Phytochemical analysis of different varieties of Sorghum biocolor in Telangana state, India

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Dharavath Bhaskara Naik^* , Bukya Bheem Rao , Ch. Uday Kumar , N. Lakshmi Bhavani

Department of Botany, University College of Science, Saifabad, Osmania University, 500004 Hyderabad-India

Corresponding Author Email: dharavathu.bhaskara@gmail.com

DOI : https://doi.org/10.5281/zenodo.7198587

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Introduction

Millet is a small herb that grows around the world as a grain or plant for human consumption. Almost 50 years ago millet was a popular grain in India. With government policies supporting rice and wheat production and consumption, millet cultivation has declined significantly. Before the Green Revolution, millets represented 40% of cultivated cereal crops. After the Green Revolution, rice replaced most cereals. In addition to changing attitudes, many organizations are supporting the development of cereals [1-2]. Educating farmers on the best techniques for growing millet is an important step. Millet is fibrous and contains magnesium, niacin (vitamin B₃), and gluten-free proteins [3]. The sorghum is not only a crop, but also a source of biological energy for alcohol, sugar, and ethanol production. In Telangana of India, the Deccan Development Society has noticed a revival of traditional seed banks. The company works with approximately 5,000 small and poor women farmers in Zahirabad, who cultivate not less than 10 crops, mainly millets, per hectare of their farm. Some of them cultivated up to 25 varieties of crops per hectare. The valuable content of fiber and its bioactive compounds help in weight loss [4-5]. Efforts are being made to include unprocessed grains in government lunch programs in public schools in Karnataka and Telangana.

S. No.	Types of Jowar	Crop Time	Yield
1	C.S.V-15	105-110	1315
2	C.S.V -27	110	12-14
3	C.S.V-31	115	11-12
4	C.S.H-36	110	16-18
5	C.S.H-39	110-115	17-18

Table 1: Types of Jowar, Crop time, and Yield information

Material and Methods

The current investigation was carried out at the laboratory in the Department of Botany, and College of Technology, Osmania University, Hyderabad, TS, India. The millet samples were provided by Acharya Jayashankar Agricultural University, Rajendra Nagar, Hyderabad (Table 1). The seeds of all the genotypes were ground to a fine powder using a grinder. The dried millet seed material was stored in paper bags. Approximately 100 grams of dry plant powder soaked in 70% methanol were extracted for 24 hours at room temperature. The next day, the material was refluxed for 3 hours at a temperature not exceeding 30 ° C, cooled, and filtered. The filtrate was then dried to dryness at a reduced temperature. The product was stored in a dryer and used for further research. Phytochemical analysis of ethanol seed extract was carried out following certain prescribed methods [6-8].

Test for Steroids

In a test tube 2ml solution of extract, 2ml chloroform, and 2ml H₂SO₄ were taken and shaken well. The organic layer turns to red and yellowish-green fluorescence to the aqueous layer confirming the steroids.

Test for Volatile oils

Odor test: Characteristic odor of extract indicates the presence of volatile oil. Solubility test: Solubility in 90% alcohol indicates the presence of volatile oil.

Test for Glycosides

In a test tube 2ml solution of extract, 1ml  glacial acetic acid, 5% FeCl₃ (3 drops), and concentrated H₂SO₄were mixed and observed.  The bluish-green color at the top and red color at the junction is due to the presence of cardiac glycosides.

Test for Saponins

To the extract, water was added and shaken for a few minutes. The formation of foam or froth is due to Saponins in the extract.

Test for Phenolic compounds like tannins

5% Ferric chloride test: To 3 ml of the methanolic seed extract, 3ml of 5% Ferric chloride solution was added and observed for blue-black color, which confirms the availability of phenolic compounds like tannins in the extract.

Test for Flavonoids

To about 0.5 grams of seed extract, ethanol is added to dissolve it. Warm the solution and filter it through filter paper. Add a few pieces of Magnesium turnings and a few drops of concentrated HCl to the filtrate. The appearance of pink or orange color is an indication of the presence of flavonoids.

Test for Alkaloids

To test the presence of alkaloids, a solution of the sample was prepared by adding a few ml of HCl to the extract. Then this solution is used for the following tests for alkaloids

Dragendorff’s test

To 2 ml of the test solution, 1 mL of Dragendorff’s reagent is added along the side of the test tube. The formation of an orange or orange-reddish-brown precipitate is an indication of the presence of alkaloids.

GC-MS Analysis

GC-MS analysis was carried out on a Shimadzu GCMSQ2010 ultra system, The injector temperature was 290°C. The samples were injected in the split mode with a split ratio of 1/23 and the injection volume was 1ul of the capillary column.

Result and Discussion

The phytochemical analysis of the methanol extract of all four genotypes of the Sorghum millet revealed the presence of steroids, glycosides, volatile oils, saponins, tannins, and flavonoids. However, alkaloids were found to be absent in all the genotypes of Sorghum under study (Table 2& Fig 1). The aromatic GC-MS chromatographic profiles of the millet chloroform extract showed various compounds in each genotype and were classified into different chemical classes. The compound Ethanne1, 1diethoxy was found prominently as the major significant constituent in all the genotypes C.S.V-39 (4.91), C.S.V-31 (3.28), C.S.V-27(1.76), C.S.V36 (1.45), C.S.V-15(0.81) followed by cyclononasiloxane, octamethyl compound. C.S.V39 contains the highest Trichlor ethane (12.97%) among all genotypes followed by the compound 3-Ethoxy1, 1,1,5,5,5, hexamethyl3trimethysiloxy as the lowest of all genotypes  (0.12). (Table3).

S. No	Consituents	C.S.V-15	C.S.V-27	C.S.V-31	C.S.V-36	C.S.V39
1	Steroids	+	+	+	+	+
2	Glycosides	+	+	+	+	+
3	Volatile oils	+	+	+	+	+
4	Saponins	+	+	+	+	+
5	Tannins	+	+	+	+	+
6	Flavonoids	+	+	+	+	+
7	Alkaloids	–	–	–	–	–

Table 2:  Phytochemical analysis of sorghum varieties.

Fig:1 Images of the Phytochemical Analysis of test samples

Table 3: Bioactive compounds of Sorghum Millet Genotypes

S.NO	COMPOUND NAME	R.TIME	MOL WEIGHT	FORMALA	CAS NO	CSV15	CSV39	CSV36	CSV31	CSV27
1	Ethane1,1, diethoxy	1.594	118.1742	C6H14O2	105-57-7	0.81	4.91	1.45	3.28	1.76
2	Cyclononasiloxane, octamethyl	0.92	66.4	C18H54O9SI9	105-57-7	0.78	0.13	0.14	0.61	0.92
3	Dextroamphetamine	0.021	135.21	C9H13N	300-62-9	–	0.78	0.46	0.21	1.46
4	Trichloro ethylene	1.483	131.38	C2HCl3	79-01-6	2.09	12.97	9.69	–	–
5	3Ethoxy 1,1,1,5,5,5, Hexamethyl 3trimethysiloxy	37.960	340.71	C17H32O4Si4	1803 0-67-6	0.39	0.12	0.15	0.53	0.39
6	1 penetene, 1,3 Diphenthyl-(trimethysiloxy)	31.899	310.5	C20H26OSi	2005-03-27	–	0.23	0.62		0.54

Discussion and Conclusion

Millet is a highly nutritious cereal and the most widely consumed in the world. In addition, these crops also contain bioactive compounds, including carbohydrates, proteins, flavonoids, and other phytochemicals [9-10]. This crop is also a new type of renewable and efficient energy crop. The production of metabolites is a response to genetic and environmental changes [11-26]. The highest protein-carbohydrate concentration found in sorghum millet suggests that cultivation should be promoted as a food crop. Ethane 1, 1, di-ethoxy were found in a remarkable amount in all genotypes followed by Cyclopentasiloxane, octametyl compound. These tests can be used as a useful tool to analyze sorghum seeds for their phenolic chemical lipid-rich components as a complete nutritional source. Finally, further research has to focus on a comprehensive profile of all the secondary metabolites of this important cereal crop.

Acknowledgment

The authors are thankful to the Department of Botany, Osmania University, Hyderabad, Telangana for their valuable help during the time of Ph.D. work and NFST for funding the work.

Consent and Ethical Approval

As per university standard guidelines, participant consent and ethical approval have been collected and preserved by the authors

Competing interests

The authors have declared that no competing interests exist.

Authors’ Contributions

This work was carried out in collaboration among all authors. All authors read and approved the final manuscript.

 

References        

1. Shyamli, P. S., Rana, S., Suranjika, S., Muthamilarasan, M., Parida, A.and Prasad, M. Genetic determinants of micronutrient traits in graminaceous crops to combat hidden hunger. Theoretical and Applied Genetics,(2021), 1-19.
2. Prasad, M. S., Madhavi, K. R., Madhav, M. S., Rambabu, R., Kumar, S. V., Sundaram, R. M., .and Kumar, R. M. Plant Germplasm Registration Notice. Indian Journal of Plant Genetic Resources,(2019), 32(3), 406-455.
3. Ramalingam, A. P., Mohanavel, W., Premnath, A., Muthurajan, R., Prasad, P. V.,andPerumal, R. Large-Scale Non-Targeted Metabolomics Reveals Antioxidant, Nutraceutical and Therapeutic Potentials of Sorghum. Antioxidants,(2021) ,10(10), 1511
4. Seal, A. N., Pratley, J. E., Haig, T., and An, M. Identification and quantitation of compounds in a series of allelopathic and non-allelopathic rice root exudates. Journal of chemical ecology,(2004), 30(8), 1647-1662.
5. Emrahi, R., Morshedloo, M. R., Ahmadi, H., Javanmard, A., & Maggi, F. Intraspecific divergence in phytochemical characteristics and drought tolerance of two carvacrol-rich Origanum vulgare subspecies: subsp. hirtum and subsp. gracile. Industrial Crops and Products, (2021)168, 113557.
6. Harborne, J. B. Phytochemical methods: A guide to modern techniques of plant analysis. Chapman and Hall Ltd, London.; Pp.(1973), 279.
7. Sofowora, A. Phytochemical Screening of Medicinal Plants and Traditional Medicine in Africa Edition. Spectrum Books Ltd., Nigeria, (1993), 150-156.
8. Trease GE, Evans WC. Pharmacognosy. W.B Scandars Company Ltd. London 14: (1989), 269- 300.
9. Ross, A. B., Shepherd, M. J., Schüpphaus, M., Sinclair, V., Alfaro, B., Kamal-Eldin, A., and Åman, P. Alkylresorcinols in cereals and cereal products. Journal of agricultural and food chemistry, (2003), 51(14), 4111-4118.
10. Sun, X. G., and Tang, M. Effect of arbuscular mycorrhizal fungi inoculation on root traits and root volatile organic compound emissions of Sorghum bicolor. South African Journal of Botany, (2013), 88, 373-379.
11. Bachheti, A., Sharma, A., Bachheti, R. K., Husen, A.and Pandey, D. P. Plant allelochemicals and their various applications. Co-Evolution of Secondary Metabolites,(2020), 441-465.
12. Xiao, Y., Li, X., Yao, L., Xu, D., Li, Y., Zhang, X., .and Guo, Y. Chemical profiles of cuticular waxes on various organs of Sorghum bicolor and their antifungal activities. Plant Physiology and Biochemistry,(2020), 155, 596-604.
13. Vasconcelos, L. C., de Souza Santos, E., de Oliveira Bernardes, C., da Silva Ferreira, M. F., Ferreira, A., Tuler, A. C., … & Praça-Fontes, M. M. (2019). Phytochemical analysis and effect of the essential oil of Psidium L. species on the initial development and mitotic activity of plants. Environmental Science and Pollution Research, 26(25), 26216-26228.
14. Hassan, O. H. A. (2020). Phytochemical screening and antibacterial activities of Sorghum bicolor leaves derived from in vitro culture. GSC Biological and Pharmaceutical Sciences, 10(1), 065-072.
15. Abba, A., Agunu, A., Abubakar, A., Abubakar, U. S., & Jajere, M. U. (2016). Phytochemical screening and antiproliferative effects of methanol extract of stem bark of Diospyros mespiliformis Hochst (Ebenaceae) against guinea corn (Sorghum bicolor) seeds radicles length. Bayero Journal of Pure and Applied Sciences, 9(1), 1-5.
16. Mpiana, P. T., Lombe, B. K., Ombeni, A. M., Tshibangu, D. S., Wimba, L. K., Tshilanda, D. D., … & Muyisa, S. K. (2013). In vitro sickling inhibitory effects and anti-sickle erythrocytes hemolysis of Dicliptera colorata CB Clarke, Euphorbia hirta L. and Sorghum bicolor (L.) Moench.
17. Ghimire, B. K., Seo, J. W., Yu, C. Y., Kim, S. H., & Chung, I. M. (2021). Comparative study on seed characteristics, antioxidant activity, and total phenolic and flavonoid contents in accessions of sorghum bicolor (L.) moench. Molecules, 26(13), 3964.
18. Mohamed, L. K., Sulieman, M. A., Yagoub, A. E. A., Mohammed, M. A., Alhuthayli, H. F., Mohamed Ahmed, I. A., … & Hassan, A. B. (2022). Changes in Phytochemical Compounds and Antioxidant Activity of Two Irradiated Sorghum (Sorghum bicolor (L.) Monech) Cultivars during the Fermentation and Cooking of Traditional Sudanese Asida. Fermentation, 8(2), 60.
19. Punia, H., Tokas, J., Malik, A., & Sangwan, S. (2021). Characterization of phenolic compounds and antioxidant activity in sorghum [Sorghum bicolor (L.) Moench] grains. Cereal Research Communications, 49(3), 343-353.
20. Oladiji, A. T., Jacob, T. O., & Yakubu, M. T. (2007). Anti-anaemic potentials of aqueous extract of Sorghum bicolor (L.) moench stem bark in rats. Journal of Ethnopharmacology, 111(3), 651-656.
21. Salazar-López, N. J., Gonzalez-Aguilar, G., Rouzaud-Sandez, O., & Robles-Sanchez, M. (2018). Technologies applied to sorghum (Sorghum bicolor L. Moench): changes in phenolic compounds and antioxidant capacity. Food Science and Technology, 38, 369-382.
22. Satish, S., Raghavendra, M. P., Mohana, D. C., & Raveesha, K. A. (2008). Antifungal activity of a known medicinal plant Mimusops elengi L. against grain moulds. Journal of Agricultural technology, 4(1), 151-165.\ Hansen, K. S., Kristensen, C., Tattersall, D. B., Jones, P. R., Olsen, C. E., Bak, S., & Møller, B. L. (2003). The in vitro substrate regiospecificity of recombinant UGT85B1, the cyanohydrin glucosyltransferase from Sorghum bicolor. Phytochemistry, 64(1), 143-151.
23. Cowan, M. F., Blomstedt, C. K., Møller, B. L., Henry, R. J., & Gleadow, R. M. (2021). Variation in production of cyanogenic glucosides during early plant development: A comparison of wild and domesticated sorghum. Phytochemistry, 184, 112645.
24. Agbangnan, P. D., Tachon, C., Bonin, H., Chrostowka, A., Fouquet, E., & Sohounhloue, D. C. (2012). Phytochemical study of a tinctorial plant of Benin traditional pharmacopoeia: The red sorghum (sorghum caudatum) of Benin. Scientific Study & Research. Chemistry & Chemical Engineering, Biotechnology, Food Industry, 13(2), 121.
25. Avato, P., Bianchi, G., & Murelli, C. (1990). Aliphatic and cyclic lipid components of sorghum plant organs. Phytochemistry, 29(4), 1073-1078.
26. Awika, J. M., Rooney, L. W., Wu, X., Prior, R. L., & Cisneros-Zevallos, L. (2003). Screening methods to measure antioxidant activity of sorghum (Sorghum bicolor) and sorghum products. Journal of agricultural and food chemistry, 51(23), 6657-6662.