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Journal of Stress Physiology & Biochemistry, Vol. 20, No. 3, 2024, pp. 29-36 ISSN 1997-0838 Original Text Copyright © 2024 by Jebisha and Begila David
ORIGINAL ARTICLE
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Preparation and Characterization of Renewable Bio-Polyol
from the Edible Seed Oil
1 Research scholar (Reg no:21213162032019), Department of Chemistry and research Centre, Scott Christian college (autonomous), Nagercoil-629 003, Tamil Nadu, India.
(Affiliated to Manonmaniam Sunderanar University, Tirunelveli-627 012, Tamil Nadu, India)
2 Research supervisor & Associate professor, Department of Chemistry & research Centre, Scott Christian college (autonomous), Nagercoil-629 003, Tamil Nadu, India.
(Affiliated to Manonmaniam Sunderanar University, Tirunelveli-627 012, Tamil Nadu, India)
*E-Mail: begilarobinfagmail. com
Polyol is an organic compound containing multiple hydroxyl groups. This study looked at the possibility of using an edible oil extract from Salvia hispanica seeds as a sustainable source for polyols and, eventually, biodiesel or polyurethane. For this, a combination of hydrogen peroxide and acetic acid was used to create new polyol from the aforementioned oil in one-step synthesis. Standard techniques such as physicochemical analysis, phytochemical and basic radical identification, FTIR and NMR were used to characterize the polyol derivative that was extracted from the oil. Antimicrobial activity of both oil and polyol were tested against certain bacteria and fungi. Spectral analysis demonstrated the formation of polyol and this result indicated the possible of using Salvia hispanica polyol as a raw material for the preparation of bio-polymers.
Key words: Salvia hispanica oil, polyol, FTIR, NMR, antimicrobial, bio-polymers
J.L. Jebisha 1, S. Begila David
2
Received March 13, 2024
The chemical industry has been prompted to explore for new sources of renewable resources as raw materials by the strong demand for products with petrochemical origins, as well as their unfavorable environmental effects and the growing scarcity of these non-renewable resources. Due to nature's enormous synthetic potential and many green chemistry principles, these raw materials have significantly aided the plastics industry's sustainable development (Eissen et al., 2002; Tian et al., 2012). Due to their availability, low toxicity, biodegradability, natural fluidity, and affordability, vegetable oils are among the most popular alternatives (Biermann et al., 2000; Guner et al., 2006; de Espinosa, et al., 2011). To create polyols, a number of vegetable oil molecules must undergo a chemical transformation. These bio-polyols are then used, among other things, to create polyurethanes (Zhang et al., 2015), polyesters (Chaudhari. et al., 2015), and epoxy (Fernandes et al., 2017). Vegetable oil polyols are fascinating substances with applications as corrosion inhibitors (Kosari et al., 2014; Yoo et al., 2012), tensioactive agents or organogelators (Stan et al., 2008), and useful monomers for the production of diverse macromolecular compounds (Ji et al., 2015). Polyols have been created using natural oils like castor (Ugarte et al., 2014), soybean (Ji et al., 2015), camelina (Balanuca et al., 2015), palm kernel (Septevani et al., 2015), jatropha (Saalah et al., 2015), and rapeseed (Zieleniewska et al., 2015; Kuranska et al., 2015).
An herbaceous plant called chia has opposing, serrated leaves that range in size from 1.5 to 3 inches long and 1 to 2 inches wide. Oval in shape, seeds are roughly 2 mm (0.08 inches) in length and 1 mm (0.04 inches) in width. The lustrous seeds contain darker irregular marks or specks on them and their coat can range in hue from cream to charcoal grey. An annual herb called chia (Salvia hispanica L.) blooms in the summer. It is about a meter tall and has opposite, petiolate, serrated leaves that range in length from 4 to 8 cm a width of 3 to 5 cm (Ayerza et al., 2010).
Therefore, this study concentrates in the obtention of new bio-based polyol from the edible oil extract from the seeds of the Salvia hispanica and also to test the
phytochemicals and basic radicals present in the oil extract. In addition, we characterize the prepared polyols using various physical properties and spectroscopic methods. The application of the prepared polyol against certain microbes were also tested.
MATERIALS AND METHODS
OIL EXTRACTION:
The extraction of oil was carried out using AOAC technique Am2-93 (Official Methods and Recommended Practices of the American Oil Chemists' Society; AOCS Press: Champaign, IL, USA, 1995). In a Soxhlet device, 5g of Salvia hispanica L. seeds were extracted with 100 ml of n-hexane as the extraction solvent. Following an 8hour period, the n-hexane was eliminated at 40°C and low pressure. After obtained, the Salvia hispanica L. oil (SHO) was kept in a refrigerator till additional research was conducted. SYNTHESIS OF POLYOL:
The synthesis of the polyols was done in accordance with the approach outlined by Monteavaro et al. (2005). 5g (5.6 mmol) of oil 9.30 mL (0.162 mol) of glacial acetic acid in 20 mL of toluene, together with a few drops of sulfuric acid, were combined in a three-necked flask that was fitted with an isobaric funnel, reflux condenser, and mechanical stirrer. At room temperature, the mixture was mechanically agitated until it was completely homogenized. Subsequently, 5.30 mL of a 30% H2O2, solution was added gradually while maintaining the temperature. Following the addition of H2O2, the mixture was heated for 12 hours to 60°C. After bringing the reaction mixture down to room temperature, surplus peroxide was removed by stirring it for 20 minutes while adding a 10% (w/v) sodium bisulfide solution. Following that, the mixture was mixed with 50 mL of ethyl ether, and the organic phase was repeatedly washed to a pH of neutral using a 10% (w/v) sodium carbonate solution. In order to extract Salvia hispanica L. (SHP), the organic phase was lastly dried over sodium sulphate and concentrated under vacuum to remove the ethyl ether. CHARACTERISATION OF OIL AND POLYOL: PHYSICO CHEMICAL ANALYSIS:
The acid value of SHP was determined by the volumetric titration method according to ASTM D1980-
87. The known quantity of sample is dissolved in neutral butanol-toluene mixture and 3-4 drop of phenolphthalein indicator added, swirling gently and titrated against the 0.1 N alcoholic KOH solution. Equation (1) was used to determine the acid value of SHP.
Acid value=RX N X 56.1 (1)
W
where B = Burette reading (ml), N = Normality of alcoholic KOH solution, W = weight of sample (g).
The hydroxyl value of SHP was determined by acetic anhydride-pyridine method according to ASTM D4274-16. The calculation of hydroxyl value was done by using Equation (2).
Hydroxyl value = (B - S) x N x 56.1 (2)
W
where B = Burette reading for blank (ml), S = Burette reading for sample (ml), N = Normality of alcoholic KOH solution, W = weight of sample (g). PHYTOCHEMICAL AND BASIC RADICAL IDENTIFICATION:
Phytochemical screening and basic radical identification were performed to asses the qualitative chemical composition of different samples of crude extracts using commonly employed precipitation and colouration reactions to identify the major and secondary metabolites. FTIR ANALYSIS:
The chemical structure of the SHO and SHP were identified by FTIR on a Bruker ATR spectrophotometer. The spectra were observed in the 600 - 4000 cm-1 wavelength range. 1H NMR SPECTROSCOPY:
The supportive confirmation of chemical structure was given by 1H nuclear magnetic resonance (NMR)spectroscopy. 1H spectra of the product were analyzed using Bruker DPX 400 MHz spectrophotometer with CDCls as solvent. ANTIMICROBIAL ACTIVITY:
Antimicrobial activity was examined using "The Kirby-Bauer Method" against the number of pathogens including both gram-positive and gram-negative bacteria and also certain fungi. The zone of inhibition of both SHO and SHP were examined against certain pathogens.
RESULTS AND DISCUSSION
PHYSICO CHEMICAL ANALYSIS:
The acid number is expressed as the number of milligrams of KOH required to neutralize the acidity of sample. The OH number is the amount of available reactive hydroxyl groups on polyol molecules. The viscosity was measured using Brookfield viscometer and the density of the polyol were also analysed. The SHP is well dissolved in Chloroform. The results of the certain physico chemical analysis were mentioned in the Table 1.
PHYTOCHEMICAL SCREENING:
The preliminary phytochemical study reveals the presence of glycosides, reducing sugars, phenolic compounds and saponins (Table 2) in the SHO. The color change in the extract were also listed in the table. BASIC RADICAL IDENTIFICATION:
Basic radical test shows the presence of bismuth, barium calcium and magnesium in the SHO were listed in the Table 3. FTIR:
The successful conversion of SHO to SHP was confirmed qualitatively by FT-IR spectroscopy. Fig. 1 and Fig. 2 shows the FTIR spectra of SHO and SHP respectively. The SHP spectra shows a broad band at 3562 cm-1 which were assigned to the presence of a hydroxyl (-OH) stretching vibration. A strong band at 3181 cm-1 was attributed to the presence of aromatic -CH-stretching and bands at 1644 cm-1 and 1400 cm-1 were assigned to the presence of amide band from protein carbonyl stretches and C-O-H bending vibrations. The appearance of new peak at 3562 cm-1 in SHP spectra confirms the formation of polyol. 1-H NMR spectra:
Fig. 3 and Fig. 4 gives the 1-H NMR spectra of the SHO and SHP respectively. The methylene protons of the aliphatic chains may be responsible for the signal seen as a triplet between 1.63 to 1.67 ppm (3H) in Fig. 4. The proton of the hydroxyl group is represented by the sharp singlet peak at 2.731 ppm (6H); the electronegative influence of the oxygen atom is responsible for the absorption shift towards the downfield. The multiplet signal at 3.8 ppm (2H) is indicative of the protons next to the acid's carbonyl
32 Preparation and Characterization of Renewable Bio-Polyol...
group. The peak shift downwards as a result of the present in the range of 5.3 to 5.3 ppm (Fig. 3) in SHO
carbonyl group's electronegative action, which de-shield vanished, indicating that the SHP structures were
the area (Dai et al., 2005). The signals corresponding to essentially unsaturated. Lastly, the hydroxylation
the toluene structure were found at 7.5-7.6ppm as reaction was verified by the lack of signals in the range
multiplet (Fig. 4). This evidence confirmed that the of 2.8 to 3.3ppm in relation to the epoxide groups (-
elimination of toluene was ineffective. Furthermore, the CH(O)CH-).
olefinic hydrogen (-CH=CH-) signal that had been
Table 1: Physico chemical analysis of polyol
Parameters Chia based polyol
Acid number (mg KOH/g) 5.386
OH number (mg KOH/g) 166.54
Density (g/cm3) 0.868
Viscosity (cps) 464
Solubility Chloroform
Table 2: Phytochemical screening of SHO
EXPERIMENT OBSERVATION INFERENCE
NaOH test No blue green color obtained Absence of anthocyanin
Extract + 2ml of NaOH
Bromine water test: Pale yellow color obtained Presence of glycosides
Extract + bromine water
1ml of + lead acetate solution No precipitate obtained Absence of phenol
5 ml of extract + 2ml of CHCls + 3 ml of conc. H2SO4 No reddish-brown precipitate obtained Absence of terpenoids
LEAD ACETATE TEST: No red precipitate Absence of tannins
Extract + 1ml of lead acetate
Extract + CHCl3 + conc. H2SO4 No purple color obtained Absence of steroids
Extract + Molisch's reagent Purple color is obtained Presence of reducing sugars
Extract + 2N HCl and remove the No white precipitate obtained Absence of alkaloids
aqueous layer and add Mayer's reagent
Alcohol + extract + ferric chloride Intense color Presence of phenolic compounds
Extract + water + shake well Foamy lather is obtained Presence of saponins
Alcohol + extract + Mg ribbon + No color change Absence of flavonoids
Conc. HCl
Alcohol + Conc. HNO3 + NH3 No reddish orange color is obtained Absence of xanthoproteins
Table 3: Basic radicals' identification
EXPERIMENT OBSERVATIONS INFERENCE
Extract + KI No golden spangles Absence of lead
Extract + NH4OH white precipitate Presence of bismuth
Extract + cupron reagent + NaOH No green color obtained Absence of copper
Extract + potassium ferrocyanide No white precipitate obtained Absence of zinc
Extract + dil. HCl + water + H2S No yellow precipitate Absence of cadmium
Extract + potassium thiocyanate No red or blue color Absence of ferric and cobalt
Extract + dil. HCl + aluminon reagent + (NH4)2CO3 No bright red precipitate is obtained Absence of aluminum
Extract + conc. HNO3 + sodium bismuthate + No pink color Absence of manganese
water
Extract + dimethyl glyoxime + NH4OH No scarlet red precipitate Absence of nickel
Extract + potassium chromate Pale yellow precipitate Presence of barium
Extract + NH4OH + ammonium oxalate White precipitate Presence of calcium
Extract + Magneson reagent + NaOH Blue precipitate Presence of magnesium
Extract +NaOH + Nesseler's reagent No reddish-brown precipitate Absence of ammonium
Table 4: Shows the zone of inhibition against certain microbes
Bacteria Zone of inhibition
SHO SHP Control (Amikacin)
Proteus mirabilis 12 mm 11 mm 18 mm
Salmonella ttypi 13 mm 14 mm 19 mm
Fungi Zone of inhibition
SHO SHP Control (nystatin)
Aspergillus niger 10 mm 8 mm 15 mm
C. tropicalis 15 mm 14 mm 13 mm
Figure 1. FTIR spectra of SHO
Figure 2. FTIR spectra of SHP
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WIM/\\\l/
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Figure 3. 1-H NMR spectra of SHO
Figure 4. 1-H NMR spectra of SHP
Figure 5. Zone of Inhibition against fungi
Figure 6. Zone of Inhibition against bacteria
Antifungal property
16 14
12 10
8 &
4
2 о
Aspergillus niger С.Tropicalis
■ SHO ■ SHP ■ Control (Nystatin)
Antibacterial property
20 18 16 14 12 10 8 6 4 2 о
Proteus mirabilis Salmonella typi
■ SHO BSHP ■ Control (Amikacin)
Figure 7. Comparative study
APPLICATION: ANTIMICROBIAL ACTIVITY:
Two bacterial cultures (Fig. 6) including Proteus mirabilis (gram positive), Salmonella typi (gram negative) and two fungal cultures (Fig. 5) including Aspergillus niger and C. tropicalis were used to check the antimicrobial potential of both SHO and SHP. Amikacin was used as a control against bacterial cultures while Nystatin was used as a control against fungal cultures. The zone of inhibition against both bacteria and fungi were listed in the Table 4. The bar graph shows the comparative study of SHO and SHP against certain microbes (Fig. 7).
CONCLUSION
We can infer from the results above that SHP was successfully prepared from the hexane extract of Salvia Hispanica and it was characterized by using FTIR and 1H NMR spectroscopy. The physico-chemical analysis of the SHP and phytochemical screening and basic radical identification of SHO were examined. The antimicrobial potential of the prepared polyol and the oil were tested against certain bacteria and fungi. Thus, the prepared bio-polyol can be used as an alternative for organic polyols which can be effectively use for the preparation of certain bio- polymers.
CONFLICTS OF INTEREST
The authors declare that they have no potential
conflicts of interest.
REFERENCES
AOAC. (1995) Official Method Cd 3d-63, Official Methods and Recommended Practices of the American Oil Chemists' Society; AOCS Press: Champaign, IL, USA,.
Ayerza, R. (2010). Effects of seed color and growing locations on fatty acid content and composition of two chia (Salvia hispanica L.) genotypes. Journal of the American Oil Chemists' Society, 87, 11611165.
Biermann, U., Friedt, W., Lang, S., Lühs, W.,
Machmüller, G., Metzger, J. O.....& Schneider, M.
P. (2000). New syntheses with oils and fats as renewable raw materials for the chemical industry. Angewandte Chemie International Edition, 39(13), 2206-2224.
Chaudhari, A., Kulkarni, R., Mahulikar, P., Sohn, D., & Gite, V. (2015). Development of PU coatings from neem oil based alkyds prepared by the monoglyceride route. Journal of the American Oil Chemists' Society, 92, 733-741.
Dai, J., Ma, S., Wu, Y., Han, L., Zhang, L., Zhu, J., & Liu, X. (2015). Polyesters derived from itaconic
acid for the properties and bio-based content enhancement of soybean oil-based thermosets. Green chemistry, 17(4), 2383-2392.
de Espinosa, L. M., & Meier, M. A. (2011). Plant oils: The perfect renewable resource for polymer science?!. European Polymer Journal, 47(5), 837852.
Eissen, M., Metzger, J. O., Schmidt, E., & Schneidewind, U. (2002). 10 years after Rio— concepts on the contribution of chemistry to a sustainable development. Angewandte Chemie International Edition, 41(3), 414-436.
Fernandes, F. C., Kirwan, K., Lehane, D., & Coles, S. R. (2017). Epoxy resin blends and composites from waste vegetable oil. European Polymer Journal, 89, 449-460.
Guner, F. S., Yagci, Y., & Erciyes, A. T. (2006). Polymers from triglyceride oils. Progress in polymer science, 31(7), 633-670.
Ji, D., Fang, Z., He, W., Luo, Z., Jiang, X., Wang, T., & Guo, K. (2015). Polyurethane rigid foams formed from different soy-based polyols by the ring opening of epoxidised soybean oil with methanol, phenol, and cyclohexanol. Industrial crops and products, 74, 76-82.
Kosari, A., Moayed, M. H., Davoodi, A., Parvizi, R., Momeni, M., Eshghi, H., & Moradi, H. (2014). Electrochemical and quantum chemical assessment of two organic compounds from pyridine derivatives as corrosion inhibitors for mild steel in HCl solution under stagnant condition and hydrodynamic flow. Corrosion Science, 78, 138150.
Kuranska, M., Prociak, A., Kirpluks, M., & Cabulis, U. (2015). Polyurethane-polyisocyanurate foams modified with hydroxyl derivatives of rapeseed oil. Industrial Crops and Products, 74, 849-857.
Monteavaro, L. L., Da Silva, E. O., Costa, A. P. O., Samios, D., Gerbase, A. E., & Petzhold, C. L. (2005). Polyurethane networks from formiated soy polyols: synthesis and mechanical
characterization. Journal of the American Oil Chemists' Society, 82(5), 365-371.
Saalah, S., Abdullah, L. C., Aung, M. M., Salleh, M. Z., Biak, D. R. A., Basri, M., & Jusoh, E. R. (2015). Waterborne polyurethane dispersions synthesized from jatropha oil. Industrial Crops and Products, 64, 194-200.
Septevani, A. A., Evans, D. A., Chaleat, C., Martin, D. J., & Annamalai, P. K. (2015). A systematic study substituting polyether polyol with palm kernel oil based polyester polyol in rigid polyurethane foam. Industrial Crops and Products, 66, 16-26.
Stan, R., Chira, N., Ott, C., Todasca, C., & Perez, E. (2008). Catanionic organogelators derived from D-sorbitol and natural fatty acids. Rev. Chim, 59(3), 273-276.
Tian, H., Tang, Z., Zhuang, X., Chen, X., & Jing, X. (2012). Biodegradable synthetic polymers: Preparation, functionalization and biomedical application. Progress in Polymer Science, 37(2), 237-280.
Ugarte, L., Saralegi, A., Fernández, R., Martín, L., Corcuera, M. A., & Eceiza, A. (2014). Flexible polyurethane foams based on 100% renewably sourced polyols. Industrial Crops and Products, 62, 545-551.
Yoo, S. H., Kim, Y. W., Chung, K., Baik, S. Y., & Kim, J. S. (2012). Synthesis and corrosion inhibition behavior of imidazoline derivatives based on vegetable oil. Corrosion Science, 59, 42-54.
Zhang, C., Madbouly, S. A., & Kessler, M. R. (2015). Biobased polyurethanes prepared from different vegetable oils. ACS applied materials & interfaces, 7(2), 1226-1233.
Zieleniewska, M., Leszczynski, M. K., Kuranska, M., Prociak, A., Szczepkowski, L., Krzyzowska, M., & Ryszkowska, J. (2015). Preparation and characterisation of rigid polyurethane foams using a rapeseed oil-based polyol. Industrial Crops and Products, 74, 887-897.
