.2 meq/kg.

Azeotropes:

The soybean meal from different azeotropic extractions was analyzed for various characters and presented in Table 20. Table 20. Characteristics of soy meal from the azeotropic extractions Component nnnhexane+ hexane+ heptane+ IPA Ethanol ethanol Residual solvents,ppm TI, mg/g Urease,change in pH units NSI, % Ash, % Oil, % Fiber, % Protein, % Fungal counts, /gx103 Bacterial counts, /gx103 Salmonella Coliforms 45/25 1.2 0 8.5 5.5 <1% 3.7 53 0.70 1.5 39/20 1.1 0 8.7 5.6 <1% 3.6 54 0.65 1.7 39/15 1.0 0.12 7.2 5.6 <1% 3.3 52 0.63 1.4 Azeotropes nWater+ Water+ Water+ hexane+ Ethanol nIPA npropanol propanol 60/12 43 55 59 1.3 0.1 8.0 5.7 <1% 3.8 51 0.73 1.3 0.9 0 4.8 6.0 <1% 3.7 55 0.34 1.0 0.8 0 4.5 6.1 <1% 3.6 54 0.42 1.1 0.9 0 5.2 5.6 <1% 3.6 52 0.32 1.2 -

The data indicate that the TI and urease were at the minimum. The protein content varied from 52% to 55%. The NSI values were in the range of 4.5-8.7. The meal was free from the microbial load.

Oil: The oil was analyzed before and after refining and the data are presented in Table 21.

Table 21. Characteristics of crude and refined soy oil with azeotropes Characteristics nhexane+ IPA nhexane+ Ethanol Azeotropes nn-hexane Water+ heptane+ +nEthanol ethanol propanol 0.42 0.15 0.7 2.2 123 99.8 0.04 0.05 Colorless 0.002 <1 0.04 0.38 0.12 0.2 2.1 380 99 0.02 0.03 Colorless 0.003 <1 0.02 0.13 0.19 0.18 2.3 10 98.7 0.04 0.04 Colorless 0.003 <1 0.04

Water+ npropanol 0.14 0.21 0.16 2.0 15 98 0.05 0.04 Colorless 0.002 <1 0.03

Water+ IPA

Crude oil Total volatile 0.50 0.47 matter, % Water, % 0.20 0.18 FFA, % 0.6 0.65 Peroxide value, 2.1 2.0 meq/kg Phosphorus, 400 250 ppm Neutral oil, % 99.5 97 Refined oil Water, % 0.04 0.03 FFA, % 0.05 0.04 Peroxide value, meq/kg Color Colorless Colorless Copper, ppm 0.002 0.003 Phosphorus, <1 <1 ppm Iron, ppm 0.04 0.03

0.20 0.17 0.06 2.1 8 99.3 0.03 0.05 Colorless 0.002 <1 0.03

The data indicate that the oils extracted with azeotropes contained more than 99% neutral oil. The phosphorus content in the crude oils was more in case of azeotropes like hexane + IPA, hexane+ethanol, heptane+n-propanol. After refining the content was reduced to <1% in all cases. The volatile matter was more with azeotropes other than water+ethanol, water+n-propanol and water+ IPA.

Utilization of soy meal:

The soy meal obtained from IPA extraction was used for utilization. The meal was first tested for its residual solvent using the overhead space technique with HPLC. The meal contained 15 ppm. The chemical composition is given in Table 22. Table 22 Chemical composition of soy meal Constituent Moisture content, wb Protein Fat Ash NSI Values, % 7.3 51 0.2 5.3 35

Chapatis: The whole-wheat flour was procured from the local market. Its composition was crude protein 11.9%, crude fat 0.5%, crude fiber 1.55%, ash 5.2% and moisture 10%. Chapatis were made from the wheat flour alone and also with 5%, 10%, 15% and 20% fortification of defatted soy flour. All mixtures had 0.5% salt. The chapattis were prepared using the method described earlier (Gandhi&Bourne 1988). For staling study the chapattis were kept at room temperature (28oC) for four hours. The nutritive profile of these chapattis is given in Table 23. Table 23 Chemical composition of soy fortified chapattis Treatments All wheat (control) 5% blend 10% blend 15% blend 20% blend Protein 11.9 14 16.5 18.2 19.9 Nutrients, % Fat 0.5 0.5 0.5 0.6 0.6 Ash 5.2 5.22 5.26 5.31 5.33

The data indicate that with the incorporation of soy meal the protein content was increased significantly from 11.9% to 19.9% at 5% to 20% of blending inferring an increase of 67% over control. The moisture content of chapatis varied from 40-46%. There were no appreciative differences in the other constituents. Since the soybeans are legumes and wheat a cereal the essential amino acid profile should be in better balance in the soy fortified chapattis. (Gandhi et al 1983). This enrichment with the soy meal will combat the protein calorie mal nutrition exists in India at affordable cost.

Textural Profile Analysis:

The textural profile parameters were measured on a Textural analyzer, TA-XT/2i with Volodkevich Bite Jaws. The operating conditions were: pretest speed 10 mm/s, test speed 2mm/s, post speed 10mm/s, time lag 5sec, strain 30% and load cell 25 kg with trigger force of 0.2N. The sample size was strips of chapatis of 10mm width and 20mm length. Twenty replications each were used. The textural profile parameters like hardness; cohesiveness, springiness and chewiness were determined and presented in Table 24. Table 24 Textural profile parameters of soy meal fortified chapatis at initial and final stage (4h) after preparation Treatment All wheat (control) 5% blend 10% blend 15% blend 20% blend Hardness Initial Final 2.2 9.38 2.57 3.38 3.45 3.60 16.02 18.51 20.70 22.67 TPP Cohesiveness, N.Cm Springiness, mm Initial Final Initial Final 0.6 0.7 0.61 0.69 0.65 0.64 0.70 0.70 0.61 0.69 0.68 0.68 0.73 0.67 0.73 0.73 0.74 0.69 0.74 0.74

Chewiness, N.Cm Initial Final 0.97 4.51 1.22 1.45 1.51 1.51 6.0 8.11 10.12 10.12

The data show that the hardness was more in the soy-fortified chapattis over all wheat chapattis ranging from 26% to 63% for the blends used. In general hardness and chewiness were increased over storage of four hours at ambient conditions in all treatments. Almost no change in cohesiveness and springiness was observed. The addition of soy meal thus affects the hardness and the chewiness in the initial stage itself. It may be probably due to enrichment with protein. These results show that some textural parameters change during the period of storage while others do not. The parameters of hardness and chewiness are the most important from the consumer point of view. The moisture content was reduced almost to 50% in all treatments at the end of experiments. Hence after four hours they were not liked. Water Absorption Capacity: The soy meal at different blend levels was tested for its water holding capacity. The data are given in Table 25. Table 25 WAC of soy meal blends Blend, % WAC, % 0 200 5 262 10 286 15 298 20 328 The water holding capacity was increased with the increase in level of blending. This adds to the functional properties of the dough so prepared.

Sweet biscuits: The soy meal was also used in the preparation of sweet biscuits. Biscuits are very convenient and inexpensive food products and are becoming very popular among both rural and urban people in India. These baked products have 6-7% protein. There is ever growing demand for high protein biscuits for therapeutic value. They can be considered for Applied Nutrition program. The enrichment of protein may be achieved through incorporation of protein rich ingredients. Among them soy meal has a great potential due to its high and good quality protein (50-52%) at affordable cost. Besides its nutritive value, it may improve the functional properties of the dough. Hence the use of soy meal in the standard sweet biscuit formula at 10%, 20%, 30% and 40% level increased the level of nutrients. The results are given in Table 26. Table 26 Nutritional status of various blends Nutrients, % Crude protein Crude fat Total carbohydrates Ash 0 6.5 26.0 61.5 4.2 10 10.0 25.0 58.9 4.4 Level of blending, % 20 30 11.2 12.2 23.0 22.0 57.4 56.1 4.6 4.8 40 14.8 21.0 55.2 5.0

The data show that the protein and ash contents were increased from 6.5% to 14.8% and 4.2% to 5.0% respectively as the level of blending increased from 10% to 40%. The fat and carbohydrate contents were decreased from 26% to 21% and 61.5% to 55.2% respectively with the incorporation of soy meal. The moisture contents of these biscuits varied from 1.7% to 2.0%. The addition of soy meal increases the protein content substantially. Since soybean is a legume and wheat a cereal, the essential amino acid profile will be in better balance. (Gandhi et al 1983). Gandhi & Bourne (1988) found similar results with soyfortified chapatis. Texture Analysis: The Texture Analyzer, TA-XT/2i measured the stickiness/work of adhesion of the dough and the bending hardness/cutting hardness of the biscuits on the fresh samples. The conditions for measurement of dough stickiness were :option-adhesive test, pretest speed 2mm/s and test speed data acquisition rate 400 pps. SMS/Chen-Hosney Dough stickiness cell (A/DESC) was used with 5kg load cell and 25mm Perspex cylinder probe (P/25P). The highest positive peak is the measure of stickiness and the positive area is the work of adhesion. The conditions for hardness measurement of biscuits by cutting were: optionreturn of start, pretest speed 3mm/s, test speed 3mm/s, post test speed 10mm/s, distance 5mm, trigger type 20 g and data acquisition rate 200 pps. Knife-edge with slotted insert (HDE/BS) blade set and 25 kg load cell was used. The samples were halved and placed centrally under knife-edge. The condition for hardness measurement of biscuits by bending/snap test were: option-return to start, pretest speed 2mm/s, test speed 0.5mm/s, post test speed 10mm/s, distance 5mm, trigger type auto 20g, data acquisition rate 400pps

and a three point bending ring (HDP/3PB) with 25 kg load cell. The bending hardness is the maximum force that these biscuits bear before they break and expressed in Newton (N). Effect on stickiness and work of adhesion: Blending of soy meal reduced the stickiness of the dough as evident from the results in Table 27. Table 27 Blending vs. stickiness and work of cohesion Parameter Moisture content, % Stickiness Work of adhesion, mm 0 68.0 0.483 0.040 Level of blending, % 10 20 30 69.1 71.0 73.3 0.466 0.62 0.431 0.113 0.416 0.162 40 76.2 0.387 0.180

The data indicate that the moisture content was increased from 68% to 76.2% and correspondingly the stickiness was reduced from 0.483 N to 0.387 N and the work of adhesion increased from 0.040 N mm to 0.180 N mm with the increase in level of soy meal. This infers that the inclusion of soy meal improves the water holding property of the dough. On the other hand the decrease in stickiness may be attributed to the decrease in total gluten content of the dough, as the soy meal has no gluten content. Effect on bending/cutting hardness: The bending/cutting hardness of the biscuits were measured and presented in Table 28. Table 28 Bending and cutting hardness of blended biscuits Treatment Control 10% soy meal 20% soy meal 30% soy meal 40% soy meal Bending 3.60 4.27 5.31 7.46 9.85 Hardness, N Cutting 6.02 10.50 18.46 20.02 23.04

The results reveal that the hardness was increased from 3.60 N to 9.85 N and the cutting hardness from 6.02 N to 23.0 N with the increase in blending. These results were expected due to the increase in protein content and decrease in fat and carbohydrates. Sensory Evaluation:

The sensory attributes like taste, color, appearance, texture and over all acceptability were determined using an informal taste panel consisting of 10 untrained panelists. The ratings were on a zero point hedonic scale ranging from 9 (like extremely) to 1(dislike extremely). Results obtained were subjected to ANOVA. The mean score values for different sensory characters are depicted in Table 29. Table 29 Mean scores of sensory panel judges for the characteristics of soy meal fortified biscuits Biscuit sample 10% blend 20% blend 30% blend 40% blend Color 7.1 6.4 6.9 6.9 Appearance 7.1 6.9 6.7 7.0 Characters Flavor Texture 6.3 6.1 6.9 6.6 6.9 6.4 7.0 7.5 Taste 7.3 7.5 7.1 8.0 Overall acceptability 7.4 6.8 7.0 7.3

The scores were given as compared to the control with out soy meal, assuming the highest acceptability for them (score of nine). The mean scores for all the quality characters and overall acceptability was more than the minimum acceptable score of five. A separate analysis of variance was done for each character: taste, color, appearance, texture and over all acceptability from every individual score of taste panel. This analysis was carried out to find out the difference among the characteristics and panelists. The results are shown in Table 30. Table 30 ANOVA of taste panel score for the characteristics of different soy meal fortified biscuits Source of variation Product Judges Error d.f 3 9 27 Color 0.89** 3.11** 0.558 Appearance 0.29** 3.33** 0.336 MSS Flavor Texture 1.5* 3.98** 0.476 2.09* 3.58** 0.536 Taste 0.29** 3.58** 0.55 Overall acceptability 0.75** 0.73** 0.499

*Significant at 5% level ** Significant at 10% level The results indicate that the differences among the treatments was significant for flavor and texture and non significant for color, appearance, taste and over all acceptability at 55 level of significance. The disagreement among the judges for all the characteristics

was not significant at 5% level of significance. Thus the data show that the source of variance among the products was only due to characteristics like flavor and texture and the variance due to judges did not exist. Soy papad: Soy fortified papads were prepared at 10%, 20%, 30% and 40% level of blending. The papads were of average size of 12 cm dia and 1mm thickness. The composition of all the blends is given in Table 31. Table 31. Constituents of soy papad Ingredients Black gram flour Papad Khara, g Soy meal, g Salt White sesame seeds Water, ml 0 100 2-3 To taste 2-3 45 10 90 Treatments- blending % 20 30 80 70 2-3 20 To taste 2-3 45 2-3 30 To taste 2-3 45

40 60 2-3 40 To taste 2-3 45

2-3 10 To taste 2-3 45

Preparation of papads: Mixed soy meal, black gram flour, papad khara, asfoetida and salt. Added water to make stiff dough. Kneaded the dough and left it for 15 min. Divided the dough in to small balls and lightly rolled on sesame seeds. Sheeted the balls in to thin papad and dried at 150oC for 15 min in an oven and packed for further use.

The papads were analyzed for various chemical constituents and shown in Table 32. Table 32 Chemical compositions of papads

Treatment Control 10% 20% 30% 40%

Moisture content, % 3.0 4.0 4.0 5.0 5.0

Protein, % 18.5 20.4 22.4 24.4 26.0

Fat, % 0.9 0.9 0.9 0.9 1.0

Ash, % 4.3 4.3 4.0 4.0 4.0

The data indicate that the protein content was increased from 18.5% to 26%. The sample with 10% soy meal was preferred over others in all the characters. Nan Khatai: Nan Khatai biscuits were also prepared with the incorporation of soy meal at different levels of blending namely 10%, 20%, 30% and 40%. The composition of recipe is given in Table 33. Table 33. Composition of Nan Khatai biscuits Constituents Refined wheat flour Soy meal Dalda, g Sugar, g Baking powder, g Baking soda, g Control 100 60 50 1 0.8 10 100 10 70 60 1 0.8 Level of blending, % 20 30 100 100 20 70 60 1 0.8 30 70 60 1 0.8 40 100 40 70 60 1 0.8

The chemical analyses of these biscuits were carried out and presented in Table 34. Table 34. Chemical composition of Nan Khatai biscuits Level of blending with soy meal, % Control 10 20 30 40 Moisture content 3.7 3.4 3.9 3.5 3.0 Constituents, % Protein Fat 10.9 15.3 19.6 26.2 30.6 28.3 27.5 25.8 24.7 23.3 Ash 4.0 4.4 4.1 4.3 4.1

The results indicate that the protein content was increased from 10.9% to 30.6%. The fat content however decreased from 28% to 25.55. The biscuits were all acceptable. Salt Biscuits:

The salt biscuits were prepared with different blends. The ingredients are shown in Table 35. Table 35 Composition of different blends Ingredients Refined wheat flour Dalda, g 42 Baking 1 powder, g Baking soda, 0.8 g Salt, g 5 Soy meal, g Cumin seed According to powder taste Control 100 10% 100 42 1 0.8 5 10 According to taste Level of blending, % 20% 30% 100 100 42 1 0.8 5 20 According to taste 42 1 0.8 5 30 According to taste 40% 100 42 1 0.8 5 40 According to taste

The biscuits were analyzed for proximate analysis and presented in Table 36. Table 36 Chemical compositions of different blends Treatment Control 10% 20% 30% 40% Moisture, % 2.5 2.5 3.0 3.0 3.0 Protein, % 10.9 15.0 21.8 26.2 30.8 Fat, % 24.0 22.5 20.0 19.0 18.0 Ash, % 4.0 4.0 4.2 4.2 4.0

The results show that the protein content was increased from 10.9% to 30.8% and the fat content decreased from 24% to 18%. All of them were acceptable.

Cake:

The cake was prepared by incorporating the soy meal at different levels. The composition of the recipe is given in Table 37. Table 37 Composition of Cake recipes Ingredients, Level of blending, % g Control 10 20 Refined 100 100 100 wheat flour Soy meal 10 20 Egg 60 60 60 Sugar 40 40 40 Dalda 40 40 40 Water 20 20 20 Baking 1 1 1 powder Baking soda 0.8 0.8 0.8 Salt 0.5 0.5 0.5

30 100 30 60 40 40 20 1 0.8 0.5

40 100 40 60 40 40 20 1 0.8 0.5

The cake so prepared was analyzed for its chemical constituents and the results are presented in Table 38. Table 38 Chemical composition of cake Treatments Control 10% 20% 30% 40% Protein, % 17.0 22.0 26.5 32.0 37.0 Fat, % 21.0 20.0 19.0 18.0 17.0 Ash, % 4.0 4.0 4.0 4.0 4.1

The data indicate that the protein content of the cake was increased from 17% to 37% with out any change in the ash content. All of them were acceptable.

Soy Mathri;

It is normally prepared from refined wheat flour. Soy meal was incorporated up to 40% level. The composition of soy mathri is given Table 39. Table 39 Constituents of soy mathri Ingredients, Refined wheat flour, g Soy meal, g Salt, g Water, ml Cooking oil, ml Ajwain seeds, g Control 100 3-4 50 300-400 2 10 90 10 3-4 50 300-400 2 Level of blending, % 20 30 80 70 20 3-4 50 300-400 2 30 3-4 50 300-400 2 40 60 40 3-4 50 300-400 2

Method: All the ingredients were mixed and a hard dough was prepared with water. Rolled out in to small rounds and pierced them with a knife. Then each one of them was fried in oil till golden brown. The chemical composition was carried out and the results are shown in Table 40. Table 40 Proximate composition of soy mathri Treatments Control 10% 20% 30% 40% Moisture, % 2.0 2.0 2.0 2.0 2.0 Protein, % 10.9 14.5 18.0 21.5 26.0 Fat, % 20.0 20.0 21.0 21.5 21.5 Ash, % 4.5 4.5 5.0 5.0 5.1 Carbohydrates, % 61.6 59.0 53.5 49.5 44.9

The data reveal that the protein content was increased from 10.9% to 26% at 40% level of blending. All the treatments were acceptable.

Soy vada:

Soy vada was prepared from the soy meal at 10% to 40% level of blending. Vada is very popular In India. Now soy meal was incorporated to increase the protein content. The composition of the recipe is presented in Table 41. Table 41 Composition of soy vada Constituents Black gram dhal, g Soy meal, g Cumin powder, g Back pepper powder, g Salt, g Baking soda, g Oil, ml Asafoetida, g Control 100 2-3 2-3 To taste 2 250 pinch 10 90 10 2-3 2-3 To taste 2 250 pinch Level of blending, % 20 30 80 70 20 2-3 2-3 To taste 2 250 pinch 30 2-3 2-3 To taste 2 250 pinch 40 60 40 2-3 2-3 To taste 2 250 pinch

Method: Urd dhal was soaked in water for 6 hours and kept aside. They were blended in a Mixer with little quantity of water. Salt, baking soda, cumin powder, soy meal, black pepper powder and asafetida were all added to the batter. They were made in to small balls and deep-fried. The products were analyzed for their chemical composition and the results are given in Table 42. Table 42 Nutritive profile of soy vada Treatment Control 10% 20% 30% 40% Moisture, % 6.5 6.5 7.2 7.5 8.0 Protein, % 24.0 26.5 28.5 30.8 34.0 Fat, % 1.5 2.1 2.8 3.0 3.5 Carbohydrates, % 63.1 60.0 56.5 53.6 49.4 Ash, % 4.9 4.9 5.0 5.1 5.1

The results indicate that the protein content was increased from 24% to 34% at 40% level of blending. The organoleptic evaluation also suggested that all these products were acceptable.

Soy vadi: Soy fortified vadis were prepared using the defatted soy dhal. The soy dhal had 4.9% moisture, 52% protein, <1% oil and 5.1% ash. The ingredients are given in Table 43. Table 43 Recipe for soy fortified vadi Ingredients Green gram dhal Defatted soy dhal Asafoetida Cumin seeds Ginger Garlic Salt Method: The green gram dhal and defatted soy dhal were soaked in 200 ml water for 8-10 hours. Both the dhals were mixed and the salt, cumin seeds, asafetida, ginger and garlic paste were added to this batter. They were mixed properly and made in to small badis manually on a plastic sheet. They were later dried in sun for three days continuously and stored in airtight containers. Values 60-100 g 10-40% blend 0.5-1.0. g 2-3 g 2 inches 3-4 cloves 2-3, g

They were analyzed for their chemical composition and shown in Table 44. Table 44 Chemical compositions of soy badis Treatments Control 10% 20% 30% 40% Moisture, % 4.2 4.0 4.1 4.0 4.3 Protein, % 23.5 25.5 28.9 30.5 34.0 Fat, % Traces Traces Traces Traces Traces Ash, % 3.0 3.6 3.8 3.9 3.9

The data thus indicate that the protein content was increased from 23.5% to 34% at 40% blend level. There were no appreciative changes in the total fat however the ash content was increased from 3.0% to 3.9%.

Soy pakoda: The soy pakoda was prepared with defatted soy dhal incorporation in the original recipe. The ingredients are given in Table 45.

Table 45 Recipe compositions Ingredients Green gram dhal Soy dhal defatted Cumin seed Onion Salt Coriander Oil Quantity 100,g 10-40% blend 2-3 g One medium size To taste For garnishing 200 ml

Method: Green gram dhal and defatted soy dhal were soaked separately in 200 ml water for 8 hours. They were ground till a coarse paste using little water. Chopped onions, green chillies, salt, coriander leaves and cumin seeds were added to the batter. Oil is added in a frying pan and heated to 180oC and drop the batter in the form of pakodas and deep-fried till golden brown. The products were organoleptically evaluated by an informal panel and found accepted. The chemical composition of the products is presented in Table 46. Table 46 Chemical constituents Treatments Control 10% 20% 30% 40% Moisture, % 9.0 9.0 8.0 8.5 8.5 Protein, % 24.0 26.5 28.8 31.5 34.0 Fat, % 2.5 2.1 2.1 2.2 2.2 Ash, % 3.5 2.8 2.5 2.3 2.1

The data indicate that the protein content was increased from 24% to 34%, however the fat content remained the same. There was significant difference in the total ash content.

Mass balance of IPA in soy oil extraction:

Experiments were conducted to calculate the mass balance of IPA used in the extraction of soy oil in batch extractor. The details are shown in the flow diagram. The ratio of solvent to soy flakes was 2:1. About 250 g of soy flakes containing 45 g oil, 180 g meal and 25 g water was taken in a batch extractor (traditional Soxhlet Apparatus). Five hundred ml of IPA was added and the extraction was done at its boiling point for 10 hours. After extraction the miscella and marc were collected separately. The miscella contained 350 ml IPA, 42.5 g oil and 0.5 g water. The marc contained 180 g meal, 2.5 g oil, 24.5 g water and 150 ml IPA. The miscella was subjected to phase separation through chilling first. The solvent phase contained 320 ml IPA and the oil phase contained 42.5 g oil, 30 ml iPA and 0.3 g water. The oil phase was further subjected to heating and about 10 ml IPA was recovered. The marc was desolventised and about 130 ml IPA was recovered. The desolventised marc contained 180 g meal, 2.5 g oil and 24.5 g water. In all about 30 ml of IPA was lost indicating a loss of 6% solvent in the process. The experiment was conducted in triplicate and the average values were computed. IPA with higher flash point (117oC) and auto ignition temperature (82.5oC) is safer than nhexane (22.8oC & 45.7oC). IPA is heavier (6.83 lb vs. 5.60 lb/gallon) and requires more energy for pumping. IPA vapors have lower density than n-hexane and are more noticeable as the boiling point is approached. Steam is often sparged in to desolventizers to remove nhexane. Nearly dry hexane is recovered by gravity where as IPA is completely miscible and retains all water introduced. IPA has a higher latent heat of vaporization. It requires two times energy for recovery. N-hexane can be removed only by vaporization however chilling the miscella separates IPA. Chilling needs less energy (16 BTU) as compared to vaporization (371 BTU). Through chilling the solvent rich phase will be at upper and the oil rich phase at lower. Losses of solvent vary with the type of solvent used. More in n-hexane, less in other alcohols and least in IPA. The quality of meal in the alcoholic extractions is better as most of the anti nutritional factors are destroyed more effectively. Though the latent heat of vaporization in case of IPA is more, a little quantity only requires heat and most of it is removed through chilling in phase separation. This is more versatile way of separating the solvent with out appreciative losses and health hazards. In the case of aqueous extractions since some quantity of water also exists in the mixture needs more time to remove water.

The comparison of n-hexane and IPA for their different characters is shown in Table 47.

Table 47 Characteristics of n-hexane and IPA Characters Formula M.W Density Flash point Auto ignition temperature Oil solubility Water solubility BP Latent heat of vaporization Specific heat Toxicity, LD 50 n-hexane C6H14 85.17 0.6603 at 20oC 22.80C 6.5oC Completely Immiscible 68.7oC 80 kcal/kg; 144 BTU/lb 0.533 0.029 g/kg Solvents IPA C3H8O 60.11 0.7854 at 20oC 117oC 28.05oC Varies with the temperature, oil type, water content etc. Completely miscible 82.4oC 160 kcal/kg; 288 BTU 0.723 5.8 g/kg

Economics of the alternative solvent system: The relative cost of the solvents of laboratory grade are given in Table 48 Table 48 Cost of the solvents used Solvent n-hexane n-heptane n-propanol IPA Ethanol Cost, Rs/2.5 liters 260 550 975 360 2000

Based on the unit cost of the solvent available at present n-hexane is the cheapest and economical followed by IPA. However this solvent needs more energy for recovery, as its latent heat of vaporization is more apparent differences in energy recovery requirements is 30-40% between n-hexane and IPA. IPA extracts more slowly than n-hexane. From the studies it stems out that the IPA seems to be a better alternate solvent among other solvents used. Based on the information on the chemical characteristics available it is less inflammable than n-hexane. The extracts of crude oil and meal are equal to those obtained with n-hexane. Use of IPA as a replacement seems to be feasible subject to confirmation by sustained trials in a scaled up pilot plant. 12.Papers Published a. Gandhi,A.P; Kotwaliwale, N; Taimini, V; Kawalkar, J; Srivastav, D.C; Parihar, V.S and Raghunadh,P.2000. Effect of incorporation of defatted soy flour on the texture of chapatis. J. Texture Studies (USA), 31(6), 609-614.

b. Gandhi,A.P; Kotwaliwale, N; Kawalkar, J; Srivastav, D.C; Parihar, Vand Raghunadh,P.2001. Effect of incorporation of defatted soy flour on the quality of sweet biscuits. J.Food Sci and Tech, 38(5),502-503. c. Gandhi,A.P; Joshi,K.C; Krishna Jha; Srivastav,D.C; Parihar,V.S; Kawalkar,J; Raghunadh, P ; Jain,S.K and Tripathi,R.N. 2003. Studies on alternative solvents-I Soybean, International Journal of Food Science and Technology (UK), 13. Approximate expenditure incurred in the project (Rs): i. Salary of Scientific staff: 1390558 ii. Salary of Technical staff: 360000 iii. Salary of Supporting staff: 21600 iv. Casual Laborers wages: 10000 Cost of laboratory equipment/chemicals/glassware: Equipment: 1550000 Chemicals: 150000 Glassware: 100000 Total: 3682188 14.Details of field/laboratory books and final materials and their location: The data registers are with the Project Leader and the reports are available in the center. The equipment, glassware and chemicals are available in the Biochemistry Laboratory. 15.Coments/suggestions of the project leader regarding possible future lines of work that may be taken up arising from the project: The use of alternative solvents at this stage is not economical. However in case of need they may be tried.

16.Signatures and name of the Investigators;

(A.P.Gandhi) 17.Signature of Project Director: (S.D.Kulkarni)

(Krishna Jha)

(K.C.Joshi)

18. Signature of the Director (with comments if any)

(Nawab Ali)